do/-l48620BI
201-14860B3
IUCLID
Data Set
Existing Chemical ~ : ID: 107-12-o
CAS No. : 107-12-o
EINECS Name : propiononitrile
: 203-464-4
EC No.
TSCA Name : Propanenitrile
Molecular Formula : C3H5N
Producer related part
: Solutia Inc.
Company
: 06.06.2003
Creation date
Substance related part
Company : Solutia Inc.
: 06.06.2003
Creation date
Status
Memo
: 06.10.2003
Printing date
: 13.11.2003
Revision date
: 30.10.2003
Date of last update
Number of pages : 58
Chapter (profile) : Chapter: 1, 2, 3,4, 5, 6, 7, 8, 10
: Reliability: without reliability, 1, 2, 3,4
Reliability (profile)
: Flags: without flag, confidential, non confidential, WGK (DE), TA-Luft (DE),
Flags (profile)
Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SIDS
l/l
Id 107-12-0
1. General Information
Date 02.10.2003
1.0.1 APPLICANT AND COMPANY INFORMATION
1.0.2 LOCATION OF PRODUCTION SITE, IMPORTER OR FORMULATOR
1.0.3 IDENTITY OF RECIPIENTS
1.0.4 DETAILS ON CATEGORY/TEMPLATE
1.1.0 SUBSTANCE IDENTIFICATION
IUPAC Name :
C(#N)CC
Smiles Code :
C3H5N
Molecular formula :
55.079
Molecular weight :
Petrol class :
11.06.2003
1.1.1 GENERAL SUBSTANCE INFORMATION
typical for marketed substance
Purity type :
organic
Substance type :
liquid
Physical status :
= 99.6 % v/v
Purity :
clear
Colour :
pungent
Odour :
11.06.2003 (32)
1.1.2 SPECTRA
1.2 SYNONYMS AND TRADENAMES
cyanoethane
10.07.2003
ethyl cyanide
10.07.2003
propanenitrile
10.07.2003
propionic nitrile
10.07.2003
2/2
Id 107-12-0
1. General Information
Date 02.10.2003
propionitrile
10.07.2003
propylnitrile
10.07.2003
1.3 IMPURITIES
1.4 ADDITIVES
1.5 TOTAL QUANTITY
1.6.1 LABELLING
1.6.2 CLASSIFICATION
1.6.3 PACKAGING
1.7 USE PATTERN
industrial
Type of use :
Category :
(2) valid with restrictions
Reliability :
11.06.2003
1.7.1 DETAILED USE PATTERN
1.7.2 METHODS OF MANUFACTURE
1.8 REGULATORY MEASURES
1.8.1 OCCUPATIONAL EXPOSURE LIMIT VALUES
1.8.2 ACCEPTABLE RESIDUES LEVELS
3/3
Id 107-12-0
1. General Information
Date 02.10.2003
1.8.3 WATER POLLUTION
1.8.4 MAJOR ACCIDENT HAZARDS
1.8.5 AIR POLLUTION
1.8.6 LISTINGS E.G. CHEMICAL INVENTORIES
EINECS
Type :
Additional information :
11.06.2003
TSCA
Type :
Additional information :
11.06.2003
1.9.1 DEGRADATION/TRANSFORMATION PRODUCTS
1.9.2 COMPONENTS
1.10 SOURCE OF EXPOSURE
1.11 ADDITIONAL REMARKS
1.12 LAST LITERATURE SEARCH
1.13 REVIEWS
4/4
Id 107-12-0
2. Physico-Chemical Data
Date 02.10.2003
2.1 MELTING POINT
= -92.8 癈
Value :
Sublimation :
other
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed, published data.
Critical study for SIDS endpoint
Flag :
11.06.2003 (29)
2.2 BOILING POINT
= 97 癈 at 1013 hPa
Value :
Decomposition :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed, published data.
Critical study for SIDS endpoint
Flag :
10.07.2003 (29)
2.3 DENSITY
relative density
Type :
= .7818 g/cm?at 20 癈
Value :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed, published data.
10.07.2003 (37)
2.3.1 GRANULOMETRY
2.4 VAPOUR PRESSURE
= 52 hPa at 20 癈
Value :
Decomposition :
other (measured)
Method :
Year :
no
GLP :
5/5
Id 107-12-0
2. Physico-Chemical Data
Date 02.10.2003
as prescribed by 1.1 - 1.4
Test substance :
Purity of the test material is 99.6%.
Remark :
(2) valid with restrictions
Reliability :
Source of data is a MSDS.
Critical study for SIDS endpoint
Flag :
29.07.2003 (32)
= 53.3 at 22 癈
Value :
Decomposition :
other (measured): not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of the material is unknown.
Remark :
Value is 40 mm Hg, which converts to 53.3 hPa.
Result :
Hazardous Substances Data Bank for Priopionitrile, dated 5/13/99.
Source :
(2) valid with restrictions
Reliability :
Primary source is peer reviewed and published.
10.07.2003 (10)
2.5 PARTITION COEFFICIENT
Partition coefficient :
= .16 at 癈
Log pow :
pH value :
other (measured)
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The value was obtained using the Shake-Flask method. The aqueous
Test condition :
phase was octanol-saturated water. The concentration of material in the
aqueous phase was measured using gas-liquid chromatography.
Purity of the test material was not mentioned.
Test substance :
(2) valid with restrictions
Reliability :
Data were from a peer reviewed, published source.
Critical study for SIDS endpoint
Flag :
13.08.2003 (31)
octanol-water
Partition coefficient :
= .35 at 20 癈
Log pow :
pH value :
other (calculated)
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-92.8 degrees
Remark :
C), boiling point (97 degrees C), vapor pressure (39 mm Hg), and water
solubility (93380 mg/l).
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
13.08.2003 (17)
2.6.1 SOLUBILITY IN DIFFERENT MEDIA
6/6
Id 107-12-0
2. Physico-Chemical Data
Date 02.10.2003
water
Solubility in :
= 93380 mg/l at 25 癈
Value :
pH value :
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
Deg. product :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of the test material was not mentioned.
Test substance :
(2) valid with restrictions
Reliability :
Data were from a peer reviewed, published source.
Critical study for SIDS endpoint
Flag :
13.08.2003 (38)
water
Solubility in :
= 119 g/l at 40 癈
Value :
pH value :
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
Deg. product :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed, published data.
15.08.2003 (37)
water
Solubility in :
= 55650 mg/l at 癈
Value :
pH value :
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
Deg. product :
other: calculated
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-92.8 degrees
Remark :
C), boiling point (97 degrees C), vapor pressure (39 mm Hg), and water
solubility (93380 mg/l).
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
15.08.2003 (18)
7/7
Id 107-12-0
2. Physico-Chemical Data
Date 02.10.2003
2.6.2 SURFACE TENSION
2.7 FLASH POINT
= 16 癈
Value :
open cup
Type :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of the test material is unknown.
Remark :
Hazardous Substances Data Bank for propionitrile, dated 5/13/99.
Source :
(2) valid with restrictions
Reliability :
Primary reference is peer-reviewed and published.
10.07.2003 (23)
2.8 AUTO FLAMMABILITY
2.9 FLAMMABILITY
2.10 EXPLOSIVE PROPERTIES
2.11 OXIDIZING PROPERTIES
2.12 DISSOCIATION CONSTANT
2.13 VISCOSITY
2.14 ADDITIONAL REMARKS
8/8
Id 107-12-0
3. Environmental Fate and Pathways
Date 02.10.2003
3.1.1 PHOTODEGRADATION
air
Type :
Sun light
Light source :
nm
Light spectrum :
based on intensity of sunlight
Relative intensity :
INDIRECT PHOTOLYSIS
OH
Sensitizer :
Conc. of sensitizer :
= .0000000000001938 cm?(molecule*sec)
Rate constant :
= 50 % after 55.2 day(s)
Degradation :
Deg. product :
other (calculated)
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-92.8 degrees
Remark :
C), boiling point (97 degrees C), vapor pressure (39 mm Hg), and water
solubility (93,380 mg/l).
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
Critical study for SIDS endpoint
Flag :
15.08.2003 (13)
3.1.2 STABILITY IN WATER
abiotic
Type :
other
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
: EPIWIN Hydrowin cannot calculate hydrolysis rate constants for nitriles.
Remark
The theoretical hydrolysis of propionitrile and several other chemicals has
been examined by Dr. Lee Wolfe at the USEPA Environmental Research
Laboratory in Athens, Georgia. The results of these analyses were
published in a report by Dr. Wolfe that could not be located. In a personal
communication, Dr. Wolfe stated that propionitrile can hydrolyze (albeit
slowly). According to a study cited in the Hazardous Substances Data
Bank, the chemical hydrolysis of the related material acetonitrile in water is
base-catalyzed (the rate constant for base catalyzed hydrolysis is 5.8X10-
3/M-hr), but the half-life at pH 7 is more than 150,000 yrs (Ellington et al.,
1988). Acetonitrile (CH3CN, CAS No. 75-05-8) is the 2-carbon analog of
the category members, possessing the same functionality, but having one
less carbon than propionitrile. Taken together, these data suggest that
hydrolysis of propionitrile at environmentally relevant pHs will occur too
slowly to be a significant means of degradation.
: (2) valid with restrictions
Reliability
Experimental results for the test material could not be located. Results are
for a related material.
07.08.2003 (16)
3.1.3 STABILITY IN SOIL
9/9
Id 107-12-0
3. Environmental Fate and Pathways
Date 02.10.2003
3.2.1 MONITORING DATA
3.2.2 FIELD STUDIES
3.3.1 TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS
fugacity model level III
Type :
other: air, water, soil and sediment
Media :
14.4 % (Fugacity Model Level I)
Air :
48.7 % (Fugacity Model Level I)
Water :
.0821 % (Fugacity Model Level II/III)
Biota :
36.9 % (Fugacity Model Level II/III)
Soil :
other
Method :
2003
Year :
Measured inputs to the program are CAS No., melting point (-92.8 degrees
Remark :
C), boiling point (97 degrees C), vapor pressure (39 mm Hg), and water
solubility (93,380 mg/l).
Henry's Law Constant (Bond Est) is estimated to be 4.06E-005 atm
Result :
m3/mole. The soil/sediment constant Koc is 8.3 as estimated by the
EPIWIN PCKOC Program (v1.66).
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
Critical study for SIDS endpoint
Flag :
15.08.2003 (15)
3.3.2 DISTRIBUTION
3.4 MODE OF DEGRADATION IN ACTUAL USE
3.5 BIODEGRADATION
aerobic
Type :
other: activated sludge
Inoculum :
1000 mg/l related to COD (Chemical Oxygen Demand)
Concentration :
6 hour(s)
Contact time :
other: biodegradable
Result :
Deg. product :
other
Method :
1960
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The test conditions state that the test was performed for 6 hours, but the
Remark :
results for 10 hours are listed.
The study results show that biodegradation occurred, but the results are
not related to TOD. Based on the TOD of the previous study (1670 mg/l),
the percentage of material that biodegraded in 10 hours was approximately
24%.
The oxygen uptake was as follows:
Result :
Time (hrs) Approximate O2 uptake (mg/l)
10 / 10
Id 107-12-0
3. Environmental Fate and Pathways
Date 02.10.2003
2 100
4 210
6 300
8 350
10 400
The study results show that 400 mg/l O2 out of a possible 1000 mg/l (COD)
was utilized (40%).
The kinetics of the metabolism of propionamide and propionic acid
suggested that propionitrile was sequentially metabolized to these
materials.
Bacteria: Activated sludges were grown in 1.5 liter pilot plants under 23
Test condition :
hour aeration. The material was allowed to settle for 1 hour. After settling,
1.0 liter of supernatant liquor was withdrawn, 1.0 liter of tap water, organics
and buffer were added, and aeration was resumed. This cycle was
repeated 7 days a week for the duration of the study.
Test material: The quantity of feed was 1000 mg/l (based on COD).
Occasionally, at the end of the aeration period, 25 ml of mixed liquor was
passed through a membrane filter, which was then dried at 103 degrees C
and reweighed to determine the amount of suspended solids. The filter
was then analyzed for COD and all forms of inorganic nitrogen.
Washed sludge, grown on propionitrile, was placed in a Warburg
Respirometer with low concentrations of several possible breakdown
products. Oxygen uptake was then measured for 6 hours. After this time,
the contents of each flask were passed through a membrane filter so that
the filtrate could be analyzed for COD and inorganic nitrogen forms.
(2) valid with restrictions
Reliability :
Purity of the test material was not given.
Critical study for SIDS endpoint
Flag :
10.08.2003 (34)
aerobic
Type :
other: activated sludge
Inoculum :
500 mg/l
Concentration :
72 hour(s)
Contact time :
= 0 (? % after 72 hour(s)
Degradation :
other: material was toxic
Result :
Deg. product :
other
Method :
1970
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
TOD for propionitrile was 1670 mg/l. Test material was toxic to all three
Result :
sludges. No biodegradation occurred.
Bacteria: The activated sludges were obtained from the municipal plant at
Test condition :
Franklin, TN, the municipal plant at Nashville TN and the plant at
Bordeaux, a suburb of Nashville. Mixed liquor from the aeration tanks was
collected in the morning, the day of the Warburg run. Each sample was
packed in ice and transported to the laboratory within 1 hour of collection.
Before the run began, the sludge sample was blended for 10 sec and the
homogenous blend was analyzed for concentration of SS (not defined, but
assumed suspended solids), using a membrane-filter technique. The
original sample was adjusted to a SS concentration of 2,500 mg/l.
Test conduct: Test material was added to a Warburg flask (125 ml) in
order to obtain a final concentration of 500 mg/l in the reaction
compartment (final volume of 20 ml). KOH (1.0 ml, 20%) was added to the
11 / 11
Id 107-12-0
3. Environmental Fate and Pathways
Date 02.10.2003
center well. A 20 ml volume of activated sludge was then introduced. The
test was performed in duplicate. Flasks were incubated for 72 hours
(constant motion) at 20 degrees C. All three sludges were tested. Oxygen
uptake curves were plotted. Respiration of the sludge alone was plotted as
the control curve.
Theoretical O2 demand (the mg/l O2 required to completely oxidize the test
material) was calculated on the basis of the test material to CO2 and water,
plus nitrate according to the following equation: [TOD = moles of O2
required to balance the equation x molecular weight of O2 x concentration
of test material/ (moles of test material required to balance the oxidation
equation x molecular weight of the test material)]. The percentage of TOD
was to be calculated as follows: % TOD = 100 x D (the difference in mg/l of
O2 uptake between substrate and control)/ TOD. The material was
considered toxic if D was less than 0.
(2) valid with restrictions
Reliability :
Purity of the test material was not given.
08.08.2003 (26)
aerobic
Type :
other: mixed microbial culture
Inoculum :
1000 mg/l
Concentration :
48 hour(s)
Contact time :
other: biodegradable
Result :
Deg. product :
other
Method :
1992
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The final protein and ammonia concentrations and pH were 10.20 mg/l,
Result :
76.9 micromoles/ml and 8.66, respectively, indicating that the mixed culture
could use this material as a growth substrate.
A mixed microbial culture (protein concentration of 0.085 mg/l) was isolated
Test condition :
from an environment contaminated with organic cyanides and
polychlorinated biphenyls. This was grown for 48 hours on phosphate
buffer (ph 7.0, 30 degrees C) containing propionitrile (1 g/l) as the sole
source of carbon and nitrogen. The final concentration of protein, ammonia
and pH were determined.
Test material was obtained from Aldrich Chemical Co. It is presumed that
Test substance :
the material has high purity.
(4) not assignable
Reliability :
The study shows that the test material was used as a substrate (and
therefore was metabolized); however, the extent to which the test material
biodegraded is difficult to determine from the study.
07.08.2003 (9)
3.6 BOD5, COD OR BOD5/COD RATIO
3.7 BIOACCUMULATION
3.8 ADDITIONAL REMARKS
12 / 12
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
4.1 ACUTE/PROLONGED TOXICITY TO FISH
flow through
Type :
Pimephales promelas (Fish, fresh water)
Species :
96 hour(s)
Exposure period :
mg/l
Unit :
= 1520
LC50 :
= 1520
EC50 :
no
Limit test :
yes
Analytical monitoring :
other: following the U.S. EPA Committee on Methods for Toxicity Tests
Method :
with Aquatic Organisms
1990
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
: None of the controls or fish exposed to measured concentrations < = 887
Result
mg/l died or had abnormal behavior. One fish exposed to 1100 mg/l died by
24 hours. All fish exposed to 2188 mg/l died by24 hours. Affected fish lost
schooling behavior, were darkly colored and lost equilibrium prior to death.
The 96 hour LC50 and EC50 values (with confidence intervals) were the
same (1520 and 1450-1580 mg/l, respectively).
The average (+/- SD) temperature, dissolved oxygen, hardness, alkalinity
and pH of the water in the test chambers were 24.4 +/- 0.63 degrees C, 7.3
+/- 0.22 mg/l, 47.0 +/- 0.44 mg/l CaCO3, 40.1 +/- 1.04 mg/l CaCO3, and 7.6
+/- 0.21. It is not known whether these variables were affected by test
material concentration.
Average and ranges of analytical concentrations of the chambers treated
with 0, 455, 700, 1080, 1660 and 2550 mg/l material were <5, 375, 610,
885, 1098 and 2184 mg/l, respectively. When corrected for recovery (99.8
%), test material concentrations were < 5.01, 375, 611, 887, 1100 and 2188
mg/l.
The mean length and weight (+/- SD) of the fish at study termination were
20.8 +/- 1.673 mm and 0.092 +/- 0.0244 g.
: Newly hatched minnows from adults reared in flow-through tanks were held
Test condition
at 25 degrees C in flowing water with a 16-hr photoperiod and were fed
brine shrimp nauplii three times daily (twice on weekends). They were
cultured in filtered Lake Superior water or dechlorinated water from the city
of Superior, WI (exact source not given) The two waters were similar in all
measured chemical parameters. This water was used for test material
dilution and all tests.
Healthy fish (32 days old) were fasted for 24 hours before treatment. They
were pooled together in one tank and randomly distributed among the
exposure chambers. Tests were initiated by adding 20 fish per treatment
(455, 700, 1080, 1660 and 2550 mg/l) and control to test chambers
containing 1.0 liter of water. Fish loading was 0.1278 g/l. The rate of
exchange was 14.4 volumes of test water per day.
Observations of fish behavior and toxic signs were made at 2-8, 24, 48, 72
and 96 hr and recorded on checklists specifically formatted to convert
observational data for approximately 100 endpoints into a numerically
coded form. Death (cessation of opercular movements and inability to
respond when prodded) was recorded at 24, 48, 72 and 96 hours. Dead
fish were removed. At study termination, individual control fish were
weighed (wet) and measured.
13 / 13
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
All test exposure chambers were sampled for test material concentration at
the beginning of the test and daily thereafter. Concentrations of test
material were analyzed using gas-liquid chromatography. All analyses
included one spike and one duplicate sample for every 6 to 12 water
samples.
Five water quality parameters were routinely measured for each test:
temperature, dissolved oxygen, total hardness, total alkalinity, and pH. The
desired test temperature was 25 +/- 1 degrees C. Daily measurements of
oxygen concentration were taken in each treatment and the control
exposure chambers if fish were present. The control and one or more
treatment chambers were sampled once for total hardness and alkalinity.
The pH was measured once in the control and in one to five of the
treatment tanks (specific times were not stated).
The estimated LC50 and EC50 values, with corresponding 95% confidence
intervals were calculated using the corrected average of the analyzed tank
concentrations and the Trimmed Spearman-Karber Method. The EC50
values were based on loss of equilibrium manifested by an inability of the
fish to remain in an upright position when swimming. The mean
concentrations used in the calculations were corrected for analytical
recoveries of spiked water samples.
: Purity of test material was 99 %.
Test substance
: (1) valid without restriction.
Reliability
The study was comparable to a guideline study.
: Critical study for SIDS endpoint
Flag
10.08.2003 (20)
static
Type :
Lepomis macrochirus (Fish, fresh water)
Species :
96 hour(s)
Exposure period :
mg/l
Unit :
< 10 measured/nominal
NOEC :
= 41 measured/nominal
LC50 :
no
Limit test :
no
Analytical monitoring :
other: APHA, Standard Methods for Examination of Water and Wastewater,
Method :
14th Ed., 1975
1981
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Although the dissolved oxygen concentration in all 3 tested solutions
Remark :
dropped below 40% saturation at 96 hours, the authors concluded that this
did not have an effect on mortality.
None of the negative control fish died. Ten percent of the fish exposed to
Result :
10 or 18 mg/l died by 96 hours. The mortality rate for fish exposed to 32
mg/l was 20% at 24 hours, 20% at 48 hours, 40% at 72 hours, and 50% at
96 hours. The mortality rate for fish exposed to 56 mg/l was 0% at 24
hours, 40% at 48 and 72 hours, and 50% at 96 hours. The mortality rate
for fish exposed to 100 mg/l was 50% at 24 hours, and 90% at 48, 72 and
96 hours. The 24, 48 and 96 hour LC50 values for propionitrile (with
confidence limits if applicable) were > 100 mg/l, 56 (43 - 78) mg/l and 41
(28 - 66) mg/l, respectively.
Negative control fish appeared normal at all observations. One fish in each
of the 10 and 18 mg/l groups was surfacing at 96 hours. All fish that did not
die after exposure to 32 mg/l appeared normal (with the exception of 1 fish
that had an illegible observation at 48 hours). All survivors exposed to 56
and 100 mg/l were observed to be surfacing at 96 hours. The weight and
14 / 14
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
length of the fish at the end of the test were 0.12 +/- 0.02 g and 19 +/- 1.5
mm, respectively.
The temperature was 22 degrees C for all water samples. The dissolved
oxygen concentration was 9.0 in each tested solution at 0 hours. Dissolved
oxygen concentration ranged from 6.0 - 8.5 at 48 hours, and 2.0 - 4.3 at 96
hours. The water containing 100 mg/l propionitrile had the lowest dissolved
oxygen concentration at 48 and 96 hours. The pH ranged from 7.1 - 7.7.
Total ammonia concentrations were < 0.1 mg/l at each measurement. All
temperatures, pH values and ammonia concentrations were within
acceptable limits. Dissolved oxygen concentrations at 96 hours were not
within acceptable limits (as defined by the protocol as 40-100% saturation).
The 24, 48 and 96 hour LC50 values for the positive control (with
confidence limits if applicable) were > 0.00014 mg/l, 0.00012 mg/l and
0.00010 (0.000075 - 0.00014) mg/l, respectively. The LC50 values for the
positive control were within the 95 % confidence limits reported in the
literature.
Test organisms: The bluegill sunfish used in the study were obtained from
Test condition :
Osage Catfisheries, Inc., Osage Beach, MO. All fish were on a 16 hour
daylight photoperiod and observed for at least 14 days prior to testing.
Fish received a standard commercial fish food daily until 48 hours prior to
testing. The mean weight and length of the fish at the end of the test were
0.12 +/- 0.02 g and 19 +/- 1.5 mm, respectively. Maximum loading was 0.8
g fish/liter of solution.
Test material: Test concentrations were prepared based on the total
compound. They were obtained by transferring appropriate aliquots from a
working standard (150 mg/ml test material in absolute ethanol) directly to
the test chambers. A preliminary, 48-hour range finding test was
conducted with 1, 10 and 100 mg/l. The definitive, 96 hour test was
conducted with 5 concentrations of test material ranging from 10 - 100
mg/l. The negative control chamber received an ethanol aliquot equivalent
to the highest amount used in the test solutions. A positive control
(Antimycin A), also was tested at concentrations ranging from 0.000014 to
0.00014 mg/l.
Test water: The well water from which the reconstituted water was
prepared contained < 0.01 ppm aluminum, copper and zinc, <0.001 ppm
arsenic, cadmium, and cobalt, 0.001 ppm chromium, 0.012 ppm iron, 0.009
ppm lead, <0.0001 ppm mercury, 0.0157 ppm nickel, and <0.3 ppb of
commonly analyzed pesticides. The water was reconstituted to contain 48
mg/l NaHCO3, 30 mg/l CaSO4, 30 mg/l MgSO4, and 2 mg/l KCl. The
hardness, alkalinity and initial pH of the water were 45 mg/l (as CaCO3), 35
mg/l (as CaCO3) and 7.3, respectively. The dissolved oxygen
concentration at the start of the test was 9.0 mg/l. The temperature of the
water was kept at 22 +/- 1 degrees C.
Test conduct: Tests were conducted in 5 gallon glass vessels containing
15 liters of reconstituted water. The test fish (10 per test concentration)
were acclimated to the dilution water for 48 hours prior to testing. They
were not fed during this acclimation period or during the test. The test
concentrations (10, 18, 32, 56 and 100 mg/l) were chosen based on the
results of a preliminary study. Two additional groups of 10 fish were
exposed to the negative or positive control. Fish were added randomly
within 30 minutes of preparation of the test solutions. All fish were
observed at 24, 48, 72 and 96 hours for mortality and abnormal behavior.
The pH, dissolved oxygen concentration, and temperature of water in the
negative control, and 10 mg/l and 100 mg/l test vessels were determined at
the beginning of the test and after 48 and 96 hours. Total ammonia
concentration of water in these 3 vessels was determined at the beginning
and end of the test.
15 / 15
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
Statistical analysis: Concentration vs. lethality data were analyzed by a
computer program which utilized the binomial, moving average and probit
tests to determine the LC50 values (and 95% confidence limits) at 24, 48
and 96 hours. The method of calculation selected for presentation (the
probit method for propionitrile and the binomial method for Actinomycin A)
was the one that gave the narrowest confidence limit.
The purity of the test material (lot # 34) was 96.1%. Contaminants were not
Test substance :
mentioned.
(2) valid with restrictions
Reliability :
Test concentrations were not analytically confirmed.
07.08.2003 (2)
static
Type :
Salmo gairdneri (Fish, estuary, fresh water)
Species :
96 hour(s)
Exposure period :
mg/l
Unit :
= 180 measured/nominal
NOEC :
= 340 measured/nominal
LC50 :
no
Limit test :
no
Analytical monitoring :
other: Committee on Methods for Toxicity Tests With Aquatic Organisms,
Method :
EPA-660/3-75-009, 1975
1981
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
A brown film was present on the surface of water containing the highest 2
Remark :
concentrations of propionitrile.
None of the negative control fish or fish exposed to 100 or 180 mg/l died.
Result :
The mortality rate for fish exposed to 320 mg/l was 10% at 24 hours, 20%
at 48 hours, 30% at 72 hours, and 40% at 96 hours. The mortality rate for
fish exposed to 560 or 1000 mg/l was 100% by 24 hours. The 24, 48 and
96 hour LC50 values for propionitrile (with confidence limits if applicable)
were 400 (320 - 560) mg/l, 380 (180 - 560) mg/l and 340 (180 - 560) mg/l,
respectively.
Negative control fish and fish exposed to 100 and 180 mg/l appeared
normal at all observations. Surfacing was observed in some of the fish
exposed to 320 mg/l at 48 hours (N = 8), 72 hours (N = 2) and 96 hours (N
= 1). An illegible abnormality was observed in one fish exposed to 320
mg/l for 24 hours. The weight and length of the fish at the end of the test
were 0.12 +/- 0.02 g and 19 +/- 1.5 mm, respectively.
The temperature was 12 degrees C for all water samples. The dissolved
oxygen concentration was 9.4 and 8.7 in each tested solution at 0 and 48
hours, respectively. Dissolved oxygen concentration ranged from 8.0 - 9.1
at 96 hours. The pH ranged from 6.8 - 7.7 in all water samples tested
(except for the water containing 1000 mg/l, which had a pH of 9.1 at 0
hours). Total ammonia concentrations were < 0.1 mg/l in the control and
100 mg/l solution at 0 hours, 0.21 mg/l in the control solution at 96 hours,
0.27 mg/l in the 1000 mg/l solution at 0 hours and the 100 mg/l solution at
96 hours, and 0.52 mg/l in the 320 mg/l solution at 96 hours. All
temperatures, pH values, and dissolved oxygen and ammonia
concentrations were within acceptable limits.
The 24, 48 and 96 hour LC50 values for the positive control (with
confidence limits if applicable) were 0.00014 (0.00010 - 0.00047) mg/l,
0.000041 (0.000032 - 0.000052) mg/l and 0.000030 (0.000024 - 0.000042)
mg/l, respectively. The LC50 values for the positive control were within the
95 % confidence limits reported in the literature.
Test organisms: The rainbow trout used in the study were obtained from
Test condition :
16 / 16
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
Spring Creek Trout Hatchery in Lewistown, Montana. All fish were on a 16
hour daylight photoperiod and observed for at least 14 days prior to testing.
Fish received a standard commercial fish food daily until 48 hours prior to
testing. The mean weight and length of the negative control fish at the end
of the test were 1.16 +/- 0.37 g and 44 +/- 3.7 mm, respectively. Maximum
loading was 0.8 g fish/liter of solution.
Test material: Test concentrations were prepared based on the total
compound. They were obtained by transferring appropriate aliquots from a
working standard (150 mg/ml test material in absolute ethanol) directly to
the test chambers. A preliminary, 48-hour range finding test was
conducted with 10 and 100 mg/l. The definitive, 96 hour test was
conducted with 5 concentrations of test material in a logarithmic series
ranging from 100 - 1000 mg/l. The negative control chamber received an
ethanol aliquot equivalent to the highest amount used in the test solutions.
A positive control (Antimycin A), also was tested at concentrations ranging
from 0.000014 to 0.00014 mg/l.
Test water: The well water from which the reconstituted water was
prepared contained < 0.01 ppm aluminum, copper and zinc, <0.001 ppm
arsenic, cadmium, and cobalt, 0.001 ppm chromium, 0.012 ppm iron, 0.009
ppm lead, <0.0001 ppm mercury, 0.0157 ppm nickel, and <0.3 ppb of
commonly analyzed pesticides. The water was reconstituted to contain 48
mg/l NaHCO3, 30 mg/l CaSO4, 30 mg/l MgSO4, and 2 mg/l KCl. The
hardness, alkalinity and initial pH of the water were 45 mg/l (as CaCO3), 35
mg/l (as CaCO3) and 7.3, respectively. The dissolved oxygen
concentration at the start of the test was 9.4 mg/l. The temperature of the
water was kept at 12 +/- 1 degrees C.
Test conduct: Tests were conducted in 5 gallon glass vessels containing
15 liters of reconstituted water. The test fish (10 per test concentration)
were acclimated to the dilution water for 48 hours prior to testing. They
were not fed during this acclimation period or during the test. The test
concentrations (100, 180, 320, 560 and 1000 mg/l) were chosen based on
the results of a preliminary study. Two additional groups of 10 fish were
exposed to the negative or positive control. Fish were added randomly
within 30 minutes of preparation of the test solutions. All fish were
observed at 24, 48, 72 and 96 hours for mortality and abnormal behavior.
The pH, dissolved oxygen concentration, temperature of water and total
ammonia concentration in the negative control, and 100 mg/l and 1000
mg/l test vessels were determined at the beginning of the test. At 48 and
96 hours, these variables were tested in control water and water containing
100 and 320 mg/l test material (with the exception that total ammonia was
not measured at 48 hours).
Statistical analysis: Concentration vs. lethality data were analyzed by a
computer program which utilized the binomial, moving average and probit
tests to determine the LC50 values (and 95% confidence limits) at 24, 48
and 96 hours. The method of calculation selected for presentation (the
binomial method) was the one that gave the narrowest confidence limit.
The purity of the test material (lot # 34) was 96.1%. Contaminants were not
Test substance :
mentioned.
(2) valid with restrictions
Reliability :
Test concentrations were not analytically confirmed.
07.08.2003 (1)
4.2 ACUTE TOXICITY TO AQUATIC INVERTEBRATES
static
Type :
Daphnia magna (Crustacea)
Species :
17 / 17
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
48 hour(s)
Exposure period :
mg/l
Unit :
= 100 measured/nominal
NOEC :
= 250 measured/nominal
EC50 :
no
Limit Test :
no
Analytical monitoring :
other: Committee on Methods for Toxicity Tests With Aquatic Organisms,
Method :
EPA-660/3-75009, 1975
1981
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
A no observable effect level of 100 mg/l was listed by the authors although
Remark :
abnormal behavior was noted in 1/20 daphnids exposed to this
concentration.
None of the controls or daphnids exposed to 100 mg/l died during the
Result :
study. One daphnid in one vessel containing 180 mg/l died between 24
and 48 hours. Three fish exposed to this concentration in another vessel
died (one by 24 hours). Therefore, the overall death rate of daphnids
exposed to 180 mg/l was 5% and 24 hours and 20% at 48 hours. Ten out
of 20 daphnids exposed to 320 mg/l (4 in one vessel and 6 in the other)
died within 24 hours, and 15 died by 48 hours (8 in one vessel and 7 in the
other). Therefore, the overall death rate of daphnids exposed to 320 mg/l
was 50% and 24 hours and 75% at 48 hours. All daphnids exposed to 560
or 1000 mg/l died within 24 hours. The 24 and 48 hour LC50 values (with
confidence limits) were 310 (270 - 350) mg/l and 250 (210 - 290) mg/l,
respectively.
In one vessel containing daphnids exposed to 100 mg/l, 1 daphnid was
observed on the surface at 48 hours. Two daphnids exposed to 320 mg/l
(in one vessel) were on the surface at 24 hours. Behavior of other
daphnids was normal.
The initial temperature, dissolved oxygen concentration and pH of the
control water were 20 degrees C, 8.8 mg/l and 8.6. The temperature,
dissolved oxygen concentration and pH of all water assayed at 48 hours
were 21 degrees C, 7.8 - 8.3 mg/l and 8.6 - 8.7. All temperatures, dissolved
oxygen concentrations and pH values were within acceptable limits.
Test organisms: The Daphnia magna used in the study were obtained from
Test condition :
an in-house culture. The adults were fed a suspension of trout chow and
alfalfa daily until 24 hours prior to testing. All daphnids were held at 20 +/- 2
degrees C, under a 16 hour daylight photoperiod. First instar daphnids (<
24 hours old) were used in the test. Test daphnids were not fed during the
study.
Test material: Test concentrations were not corrected for sample purity. A
primary standard of 20 mg/ml was prepared in water. Appropriate volumes
of this standard were added to test water to obtain test concentrations.
Test water: The water used in the study was from a deep well source. It
contained < 0.01 ppm aluminum, copper and zinc, <0.001 ppm arsenic,
cadmium, and cobalt, 0.001 ppm chromium, 0.012 ppm iron, 0.009 ppm
lead, <0.0001 ppm mercury, 0.0157 ppm nickel, and <0.3 ppb of commonly
analyzed pesticides. The hardness, alkalinity, conductivity, dissolved
oxygen concentration and initial pH of the well water were 255 ppm (as
CaCO3), 368 ppm (as CaCO3), 50 micromhos/ cm, 9.2 ppm, and 7.8,
respectively. The temperature of the water was kept at 20 +/- 1 degrees C.
Test conduct: Tests were conducted in 250 ml glass beakers containing
200 ml of well water. The test organisms (10 per test concentration) were
added randomly to the test water within 30 minutes of addition of test
material. The test concentrations (100, 180, 320, 560 and 1000 mg/l) were
18 / 18
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
based on the results of a preliminary study performed with 1, 10 and 100
mg/l. One additional group of 10 organisms was exposed to water only
served as a control. Each condition was tested in duplicate. All organisms
were observed initially and after 24 and 48 hours of exposure for mortality
and abnormal behavior (surfacing or loss of equilibrium). The pH, dissolved
oxygen concentration and temperature of the control water were
determined at the beginning and end of the study. Water containing 100,
320 and 1000 mg/l was analyzed for pH, dissolved oxygen concentration
and temperature at the end (but not the beginning) of the study.
Statistical analysis: Concentration vs. lethality data were analyzed by a
computer program which utilized the binomial, moving average and probit
tests to determine the LC50 value (and 95% confidence limit) at 24 and 48
hours. The method of calculation selected for presentation (probit) was the
one that gave the narrowest confidence limit.
The purity of the test material (lot # 34) was 96.1%. Contaminants were not
Test substance :
mentioned.
(2) valid with restrictions
Reliability :
Test concentrations were not analytically confirmed.
Critical study for SIDS endpoint
Flag :
01.08.2003 (3)
4.3 TOXICITY TO AQUATIC PLANTS E.G. ALGAE
Scenedesmus subspicatus (Algae)
Species :
biomass
Endpoint :
96 hour(s)
Exposure period :
mg/l
Unit :
= 789.303 calculated
EC50 :
other
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-92.8 degrees
Remark :
C), boiling point (97 degrees C), vapor pressure (39 mm Hg), and water
solubility (93,380 mg/l).
This is a supporting study for the SIDS endpoint.
(2) valid with restrictions
Reliability :
15.08.2003 (14)
Selenastrum capricornutum (Algae)
Species :
other: biomass and growth rate
Endpoint :
72 hour(s)
Exposure period :
mg/l
Unit :
= 87.8 measured/nominal
NOEC :
> 87.8 measured/nominal
EC50 :
yes
Limit test :
yes
Analytical monitoring :
other: OECD: TG-201 and EEC/Annex V C.3
Method :
1999
Year :
yes
GLP :
other TS
Test substance :
No protocol deviations were noted. The EbC50 (0-72 hr) and the ErC50 (0
Remark :
72 hr) were inestimable as greater than 50% inhibition in growth and/or
biomass was not achieved. The significant loss (up to 80.7% over the
course of the study) in test material was attributed to volatilization.
19 / 19
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
This is a supporting study for the SIDS endpoint for propionitrile.
Algae exposed to test material exhibited normal growth with respect to
Result :
control. No deformed cells were noted. At the end of the test, the mean
cell density in treated cultures was 1.365 x 10E6 cells /ml (compared to
1.356 x 10E6 cells in control).
The average concentrations of material in the test flasks at the beginning of
the test and after 72 hours were 200.68 and 38.65 mg/l, respectively.
Approximately 80.74% of the material was lost over the course of the
experiment. The mean concentration was 87.82 mg/l. This concentration
was listed as the NOEC.
Results of the photostability tests were similar to those in flasks containing
test material and algae. The control solutions that were exposed to light or
were in the dark exhibited 78.11% and 69.35% losses of test material.
The mean temperature and illumination were 24 degrees C and 746 foot
candles (range 744 - 748 foot-candles) throughout the test. The pH ranged
from 7.42 - 7.88. The shaker speed was maintained at 100 rpm.
The test was considered to be valid since the mean cell concentration in
control cultures increased by a factor of 136-fold within 72 hours.
A 4-day culture of Selenastrum capricornutum SF-3148 (passage 5 in liquid
Test condition :
algal medium) was used as the test algae. Several passages were
performed prior to the test to confirm exponential growth.
Test medium: Sterile growth medium was prepared using high quality
distilled water. The pH of the medium was measured and adjusted to 7.5
(+/- 0.1) using 0.01N NaOH.
Test material stock solution: Test material (0.156 ml) was added to 600.0 g
of algal growth medium (to produce a nominal concentration of 200 mg/l).
The solution was immediately capped and stirred for 1-2 minutes. An
aliquot of the solution was removed for analysis of concentration at time 0.
Test conduct: All steps were carried out aseptically in a hood to prevent
contamination. Test vessels were sterile 250 ml Erlenmeyer flasks. Test
material stock solution (100 ml) was added to 5 flasks and test medium that
did not contain test material was added to 3 flasks. Algae (515 microliters
of algal stock culture to achieve an initial cell density of 1 x 10E4 cells/ml)
were added to 3/5 flasks that contained test material and the three that did
not. The two flasks that contained test material but were not inoculated
served as photostability controls. One of the flasks was exposed to light
and one was wrapped in foil to shield it from light. All flasks were secured
with foam stoppers and transferred to a shaking incubator (24 degrees C,
100 rpm). They were illuminated at an average of 746.2 footcandles
throughout the study.
Temperature, light intensity, and shaker speed (rpm) were assessed at the
0, 24, 48, and 72 hours. Concentrations of test material in the flasks that
contained algae also were assessed at these times. The pH and was
assessed at time 0 and after 72 hours. Concentrations of test material in
the photostability controls also were measured at 0 and 72 hours.
Concentrations of test material were analyzed using gas chromatography
with flame ionization detection (GC/FID).
The exposure concentration was calculated as the geometric mean of the
test concentrations analyzed at the 4 time points. Cell counts were
performed after 24, 48 and 72 hours of exposure using a calibrated Coulter
Counter. The mean algal cell count for the test and control curves was
calculated. Two measures of growth [biomass (area under the growth
20 / 20
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
curve) and growth rate] were used to determine the effect of the material
on algae. The concentrations that produced a 50% inhibition of biomass
(EbC50) and growth rate (ErC50) relative to control were to be calculated
by fitting linear regression models to the data.
The test was considered valid if the mean cell concentration in the control
cultures increased by a factor of at least 16 within 72 hours. exhibited
78.11% and 69.35% losses of test material.
The mean temperature and illumination were 24 degrees C and 746 foot
candles (range 744 - 748 foot-candles) throughout the test. The pH ranged
from 7.42 - 7.88. The shaker speed was maintained at 100 rpm.
The test was considered to be valid since the mean cell concentration in
control cultures increased by a factor of 136-fold within 72 hours.
The test material was isobutyronitrile (CAS No. 78-82-0). Purity was 99.9%.
Test substance :
The results of this study indicate that the test substance would not be
Conclusion :
classified according to the European Union's labeling directive and would
correspond to a "low concern level" according to the U.S. EPA's
assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD-study conducted under GLP
assurances.
07.08.2003 (11)
Selenastrum capricornutum (Algae)
Species :
other: biomass and growth rate
Endpoint :
72 hour(s)
Exposure period :
mg/l
Unit :
= 133.4 measured/nominal
NOEC :
> 133.4 measured/nominal
EC50 :
yes
Limit test :
yes
Analytical monitoring :
other: OECD: TG-201 and EEC/Annex V C.3
Method :
1999
Year :
yes
GLP :
other TS
Test substance :
Results of a pilot study conducted prior to this test indicated that a limit test
Remark :
design would be appropriate for the material.
The EbC50 (0-72 hr) and the ErC50 (0-72 hr) were inestimable as greater
than 50% inhibition in growth and/or biomass was not achieved. No
protocol deviations were noted.
This is a supporting study for the SIDS endpoint for propionitrile.
Algae exposed to test material exhibited normal growth with respect to
Result :
control. At the end of the test, the mean cell density in treated cultures was
9.5 x 10E5 cells /ml (compared to 9.0 x 10E5 cells in control).
The average concentrations of material in the test flasks at the beginning of
the test and after 72 hours were 206.0 and 85.7 mg/l, respectively. The
mean concentration was 133.4 mg/l. This concentration was listed as the
NOEC, EbC50 and ErC50.
Results of the photostability tests were similar to those in flasks containing
test material and algae. The control solutions that were exposed to light or
were in the dark exhibited 58% and 50% losses of test material.
The mean temperature and illumination were 24 degrees C and 747 foot
candles (+/- 5.5 foot-candles) throughout the test. The pH ranged from 7.4
- 7.6.
21 / 21
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
The test was considered to be valid since the mean cell concentration in
control cultures increased by a factor of 90.2-fold within 72 hours.
Test Organisms: A 4-day culture of Selenastrum capricornutum SF-3148
Test condition :
(passage 3 in liquid algal medium) was used as the test algae. Several
passages were performed prior to the test to confirm exponential growth.
The density of cells in the stock culture was 2.58 x 10E6 cells/ml prior to
use.
Test medium: Sterile growth medium was prepared using high quality
distilled water. The pH of the medium was measured and adjusted to 7.5
(+/- 0.1) using 0.1N NaOH prior to use.
Test material stock solution: Approximately 0.151 ml (120 mg) of the test
material was added to 600 ml of algal growth medium with a gas tight
Hamilton syringe (to produce a nominal concentration of 200 mg/l). The
solution was stirred for approximately 1 minute. An aliquot (1.0) of the
solution was removed for analysis of concentration.
Test conduct: All steps were carried out aseptically in a hood to prevent
contamination. Test vessels were sterile 250 ml Erlenmeyer flasks. Test
material stock solution (100 ml) was added to 5 flasks and test medium that
did not contain test material was added to 3 flasks. Algae (388 microliters
of algal stock culture to achieve an initial cell density of 1 x 10E4 cells/ml)
were added to 3/5 flasks that contained test material and the three that did
not. The two flasks that contained test material but were not inoculated
served as photostability controls. One of the flasks was exposed to light
and one was wrapped in foil to shield it from light. All flasks were secured
with foam stoppers and transferred to a shaking incubator (24 degrees C,
100 rpm). They were illuminated at 747 (+/- 5.5) footcandles throughout the
study.
Temperature, light intensity, and shaker speed (rpm) were assessed at the
0, 24, 48, and 72 hours. Concentrations of test material in the flasks that
contained algae also were assessed at these times. The pH and was
assessed at time 0 and after 72 hours. Concentrations of test material in
the photostability controls also were measured at 0 and 72 hours.
Concentrations of test material were analyzed using gas chromatography
with flame ionization detection (GC/FID). The exposure concentration was
calculated as the geometric mean of the test concentrations analyzed at
the 4 time points. Cell counts were performed after 24, 48 and 72 hours of
exposure using a calibrated Coulter Counter. The mean algal cell count for
the test and control curves. Two measures of growth [biomass (area under
the growth curve) and growth rate] were used to determine the effect of the
material on algae. The concentrations that produced a 50% inhibition of
biomass (EbC50) and growth rate (ErC50) relative to control were to be
calculated by fitting linear regression models to the data.
The test was considered valid if the mean cell concentration in the control
cultures increased by a factor of at least 16 within 72 hours.
The test material was butyronitrile (CAS No. 109-74-0). Purity was 99.9%
Test substance :
(GC/FID).
The results of this study indicate that the test substance would not be
Conclusion :
classified according to the European Union's labeling directive and would
correspond to a "low concern level" according to the U.S. EPA's
assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD-study conducted under GLP
assurances.
07.08.2003 (12)
22 / 22
Id 107-12-0
4. Ecotoxicity
Date 02.10.2003
4.4 TOXICITY TO MICROORGANISMS E.G. BACTERIA
4.5.1 CHRONIC TOXICITY TO FISH
4.5.2 CHRONIC TOXICITY TO AQUATIC INVERTEBRATES
4.6.1 TOXICITY TO SEDIMENT DWELLING ORGANISMS
4.6.2 TOXICITY TO TERRESTRIAL PLANTS
4.6.3 TOXICITY TO SOIL DWELLING ORGANISMS
4.6.4 TOX. TO OTHER NON MAMM. TERR. SPECIES
4.7 BIOLOGICAL EFFECTS MONITORING
4.8 BIOTRANSFORMATION AND KINETICS
4.9 ADDITIONAL REMARKS
23 / 23
Id 107-12-0
5. Toxicity
Date 02.10.2003
5.0 TOXICOKINETICS, METABOLISM AND DISTRIBUTION
: In vivo
In Vitro/in vivo
: Excretion
Type
: rat
Species
Number of animals
:3
Males
Females : 3
Doses
: approximately 1.65 mg 14C labeled priopionitrile
Males
Females : approximately 1.65 mg 14C labeled priopionitrile
Vehicle :
: gavage
Route of administration
Exposure time :
Product type guidance :
Decision on results on acute tox. tests :
Adverse effects on prolonged exposure :
: 1st:
Half-lives
2nd:
3rd:
Toxic behaviour :
Deg. product :
: other
Method
: 1987
Year
: no data
GLP
: as prescribed by 1.1 - 1.4
Test substance
The overall 14C recovery was 92.05 %. The majority of the material was
Remark :
eliminated in exhaled air or urine by 24 hours. An average of 27.05% of
test material was detected in the Aquasol-2 bubbler (represents volatile
organics) 0.5 hours after administration of 14C-labeleled propionitrile by
gavage. By 3 hours, recovery in the 1N sodium hydroxide bubbler (material
that had been exhaled as either 14CO2 or free cyanide) ranged from 38.54
to 49.25%. At 24 hours, the total 14C recovery in the urine (which
represented a parent metabolite or thiocyanate) ranged from 0.76 to
5.83%. At 72 hours, a small percentage (< 2%) was found in the liver and
kidneys.
Purity of the 14C labeled propionitrile was 98.4%. The specific activity was
Test substance :
4.0 mCi/mmol.
The material is rapidly absorbed from the GI tract and eliminated through
Conclusion :
expired air as parent material, free CO2 or free cyanide.
(4) not assignable
Reliability :
The study was given a reliability rating of 4 because it was not reviewed in
detail.
10.08.2003 (7)
In vivo
In Vitro/in vivo :
Excretion
Type :
rat
Species :
Number of animals
3
Males :
3
Females :
Doses
5.74 mg/kg, 7.9 microcuries
Males :
7.35 mg/kg, 7.9 microcuries
Females :
Vehicle :
gavage
Route of administration :
Exposure time :
Product type guidance :
24 / 24
Id 107-12-0
5. Toxicity
Date 02.10.2003
Decision on results on acute tox. tests :
Adverse effects on prolonged exposure :
: 1st:
Half-lives
2nd:
3rd:
Toxic behaviour :
Deg. product :
: other
Method
: 1981
Year
: no data
GLP
: as prescribed by 1.1 - 1.4
Test substance
The total 14C-activity eliminated in 72 hours was 93.98 % and 74.73% of
Remark :
the administered dose in males and females, respectively. After 72 hours,
the 14C-activity found in the urine of 3 males and 3 females averaged 6.2%
and 4.4% of dose, respectively. The cumulative 14C-activity after 72 hours
found in the feces averaged 0.9% and 1.1% of the dose in males and
females, respectively. The percentage of the dose recovered in whole
blood cells was 0.38% and 0.31% for males and females, respectively. The
majority of the 14C-activity (81.31 % and 63.26% of the administered dose
for males and females, respectively) was in respired air trapped in a
sodium hydroxide bubbler (which represents CO2 or free cyanide). The
amount of volatile material trapped in Aquasol-2 (which represented volatile
organics) was 5.55% and 5.93% of dose for males and females,
respectively.
It was thought that the lower recovery in females was due to loss of 78 ml
of NaOH from the bubbler for one rat. Assuming that this was not lost, the
amount of material in the bubbler would have been 76.25 % of the dose.
The total amount of 14C recovered in this animal would be 85.59%, which
is in agreement with the average total amount of material recovered from
the other 2 females (84.3%).
Purity of the 14C labeled priopionitrile was 97.5%.
Test substance :
(4) not assignable
Reliability :
The study is given a reliability rating of 4 because it was not reviewed in
detail.
05.08.2003 (8)
5.1.1 ACUTE ORAL TOXICITY
LD50
Type :
= 40 mg/kg bw
Value :
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
60
Number of animals :
Vehicle :
25.1, 31.6, 39.8, 50.1, 63.1 and 79.4 mg/kg bw
Doses :
other
Method :
1980
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Whether the study was done according to GLP was not stated in the study
Remark :
documents. However, an accompanying letter from the company that
commissioned the study requested that the study should be performed
according to GLP. Therefore, it is assumed that it was.
The numbers of animals that died after each of the doses is as follows:
Result :
Dose (mg/kg) Number of Deaths (all within 1 day)
25 / 25
Id 107-12-0
5. Toxicity
Date 02.10.2003
25.1 1/5 males, 0/5 females
31.6 2/5 males, 1/5 females
39.8 4/5 males, 1/5 females
50.1 5/5 males, 2/5 females
63.1 3/5 males, 5/5 females
79.4 5/5 males, 4/5 females
The LD50 values [with 95% confidence limits (CL)] and slopes of the
curves were as follows:
Sex LD50 (mg/kg) 95 % CL (mg/kg) slope
M/F 40 35 - 46 4.9
M 38 28 - 51 3.5
F 52 43 - 63 5.6
Signs of toxicity included increasing weakness and collapse (both sexes),
and tremors (in 2 males). Necropsies of animals that died revealed
hemorrhagic lungs and liver, discoloration of liver, kidneys and spleen, and
acute gastrointestinal inflammation. Average weights of survivors at days 7
and 14 were higher than initial weights. Viscera appeared normal in
survivors.
Thirty rats/sex were fasted overnight and divided into 6 groups of 5
Test condition :
rats/sex. These groups were given a single oral dose (undiluted) of
propionitrile at 25.1, 31.6, 39.8, 50.1, 63.1 or 79.4 mg/kg. The average
initial weights of males and females were 200 and 184 g, respectively.
Animals were observed for 14 days, and weighed on days 7 and 14. It is
presumed that they were observed daily. Animals that died were
necropsied upon discovery. Survivors were euthanized and necropsied on
day 14. The LD50 value, 95% confidence interval and slope of the curve
were calculated by an unknown method.
The approximate composition of the test material was 97% propionitrile, 1
Test substance :
3% acrylonitrile, 1% adiponitrile and 2% water.
(2) valid with restrictions
Reliability :
The method of calculating the LC50 value was not listed.
13.08.2003 (39)
LD50
Type :
Value :
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
60
Number of animals :
Vehicle :
50.1, 63.1, 79.4, 100, 126, 158 mg/kg (males); 158, 200, 251, 316 mg/kg
Doses :
(females)
other
Method :
1979
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The numbers of animals that died after each of the doses is as follows:
Result :
Dose (mg/kg) Number of Deaths (all in 1 - 3 days)
50.1 0/5 males
63.1 2/5 males
79.4 4/5 males
100 3/5 males
26 / 26
Id 107-12-0
5. Toxicity
Date 02.10.2003
126 3/5 males
158 4/5 males, 0/5 females
200 1/5 females
251 1/5 females
316 4/5 females
The LD50 values [with 95% confidence limits (CL)] and slopes of the
curves were as follows:
Sex LD50 (mg/kg) 95 % CL (mg/kg) slope
M 75 58 - 98 3.9
F 270 235 - 310 9.0
Signs of toxicity in both sexes included weight loss (one to three days in
survivors), increasing weakness, ocular discharge, tremors, convulsions,
collapse and death. Diarrhea and dyspnea were also observed in females.
Animals that died had hemorrhagic lungs, liver hyperemia and/or
discoloration (in some instances), and gastrointestinal inflammation (acute
in some instances). Viscera appeared normal in survivors.
Thirty males and 20 females were divided into 6 groups of 5 rats/sex.
Test condition :
Males were given a single oral dose (undiluted) of propionitrile at 50.1,
63.1, 79.4, 100, 126, or 158 mg/kg, and females were given 158, 200, 251
or 316 mg/kg. The average initial weights of males and females were 237.5
and 242.5 g, respectively. Animals were observed for 14 days. It is
presumed that they were observed daily. Animals that died were
necropsied upon discovery. Survivors were euthanized and necropsied on
day 14. The LD50 value, 95% confidence interval and slope of the curve
for each sex were calculated by an unknown method.
A data sheet containing information about the test material listed the purity
Test substance :
to be 90+%. Impurities included 0.05% acrylonitrile, < 0.1% water, and
0.16% adiponitrile. Additional impurities (other than an illegible one at 510
ppm) were not listed.
(2) valid with restrictions
Reliability :
Test conditions are not described in detail.
10.08.2003 (40)
5.1.2 ACUTE INHALATION TOXICITY
LC50
Type :
= 3.3 mg/l
Value :
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
60
Number of animals :
Vehicle :
1.58, 2.51, 3.98, 6.31, 10.0, 15.8 mg/l
Doses :
4 hour(s)
Exposure time :
other
Method :
1978
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The NOAEL in ppm can be calculated using the following equation [ppm =
Remark :
3300 mg/m3 x 24.45/55.09(MW)]. For this experiment, the value is 1464.6.
The numbers of animals that died after exposure to propionitrile is as
Result :
follows:
27 / 27
Id 107-12-0
5. Toxicity
Date 02.10.2003
Concentration (mg/l) Number of Deaths
1.58 0/5 males, 0/5 females (0/10)
2.51 5/5 males 0/5 females (5/10)
3.98 5/5 males, 0/5 females (5/10)
6.31 5/5 males, 3/5 females (8/10)
10.0 5/5 males, 5/5 females (10/10)
15.8 5/5 males, 5/5 females (10/10)
All deaths occurred within 4 - 24 hours (with the exception of 1 male
exposed to 3.98 mg/l, that died after 13 days).
The LD50 value for males and females combined [with 95% confidence
limits (CL)] was 3.3 (2.6 - 4.3) mg/l. The slope of the curve was 4.1. These
values were not calculated for males and females individually.
Signs of toxicity included salivation, lethargy, increasing weakness, tremors
and convulsions, and collapse. Animals that died had hemorrhagic lungs,
discolored liver (mottled in some) and acute gastrointestinal inflammation.
Necropsies of survivors were normal.
Sixty rats (30/sex) were divided into 6 groups (5/sex/group). Each group
Test condition :
was exposed to 1.58, 2.51, 3.98, 6.31, 10.0 or 15.8 mg/l by inhalation.
During the test, males and females receiving the same concentration were
placed together in 9 inch x 16 inch x 7 inch cages that were suspended in
the middle of 210 liter drum-like chambers that were equipped with
circulating fans. Test material was introduced into the chamber through a
port equipped with a needle and syringe. No supplementary air was
introduced. The average temperature and relative humidity inside the
chambers were 24-25 degrees C and 70%, respectively.
Animals were observed for signs of toxicity during exposure and for 14
subsequent days. Animals that died were necropsied upon discovery. On
day 14, survivors were euthanized and necropsied.
The LC50 value, 95% confidence limits and slope of the curve were
calculated according to the method of DeBeer (reference was not given).
The purity of the material was not listed. However, in a study conducted by
Test substance :
the same laboratory in 1979, a data sheet containing information about the
test material listed the purity to be 90+%. Impurities included 0.05%
acrylonitrile, < 0.1% water, and 0.16% adiponitrile. Additional impurities
(other than an illegible one at 510 ppm) were not listed.
(2) valid with restrictions
Reliability :
Basic data and methodologies are given.
Critical study for SIDS endpoint
Flag :
10.08.2003 (41)
LC100
Type :
= 90.3 mg/l
Value :
rat
Species :
Sprague-Dawley
Strain :
male
Sex :
6
Number of animals :
Vehicle :
90.3 mg/l
Doses :
1.25 hour(s)
Exposure time :
other
Method :
1979
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
All six rats exposed to 90.3 mg/l for 1.25 hours died. The NOAEL in ppm
Remark :
can be calculated using the following equation [ppm = 90300 mg/m3 x
28 / 28
Id 107-12-0
5. Toxicity
Date 02.10.2003
24.45/55.09(MW)]. For this experiment, the value is 40076.87.
A data sheet containing information about the test material listed the purity
Test substance :
to be 90+%. Impurities included 0.05% acrylonitrile, < 0.1% water, and
0.16% adiponitrile. Additional impurities (other than an illegible one at 510
ppm) were not listed.
(4) not assignable
Reliability :
The study was given a reliability rating of 4 because it was not reviewed in
detail.
10.08.2003 (40)
5.1.3 ACUTE DERMAL TOXICITY
LD50
Type :
= 40 mg/kg bw
Value :
rabbit
Species :
New Zealand white
Strain :
male/female
Sex :
40
Number of animals :
Vehicle :
12.5, 25, 50 and 100 mg/kg
Doses :
other: 40 CFR, Part 163.81-2
Method :
1981
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The numbers of animals that died after each of the doses is as follows:
Result :
Dose (mg/kg) Number (and time) of Deaths
12.5 0/10
25 0/10
50 9/10 (4/5 M, 5/5 F), 3-23 hr
100 10/10, 2-7.5 hr
The LD50 values [with 95% confidence limits (CL)] were as follows:
Sex LD50 (mg/kg) 95 % CL (mg/kg)
M/F 40 29 - 51
M 43 24 - 62
F 35 (could not be calculated)
Signs of toxicity included ataxia, convulsions, tremors, respiratory
abnormalities (hyperpnea, hypopnea, dyspnea and arrythmic respiration),
hypoactivity, prostration, hypothermia, nasal or ocular discharge, fecal
staining, nasal discharge, soft stool, reddened nictitating membrane or
reduced food consumption. Most animals in the 12.5 and 25 mg/kg dose
groups were free of significant abnormalities from days 3 to 14.
Necropsies of animals that died revealed reddening of the nictitating
membrane, mottling of the liver, GI abnormalities (reddened walls, black
foci in the mucosa, black material adhered to the mucosa), and red
interstitial walls or intestinal contents. Necropsies of survivors were normal.
Average weights of survivors at days 7 and 14 were generally higher than
initial weights; however a few animals exhibited slight weight losses (0.1 or
29 / 29
Id 107-12-0
5. Toxicity
Date 02.10.2003
0.2 kg). No dose-related differences in weight gain were apparent. Six
and 3 of the animals given 12.5 and 25 mg/kg (respectively) had no dermal
irritation. The other animals in these groups had only slight or very slight
erythema without edema.
Forty animals were acclimated for 38 days, then randomized to four groups
Test condition :
of 5/sex. They were given food and water ad libitum. Animals considered
unsuitable due to poor health or outlying weights were excluded. Pretest
body weights were 2.6 - 3.3 kg for males and 2.5 - 3.6 kg for females.
Approximately 18 hours prior to treatment, the hair on each rabbit was
closely clipped from the trunk (dorsal and ventral surface and sides from
scapular to pelvic area). Skin was not abraded. Test material (12.5, 25.0,
50.0 or 100 mg/kg) was applied directly onto the exposed skin and was
spread evenly over the entire area. A layer of 8-ply gauze was then
wrapped around the animal to cover the application site. This was
wrapped in an impervious plastic sleeve, which was secured with masking
tape. Elizabethan collars were placed on all animals.
Wrappings were removed after 24 hours, and the test site was wiped free
of test material. Skin was scored for irritation according to the method of
Draize 30 minutes after removal of the dressing. Animals were observed
for toxicity at 1, 2, and 4 hours and daily thereafter for 14 days. Animals
were weighed prior to treatment and 7 and 14 days after treatment.
Animals that did not survive for 14 days were weighed and examined
grossly at the time they were found dead. All animals surviving to day 14
were euthanized at this time and examined grossly.
The LD50 values (and 95% confidence intervals) for males, females and
both sexes together were calculated using logarithmic-probit graph paper.
The purity of the test material was 94.5%. It also contained 1.1%
Test substance :
adiponitrile, < 0.1% acrylonitrile, < 0.1% water, < 0.1 % solids and 0.07%
para nitrosodiphenylamine.
(1) valid without restriction
Reliability :
The test was performed according to an established guideline.
Documentation is thorough.
10.08.2003 (4)
LD50
Type :
= 56 mg/kg bw
Value :
rabbit
Species :
New Zealand white
Strain :
male/female
Sex :
40
Number of animals :
Vehicle :
12.5, 25, 50 and 100 mg/kg
Doses :
other: 40 CFR, Part 163.81-2
Method :
1981
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The numbers of animals that died after each of the doses is as follows:
Result :
Dose (mg/kg) Number (and time) of Deaths
12.5 0/10
25 0/10
50 4/10 (3/5 M, 1/5 F), 3.5 - 21.5 hr
100 9/10 (5/5 M, 4/5 F), 4-21 hr
The LD50 values [with 95% confidence limits (CL)] were as follows:
30 / 30
Id 107-12-0
5. Toxicity
Date 02.10.2003
Sex LD50 (mg/kg) 95 % CL (mg/kg)
M/F 56 40 - 72
M 45 19 - 71
F 70 44 - 96
Signs of toxicity included ataxia, convulsions, respiratory abnormalities
(hypopnea, dyspnea and gasping), hypoactivity, prostration, nasal or ocular
discharge, fecal staining or reduced food consumption. Necropsies of
animals that died revealed reddening of the nictitating membrane, red or
black foci or patches in the gastric mucosa, discoloration of liver or kidneys,
red material in the GI tract or thickening of the abdominal wall. Necropsies
of survivors were normal. Average weights of survivors at days 7 and 14
were higher than initial weights and were not affected by dose of test
material. Approximately one half of the survivors at 24 hours had no
dermal irritation. The others had only slight or very slight erythema without
edema.
Forty animals were acclimated for 35 days, then randomized to four groups
Test condition :
of 5/sex. They were given food and water ad libitum. Animals considered
unsuitable due to poor health or outlying weights were excluded. Pretest
body weights were 2.7 - 3.2 kg for males and 2.6 - 3.4 kg for females.
Approximately 18 hours prior to treatment, the hair on each rabbit was
closely clipped from the trunk (dorsal and ventral surface and sides from
scapular to pelvic area). Skin was not abraded. Test material (12.5, 25.0,
50.0 or 100 mg/kg) was applied directly onto the exposed skin and was
spread evenly over the entire area. A layer of 8-ply gauze was then
wrapped around the animal to cover the application site. This was
wrapped in an impervious plastic sleeve, which was secured with masking
tape. Elizabethan collars were placed on all animals.
Wrappings were removed after 24 hours, and the test site was wiped free
of test material. Skin was scored for irritation according to the method of
Draize 30 minutes after removal of the dressing. Animals were observed
for toxicity at 1, 2, and 4 hours and daily thereafter for 14 days. Animals
were weighed prior to treatment and 7 and 14 days after treatment.
Animals that did not survive for 14 days were weighed and examined
grossly at the time they were found dead. All animals surviving to day 14
were euthanized at this time and examined grossly.
The LD50 values (and 95% confidence intervals) for males, females and
both sexes together were calculated using logarithmic-probit graph paper.
The purity of the test material was not listed. It was used as received from
Test substance :
Monsanto. The density was listed as 0.7978 g/ml.
(2) valid with restrictions
Reliability :
Purity of the material and documentation of GLP were not provided.
10.08.2003 (6)
LD50
Type :
= 90 mg/kg bw
Value :
rabbit
Species :
New Zealand white
Strain :
male/female
Sex :
40
Number of animals :
Vehicle :
12.5, 25, 50 and 100 mg/kg
Doses :
other: 40 CFR, Part 163.81-21
Method :
1981
Year :
no data
GLP :
31 / 31
Id 107-12-0
5. Toxicity
Date 02.10.2003
as prescribed by 1.1 - 1.4
Test substance :
The numbers of animals that died after each of the doses is as follows:
Result :
Dose (mg/kg) Number (and time) of Deaths
12.5 0/10
25 0/10
50 1/10 (0/5 M, 1/5 F), 23.5 hr
100 6/10 (3/5 M, 3/5 F), 2.5-23.5 hr
The LD50 values [with 95% confidence limits (CL)] were as follows:
Sex LD50 (mg/kg) 95 % CL (mg/kg)
M/F 90 65 - 115
M 90 49 - 131
F 85 35 - 135
Signs of toxicity included ataxia, convulsions, tremors, respiratory
abnormalities (hyperpnea and dyspnea), hypoactivity, prostration, red eyes,
nasal discharge, soft stool, fecal staining or reduced food consumption.
Most animals in the 12.5 and 25 mg/kg dose groups and most survivors in
the 50 and 100 mg/kg dose groups were free of significant abnormalities
from days 2 to 14. Necropsies of animals that died revealed reddening of
the nictitating membrane, pale liver, and GI abnormalities (reddened walls,
black foci in the mucosa, and/or white film on the mucosa). Necropsies of
survivors were normal.
Average weights of survivors at days 7 and 14 were higher than initial
weights and were not affected by dose of test material. Approximately one
half of the survivors at 24 hours had no dermal irritation. The others had
only slight or very slight erythema, generally without edema.
Forty animals were acclimated for 31 days, then randomized to four groups
Test condition :
of 5/sex. They were given food and water ad libitum. Animals considered
unsuitable due to poor health or outlying weights were excluded. Pretest
body weights were 2.5 - 3.1 kg for males and 2.7 - 3.4 kg for females.
Approximately 25 hours prior to treatment, the hair on each rabbit was
closely clipped from the trunk (dorsal and ventral surface and sides from
scapular to pelvic area). Skin was not abraded. Test material (12.5, 25.0,
50.0 or 100 mg/kg) was applied directly onto the exposed skin and was
spread evenly over the entire area. A layer of 8-ply gauze was then
wrapped around the animal to cover the application site. This was
wrapped in an impervious plastic sleeve, which was secured with masking
tape. Elizabethan collars were placed on all animals.
Wrappings were removed after 24 hours, and the test site was wiped free
of test material. Skin was scored for irritation according to the method of
Draize 30 minutes after removal of the dressing. Animals were observed
for toxicity at 1, 2, and 4 hours and thereafter for 14 days. Animals were
weighed prior to treatment and 7 and 14 days after treatment. Animals that
did not survive for 14 days were weighed and examined grossly at the time
they were found dead. All animals surviving to day 14 were euthanized at
this time and examined grossly.
The LD50 values (and 95% confidence intervals) for males, females and
both sexes together were calculated using logarithmic-probit graph paper.
The purity of the test material was not listed. It was used as received from
Test substance :
Eastman Kodak. The density was listed as 0.7885 g/ml.
(2) valid with restrictions
Reliability :
32 / 32
Id 107-12-0
5. Toxicity
Date 02.10.2003
Purity of the material and documentation of GLP were not provided.
10.08.2003 (5)
5.1.4 ACUTE TOXICITY, OTHER ROUTES
5.2.1 SKIN IRRITATION
5.2.2 EYE IRRITATION
5.3 SENSITIZATION
5.4 REPEATED DOSE TOXICITY
Sub-chronic
Type :
rat
Species :
male/female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
14 weeks (total of 63 exposure days)
Exposure period :
6 hours/day, 5 days per week
Frequency of treatm. :
Post exposure period :
60, 120, 209 ppm
Doses :
yes
Control group :
< 60 ppm
NOAEL :
= 60 ppm
LOAEL :
other
Method :
1984
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Test material concentrations: The nominal concentrations (+/- SD) were
Result :
56.0 +/- 8.8, 119.1 +/- 16.9 and 203.0 +/- 19.6. Corresponding analytical
concentrations were 60.2 +/- 1.0, 120.3 +/- 1.1 and 209.0 +/- 1.3 ppm.
Since the nominal analytical ratios were close to 1, the material was a true
vapor. There was no indication that the test material was unstable.
Distribution was uniform (> 96%) for each of the exposure concentrations.
Airflow, temperature and relative humidity ranged from 1719-1765 l/min,
21.0 - 26.1 degrees C and 17-50%, respectively.
Effects at all exposure concentrations: Signs of toxicity (labored breathing,
nasal discharge, salivation, discharge from the eyes, hypoactivity and/or
alopecia) were observed in all exposed groups. Incidences of these signs
increased in a dose-dependent manner. Males and/or females in all
exposed groups had significant decreases in red blood cells and
hemoglobin values. Urine thiocyanate concentrations increased in all
exposed groups, with concentrations from animals exposed to 120 ppm
similar to or higher than those exposed to 210 ppm. However, since a
dose-dependent diuresis occurred, the total amount of urine thiocyanate
present (concentration x urine volume) increased with increasing
concentrations.
Effects at 209 ppm: Three males died or were killed in extremis (2
between exposures 2 and 3 and one between exposures 18 and 19).
Arched back, ataxia, tremors or convulsions, biting, pawing or rubbing chin
33 / 33
Id 107-12-0
5. Toxicity
Date 02.10.2003
against cage, irritation of the conjunctiva and breathing difficulties were
noted in a few animals during exposure. Males and females exhibited
significant decreases in body weight throughout the study (P <= 0.01).
Average final body weights of males and females were lower than controls
(377.5 g in exposed males vs. 435.4 g in controls and 255.0 g in exposed
females vs. 276.6 g in controls). Absolute and/or relative heart, liver, spleen
and kidney weights were increased in males and/or females. Absolute
testes weights were decreased in males. Mean corpuscular hemoglobin
concentrations (males only) were lower than control (all P <= 0.05). Serum
alkaline phosphatase (males and females), SGOT (males only) and SGPT
(males only) were increased, and BUN concentrations were decreased. No
gross lesions were observed at termination. There was mildly increased
hemosiderin deposition in the spleens of 10/15 females. Other microscopic
changes were considered to be spontaneous.
Effects at 120 ppm: Ataxia was observed during exposure in 2 females.
Males had significant body weight decreases (P < = 0.05) at two time
points. Absolute and/or relative liver weights were increased in males and
females and absolute and/or relative spleen weights were increased in
males. Mean corpuscular hemoglobin concentrations (males only, P < =
0.05) were lower than control. No gross lesions were observed at
termination. There was mildly increased hemosiderin deposition in the
spleens of 11/15 females. Other microscopic changes were considered to
be spontaneous.
Effects at 60 ppm: Absolute and relative spleen weights were increased in
males. Serum thiocyanate concentrations were marginally increased in
males and females.
Animals: Rats were acclimated for at least 10 days prior to use. Two days
Test condition :
prior to the start of the study, males weighed 174-200 g and females
weighed 132-145 g. On the first day of the study, the animals were 43
days old. Animals were randomly allocated by body weight into 4 groups of
15 animals/sex/group. Animals were individually housed in suspended
mesh cages and given food and water ad libitum (except during exposure).
Animal rooms were maintained at 70-74 degrees C and 35-60% relative
humidity, with a 12 hour light/dark cycle.
Exposure conditions: Exposures (6 hr/day, 5 days/week) occurred in 10
m3 Rochester-style stainless steel and glass inhalation chambers. Rats
were placed individually in wire mesh cages that were suspended in the
chambers by 3-tiered racks. Males were placed on one side and females
on the other. The concentrations of material in the chambers (20, 120 or
210 ppm) were controlled by either adjusting the nitrogen flow though the
propionitrile in the bubblers or by changing the amount of test material in
the bubblers. The bubbler was connected to a side port in the vertical
particle-size separator, which was in turn connected to the air inlet at the
top of the inhalation chamber. One bubbler was used in each of the
generation systems. Airflow was maintained at a constant flow of 1727
liters/min. Nominal concentration measurements were determined daily for
each chamber following exposure, by dividing the amount of test material
delivered to the chamber (the difference between the pre- and post-
exposure weights) over the 6-hr exposure period by the total air volume
during the same period. Concentrations of test material in the chambers
were measured 4 times daily using a Miran 1A General Purpose Gas
Analyzer. Additional samples of atmosphere from 9 specified locations in
each chamber were also taken at 3 different times to determine if the vapor
was distributed uniformly.
Test conduct: Animals were observed for clinical signs between the second
and fifth hour of each exposure. Estimations of the percentages of animals
exhibiting hypoactivity, eye irritation and breathing difficulties were made.
All animals were individually examined for gross signs of toxicity preceding
34 / 34
Id 107-12-0
5. Toxicity
Date 02.10.2003
and following each exposure and checked for mortality. Each animal was
weighed and given a thorough examination for gross signs of toxicity on a
weekly basis.
Animals were euthanized after 14 total weeks on the study. Terminal body
weights were obtained (following an overnight fast). Blood and urine were
collected. Whole blood was treated with an anticoagulant and was
analyzed for total and differential erythrocyte count, total leukocyte count,
platelet count, hematocrit, hemoglobin, and red blood cell indices (mean
corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular
hemoglobin concentration). Serum was analyzed for albumin, globulin, total
protein, blood urea nitrogen, total bilirubin, glucose, glutamic pyruvic
transaminase (SGPT), alkaline phosphatase, glutamic oxaloacetic
transaminase (SGOT), T3, T4, thiocyanate and lactate dehydrogenase.
Urine was analyzed for the presence of thiocyanates.
Detailed necropsies were conducted on all rats that died during the course
of the study, those that were killed moribund, and those that survived to
study termination. The adrenal glands (both together), testes (with
epididymides, heart, kidneys, liver, pituitary and spleen were weighed. The
aforementioned organs and the following tissues were fixed in 10% neutral
formalin: abdominal aorta, bone and bone marrow (femur), brain,
esophagus, ovaries, colon, ileum, lung, lymph nodes (mesenteric),
mammary gland, nasal turbinates, pancreas, thyroid/parathyroid, prostate,
salivary gland, sciatic nerve, skeletal muscle, skin, spinal cord, stomach,
thymus, trachea, urinary bladder, uterus (with cervix) and gross lesions.
Eyes (with optic nerve) were fixed in a solution of 2% glutaraldehyde and
10% neutral buffered formalin. Tissues were processed, embedded in
paraffin, cut at five microns, stained with hematoxylin and examined
microscopically.
Statistical analyses: In life and terminal body weights and organ weight
data were analyzed using Dunnett's test. Organ to body weight ratios were
analyzed using the Mann-Whitney test, with the Bonferroni inequality. Data
for frequencies of microscopic lesions were evaluated with the Fisher's
exact test with the Bonferroni inequality. Hematological and serum and
urine chemistry variables were examined using Dunnett's test.
The purity of the test material was 96%. Impurities were not listed.
Test substance :
(2) valid with restrictions
Reliability :
The study is comparable to a guideline study; however, a NOAEL was not
established.
Critical study for SIDS endpoint
Flag :
10.08.2003 (35)
Sub-acute
Type :
rat
Species :
male/female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
18 days
Exposure period :
6 hr/day, 5 days per week
Frequency of treatm. :
none
Post exposure period :
44, 115, 329 ppm
Doses :
yes
Control group :
= 115 ppm
NOAEL :
= 329 ppm
LOAEL :
other
Method :
1981
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Thirteen males and one female died. High dose animals exhibited
Result :
35 / 35
Id 107-12-0
5. Toxicity
Date 02.10.2003
decreases in mean body weight, ruffled fur, irritability, tremors, and
discharges around the nose and eyes. Hematological measurements were
also depressed in high dose animals. No signs of toxicity or changes in
hematological measurements were observed in animals exposed to 44 or
115 ppm. There was no effect of treatment on clinical chemistries or
urinalyses. No treatment-related gross or microscopic lesions were
observed.
Groups of 18 animals/sex were exposed to 0 (control) 44, 115 or 329 ppm,
Test condition :
6 hr/day for 18 days.
Purity of the test material was 94.87%.
Test substance :
(4) not assignable
Reliability :
The study was given a reliability rating of 4 because it was not reviewed in
detail.
10.08.2003 (30)
5.5 GENETIC TOXICITY `IN VITRO`
Mammalian cell gene mutation assay
Type :
L5178Y Mouse Lymphoma Cells
System of testing :
2143 to 5000 micrograms/ml
Test concentration :
> 5000 micrograms/ml
Cytotoxic concentr. :
without
Metabolic activation :
negative
Result :
other
Method :
1982
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
An initial toxicity test showed that complete toxicity was not reached at
Remark :
5000 micrograms/ml.
First study: After the 2 day expression period, ten cultures (2 per
Result :
concentration) were cloned at 2714, 3286, 3857, 4429, and 5000
micrograms/ml. All cultures exhibited mutant frequencies similar to
controls (ranged from 0.2 - 0.4 per 10E4 surviving cells) and the total
growth ranged from 61 to 106%.
Second study: After the 2 day expression period, twelve cultures (2 per
concentration) were cloned at 2143, 2714, 3286, 3857, 4429, and 5000
micrograms/ml. All cultures exhibited mutant frequencies similar to
controls (ranged from 0.2 - 0.3 per 10E4 surviving cells) and the total
growth ranged from 69 to 94%.
Based on the combined results of both assays, none of the test
concentrations induced a positive response (all p values were > 0.01) .
The linear component of the dose-response curve also was not statistically
significant.
The test was considered valid, as the control frequencies were within the
required range (0.2 - 0.3 per 10E4 surviving cells in the two studies) and
EMS induced 9.2 and 7.6 mutants per 10E4 surviving cells at 0.5
microliters/ml and 28.4 and 24.5 mutants per 10E4 surviving cells at 0.5
microliters/ml and 1.0 microliters/ml, in the 2 studies, respectively.
Cell preparation: Prior to use in the test, L5178Y cells that were actively
Test condition :
growing in culture were cleansed as described by Clive et al. (Mutat Res
31:17-29, 1975). Three ml of thymidine, hypoxanthine, methotrexate and
glycine (THMG) stock solution was added to 100 ml cell suspension
containing 0.1 x 10E6 cells/ml. The culture was gassed with 5% CO2 in air
and placed in a shaking incubator (125 rpm, 37 degrees C). After 24
hours, the THMG was removed by pelletizing the cells and decanting off
the supernatant. The cells were rinsed in 20 ml of F10 P (Fisher's Media for
36 / 36
Id 107-12-0
5. Toxicity
Date 02.10.2003
leukemic cells with 10% heat inactivated horse serum) and reinstated in
culture at 3 x 10E 4 cells/ml in 100 ml of F10 P plus 1 ml of THMG stock
solution.
Test conduct: The test material was solubilized and diluted to produce
dose levels from 2714 to 5000 micrograms/ml. Each concentration was
tested in duplicate, and the test was performed twice. An additional
concentration of 2143 micrograms/ml was tested in one of the experiments.
All stock solutions were freshly prepared. Four ml of FoP was added to
each tube. This yielded a final cell suspension of 0.6 x 10E6 cells/ml. Two
control tubes received solvent only (DMSO). Positive controls were treated
with ethyl methane sulfate (1.0 and 0.5 microliters/ml). All tubes were
gassed with 5% CO2 in air and placed on a roller drum apparatus for 4
hours at 37 degrees C. All steps were carried out under amber lighting and
the cells were incubated in the dark for 4 hours. After 4 hours, the cells
were washed twice with 10 ml of F10P, then resuspended in 20 ml of F10P
gassed with 5% CO2 in air, and replaced on a roller drum apparatus at 37
degrees C.
After the initial exposure, the cells were incubated for 2 days with a cell
population adjustment (to 0.3 x 10E6 cells/ml) at 24 and 48 hours. At the
end of this expression period, cells (a 2 x 10E-2 dilution of 1.5 x 10E6
cells/ml) were placed in 100 ml cloning medium containing 0.37% noble
agar, and incubated in a shaking incubator at 37 degrees C.
Trifluorothymidine (TFT; final concentration of 3 micrograms/ml) was added
to one of the duplicate flasks per concentration. After 15 minutes, the flasks
were removed, and 33 ml of the cell suspension from each flask was
pipetted into 3 100 mm Petri plates. To accelerate the gelling process, the
plates were stored at 4 degrees C for 20 min. The plates were then
removed and incubated at 37 degrees C in a humidified 5% CO2
atmosphere for 10-12 days.
After the incubation period, the plates were scored for total number of
colonies per plate. The plates that did not contain TFT served as viability
controls. Each plate was counted 3 times by an automatic colony counter,
and the median count was recorded. The mutation frequency was
determined by dividing the average number of colonies in the 3 TFT plates
by the average number of colonies x 10E4 in the 3 viability control plates
and multiplying the quotient by 2.
Validity criteria: The test was considered acceptable if the positive control
induced at least a 2-fold increase in the frequency of mutants with respect
to the solvent control and resulted in a viability rate of 10-80%. The solvent
control frequency also had to be 0.2 - 1 per 10E 4 cells, the plating
efficiency of the control at least 50%, and the material tested to either 500
micrograms/ml, or at the limit of 10% viability or solubility. A test was
considered positive if at least 2 concentrations that caused no more than
90% toxicity caused 2-fold increases in the frequency of mutants with
respect to solvent controls and the response was concentration-dependent
(p < 0.01) for at least 2 concentrations that did not cause > 90%
cytotoxicity. A test was considered negative if none of the concentrations
caused a significant increase in the frequency of mutations (p > 0.01) and
the linear component of the dose-response curve was not significant (p >
0.1) for test concentrations resulting in at least 10% relative total growth.
The test material was propionitrile, used as received from Kodak. The
Test substance :
purity was listed as 97.8%. Impurities included adiponitrile (0.3%),
paranitrosophenylamine (0.1%), water (0.1%), acrylonitrile (<0.1%), and
solids (<0.1%).
(1) valid without restriction
Reliability :
The study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (27)
37 / 37
Id 107-12-0
5. Toxicity
Date 02.10.2003
Mammalian cell gene mutation assay
Type :
L5178Y Mouse Lymphoma Cells
System of testing :
100 to 1457 micrograms/ml
Test concentration :
1457 micrograms/ml
Cytotoxic concentr. :
without
Metabolic activation :
positive
Result :
other
Method :
1982
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
An initial toxicity test showed that complete toxicity was caused by 5000
Remark :
micrograms/ml. Based on this test, the maximum concentration tested in
the mutagenesis study was 1186 micrograms/ml. In the second
mutagenicity test, one plate at 100 micrograms/ml and one solvent control
were lost due to human error. This did not have a bearing on the outcome
of the test.
First study: After the 2 day expression period, ten cultures (2 per
Result :
concentration) were cloned at 100, 371, 643, 914, 1186 and 1457
micrograms/ml. All cells incubated with 1457 micrograms/ml died. The
cultures cloned at 643, 914 and 1186 micrograms/ml exhibited mutant
frequencies greater than 2 times the average number in the vehicle
controls (1.0, 1.7 - 1.9, and 2.8 - 4.0 per 10E4 surviving cells, respectively).
The % total growth of these cultures ranged from 3 to 37% (toxicity
increased with increasing concentration). The four remaining cultures
exhibited mutant frequencies similar to solvent controls, with a total growth
range of 71% to 87%.
Second study: After the 2 day expression period, ten cultures (2 per
concentration) were again cloned at 100, 371, 643, 914, 1186 and 1457
micrograms/ml. All cells incubated with 1457 micrograms/ml died. The
cultures cloned at 643, 914 and 1186 micrograms/ml exhibited mutant
frequencies greater than 2 times the average number in the vehicle
controls (1.0, 1.6 - 1.9, and 2.2 - 2.4 per 10E4 surviving cells, respectively).
The % total growth of these cultures ranged from 4 to 38% (toxicity
increased with increasing concentration). The four remaining cultures
exhibited mutant frequencies similar to solvent controls, with a total growth
range of 60% to 83%.
The combined statistical analysis of the 2 studies indicated a positive
response. The mean mutation frequencies of cultures cloned at 643 and
914 micrograms/ml (each with greater than 10% relative growth), are
significantly greater than that of the solvent control (p < 0.01). The linear
component of the dose-response relationship also was statistically
significant (p < 0.01) and exhibited a positive slope for test concentrations
with at least 10% relative total growth.
The test was considered valid, as the control frequencies were within the
required range (0.3 - 0.4 per 10E4 surviving cells in the two studies) and
EMS induced 9.2 and 7.3 mutants per 10E4 surviving cells at 0.5
microliters/ml and 28.4 and 38.4 mutants per 10E4 surviving cells at 0.5
microliters/ml and 1.0 microliters/ml, in the two studies, respectively.
Cell preparation: Prior to use in the test, L5178Y cells that were actively
Test condition :
growing in culture were cleansed as described by Clive et al. (Mutat Res
31:17-29, 1975). Three ml of thymidine, hypoxanthine, methotrexate and
glycine (THMG) stock solution was added to 100 ml cell suspension
containing 0.1 x 10E6 cells/ml. The culture was gassed with 5% CO2 in air
and placed in a shaking incubator (125 rpm, 37 degrees C). After 24
hours, the THMG was removed by pelletizing the cells and decanting off
the supernatant. The cells were rinsed in 20 ml of F10 P (Fisher's Media for
leukemic cells with 10% heat inactivated horse serum) and reinstated in
38 / 38
Id 107-12-0
5. Toxicity
Date 02.10.2003
culture at 3 x 10E 4 cells/ml in 100 ml of F10 P plus 1 ml of THMG stock
solution.
Test conduct: The test material was solubilized and diluted to produce
dose levels from 100 to 1457 micrograms/ml. Each concentration was
tested in duplicate, and the test was performed twice. All stock solutions
were freshly prepared. Four ml of FoP was added to each tube. This
yielded a final cell suspension of 0.6 x 10E6 cells/ml. Two control tubes
received solvent only (DMSO). Positive controls were treated with ethyl
methane sulfate (1.0 and 0.5 microliters/ml). All tubes were gassed with
5% CO2 in air and placed on a roller drum apparatus for 4 hours at 37
degrees C. All steps were carried out under amber lighting and the cells
were incubated in the dark for 4 hours. After 4 hours, the cells were
washed twice with 10 ml of F10P, then resuspended in 20 ml of F10P
gassed with 5% CO2 in air, and replaced on a roller drum apparatus at 37
degrees C.
After the initial exposure, the cells were incubated for 2 days with a cell
population adjustment (to 0.3 x 10E6 cells/ml) at 24 and 48 hours. At the
end of this expression period, cells (a 2 x 10E-2 dilution of 1.5 x 10E6
cells/ml) were placed in 100 ml cloning medium containing 0.37% noble
agar, and incubated in a shaking incubator at 37 degrees C.
Trifluorothymidine (TFT; final concentration of 3 micrograms/ml) was added
to one of the duplicate flasks per concentration. After 15 minutes, the flasks
were removed, and 33 ml of the cell suspension from each flask was
pipetted into 3 100 mm Petri plates. To accelerate the gelling process, the
plates were stored at 4 degrees C for 20 min. The plates were then
removed and incubated at 37 degrees C in a humidified 5% CO2
atmosphere for 10-12 days.
After the incubation period, the plates were scored for total number of
colonies per plate. The plates that did not contain TFT served as viability
controls. Each plate was counted 3 times by an automatic colony counter,
and the median count was recorded. The mutation frequency was
determined by dividing the average number of colonies in the 3 TFT plates
by the average number of colonies x 10E4 in the 3 viability control plates
and multiplying the quotient by 2.
Validity and positive assay criteria: The test was considered acceptable if
the positive control induced at least a 2-fold increase in the frequency of
mutants with respect to the solvent control and resulted in a viability rate of
10-80%. The solvent control frequency also had to be 0.2 - 1 per 10E 4
cells, the plating efficiency of the control at least 50%, and the material
tested to either 500 micrograms/ml, or at the limit of 10% viability or
solubility. A test was considered positive if at least 2 concentrations that
caused no more than 90% toxicity caused 2-fold increases in the frequency
of mutants with respect to solvent controls and the response was
concentration-dependent (p< 0.01) for at least 2 concentrations that did not
cause > 90% cytotoxicity.
The test material was "propionitrile tails", used as received from Monsanto.
Test substance :
This contained propionitrile (97.0%), adiponitrile (1.2%),
paranitrosophenylamine (0.12%), water (0.04%), and acrylonitrile (0.02%).
(1) valid without restriction
Reliability :
The test is comparable to a guideline study.
10.08.2003 (28)
Ames test
Type :
S. typhimurium strains TA98, TA100, TA1535 and TA1538
System of testing :
up to 10,000 micrograms per plate
Test concentration :
> 10,000 micrograms per plate
Cytotoxic concentr. :
with and without
Metabolic activation :
negative
Result :
39 / 39
Id 107-12-0
5. Toxicity
Date 02.10.2003
other
Method :
1977
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The authors stated that the test was negative, even though there was a
Remark :
positive response with metabolic activation at one concentration in one
strain.
Experiment 1: There was no effect of treatment with test material on the
Result :
number of revertants in any strain in the absence or presence of S-9. The
numbers of revertants per plate in control strains TA98, TA100, TA1535
and TA1538 without microsomes were 13, 42, 11 and 11, respectively.
The numbers of revertants in strains TA98, TA100, TA1535 and TA1538
treated with 5 to 1000 micrograms/plate test material without microsomes
ranged from 3-12, 24-47, 6-19 and 5-9, respectively. The numbers of
revertants per plate in control strains TA98, TA100, TA1535 and TA1538
with microsomes were 47, 47, 11 and 40, respectively. The numbers of
revertants in strains TA98, TA100, TA1535 and TA1538 treated with 5 to
1000 micrograms/plate test material with microsomes ranged from 33-50,
24-44, 7-13 and 12-39, respectively. The positive controls induced 11, 104
and 26 revertants per plate in strains TA98, TA100 and TA1538 without
microsomes and 358, 832 and 202 revertants/plate with microsomes.
Experiment 2: There was no effect of treatment with test material on the
number of revertants in any strain in the absence of S-9. The numbers of
revertants per plate in control strains TA98, TA100, TA1535 and TA1538
without microsomes were 7, 37, 7 and 8, respectively. The numbers of
revertants in strains TA98, TA100, TA1535 and TA1538 treated with 2500
to 10000 micrograms/plate test material without microsomes ranged from
4-7, 3-36, 7-12 and 4-11, respectively. The numbers of revertants per plate
in control strains TA98, TA100, TA1535 and TA1538 with microsomes
were 36, 34, 5 and 26, respectively. The numbers of revertants in strains
TA98, TA100, and TA1538 treated with 2500 to 10000 micrograms/plate
test material with microsomes ranged from 28-41, 25-31, and 19-31,
respectively. In strain 1535 in the presence of microsomes, 4, 555, 7 and 9
revertants/plate were found after incubation with 2500, 500, 7500 and
10000 micrograms/plate. The response at 5000 micrograms/plate
appeared to be an aberration, since it was not dose-dependent. The
positive controls induced 15, 1011 and 20 revertants per plate in strains
TA98, TA100 and TA1538 without microsomes and 331, 1395 and 299
revertants/plate with microsomes.
The enzyme controls and histidine response check all gave predicted
responses.
Two plate tests were performed - one with 5, 10, 25, 50, 100, 500 and 1000
Test condition :
micrograms/plate and another with 2500, 5000, 7500 and 10000
micrograms/plate. The vehicle for the test material was water. Both tests
were conducted in the presence and absence of liver microsome
preparations from Aroclor-induced rats (sex not stated). The positive
controls for strains TA98, TA100, TA1535 and TA1538 were
benzo(a)pyrene, 2-aminoanthracene, diethyl sulfate, and 2
aminoanthracene, respectively (doses were not stated). Enzyme controls
and histidine response checks also were performed.
(4) not assignable
Reliability :
Purity of the test material was not given. Only 4 strains were tested.
Numerical results for the positive control in strain TA1535 were not given
(only listed as a positive sign). Documentation is limited. It appears that
the assay was not performed in triplicate. The positive control for TA98
without microsomes did not induce an increase in revertants in the first
experiment. Criteria for a positive test were not mentioned. The test that
gave a positive result at one concentration should have been repeated.
10.08.2003 (19)
40 / 40
Id 107-12-0
5. Toxicity
Date 02.10.2003
Unscheduled DNA synthesis
Type :
Primary rat liver cell cultures
System of testing :
up to 2500 micrograms/ml
Test concentration :
5000 micrograms/ml
Cytotoxic concentr. :
Metabolic activation :
negative
Result :
other
Method :
1985
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The hepatocytes would be expected to metabolize the material, due to the
Remark :
presence of cytochrome p450 and other enzymes in the cells.
A positive control (2-acetylaminofluorene) which is known to induce
unscheduled DNA synthesis after metabolic conversion to its active form
was positive in the test.
Purity of the test material was 97 +/- 1%.
Test substance :
(4) not assignable
Reliability :
The study is given a reliability rating of 4 because it was not reviewed in
detail.
07.08.2003 (33)
5.6 GENETIC TOXICITY `IN VIVO`
Cytogenetic assay
Type :
rat
Species :
male/female
Sex :
Sprague-Dawley
Strain :
gavage
Route of admin. :
6, 24 or 48 hours
Exposure period :
0, 100 and 200 mg/kg
Doses :
negative
Result :
other
Method :
1985
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The doses used in this study were based on a preliminary study performed
Remark :
with 30, 100, 300, 500 and 750 mg/kg, which showed abnormal clinical
signs and loss of body weight at doses > = 100 mg/kg, and no effect on
mitotic index at 100 or 300 mg/kg. Initially, 200 mg/kg was chosen as the
dose for both males and females. However, since 9/15 males administered
200 mg/kg died within 24 hours, the dose in males was reduced to 100
mg/kg.
The number of metaphases analyzed in control animals at each time point
Result :
ranged from 475-500 cells (from a total of 10 animals). Two hundred and
fifty metaphases from 5 animals were analyzed at each time point for
animals treated with propionitrile (with the exception of 100 cells isolated at
48 hours from 2 females treated with 200 mg/kg). There was no effect of
treatment with propionitrile on the frequency of chromosomal aberrations (0
- 0.4 % aberrant cells per group in treated vs. 0 - 0.2 % in controls and 0 -
0.004 aberrations per cell in treated vs. 0 - 0.002 in control), mean
chromosome number (treated ranged from 41.74 - 41.84 and control
ranged from 41.75 - 41.83) or mitotic index (treated ranged from 0.52 - 2.40
and control ranged from 1.20 - 2.48) at any time point. A significant
increase in the percentage of aberrant cells (25.52 in treated vs. 0 in
control) and average number of aberrations per cell (1.401 in treated vs. 0
41 / 41
Id 107-12-0
5. Toxicity
Date 02.10.2003
in control), and a decrease in mitotic index (0.28 vs. 1.20 in control) was
observed in cells from animals treated with cyclophosphamide (the positive
control).
Three females treated with 200 mg/kg died. All males and females treated
with propionitrile exhibited signs of toxicity. Males exhibited depression,
red stains on nose/eyes, soft feces, dilated pupils and urine stains, and
females exhibited depression, cold to touch, red stains on nose/eyes,
wheezing, urine stains and tremors. Reduced body weights were observed
in treated males at 24 hours and females at 24 and 48 hours.
Eighty one Sprague-Dawley rats/sex (approximately 46-51 days old) were
Test condition :
acclimated for 19 days prior to treatment. Food and water was supplied ad
libitum. Eighty animals/sex were randomized and 35 per sex were assigned
to the study. A single dose of test material (10 ml/kg, corrected for 100%
purity) was administered by oral gavage to 2 groups of 15 male rats each
at 0 (corn oil vehicle control) or 100 mg/kg and 2 groups of 15 females at 0
(vehicle control) or 200 mg/kg. Five animals per sex were euthanized at
approximately 6, 24 and 48 hours after administration of test material. An
additional 5 animals per sex were given the positive control
cyclophosphamide (40 mg/kg) and euthanized 24 hours after treatment.
Animals were observed twice daily for general appearance, behavior, and
clinical signs. Body weights of all animals were recorded just prior to
administration of test material and just prior to colchicine administration (for
animals that were to be killed 24 and 48 hours after treatment).
Approximately 4, 22 and 46 hours after test material was given, the
appropriate groups of animals received a single intraperitoneal injection of
colchicine (2.0 mg/kg body weight, dosing factor 5 ml/kg) to inhibit mitosis
and arrest cells in metaphase. The colchicine was dissolved in Hank's
Balanced Salt Solution (HBSS). Animals were euthanized approximately 2
hours after colchicine injection.
Bone marrow cells were collected from both femurs of each animal by
aspiration into 5 ml of HBSS heated to 37 degrees C. The aspirate was
spun in a centrifuge for 5 minutes at approximately 1100 rpm. The
supernatant was decanted and 5.0 ml of preheated 0.075 M KCl was
added to each tube. After 25 minutes, five drops of freshly prepared
fixative (methanol:acetic acid, 3:1) were added to each tube. The tubes
were capped, inverted and spun in a centrifuge for 5 minutes at 1100 rpm.
The cells were resuspended in 5 ml of fixative, and again spun in a
centrifuge for 5 min at 1100 rpm. This procedure was repeated two more
times, and the cells were suspended in fresh fixative and refrigerated. After
chilling, the cells were spun in a centrifuge at 1100 rpm for 5 minutes, the
supernatant was decanted, and the cells were resuspended in 0.5 - 2 ml
fresh fixative. Several drops of the final cell suspension were dispersed
onto microscope slides and air-dried. Two to four slides were made per
animal and were marked with the animal's identification number.
The cells on the slides were stained in fresh Giemsa for 10 minutes, rinsed
twice with distilled water, air-dried, and mounted with glass coverslips.
Code numbers were then blindly assigned to the slides. The slides were
not decoded until all had been analyzed.
At least 50 cells in metaphase were analyzed per animal (if possible).
Otherwise, as many spreads as possible were analyzed. The slides were
scanned with a low power objective (10 or 25 X) and the chromosomes
were analyzed with a high power oil immersion lens (100X). Only those
cells in metaphase were analyzed for cytogenetic abnormalities. The
following items were recorded for each animal: numbers and types of
42 / 42
Id 107-12-0
5. Toxicity
Date 02.10.2003
chromosome aberrations (chromatid and chromosome breaks, chromatid
and chromosome gaps, exchanges, cells with >= 10 aberrations, and
pulverized cells), mitotic index, chromosome number for each metaphase
and the vernier location of each metaphase containing damage.
The mean mitotic indices, chromosome numbers, percent aberrant cells
and the mean number of aberrations per cell for each group were
statistically compared using the Kruskal-Wallis nonparametric analysis of
variance and nonparametric pairwise group comparisons. Body weight
data were analyzed by the analysis of covariance. All tests were evaluated
at the one-tailed 95% confidence interval (p < 0.05).
Purity of the test material was 95.7 %. Impurities included 3.0%
Test substance :
acrylonitrile, 0.9% adiponitrile, and 0.1% solids (not identified).
(2) valid with restrictions
Reliability :
Only two animals were available for analysis at the highest dose tested
(200 mg/kg).
07.08.2003 (21)
5.7 CARCINOGENICITY
5.8.1 TOXICITY TO FERTILITY
Fertility
Type :
rat
Species :
female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
21 to 33 days (depending on day of mating)
Exposure period :
6 hr/day, 7 days/week
Frequency of treatm. :
Premating exposure period
0 days
Male :
21 days
Female :
to gestation days 13-15
Duration of test :
No. of generation :
studies
60, 120 and 210 ppm
Doses :
yes
Control group :
= 60 ppm
NOAEL parental :
= 210 ppm
other: NOAEL :
Reproductive Toxicity
other
Method :
1984
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Exposure concentrations: The average mean daily analytical exposure
Result :
concentrations (60.1, 120.2 and 209.2 ppm) were within 1% of the target
levels (60, 120 and 210 ppm, respectively). Mean temperatures and
humidities ranged from 22-25 degrees C and 26-29%, respectively.
Signs of toxicity: None of the animals died. There was no effect of test
material on body weight. Animals exposed to 210 ppm exhibited arched
back (N = 4 on days 1-10 and N=2 on days 11-20), lacrimation (N = 2 on
days 1-10 and N = 1 on days 21-30), salivation (N= 15 on days 1-10, N =
22 on days 11-20 and N = 21 on days 21-30) hypoactivity (N = 13 on days
1-10, N = 5 on days 11-20 and N = 3 on days 21-30), staining of facial fur
(N = 2 on days 1-10, N = 4 on days 11-20 and N = 4 on days 21-30) and
red nasal encrustation (N = 1 on days 1-10, N = 5 on days 11-20 and N = 5
43 / 43
Id 107-12-0
5. Toxicity
Date 02.10.2003
on days 21-20) after exposure. Animals exposed to 120 ppm also exhibited
salivation (N = 6 on days 11-20 and N = 4 on days 21-30, staining of facial
fur (N = 7 on days 1-10, N = 5 on days 11-10 and N = 2 on days 21-30)
and red nasal encrustation (N = 2 on days 1 1-0, N = 8 on days 11-20 and
N = 6 on days 21-30). A few animals in the 60 ppm group also exhibited
red nasal encrustation (N = 1 on days 1-10, N = 3 on days 11-20 and N = 1
on days 21-30) and staining of facial fur (N = 1 on days 1-10, N = 3 on days
11-20 and N = 1 on days 21-20). One control animal had stained facial fur
on days 21-30 and another had red nasal encrustation on days 1-10. Signs
of toxicity generally abated by the morning after exposure. Alopecia was
observed in animals (N = 2 controls, N = 3 low dose, N = 5 mid dose, N =
9) at one or more of their weekly physical examinations.
The only remarkable findings at gross necropsy were bilateral uterine
hydrometra in one animal exposed to 210 ppm and hydrometra in the left
uterine horn of one animal exposed to 120 ppm.
Fertility: There was no effect of treatment on fertility. Efficiency of mating
(32.0%, 32.0%, 30.7% and 25.0% in the control, low, mid and high dose
groups) and pregnancy rate (100%, 95.8%, 100% and 91.3% in the
respective groups) were comparable between groups. There was no
difference in the numbers of live implants (ranged from 13.4 - 13.9),
resorptions (ranged from 0.6 - 0.8), nidations (ranged from 14.1 - 14.5),
corpora lutea (ranged from 13.0 - 15.2), preimplantation loss (4-8%) and
postimplantation loss (4-6%). Evaluation of the vaginal smears of 2 females
that did not copulate showed one that did not cycle (but was pregnant at
necropsy), and another that only went through the cycling stage of
proestrus.
Animals: Virgin female Sprague Dawley rats (43 days old upon receipt) and
Test condition :
virgin male Sprague Dawley rats (50 days old upon receipt) were
acclimated for one week and examined for general health and the
presence of pinworms and ectoparasites. Body weights of ten females and
ten males that were taken upon receipt were 128-144 g and 178-233 g,
respectively. No significant health problems were noted during the
acclimation period, and the animals were released for study. Males and
females were individually caged (except during mating). Food and water
were available ad libitum (except food was not available to females during
exposure). Animals were housed in a room maintained at 72 +/- 2 degrees
F and 40-60% relative humidity, under a 12 hr light/dark cycle.
Exposures: Exposures (6 hr/day, 7 days/week) occurred in 10 m3
Rochester-style stainless steel and glass inhalation chambers. Due to
inclement weather and building equipment failures, 2 exposures (days 2
and 16) were only for 4 hours and one exposure (day 1) was for 5 hours.
Atmospheres of propionitrile vapor were generated using bubbler systems.
The concentrations of material in the chambers were controlled by either
adjusting the nitrogen flow though the propionitrile in the bubblers or by
changing the amount of test material in the bubblers. Concentrations of
test material in the chambers were measured 4 times daily (except during
day 16) using a Miran 1A General Purpose Gas Analyzer. The nominal
concentration of material, temperature and humidity also were measured
(at intervals that were not listed).
Study conduct: Twenty four females per group were assigned to be
exposed to target concentrations of 0, 60, 120 or 210 ppm test material.
Exposure began when females were 63 days old. Animals were observed
during exposure for signs of toxicity. After 21 days of exposure (which was
sufficient to cover 3-4 estrus cycles), females were randomly mated (1:1) to
an untreated male that had been assigned to the corresponding treatment
group (30 males were assigned per group). At night, after exposure,
females were caged with their assigned male until copulation was
confirmed (by presence of a copulatory plug or sperm in vaginal smear) or
44 / 44
Id 107-12-0
5. Toxicity
Date 02.10.2003
5 nights without confirmed copulation. Females that failed to mate with the
assigned male were mated with another male that had copulated with
another female in the same group. Nightly co-housing with the second
male occurred until copulation was confirmed (or for a maximum of 7
nights). The day on which copulation was confirmed was considered
gestation day 0. Exposure of females continued until copulation was
confirmed or a maximum of 12 nights of cohabitation with males without
signs of copulation. Vaginal smears were taken on 5 consecutive days for
females that did not exhibit copulation.
After assignment to the study, males and females were weighed once per
week. Mated females were weighed on gestation days 0 and 13. Females
were given a thorough physical examination once per week and were
observed for clinical signs of toxicity before and after exposures. All
animals were checked twice daily for mortality and gross abnormalities.
Females were killed on gestation day 13 (or the nearest working day after
gestation day 13, up to gestation day 15). Females without confirmed
copulation ware euthanized in the second week after the last day of co-
housing. Each female was given an external examination and weighed.
The tissues and organs of the thoracic and abdominal cavities were
examined for gross lesions. Pregnancy status was determined, nidation
sites and were classified, and corpora lutea were counted. The ovaries
and uteri of females were preserved in 10% neutral buffered formalin.
Males were killed after mating and were not examined.
Statistical analyses: Body weight data were analyzed using Dunnett's test.
Mating and pregnancy rate data were analyzed by a Fisher's exact test and
an uncorrected chi-square test. Other data were analyzed by the Mann-
Whitney U test. The Bonferroni inequality was assumed when comparing
multiple treatments to control values for all tests except Dunnett's test. The
critical level for significance was p < 0.05.
Purity of the test material was 96.1%. Impurities included acrylonitrile
Test substance :
(0.1%), adiponitrile (0.7%), p-nitrosodiphenylamine (0.08%) and water (<
0.1%). Analyses indicated no significant decomposition of the test material
over the course of the study.
The authors concluded that the incidences of red nasal encrustation in the
Conclusion :
low dose animals, alopecia in the mid and high dose animals and staining
of facial fur in all treated groups were too low to be definitely related to
administration of test material. There was no effect of treatment on fertility
of females.
(1) valid without restriction
Reliability :
Study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (24)
Fertility
Type :
rat
Species :
male
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
46 to 57 days (depending on day of mating)
Exposure period :
6 hours/day, 5 days/week
Frequency of treatm. :
Premating exposure period
46 days
Male :
0 days
Female :
to gestation day 13-15
Duration of test :
No. of generation :
studies
60, 120 and 210 ppm
Doses :
yes
Control group :
= 60 ppm
NOAEL parental :
45 / 45
Id 107-12-0
5. Toxicity
Date 02.10.2003
= 210 ppm
other: NOAEL :
Reproductive Toxicity
other
Method :
1985
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Exposure concentrations: The average mean daily analytical exposure
Result :
concentrations (60.2, 120.4 and 208.9 ppm) were within 1% of the target
levels (60, 120 and 210 ppm, respectively). Mean temperatures and
humidities ranged from 22-25.5 degrees C and 24-27%, respectively.
Signs of toxicity: One of animals exposed to 210 ppm died after 2 days of
exposure. On the previous day, this animal exhibited labored breathing,
hypoactivity, poor control of the hind limbs, difficulty in standing, body
tremors and involuntary movements. No unusual findings were observed
at necropsy.
Body weights of males exposed to 210 ppm were approximately 6-9%
lower than those of the control group during most of the exposure period,
and remained lower than control (but were not significantly different) until
the end of the study.
Animals exposed to 210 ppm exhibited signs of toxicity such as arched
back (N = 8 on days 1-10, N = 3 on days 11-20 and 51-57, and N = 5 on
days 41-50), hypoactivity (N = 12-15 at each 10-day interval up to day 50,
and N = 4 from days 51-57), labored breathing (N = 10 on days 1-10, N = 3
on days 11-20 and 31-40, N = 5 on days 21-30 and N = 1 on days 51-57),
and salivation (N = 3 on days 1-10, and N = 10 - 12 at all other intervals). A
few high dose animals (individual numbers were not stated) also exhibited
abnormal behavior such as grinding of teeth, head bobbing, body tremors,
involuntary movements, and pawing at the cage. A few of the animals
exposed to 120 ppm exhibited salivation (N = 3-8 at all intervals) and
hypoactivity (N = 3 at days 11-20). Signs of toxicity generally abated by the
morning after exposure. Alopecia was observed in animals (N = 1 control,
N = 2 low dose, N = 1 mid dose, N = 5 high dose) at one or more of their
weekly physical examinations. No unusual treatment-related signs were
observed in rats exposed to 60 ppm. The only remarkable finding at gross
necropsy was a small right testis in one animal exposed to 120 ppm.
Fertility: There was no effect of treatment on male fertility. Efficiency of
mating (34.4%, 30.6%, 29.8% and 27.1% in the control, low, mid and high
dose groups) and pregnancy rate (90.5%, 97.6%, 90.0% and 97.4% in the
respective groups) were comparable between groups. There was no
difference in the numbers of live implants (ranged from 12.7 - 13.9),
resorptions (ranged from 0.7 - 1.1), nidations (ranged from 13.8 - 14.9),
corpora lutea (ranged from 13.1 - 15.2), preimplantation loss (4-8%) and
postimplantation loss (5-10%).
Animals: Virgin female Sprague Dawley rats (28 days old upon receipt) and
Test condition :
virgin male Sprague Dawley rats (50 days old upon receipt) were
acclimated for one week and examined for general health and the
presence of pinworms and ectoparasites. Body weights of fifteen females
and ten males that were taken upon receipt were 155-181 g and 80-103 g,
respectively. No significant health problems were noted during the
acclimation period, and the animals were released for study. Males and
females were individually caged (except during mating). Food and water
were available ad libitum (except food was not available to males during
exposure). Animals were housed in a room maintained at 72 +/- 2 degrees
F and 40-60% relative humidity, under a 12 hr light/dark cycle.
Exposures: Exposures (6 hr/day, 5 days/week) occurred in 10 m3
Rochester-style stainless steel and glass inhalation chambers. A
46 / 46
Id 107-12-0
5. Toxicity
Date 02.10.2003
scheduled exposure day was cancelled due in clement weather. A new
exposure day (exposure day 41) was used in its place. Due to inclement
weather and building equipment failures, 2 exposures (days 33 and 43)
were only for 4 hours and one exposure (day 32) was for 5 hours.
Atmospheres of propionitrile vapor were generated using bubbler systems.
The concentrations of material in the chambers were controlled by either
adjusting the nitrogen flow though the propionitrile in the bubblers or by
changing the amount of test material in the bubblers. Concentrations of
test material in the chambers were measured 4 times daily (except during
day 43) using a Miran 1A General Purpose Gas Analyzer. The nominal
concentration of material, temperature and humidity also were measured
(at intervals that were not listed).
Study conduct: Fifteen males per group were assigned to be exposed to
target concentrations of 0, 60, 120 or 210 ppm test material. Exposure
began when males were 43 days old. Mating was initiated when males and
females were 16 and 12 weeks old, respectively. At this time, males had
been 69 days on the study (which was sufficient to cover the
spermatogenesis cycle of the rat), and had 46 days of exposure. Males
were randomly mated (1: 1) with three untreated females (consecutively)
that had been assigned to the corresponding treatment group (45 females
were assigned per group). Exposure of males continued until the day after
the last mating opportunity (57 exposure days). At night, after exposure,
males were caged with their assigned female until copulation was
confirmed (by presence of a copulatory plug or sperm in vaginal smear) or
5 nights without confirmed copulation. The day on which copulation was
confirmed was considered gestation day 0.
After assignment to the study, males and females were weighed once per
week. Mated females were weighed on gestation days 0 and 13. Males
were given a thorough physical examination once per week and were
observed for clinical signs of toxicity before and after exposures. All
animals were checked twice daily for mortality and gross abnormalities
(except for one day prior to mating when inclement weather permitted
observations).
One half of the males of each group were euthanized on each of the 2
consecutive days at the end of the study. They had not been exposed to
propionitrile for about 2 weeks prior to termination. Each male was given
an external examination and weighed. The tissues and organs of the
thoracic, scrotal and abdominal cavities were examined for gross lesions
and the testes, epididymides, prostate glands and seminal vesicles were
preserved in 10% neutral buffered formalin. Females that were not mated
with males were euthanized and were not examined.
Mated females were euthanized on gestation day 13 (or the nearest
workday up to gestation day 15). Females that were co-housed with males
without confirmed copulation were euthanized during the second week
after the last day of co-housing. Gross necropsies were performed on
females that had copulated and those that had not. The tissues and organs
of the thoracic and abdominal cavities were examined. Pregnancy status
was determined, nidation sites and were classified, and corpora lutea were
counted.
Statistical analyses: Body weight data were analyzed using Dunnett's test.
Mating and pregnancy rate data were analyzed by a Fisher's exact test and
an uncorrected chi-square test. Other data were analyzed by the Mann-
Whitney U test. The Bonferroni inequality was assumed when comparing
multiple treatments to control values for all tests except Dunnett's test. The
critical level for significance was p < 0.05.
Purity of the test material was 96.1%. Impurities included acrylonitrile
Test substance :
(0.1%), adiponitrile (0.7%), p-nitrosodiphenylamine (0.08%) and water (<
47 / 47
Id 107-12-0
5. Toxicity
Date 02.10.2003
0.1%). Analyses indicated no significant decomposition of the test material
over the course of the study.
There was no effect of treatment on fertility of males.
Conclusion :
(1) valid without restriction
Reliability :
Study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (25)
5.8.2 DEVELOPMENTAL TOXICITY/TERATOGENICITY
rat
Species :
female
Sex :
Sprague-Dawley
Strain :
gavage
Route of admin. :
Days 6 through 19 of gestation
Exposure period :
daily
Frequency of treatm. :
to Day 20 of gestation
Duration of test :
20, 40, 80 mg/kg/day
Doses :
yes, concurrent vehicle
Control group :
= 40 mg/kg bw
NOAEL maternal tox. :
= 80 mg/kg bw
NOAEL teratogen. :
= 40 mg/kg bw
NOAEL Fetotoxicity :
= 40 mg/kg bw
NOAEL Embryotoxicity :
other
Method :
1981
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Doses used in this study were chosen based on the results of a pilot study
Remark :
(International Research and Developmental Corporation study IR-79-163,
dated December 22, 1980) conducted with 5, 10, 20, 37.5 and 75
mg/kg/day. In this study, 75 mg/kg caused a moderate to severe decrease
in maternal weight gain. Two out of 5 dams treated with 75 mg/kg/day also
had an increased number of resorptions (16 early in one animal and 7 late
in the other).
In the main study, the number of fetuses and litters with sternebrae 5
and/or 6 unossified in the mid dose group (55 and 35, respectively) was
"nearly comparable" to historical controls (3.8 - 23.7 in 13.0 - 73.9 litters).
Since this was the only change observed in the mid dose fetuses, it is not
considered to be indicative of fetotoxicity. Therefore, this dose was chosen
as the NOAEL for fetotoxicity.
The NOAEL for teratology was established by study personnel. The
summary preparer established the NOAELs for maternal toxicity,
fetotoxicity and embryotoxicity.
Maternal: One rat in the high dose group died of an undetermined cause
Result :
on gestation day 9. Survival was 100% in the other groups. There was a
slight to moderate reduction in mean maternal body weight gain over the
entire treatment period for the high dose group (118 g in treated vs. 134 g
in control). This decrease was predominantly due to decreased body
weight gain from days 6 to 9 (2 g in treated vs. 9 g in control). The mean
maternal adjusted body weight gain (body weight minus the uterus and
contents) in the high dose group also was slightly reduced during the
gestation period (52 g in treated vs. 60 g in control). Weights of animals in
other treated groups were similar to controls.
There was no effect of treatment on the number of pregnancies, mean
number of viable fetuses (all were viable), late resorptions, total
implantations, or corpora lutea. There was a significant increase in the
number of early resorptions (2.0 +/- 2.63 in treated vs. 0.7 +/- 0.81 in
48 / 48
Id 107-12-0
5. Toxicity
Date 02.10.2003
control) and a corresponding increase in the number of postimplantation
losses in the high dose group. None of the animals aborted.
Fetal: There was no effect of treatment on fetal sex ratio. Average mean
fetal body weight of fetuses from high dose animals were significantly less
than controls (3.0 +/- 0.40 g in treated vs. 3.5 +/- 0.28 g in control). The
number of control, 20 mg/kg/day, 40 mg/kg/day and 80 mg/kg/day fetuses
(and litters) with malformations were 5 (3), 1 (1), 1 (1) and 0 (0),
respectively. One fetus in the low dose group and one in the mid dose
group had a diaphragmatic hernia. An increase in the number of fetuses
and litters with sternebrae 5 and/or 6 unossified was noted in the mid dose
group (55 and 35, respectively) and high dose group (92 and 66.7,
respectively) when compared to the study control (21 and 14, respectively).
An increase in the number of fetuses and litters with sternebrae 1 and/or 2,
3, and 4 unossified was also found in the high dose group (8 in 2 treated
litters vs. 1 in control).
Rats: One hundred virgin female COBS CD rats (approximately 14 weeks
Test condition :
old) were used in the study. They were acclimated for at least 10 days
prior to mating. One female was mated with one male. Copulation was
verified by the presence of a copulatory plug or sperm in a vaginal smear.
The day that evidence of mating was detected was designated day 0 of
gestation. Mated females were assigned in a block design to a vehicle
control group and 3 treatment groups consisting of 25 rats each.
Test material: Dosing solutions of propionitrile were prepared daily at
concentrations that permitted administration of 20, 40 and 80 mg/kg/day at
a constant volume of 10 ml/kg. The material was dissolved in distilled
water and shaken by hand to ensure dissolution. The test material was
administered orally by gavage as a single daily dose on days 6 through 19
of gestation. The control group received distilled water at 10 ml/kg.
Individual dosages were determined using body weights that were taken on
gestation day 6.
Maternal observations: Prior to treatment, the dams were observed daily
for mortality and overt changes in appearance and behavior. They were
also observed daily from days 6 through 20 of gestation. Individual body
weights were recorded on gestation days 0, 6, 9, 12, 16 and 20. On
gestation day 20, all survivors were euthanized. The uterus was excised
and weighed, and the number and location of viable and nonviable fetuses,
early and late resorption and total number of implantations and corpora
lutea were recorded. The abdominal and thoracic cavities and organs of
the dams were examined grossly. Any dam that did not survive to
scheduled termination also was necropsied. Maternal tissues were
preserved when deemed necessary according to gross findings. Uteri from
females that appeared nongravid were opened and placed in 10%
ammonium sulfide solution for confirmation of pregnancy.
Fetal observations: All fetuses (total of 326, 302, 319 and 272 in the
control, low, mid and high dose groups) were individually weighed and
examined for external malformations and variations. Each fetus was
sexed. Approximately one-half of the fetuses were placed in Bouin's
fixative for subsequent visceral examination by razor-blade sectioning. The
other fetuses were fixed in alcohol, macerated in potassium hydroxide and
stained with Alizarin Red S for subsequent skeletal examination.
Statistical analyses: The sex distribution and number of litters with
malformations in treated animals were compared to controls using a Chi-
square test with Yates' correction for 2 x 2 contingency tables and/or
Fisher's exact probability test. The numbers of early and late resorptions
and postimplantation losses were compared by the Mann-Whitney U-test.
The mean number of viable fetuses, total implantations, corpora lutea and
mean fetal body weights were compared by Bartlett's test for homogeneity
49 / 49
Id 107-12-0
5. Toxicity
Date 02.10.2003
of variance, analysis of variance (one-way), and the appropriate t-test (for
equal or unequal variances). Dunnett's multiple comparison tables were
used to judge significance of differences. The level of significance was p <
0.05.
The purity of the test material was > 90%.
Test substance :
(1) valid without restriction
Reliability :
Significant differences between weights and weight gains of treated dams
and controls were not designated.
Critical study for SIDS endpoint
Flag :
10.08.2003 (22)
5.8.3 TOXICITY TO REPRODUCTION, OTHER STUDIES
5.9 SPECIFIC INVESTIGATIONS
Mechanistic Studies
Endpoint :
Study descr. in chapter :
Reference :
Type :
mouse
Species :
male
Sex :
CD-1
Strain :
i.p.
Route of admin. :
No. of animals :
other
Method :
1981
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
n-Butyronitrile also was tested in this study. The results with this material
Remark :
were similar to those of propionitrile.
This study is considered to be valid without restriction. The study was
conducted and documented in a thorough manner.
In the first study, the mortality rate for animals treated only with 45 mg/kg
Result :
propionitrile was 9/10. Co-treatment with sodium nitrite reduced the rate to
5/9. None of the 10 animals that were co-treated with sodium thiosulfate
died.
In the carbon tetrachloride study, the mortality rate for animals treated only
with 45 mg/kg propionitrile was 8/10. None of the animals co-treated with
carbon tetrachloride died.
Cyanide concentrations in liver and brain of mice co-treated with sodium
thiosulfate or carbon tetrachloride were significantly less than those of mice
treated with propionitrile alone. In mice receiving propionitrile only, 26.7 +/-
8.0 (mean +/- SD) and 12.8 +/- 5.4 nmol/g cyanide were found in liver and
brain, respectively. In mice receiving propionitrile plus sodium thiosulfate,
2.0 +/- 0.7 and 0.8 +/- 0.8 nmol/g cyanide were found in liver and brain,
respectively. In mice receiving propionitrile plus carbon tetrachloride, 0.9
+/- 1.1 and 1.5 +/- 1.4 nmol/g cyanide were found in liver and brain,
respectively.
Male CD mice (30 g) were divided into 3 groups of ten animals each. One
Test condition :
group received 45 mg/kg i.p. propionitrile only, another received i.p.
injections of 75 mg/kg sodium nitrite (a cyanide antagonist) 20 minutes
before and 100 minutes after i.p. injection of 45 mg/kg propionitrile, and
another received i.p. injections of 1 g/kg sodium thiosulfate (a cyanide
antagonist) 20 minutes before and 80 and 180 minutes after i.p. injection of
50 / 50
Id 107-12-0
5. Toxicity
Date 02.10.2003
45 mg/kg propionitrile.
Two other groups of 10 mice received either 0.2 ml of vegetable oil or 0.2
ml of 20% carbon tetrachloride (a hepatotoxic dose) in vegetable oil
subcutaneously, 24 hours before i.p. treatment with 45 mg/kg propionitrile.
In both experiments, animals were observed for 7 days. Mortality data were
analyzed statistically by the chi-square test. The criterion for significance
was p < 0.05.
In an additional study, the concentrations of cyanide in liver and brain were
determined in a) 5 mice treated only with 28 mg/kg propionitrile (i.p.), b) 5
mice given 1 g/kg sodium thiosulfate 20 minutes before and 80 minutes
after 28 mg/kg propionitrile (i.p.), and c) 5 mice given 0.2 ml of 20% carbon
tetrachloride subcutaneously 24 hours before i.p. treatment with 28 mg/kg
propionitrile. All mice were killed 2.5 hours after propionitrile injection (if
still alive at this time). The livers and brains were excised as soon as
possible after death, quick-frozen and weighed. Cyanide concentrations
were determined by the method of Bruce et al. (Anal Chem 27: 1346-1347,
1955). Results were analyzed using an unpaired t-test.
The purity of the test material was 99%. No free cyanide was found in
Test substance :
solutions made in distilled, deionized water.
Propionitrile is activated by the liver to release cyanide, which is
Conclusion :
responsible for acute toxicity.
05.08.2003 (36)
5.10 EXPOSURE EXPERIENCE
5.11 ADDITIONAL REMARKS
51 / 51
Id 107-12-0
6. Analyt. Meth. for Detection and Identification
Date 02.10.2003
6.1 ANALYTICAL METHODS
6.2 DETECTION AND IDENTIFICATION
52 / 52
Id 107-12-0
7. Eff. Against Target Org. and Intended Uses
Date 02.10.2003
7.1 FUNCTION
7.2 EFFECTS ON ORGANISMS TO BE CONTROLLED
7.3 ORGANISMS TO BE PROTECTED
7.4 USER
7.5 RESISTANCE
53 / 53
Id 107-12-0
8. Meas. Nec. to Prot. Man, Animals, Environment
Date 02.10.2003
8.1 METHODS HANDLING AND STORING
8.2 FIRE GUIDANCE
8.3 EMERGENCY MEASURES
8.4 POSSIB. OF RENDERING SUBST. HARMLESS
8.5 WASTE MANAGEMENT
8.6 SIDE-EFFECTS DETECTION
8.7 SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER
8.8 REACTIVITY TOWARDS CONTAINER MATERIAL
54 / 54
Id 107-12-0
9. References
Date 02.10.2003
(1) Analytical Biochemistry (ABC) Laboratories Inc. Static Acute Toxicity Report #27354. Acute
toxicity of propionitrile (AB-81-089) to rainbow trout (Salmo gairdneri), April 22, 1981
(unpublished study).
(2) Analytical Biochemistry (ABC) Laboratories Inc. Static Acute Toxicity Report #27355. Acute
toxicity of propionitrile (AB-81-090) to bluegill sunfish (Lepomis macrochirus), April 28,
1981 (unpublished study).
(3) Analytical Biochemistry (ABC) Laboratories Inc. Static Acute Toxicity Report #27356. Acute
toxicity of propionitrile (Lot No. 34)(AB-81-091) to Daphnia magna, April 24, 1981
(unpublished study).
(4) Biodynamics Inc. 1981. Acute dermal toxicity study in rabbits (unpublished study). Test
Material: Decatur Propionitrile. Project No 6860-81, BD-81-359, dated December 31, 1981.
(5) Biodynamics Inc. 1981. Acute dermal toxicity study in rabbits (unpublished study). Test
Material: Eastman Kodak Propionitrile. Project No 6861-81, BD-81-360, dated December
31, 1981.
(6) Biodynamics Inc. 1981. Acute dermal toxicity study in rabbits (unpublished study). Test
Material: Seal Sands Propionitrile. Project No 6859-81, BD-81-358, dated December 31,
1981.
(7) Biodynamics Inc. 1987. Absorption, distribution and elimination of 14C-propionitrile in rats
following a single oral dose. Unpublished project number 84425 (BD-84-299), dated July
16, 1987.
(8) Biodynamics Inc. The absorption, distribution and elimination of 14C-labeled propionitrile in
the rat. Unpublished project 79095 (No. BO-79-351), dated September 30, 1981.
(9) Chapatwala KD, Babu GRV, Nawaz MS. 1992. Degradation of acetonitrile and biphenyl
compounds by a mixed microbial culture. Environ Toxicol and Chem 11: 1145-1151.
(10) Clayton GD and Clayton FE (eds). Patty's Industrial Hygiene and Toxicology, 3rd Ed., New
York, Wiley and Sons, 1981-1982
(11) Eastman Kodak Company, Environmental Sciences Section, Health and Environment
Laboratories. Isobutyronitrile: A Growth Inhibition Test with the Alga, Selenastrum
capricornutum (unpublished study). Study No. EN-512-907253-A, August 30, 1999.
(12) Eastman Kodak Company, Environmental Sciences Section, Health and Environment
Laboratories. n-Butyronitrile: A Growth Inhibition Test with the Alga, Selenastrum
capricornutum (unpublished study). Study No. EN-512-900741-A, January 28, 2000.
(13) EPIWIN Aop Program (v1.90).
(14) EPIWIN ECOSAR Program (v0.99).
(15) EPIWIN Fugacity Model Level III.
(16) EPIWIN Hydrowin Program (v1.67).
(17) EPIWIN Kowwin Program (v1.66).
(18) EPIWIN Wskow Program (v1.40)
55 / 55
Id 107-12-0
9. References
Date 02.10.2003
(19) Flowers LJ. 1977. Mutagenicity plate assay: propionitrile. Unpublished Monsanto project
No LF-77-137, dated June 14, 1977.
(20) Geiger DL, Call DJ, Brooke LT (eds). 1990. Acute toxicities of organic chemicals to
fathead minnows (Pimephales-Promelas). Vol V. Superior WI: University of Wisconsin-
Superior. p. 51.
(21) Hazleton Biotechnologies Corporation. 1985. In vivo bone marrow chromosome study in
rats: propionitrile (unpublished study). Revised final report HL-84-219, dated April 17,
1985.
(22) International Research and Development Corporation. 1981. Propionitrile teratology study
in rats (IR-79-164). Unpublished study dated January 30, 1981.
(23) ITII. Toxic and Hazardous Industrial Chemicals Safety Manual. Tokyo Japan: The
International Technical Information Institute, 1988.
(24) Kier LD. 1984. Female fertility study of Sprague-Dawley rats exposed by the inhalation
route to propionitrile. Unpublished Monsanto Report No MSL-4438, dated December 31,
1984.
(25) Kier LD. 1984. Male fertility study of Sprague-Dawley rats exposed by the inhalation route
to propionitrile. Unpublished Monsanto Report No MSL-4422, dated December 17, 1984.
(26) Lutin PA. 1970. Removal of organic nitriles from wastewater systems. J Water Pollut
Control Fed 42: 1632-42.
(27) Microbiological Associates. 1982. Evaluation of test article propionitrile (MRI #708) for
mutagenic potential employing the L5178Y TK+/- mutagenesis assay (unpublished study).
Study number 065-544-708-7, dated April 2, 1982.
(28) Microbiological Associates. 1982. Evaluation of test article propionitrile tails (MRI #710) for
mutagenic potential employing the L5178Y TK+/- mutagenesis assay (unpublished study).
Study number 065-545-710-7, dated April 2, 1982.
(29) Riddick JA, Bunger WB, Sackano TK. 1986. Organic Solvents: Physical Properties &
Methods of Purification. In: Techniques of Chemistry, Vol. II (4th Ed). NY:Wiley
Interscience, p. 583-7.
(30) Roloff MV. 1981. Subacute inhalation toxicity of propionitrile administered for four weeks to
Sprague-Dawley rats (unpublished study). Monsanto report number MSL-1690, dated
June 25, 1981.
(31) Sangster J. 1989. Octanol-water partition coefficients of simple organic compounds. J Phys
Chem Ref Data 18:1111-1230.
(32) Solutia, Inc. Material Safety Data Sheet for propionitrile, refined grade, dated March 14,
2003.
(33) SRI International. Evaluation of the potential of propionitrile to induce unscheduled DNA
synthesis in primary rat hepatocyte cultures (unpublished study). SRI Project LSC-7795,
dated July, 1985.
(34) Symons JM, McKinney RE, Smith RM, Donovan EJ, Jr. 1960. Degradation of nitrogen-
containing organic compounds by activated sludge. Int J Air and Water Poll 1/2:115-138.
(35) Velasquez DJ and Thake DC. 1984. Three-month toxicity study of propionitrile vapor
administered to male and female Sprague-Dawley rats by inhalation. Unpublished
Monsanto Report No MSL-4113, dated October 1, 1984.
56 / 56
Id 107-12-0
9. References
Date 02.10.2003
(36) Willhite CC and Smith RP. 1981. The role of cyanide liberation in the acute toxicity of
aliphatic nitriles. Toxicol Appl Pharmacol 59: 589-602.
(37) Windholz M, et al. The Merck Index - An Encyclopedia of Chemicals, Drugs and
Biologicals. 10th Edition. Rahway, NJ: Merck & Co., Inc., 1983.
(38) Yaws C et al. 1992. Water Solubility Data (Chapter 10). In : Thermodynamics and Physical
Property Data, Houston TX, Gulf Publ. Co.
(39) Younger Laboratories Incorporated. Nonclinical laboratory study final report - test material
propionitrile tails from seal sands (unpublished study). Project Number Y-80-77, final report
dated October 6, 1980.
(40) Younger Laboratories Incorporated. Toxicity studies on propionitrile (unpublished). Project
Number Y-79-86, final report dated July 31, 1979.
(41) Younger Laboratories Incorporated. Toxicological investigation of propionitrile
(unpublished). Monsanto Project Number Y-78-131, dated August 3, 1978.
57 / 57
Id 107-12-0
10. Summary and Evaluation
Date 02.10.2003
10.1 END POINT SUMMARY
10.2 HAZARD SUMMARY
10.3 RISK ASSESSMENT
58 / 58
201-14860B1
IUCLID
Data Set
: ID: 109-74-o
Existing Chemical '
: 109-74-O
CAS No.
: Butyronitrile
EINECS Name
TSCA Name : Butanenitrile
Molecular Formula : C4H7N
Producer related part
: Eastman Chemical Company
Company
Creation date : 24.06.2003
Substance related part
: Eastman Chemical Company
Company
: 24.06.2003
Creation date
Status
Memo
: 06.10.2003
Printing date
: 13.11.2003
Revision date
Date of last update : 06.10.2003
Number of pages : 43
: Chapter: 1, 2, 3,4, 5, 6, 7, 8, 10
Chapter (profile)
: Reliability: without reliability, 1, 2, 3,4
Reliability (profile)
: Flags: without flag, confidential, non confidential, WGK (DE), TA-Luft (DE),
Flags (profile)
Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SIDS
l/l
Id 109-74-0
1. General Information
Date 02.10.2003
1.0.1 APPLICANT AND COMPANY INFORMATION
1.0.2 LOCATION OF PRODUCTION SITE, IMPORTER OR FORMULATOR
1.0.3 IDENTITY OF RECIPIENTS
1.0.4 DETAILS ON CATEGORY/TEMPLATE
1.1.0 SUBSTANCE IDENTIFICATION
IUPAC Name :
C(N#)CCC
Smiles Code :
C4H7N
Molecular formula :
69.11
Molecular weight :
Petrol class :
15.08.2003
1.1.1 GENERAL SUBSTANCE INFORMATION
typical for marketed substance
Purity type :
organic
Substance type :
liquid
Physical status :
>= % w/w
Purity :
clear
Colour :
Odour :
(2) valid with restrictions
Reliability :
28.07.2003
1.1.2 SPECTRA
1.2 SYNONYMS AND TRADENAMES
1-Cyanopropane
Butanenitrile
Butyric acid nitrile
n Butyl nitrile
2/2
Id 109-74-0
1. General Information
Date 02.10.2003
n-Butyronitrile
n-Propyl cyanide
Propyl cyanide
1.3 IMPURITIES
1.4 ADDITIVES
1.5 TOTAL QUANTITY
1.6.1 LABELLING
1.6.2 CLASSIFICATION
1.6.3 PACKAGING
1.7 USE PATTERN
industrial
Type of use :
Chemical industry: used in synthesis
Category :
Chemical intermediate
Remark :
02.07.2003 (5)
1.7.1 DETAILED USE PATTERN
1.7.2 METHODS OF MANUFACTURE
1.8 REGULATORY MEASURES
1.8.1 OCCUPATIONAL EXPOSURE LIMIT VALUES
1.8.2 ACCEPTABLE RESIDUES LEVELS
1.8.3 WATER POLLUTION
3/3
Id 109-74-0
1. General Information
Date 02.10.2003
1.8.4 MAJOR ACCIDENT HAZARDS
1.8.5 AIR POLLUTION
1.8.6 LISTINGS E.G. CHEMICAL INVENTORIES
1.9.1 DEGRADATION/TRANSFORMATION PRODUCTS
1.9.2 COMPONENTS
1.10 SOURCE OF EXPOSURE
1.11 ADDITIONAL REMARKS
1.12 LAST LITERATURE SEARCH
1.13 REVIEWS
4/4
Id 109-74-0
2. Physico-Chemical Data
Date 02.10.2003
2.1 MELTING POINT
= -112 癈
Value :
Sublimation :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed published data.
Critical study for SIDS endpoint
Flag :
28.07.2003 (29)
2.2 BOILING POINT
= 117.5 癈 at 1016 hPa
Value :
Decomposition :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Value is boiling point at 760 mm Hg. Purity of material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed published data.
Critical study for SIDS endpoint
Flag :
28.07.2003 (29)
2.3 DENSITY
relative density
Type :
= .7954 g/cm?at 15 癈
Value :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of the material is unknown.
Remark :
(2) valid with restrictions
Reliability :
Source is peer-reviewed published data.
28.07.2003 (29)
2.3.1 GRANULOMETRY
2.4 VAPOUR PRESSURE
= 26 hPa at 25 癈
Value :
Decomposition :
other (measured): not specified
Method :
Year :
no data
GLP :
5/5
Id 109-74-0
2. Physico-Chemical Data
Date 02.10.2003
as prescribed by 1.1 - 1.4
Test substance :
Purity of the material is unknown. Data were obtained from Hazardous
Remark :
Substances Data Bank Number: 5013. Last revision date: 9/21/1999.
Value is 19.5 mmHg
Result :
(2) valid with restrictions
Reliability :
Primary source is peer-reviewed published data.
Critical study for SIDS endpoint
Flag :
28.07.2003 (4)
2.5 PARTITION COEFFICIENT
Partition coefficient :
= .53 at 癈
Log pow :
pH value :
other (measured)
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The value obtained from the experiment was 0.53. A value of 0.60 with a
Result :
0.10 confidence limit also was listed.
The test was performed at ambient temperature (20-25 degrees C). The
Test condition :
value was obtained using the Shake-Flask method. The aqueous phase
was octanol-saturated water. The concentration of material in the aqueous
phase was measured using gas-liquid chromatography.
Purity of the test material was not mentioned.
Test substance :
(2) valid with restrictions
Reliability :
Data were from a peer reviewed, published source.
Critical study for SIDS endpoint
Flag :
13.08.2003 (25)
octanol-water
Partition coefficient :
= .837 at 25 癈
Log pow :
=7
pH value :
other (calculated)
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-112 degrees
Remark :
C), boiling point (117.5 degrees C), vapor pressure (19.5 mm Hg), and
water solubility (33,000 mg/l).
(2) valid with restrictions
Reliability :
Approved model to calculate log Kow.
13.08.2003 (17)
2.6.1 SOLUBILITY IN DIFFERENT MEDIA
water
Solubility in :
= 33000 mg/l at 25 癈
Value :
pH value :
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
6/6
Id 109-74-0
2. Physico-Chemical Data
Date 02.10.2003
Deg. product :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of material is unknown. Data obtained from Hazardous Substances
Remark :
Data Bank Number: 5013. Last revision date: 9/21/1999.
Moderate (10-100 g/L)
Result :
(2) valid with restrictions
Reliability :
Primary source is peer-reviewed published data.
Critical study for SIDS endpoint
Flag :
15.08.2003 (23)
water
Solubility in :
= 27840 at 25 癈
Value :
pH value :
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
Deg. product :
other:calculated
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-112 degrees
Remark :
C), boiling point (117.5 degrees C), vapor pressure (19.5 mm Hg), and
water solubility (33,000 mg/l).
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
15.08.2003 (19)
2.6.2 SURFACE TENSION
2.7 FLASH POINT
2.8 AUTO FLAMMABILITY
2.9 FLAMMABILITY
2.10 EXPLOSIVE PROPERTIES
2.11 OXIDIZING PROPERTIES
2.12 DISSOCIATION CONSTANT
7/7
Id 109-74-0
2. Physico-Chemical Data
Date 02.10.2003
2.13 VISCOSITY
2.14 ADDITIONAL REMARKS
8/8
Id 109-74-0
3. Environmental Fate and Pathways
Date 02.10.2003
3.1.1 PHOTODEGRADATION
air
Type :
Sun light
Light source :
nm
Light spectrum :
based on intensity of sunlight
Relative intensity :
INDIRECT PHOTOLYSIS
OH
Sensitizer :
Conc. of sensitizer :
= .000000000000498 cm?(molecule*sec)
Rate constant :
= 50 % after 21.5 day(s)
Degradation :
Deg. product :
other (calculated)
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-112 degrees
Remark :
C), boiling point (117.5 degrees C), vapor pressure (19.5 mm Hg), and
water solubility (33,000 mg/l).
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
Critical study for SIDS endpoint
Flag :
07.08.2003 (14)
3.1.2 STABILITY IN WATER
abiotic
Type :
other
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
: EPIWIN Hydrowin cannot calculate hydrolysis rate constants for nitriles.
Remark
The theoretical hydrolysis of the related material propionitrile and several
other chemicals has been examined by Dr. Lee Wolfe at the USEPA
Environmental Research Laboratory in Athens, Georgia. The results of
these analyses were published in a report by Dr. Wolfe that could not be
located. In a personal communication, Dr. Wolfe stated that propionitrile can
hydrolyze (albeit slowly). According to a study cited in the Hazardous
Substances Data Bank, the chemical hydrolysis of the related material
acetonitrile in water is base-catalyzed (the rate constant for base catalyzed
hydrolysis is 5.8X10-3/M-hr), but the half-life at pH 7 is more than 150,000
yrs (Ellington et al., 1988). Acetonitrile (CH3CN, CAS No. 75-05-8) is the
2-carbon analog of the category members, possessing the same
functionality, but having one less carbon than propionitrile. Taken together,
these data suggest that hydrolysis of butyronitrile at environmentally
relevant pHs will occur too slowly to be a significant means of degradation.
: (2) valid with restrictions
Reliability
Experimental results for the test material could not be located. Results are
for a related material.
28.07.2003 (16)
3.1.3 STABILITY IN SOIL
9/9
Id 109-74-0
3. Environmental Fate and Pathways
Date 02.10.2003
3.2.1 MONITORING DATA
3.2.2 FIELD STUDIES
3.3.1 TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS
fugacity model level III
Type :
other: air, water, soil and sediment
Media :
15 % (Fugacity Model Level I)
Air :
47.9 % (Fugacity Model Level I)
Water :
.0821 % (Fugacity Model Level II/III)
Biota :
37 % (Fugacity Model Level II/III)
Soil :
other: calculated
Method :
2003
Year :
Measured inputs to the program are CAS No., melting point (-112 degrees
Remark :
C), boiling point (117.5 degrees C), vapor pressure (19.5 mm Hg), and
water solubility (33,000 mg/l).
EPIWIN Henry Program (v3.10) estimates a Henry's Law Constant of
Result :
5.38E-005 atm-m3/mol. EPIWIN Pckocwin Program (v1.66) estimates a
Koc of 15.3.
(2) valid with restrictions
Reliability :
Approved model for estimated environmental transport values.
Critical study for SIDS endpoint
Flag :
15.08.2003 (18)
3.3.2 DISTRIBUTION
3.4 MODE OF DEGRADATION IN ACTUAL USE
3.5 BIODEGRADATION
aerobic
Type :
Inoculum :
Deg. product :
other: Directive 92/69/EEC, C.5 (BOD5) and Directive 92/69/EEC, C.6
Method :
(COD)
1998
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Refer to Section 3.6 below.
Remark :
(1) valid without restriction
Reliability :
Critical study for SIDS endpoint
Flag :
02.07.2003
aerobic
Type :
other: activated sludge
Inoculum :
500 mg/l
Concentration :
72 hour(s)
Contact time :
Deg. product :
other
Method :
1970
Year :
10 / 10
Id 109-74-0
3. Environmental Fate and Pathways
Date 02.10.2003
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
TOD for n-butyronitrile was 1679 mg/l. The test material was degraded by
Result :
Franklin sludge (10.5% TOD at 72 hours) and Nashville sludge (0.8 and
1.0% TOD at 24 and 72 hours, respectively). The test material was toxic to
Bordeaux sludge.
Bacteria: The activated sludges were obtained from the municipal plant at
Test condition :
Franklin, TN, the municipal plant at Nashville TN and the plant at
Bordeaux, a suburb of Nashville. Mixed liquor from the aeration tanks was
collected the morning of the day the Warburg run began. Each sample
was packed in ice and transported to the laboratory within 1 hour of
collection. Before the run began, the sludge sample was blended for 10
sec and the homogenous blend was analyzed for concentration of SS (not
defined, but assumed suspended solids), using a membrane-filter
technique. The original sample was adjusted to a SS concentration of
2,500 mg/l.
Test conduct: Test material was added to a Warburg flask (125 ml) in
order to obtain a final concentration of 500 mg/l in the reaction
compartment (final volume of 20 ml). KOH (1.0 ml, 20%) was added to the
center well. A 20 ml volume of activated sludge was then introduced. The
test was performed in duplicate. Flasks were incubated for 72 hours
(constant motion) at 20 degrees C. All three sludges were tested. Oxygen
uptake curves were plotted. Respiration of the sludge alone was plotted as
the control curve.
Theoretical O2 demand (the mg/l O2 required to completely oxidize the test
material) was calculated on the basis of the test material to CO2 and water,
plus nitrate according to the following equation: (TOD = moles of O2
required to balance the equation x molecular weight of O2 x concentration
of test material/ (moles of test material required to balance the oxidation
equation x molecular weight of the test material). The percentage of TOD
was to be calculated as follows: % TOD = 100 x D (the difference in mg/l of
O2 uptake between substrate and control)/ TOD. The material was
considered toxic if D was less than 0.
(2) valid with restrictions
Reliability :
Purity of the test material was not given
07.08.2003 (22)
aerobic
Type :
other: mixed microbial culture
Inoculum :
1000 mg/l related to
Concentration :
related to
48 hour(s)
Contact time :
other: biodegradable
Result :
Deg. product :
other
Method :
1992
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The final protein and ammonia concentrations and pH were 10.10 mg/l,
Result :
74.6 micromoles/ml and 8.69, respectively, indicating that the mixed culture
could use this material as a growth substrate.
A mixed microbial culture (protein concentration of 0.085 mg/l) was isolated
Test condition :
from an environment contaminated with organic cyanides and
polychlorinated biphenyls. This was grown for 48 hours on phosphate
buffer (pH 7.0, 30 degrees C) containing propionitrile (1 g/l) as the sole
source of carbon and nitrogen. The final concentration of protein, ammonia
and pH were determined.
Test material was obtained from Aldrich Chemical Co. It is presumed that
Test substance :
11 / 11
Id 109-74-0
3. Environmental Fate and Pathways
Date 02.10.2003
the material has high purity.
(4) not assignable
Reliability :
The study shows that the test material was used as a substrate (and
therefore was metabolized); however, the extent to which the test material
biodegraded is difficult to determine from the study.
07.08.2003 (1)
3.6 BOD5, COD OR BOD5/COD RATIO
BOD5
Directive 92/69/EEC, C.5
Method :
1998
Year :
yes
GLP :
COD
Directive 92/69/EEC, C.6
Method :
1998
Year :
= 2130 mg/g substance
COD :
yes
GLP :
RATIO BOD5 / COD
= .47
BOD5/COD :
Method is similar to OECD: TG-301C: Modified MITI Test. The study is the
Remark :
critical study for the biodegradation endpoint.
BOD analysis: The values obtained for the 3 different concentrations were
Result :
1.00, 1.01 and 1.03 g BOD/g test material at 5 days and 1.76, 2.07 and
1.86 g BOD/g test material at 20 days. The average BOD5 and BOD20
values were 1.0 grams BOD/gram of test substance and 1.9 grams
BOD/gram of test substance, respectively.
COD analysis: The values obtained for the 3 replicates were 2.139, 2.064
and 2.184 g COD/g test material. The average value was 2.13 g COD/g
test material. The percent recovery of the reference sample was 93.5%.
BOD5/COD: The BOD5/COD ratio is 0.47 (1.0/2.13).
Test conditions: COD Determination: A 0.50 N potassium dichromate
Test condition :
solution was used to standardize the ferrous ammonium sulfate titrant.
Mercuric sulfate was added to minimize chloride interference (if any). Three
separate replicates were tested. A potassium phthalate standard was
analyzed as a positive control. The test was considered valid if the
recovery of the standard fell between the limits of 83.47 - 116.06%. The
COD was calculated by subtracting the amount of titrant needed for the
sample from the amount of titrant needed for a blank. This result was
multiplied by the normality of the titrant and the equivalent weight of
oxygen. The product was then divided by the sample weight.
BOD Determination: The test was performed according to an Eastman
Kodak Company protocol [Need to have protocol for details]. Three
separate concentrations (0.000195%, 0.000397% and 0.000592%) were
tested. The 5-day BOD was calculated by subtracting the final dissolved
oxygen reading and the 5-day seeded dilution water drop from the initial
dissolved oxygen reading. The 20-day BOD was calculated by subtracting
the average 20-day seeded dilution water drop from the total dissolved
oxygen drop over 20 days. The results were multiplied by 100. The
products were divided by the product of the percent concentration of the
stock solution in the BOD bottle and the concentration of test chemical in
the stock solution. The products were divided by 1,000,000. The results
were in units of grams of BOD per gram of test substance for the 5-day (or
20-day) incubation. The test was considered to be valid if the average BOD
of glucose-glutamic acid standards was 198 mg/l +/- 30 mg/l.
12 / 12
Id 109-74-0
3. Environmental Fate and Pathways
Date 02.10.2003
BOD5/COD ratio: This ratio was calculated by dividing the average 5-day
BOD value by the average COD value.
Purity was 99.94%.
Test substance :
The test material is not considered to be "Readily Biodegradable" based on
Conclusion :
a BOD5/COD ratio of <0.5 (1000/2130 = 0.47)
(1) valid without restriction
Reliability :
This was a well-documented guideline study conducted under GLP
assurances.
10.08.2003 (6) (7)
3.7 BIOACCUMULATION
3.8 ADDITIONAL REMARKS
13 / 13
Id 109-74-0
4. Ecotoxicity
Date 02.10.2003
4.1 ACUTE/PROLONGED TOXICITY TO FISH
static
Type :
Pimephales promelas (Fish, fresh water)
Species :
96 hour(s)
Exposure period :
mg/l
Unit :
= 107 measured/nominal
NOEC :
measured/nominal
LC0 :
> 107 measured/nominal
LC50 :
yes
Limit test :
yes
Analytical monitoring :
other: OECG:TG-203 and EEC/Annex V C.1.
Method :
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
No significant protocol deviations were noted that would affect study
Remark :
results. The fact that the temperature (21 - 22 degrees C) was slightly
higher than that specified in the protocol (20 +/- 1 degrees C) was not
considered to have adversely affected the study.
No mortality occurred and all fish exhibited normal behavior and
Result :
appearance. No precipitation of test material was observed. The mean
concentration of test material was 107 mg/l. The analyzed percent loss of
the test material ranged from 0 - 9.1%. The temperatures of all solutions
ranged from 21 - 22 degrees throughout the test. The pH and dissolved
oxygen values ranged from 7.9 - 8.4 and 7.2 - 8.7 mg/l, respectively. The
temperature, pH and dissolved oxygen values were considered to be
acceptable for the organisms used in the test. The test was considered to
be valid.
Organisms: Juvenile fathead minnows were acclimated to test water for at
Test condition :
least two weeks prior to testing. They were randomized to 6 sets of 7 fish
each. Two sets of minnows (7/set) were killed before the start of the test to
determine average wet weight (0.35 and 0.37 g/set) and mean standard
length (2.97 cm/set).
Test water: The water was pumped from Lake Ontario into a large
underground storage vessel. Water from this vessel was subsequently
pumped into the laboratory where it passed through polystyrene filter
tubes, activated carbon filter tubes, and another set of polystyrene filter
tubes. The filtered water stream was treated with sodium thiosulfate to
further reduce trace levels of residual chlorine. The water was then heated
to 20 +/- 2 degrees C and distributed to an open basin for seasoning prior
to use. Representative values for hardness and total alkalinity (both as
CaCO3) were 120.0 and 87.4 mg/l, respectively.
Test material: A stock solution of test material was prepared in a 200 ml
volumetric flask containing test water. The exposure solutions were
prepared at a nominal concentration of 120 mg/l by adding the appropriate
amount of stock solution to glass vessels (30.5 cm Pyrex seamless,
cuboidal chromatography jars) containing 20 liters of test water. The
solutions in each test vessel were stirred with a stir rod prior to adding fish.
Duplicate test and dilution water control vessels were prepared.
Test conduct: Immediately after stirring, fish were placed into each of the
replicate test and control vessels (7 per vessel). Glass lids were placed on
top of each test vessel and sealed with Parafilm. Biological loading within
test vessels was kept below 1.0 g wet weight/l test solution.
The vessels were placed in a certified hood under 8.5 hours of fluorescent
lighting/day. Animals were observed for mortality and signs of stress at 0,
14 / 14
Id 109-74-0
4. Ecotoxicity
Date 02.10.2003
24, 48, 72 and 96 hours. Temperature, dissolved oxygen concentration and
pH of each solution also were measured at these times. Concentrations of
test material in the test vessels at 0 and 96 hours were analyzed by
GC/MS. The geometric mean of the concentrations was calculated. Since
no mortality was observed, statistical analyses were not performed.
The test was considered valid if control mortality was <= 10%, dissolved
oxygen did not fall below 60% of the initial oxygen level, the temperature
was 20 +/- 1 degrees C and there were no abnormal occurrences that
could influence the outcome.
Purity was 99.94%
Test substance :
The LC50 value indicates that the test substance would not be classified
Conclusion :
according to the European Union's labeling directive and would correspond
to a "low concern level" according to the U.S. EPA's assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
07.08.2003 (10)
4.2 ACUTE TOXICITY TO AQUATIC INVERTEBRATES
static
Type :
Daphnia magna (Crustacea)
Species :
48 hour(s)
Exposure period :
mg/l
Unit :
= 110 measured/nominal
NOEC :
measured/nominal
EC0 :
> 110 measured/nominal
EC50 :
yes
Limit Test :
yes
Analytical monitoring :
other: OECD: TG-202 and EEC/Annex V C.2
Method :
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Nine organisms (instead of 10) were used in one test vessel. No other
Remark :
protocol deviations were noted.
One out of 10 daphnids in one study was immobile after 48 hours of
Result :
exposure to test material. This was not considered to be significant (< =
10%). All other daphnids exposed to test article exhibited behavior
comparable to controls. No precipitation of test material was observed.
The geometric mean of the test concentration at 0 and 48 hours was 110
mg/l. The analyzed percent loss of the test material was 0%. The
temperatures of all solutions were maintained at 22 degrees throughout the
test. The pH and dissolved oxygen values ranged from 8.2 - 8.4 and 8.6 -
8.7 mg/l, respectively. The temperature, pH and dissolved oxygen
values were considered to be acceptable for the organisms used in the
test. The test was considered to be valid.
Organisms: Adult Daphnia magna were reared within the testing facility in
Test condition :
100 l culturing flasks. Gravid daphnids used to produce test animals were
obtained from rearing tanks that had been established for at least two
weeks. Prior to the study, approximately 100 gravid daphnids were
transferred by net into two glass bowls containing test water and food. After
18 hours in the bowls, all adult daphnids were removed. Neonates were
collected by pipette and transferred directly into exposure vessels. A total
of 10 daphnids were placed into each of the replicate test and control
vessels.
Test water: The water was pumped from Lake Ontario into a large
15 / 15
Id 109-74-0
4. Ecotoxicity
Date 02.10.2003
underground storage vessel. Water from this vessel was subsequently
pumped into the laboratory where it passed through polystyrene filter
tubes, activated carbon filter tubes, and another set of polystyrene filter
tubes. The filtered water stream was treated with sodium thiosulfate to
further reduce trace levels of residual chlorine. The water was then heated
to 20 +/- 2 degrees C and distributed to an open basin for seasoning prior
to use. Representative values for hardness and total alkalinity (both as
CaCO3) were 120.0 and 87.4 mg/l, respectively.
Test material: A stock solution of test material was prepared in a 200 ml
volumetric flask containing test water. The exposure solutions were
prepared at a nominal concentration of 120 mg/l by adding the appropriate
amount of stock solution to glass vessels (300 ml Pyrex glass, lipless
beakers) containing 20 L of test water. The solutions in each test vessel
were stirred with a stir rod prior to adding daphnids. Duplicate test and
dilution water control vessels were prepared.
Test conduct: Immediately after stirring, daphnids were placed into each of
the replicate test and control vessels 10 per vessel). Watch glasses were
placed on top of each test vessel and sealed with Parafilm. The vessels
were placed in a certified hood under 8.5 hours of fluorescent
lighting/day. Animals were observed for mortality and signs of stress at 0,
24 and 48 hours. Temperature, dissolved oxygen concentration, and pH
of each solution were measured at 0 (prior to adding organisms) and 48
hours. Concentrations of test material in the test vessels at 0 and 48 hours
were analyzed by GC/MS. The geometric mean of the concentrations was
calculated. Since no mortality was observed, statistical analyses were not
performed.
The test was considered valid if control mortality was <= 10%, dissolved
oxygen did not fall below 60% of the initial oxygen level, the temperature
was 20 +/- 2 degrees C, test daphnids in the control groups were not
trapped at the surface of the water and there were no abnormal
occurrences that could influence the outcome.
Purity was 99.94%
Test substance :
The 48-hour EC50 value indicates that the test substance would not be
Conclusion :
classified according to the European Union's labeling directive and would
correspond to a "low concern level" according to the U.S. EPA's
assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
07.08.2003 (9)
4.3 TOXICITY TO AQUATIC PLANTS E.G. ALGAE
Selenastrum capricornutum (Algae)
Species :
other: biomass and growth rate
Endpoint :
72 hour(s)
Exposure period :
mg/l
Unit :
= 133.4 measured/nominal
NOEC :
> 133.4 measured/nominal
EC50 :
yes
Limit test :
yes
Analytical monitoring :
other: OECD: TG-201 and EEC/Annex V C.3
Method :
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
16 / 16
Id 109-74-0
4. Ecotoxicity
Date 02.10.2003
Results of a pilot study conducted prior to this test indicated that a limit test
Remark :
design would be appropriate for the material.
The EbC50 (0-72 hr) and the ErC50 (0-72 hr) were inestimable as greater
than 50% inhibition in growth and/or biomass was not achieved. No
protocol deviations were noted.
Algae exposed to test material exhibited normal growth with respect to
Result :
control. At the end of the test, the mean cell density in treated cultures was
9.5 x 10E5 cells /ml (compared to 9.0 x 10E5 cells in control).
The average concentrations of material in the test flasks at the beginning of
the test and after 72 hours were 206.0 and 85.7 mg/l, respectively. The
mean concentration was 133.4 mg/l. This concentration was listed as the
NOEC, EbC50 and ErC50.
Results of the photostability tests were similar to those in flasks containing
test material and algae. The control solutions that were exposed to light or
were in the dark exhibited 58% and 50% losses of test material.
The mean temperature and illumination were 24 degrees C and 747 foot
candles (+/- 5.5 foot-candles) throughout the test. The pH ranged from 7.4
- 7.6.
The test was considered to be valid since the mean cell concentration in
control cultures increased by a factor of 90.2-fold within 72 hours.
Test Organisms: A 4-day culture of Selenastrum capricornutum SF-3148
Test condition :
(passage 3 in liquid algal medium) was used as the test algae. Several
passages were performed prior to the test to confirm exponential growth.
The density of cells in the stock culture was 2.58 x 10E6 cells/ml prior to
use.
Test medium: Sterile growth medium was prepared using high quality
distilled water. The pH of the medium was measured and adjusted to 7.5
(+/- 0.1) using 0.1N NaOH.
Test material stock solution: Approximately 0.151 ml (120 mg) of the test
material was added to 600 ml of algal growth medium with a gas tight
Hamilton syringe (to produce a nominal concentration of 200 mg/l). The
solution was stirred for approximately 1 minute. An aliquot (1.0) of the
solution was removed for analysis of concentration.
Test conduct: All steps were carried out aseptically in a hood to prevent
contamination. Test vessels were sterile 250 ml Erlenmeyer flasks. Test
material stock solution (100 ml) was added to 5 flasks and test medium that
did not contain test material was added to 3 flasks. Algae (388 microliters
of algal stock culture to achieve an initial cell density of 1 x 10E4 cells/ml)
were added to 3/5 flasks that contained test material and the three that did
not. The two flasks that contained test material but were not inoculated
served as photostability controls. One of the flasks was exposed to light
and one was wrapped in foil to shield it from light. All flasks were secured
with foam stoppers and transferred to a shaking incubator (24 degrees C,
100 rpm). They were illuminated at 747 (+/- 5.5) footcandles throughout the
study.
Temperature, light intensity, and shaker speed (rpm) were assessed at the
0, 24, 48, and 72 hours. Concentrations of test material in the flasks that
contained algae also were assessed at these times. The pH and was
assessed at time 0 and after 72 hours. Concentrations of test material in
the photostability controls also were measured at 0 and 72 hours.
Concentrations of test material were analyzed using gas chromatography
with flame ionization detection (GC/FID). The exposure concentration was
calculated as the geometric mean of the test concentrations analyzed at
17 / 17
Id 109-74-0
4. Ecotoxicity
Date 02.10.2003
the 4 time points. Cell counts were performed after 24, 48 and 72 hours of
exposure using a calibrated Coulter Counter. The mean algal cell count for
the test and control curves. Two measures of growth [biomass (area under
the growth curve) and growth rate] were used to determine the effect of the
material on algae. The concentrations that produced a 50% inhibition of
biomass (EbC50) and growth rate (ErC50) relative to control were to be
calculated by fitting linear
egression models to the data.
The test was considered valid if the mean cell concentration in the control
cultures increased by a factor of at least 16 within 72 hours.
Purity was 99.9% (GC/FID).
Test substance :
The results of this study indicate that the test substance would not be
Conclusion :
classified according to the European Union's labeling directive and would
correspond to a "low concern level" according to the U.S. EPA's
assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD-study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
07.08.2003 (8)
other algae: green algae
Species :
Endpoint :
96 hour(s)
Exposure period :
mg/l
Unit :
= 364.857 calculated
EC0 :
Limit test :
no
Analytical monitoring :
other: calculated
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program are CAS No., melting point (-112 degrees
Remark :
C), boiling point (117.5 degrees C), vapor pressure (19.5 mm Hg), and
water solubility (33,000 mg/l). The class of compound for estimation in the
model was neutral organic.
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
07.08.2003 (15)
4.4 TOXICITY TO MICROORGANISMS E.G. BACTERIA
4.5.1 CHRONIC TOXICITY TO FISH
4.5.2 CHRONIC TOXICITY TO AQUATIC INVERTEBRATES
4.6.1 TOXICITY TO SEDIMENT DWELLING ORGANISMS
4.6.2 TOXICITY TO TERRESTRIAL PLANTS
18 / 18
Id 109-74-0
4. Ecotoxicity
Date 02.10.2003
4.6.3 TOXICITY TO SOIL DWELLING ORGANISMS
4.6.4 TOX. TO OTHER NON MAMM. TERR. SPECIES
4.7 BIOLOGICAL EFFECTS MONITORING
4.8 BIOTRANSFORMATION AND KINETICS
4.9 ADDITIONAL REMARKS
19 / 19
Id 109-74-0
5. Toxicity
Date 02.10.2003
5.0 TOXICOKINETICS, METABOLISM AND DISTRIBUTION
5.1.1 ACUTE ORAL TOXICITY
LD50
Type :
= 50 - 100 mg/kg bw
Value :
rat
Species :
other: unknown
Strain :
no data
Sex :
21
Number of animals :
other: corn oil
Vehicle :
25-3200 mg/kg bw
Doses :
other
Method :
1960
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Whether any deaths occurred at 50 mg/kg was not listed. Mortality was
Result :
noted in all animals at dose of 100 mg/kg and above. Death was noted to
have occurred between 2 ?hours and 1 day. Clinical signs included:
moderate to very weak, severe tremors, vasodilatation, rough coat, sides
caved in, labored respiration, and convulsive movements progressing to
convulsions. Surviving animals were noted to have gained weight. There
was no gross evidence of adverse pathology in rats treated with 50 mg/kg.
A total of 21 rats were orally administered doses of n-butyronitrile ranging
Test condition :
from 25-3200 mg/kg. Material was administered undiluted and at 10% in
corn oil vehicle. Animals were monitored for clinical observations and
weight change for 14 days after exposure. Two rats that were given 50
mg/kg were killed 4 days after administration and the liver and kidneys
were examined for micropathology.
The boiling point of the test material was 116.4 -117.3 degrees C and the
Test substance :
index of refraction at 25 degrees was 1.3820.
Material is considered highly toxic.
Conclusion :
(2) valid with restrictions
Reliability :
Basic data are given. Purity of the test material is unknown.
07.08.2003 (12)
LD50
Type :
= 111 mg/kg bw
Value :
rat
Species :
other: Carworth-Wistar
Strain :
male
Sex :
Number of animals :
Vehicle :
Doses :
other
Method :
1962
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The LD50 value was 0.14 (0.10 - 0.19) ml/kg. Based on a density of
Result :
0.7954 (at 15 degrees C), the LD50 value in mg/kg is 111.
Test material was given by gavage to groups of 5 nonfasted rats (4-5
Test condition :
weeks old, 90-120 g). Dosages (not listed) were arranged in a logarithmic
series differing by a factor of 2. The material was administered in a
suitable vehicle (water, corn oil, 1% Tergitol Penetrant 7, or semi-solid
agar). The number of deaths was monitored over 14 days. The LD50
20 / 20
Id 109-74-0
5. Toxicity
Date 02.10.2003
value and its confidence interval was estimated by the method of
Thompson (Bacteriol Rev 11:115, 1947) using the tables of Weil
(Biometrics 8:249, 1952).
(2) valid with restrictions
Reliability :
Purity of the material, test concentrations and the number of deaths at each
concentration were not mentioned.
06.08.2003 (26)
5.1.2 ACUTE INHALATION TOXICITY
other: LC10
Type :
= 1848 ppm
Value :
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
40
Number of animals :
Vehicle :
1972, 4421, 6296 and 8261 ppm
Doses :
1 hour(s)
Exposure time :
OECD Guide-line 403 "Acute Inhalation Toxicity"
Method :
1987
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
LC10 value: LC10 (+/- 95% confidence interval) = 1848 (695-2656) ppm
Result :
(both sexes combined)
Exposure concentrations: Actual concentrations were 1972 +\- 57, 4421 +\
161, 6296 +\- 110, and 8261 +\- 204 ppm. No aerosol was present. Overall
mean chamber temperature was maintained at 21 degrees C and relative
humidity varied from 52 - 56%.
Deaths at each dose:
1972 ppm: 1/10; 1 male on Day 1
4421 ppm: 7/10; 5 males and 2 females on Day 1
6296 ppm: 9/10; 5 males and 4 females on Day 1
8261 ppm: 9/10; 5 males and 4 females on Day 1
Remarks: Clinical observations noted at 2000 ppm included gait
disturbance, poor condition, and sialorrhea (only seen in the one male that
died). One 2000 ppm-exposed female was lethargic and had sialorrhea.
Lethargy was noted prior to the death of all males at higher exposure
levels. At 4000 ppm and above, females exhibited lethargy, poor body
condition, sialorrhea, and porphyrin-like discharge around the nose and
face. These clinical signs were seen during or just after exposure
cessation. Their severity was dose-related, and they resolved after 24
hours. The mean body weight increased in all animals during the 14-day
observation period. No compound-related gross pathology was noted in
animals that died spontaneously or in animals terminated on Day 14.
Rats [CRL:CD(SD)BR] were exposed to target vapor concentrations of
Test condition :
2000, 4000, 6000 and 8000 ppm (5/sex/concentration). Males weighed
182-217 g and females weighed 171-194 g at the start of the study. Food
and water were available ad libitum (except during exposure).
Exposures were conducted in 420 l stainless steel and glass inhalation
chambers. Chambers were maintained under negative pressure (-0.5"
water) and at 12 air changes per hour. Vapors were generated by metering
the test material dropwise into a heated glass bead-packed column
supplied with metered dried oil-free compressed air. Chamber vapor
concentrations were determined 4-5 times per hour with an infrared
21 / 21
Id 109-74-0
5. Toxicity
Date 02.10.2003
analyzer equipped for automated sampling and analyses. Temperature
and humidity were measured twice per hour. Once during exposure, the
concentration of background nongaseous material was measured in the
6000 ppm chamber relative to a chamber containing air in order to insure
that exposures were to vapor and not aerosol. Distribution of test material
was determined by measuring the concentrations in 10 different positions
throughout the chamber and comparing them to a fixed reference position.
Subsequently, the chamber vapor concentration was measured from the
fixed reference position.
Exposure was for 1-hour in duration followed by a 14-day observation
period. Rats visible through the chamber windows were observed for signs
of toxicity during exposure. Each rat was removed from its cage before and
after exposure and twice daily thereafter (on each workday) and examined.
Animals that died during the study were necropsied as soon as possible
after discovery of death. Survivors were fasted for 16 hours prior to
euthanization and necropsy on Day 15. The following tissues were
examined: trachea, lungs, heart, liver, esophagus, stomach, duodenum,
jejunum, ileum , cecum, colon, pancreas, liver, salivary glands, kidneys,
urinary bladder, pituitary gland, adrenal, thyroid, parathyroid, thymus,
spleen, mesenteric lymph nodes, bone marrow (femoral), brain, testes,
epididiymides, male accessory sex glands, ovaries, vagina, uterus, and
Fallopian tubes. Mortality data were evaluated by Probit analysis.
Purity was 99.5%
Test substance :
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
07.08.2003 (13)
LC50
Type :
> 1147 ppm
Value :
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
10
Number of animals :
Vehicle :
1147 ppm (males), 1220 ppm (females)
Doses :
1 hour(s)
Exposure time :
other: DOT Final Rule 49 CFR (parts 172 and 173)
Method :
1986
Year :
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The study was conducted according to GLP with the exceptions that
Remark :
stability was not determined, reserve samples were not collected and the
degree of absorption was not determined (because it was not applicable).
All animals survived to the end of the study. Mean vapor concentrations of
Result :
the test material were 1147 +/- 24 and 1220 +/- 28 ppm for male and
female rats, respectively. The saturated vapor concentration at 20 degrees
C was 28,553 ppm. The temperature and relative humidity of the room
ranged from 68-70 degrees F and 55-60%, respectively. An aerosol was
not present.
Groups of 5 male (231-242 g) and 5 female (212-224 g) rats
Test condition :
[CRL:CD(SD)BR] were exposed to a target vapor concentration of 1200
ppm for 1 hour. Feed and water were available ad libitum (except during
exposure). Rats were observed for mortality during exposure and twice
daily on subsequent workdays for 14 days.
Chamber atmospheres were produced by passing dried, oil-free
compressed air through a glass bead-packed distilling column into which
the test material was pumped dropwise. The exposure began when the
22 / 22
Id 109-74-0
5. Toxicity
Date 02.10.2003
vapor concentration reached 1000 ppm. Males and females were exposed
in separate 20 l bell jars. Four samples of the atmosphere were analyzed
for n-butyronitrile concentration with an infrared analyzer. Two samples of
atmosphere were analyzed with a particle analyzer to insure that an
aerosol was not present. The temperature was monitored 4 times during
exposure.
Purity of the test material was 99.5% (GC-MS). 4-heptanone (0.5%) was
Test substance :
the only trace component identified.
The material is not subject to regulation under 49 CFR Parts 172 and 173,
Conclusion :
since the LC50 value was greater than 1000 ppm.
(1) valid without restriction
Reliability :
The study is comparable to an OECD guideline, GLP study.
07.08.2003 (11)
5.1.3 ACUTE DERMAL TOXICITY
Type :
= 398 mg/kg bw
Value :
rabbit
Species :
New Zealand white
Strain :
male
Sex :
Number of animals :
Vehicle :
Doses :
other: one-day cuff method of Draize
Method :
1962
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The LD50 value (with confidence interval) was 0.50 (0.37 - 0.68) ml/kg.
Result :
Based on a density of 0.7954 (at 15 degrees C), the LD50 value in mg/kg is
398.
Fur was clipped from the entire trunk of rabbits (2.5 to 3.5 kg) and the
Test condition :
material was applied beneath an impervious plastic film. Groups of 4
animals were exposed to various concentrations (not listed). The animals
were immobilized during the 24-hour contact period. The film was then
removed and the animals were caged for the subsequent 14 day
observation period. The LD50 value and its confidence interval was
estimated by the method of Thompson (Bacteriol Rev 11:115, 1947) using
the tables of Weil (Biometrics 8:249, 1952).
(2) valid with restrictions
Reliability :
Purity of the material, test concentrations and the number of deaths at each
concentration were not mentioned.
06.08.2003 (26)
5.1.4 ACUTE TOXICITY, OTHER ROUTES
5.2.1 SKIN IRRITATION
5.2.2 EYE IRRITATION
5.3 SENSITIZATION
23 / 23
Id 109-74-0
5. Toxicity
Date 02.10.2003
5.4 REPEATED DOSE TOXICITY
Sub-chronic
Type :
rat
Species :
male/female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
14 weeks (total of 63 exposure days)
Exposure period :
6 hours/day, 5 days per week
Frequency of treatm. :
none
Post exposure period :
60, 120, 209 ppm
Doses :
yes
Control group :
< 60 ppm
NOAEL :
= 60 ppm
LOAEL :
other
Method :
1984
Year :
no data
GLP :
other TS
Test substance :
Test material concentrations: The nominal concentrations (+/- SD) were
Result :
56.0 +/- 8.8, 119.1 +/- 16.9 and 203.0 +/- 19.6. Corresponding analytical
concentrations were 60.2 +/- 1.0, 120.3 +/- 1.1 and 209.0 +/- 1.3 ppm.
Since the nominal analytical ratios were close to 1, the material was a true
vapor. There was no indication that the test material was unstable.
Distribution was uniform (> 96%) for each of the exposure concentrations.
Airflow, temperature and relative humidity ranged from 1719-1765 l/min,
21.0 - 26.1 degrees C and 17-50%, respectively.
Effects at all exposure concentrations: Signs of toxicity (labored breathing,
nasal discharge, salivation, discharge from the eyes, hypoactivity and/or
alopecia) were observed in all exposed groups. Incidences of these signs
increased in a dose-dependent manner. Males and/or females in all
exposed groups had significant decreases in red blood cells and
hemoglobin values. Urine thiocyanate concentrations increased in all
exposed groups, with concentrations from animals exposed to 120 ppm
similar to or higher than those exposed to 210 ppm. However, since a
dose-dependent diuresis occurred, the total amount of urine thiocyanate
present (concentration x urine volume) increased with increasing
concentrations.
Effects at 209 ppm: Three males died or were killed in extremis (2
between exposures 2 and 3 and one between exposures 18 and 19).
Arched back, ataxia, tremors or convulsions, biting, pawing or rubbing chin
against cage, irritation of the conjunctiva and breathing difficulties were
noted in a few animals during exposure. Males and females exhibited
significant decreases in body weight throughout the study (P <= 0.01).
Average final body weights of males and females were lower than controls
(377.5 g in exposed males vs. 435.4 g in controls and 255.0 g in exposed
females vs. 276.6 g in controls). Absolute and/or relative heart, liver, spleen
and kidney weights were increased in males and/or females. Absolute
testes weights were decreased in males. Mean corpuscular hemoglobin
concentrations (males only) were lower than control (all P <= 0.05). Serum
alkaline phosphatase (males and females), SGOT (males only) and SGPT
(males only) were increased, and BUN concentrations were decreased. No
gross lesions were observed at termination. There was mildly increased
hemosiderin deposition in the spleens of 10/15 females. Other microscopic
changes were considered to be spontaneous.
Effects at 120 ppm: Ataxia was observed during exposure in 2 females.
Males had significant body weight decreases (P < = 0.05) at two time
points. Absolute and/or relative liver weights were increased in males and
24 / 24
Id 109-74-0
5. Toxicity
Date 02.10.2003
females and absolute and/or relative spleen weights were increased in
males. Mean corpuscular hemoglobin concentrations (males only, P < =
0.05) were lower than control. No gross lesions were observed at
termination. There was mildly increased hemosiderin deposition in the
spleens of 11/15 females. Other microscopic changes were considered to
be spontaneous.
Effects at 60 ppm: Absolute and relative spleen weights were increased in
males. Serum thiocyanate concentrations were marginally increased in
males and females.
Animals: Rats were acclimated for at least 10 days prior to use. Two days
Test condition :
prior to the start of the study, males weighed 174-200 g and females
weighed 132-145 g. On the first day of the study, the animals were 43
days old. Animals were randomly allocated by body weight into 4 groups of
15 animals/sex/group. Animals were individually housed in suspended
mesh cages and given food and water ad libitum (except during exposure).
Animal rooms were maintained at 70-74 degrees C and 35-60% relative
humidity, with a 12 hour light/dark cycle.
Exposure conditions: Exposures (6 hr/day, 5 days/week) occurred in 10
m3 Rochester-style stainless steel and glass inhalation chambers. Rats
were placed individually in wire mesh cages that were suspended in the
chambers by 3-tiered racks. Males were placed on one side and females
on the other. The concentrations of material in the chambers (20, 120 or
210 ppm) were controlled by either adjusting the nitrogen flow though the
propionitrile in the bubblers or by changing the amount of test material in
the bubblers. The bubbler was connected to a side port in the vertical
particle-size separator, which was in turn connected to the air inlet at the
top of the inhalation chamber. One bubbler was used in each of the
generation systems. Airflow was maintained at a constant flow of 1727
liters/min. Nominal concentration measurements were determined daily for
each chamber following exposure, by dividing the amount of test material
delivered to the chamber (the difference between the pre- and post-
exposure weights) over the 6-hr exposure period by the total air volume
during the same period. Concentrations of test material in the chambers
were measured 4 times daily using a Miran 1A General Purpose Gas
Analyzer. Additional samples of atmosphere from 9 specified locations in
each chamber were also taken at 3 different times to determine if the vapor
was distributed uniformly.
Test conduct: Animals were observed for clinical signs between the second
and fifth hour of each exposure. Estimations of the percentages of animals
exhibiting hypoactivity, eye irritation and breathing difficulties were made.
All animals were individually examined for gross signs of toxicity preceding
and following each exposure and checked for mortality. Each animal was
weighed and given a thorough examination for gross signs of toxicity on a
weekly basis.
Animals were euthanized after 14 total weeks on the study. Terminal body
weights were obtained (following an overnight fast). Blood and urine were
collected. Whole blood was treated with an anticoagulant and was
analyzed for total and differential erythrocyte count, total leukocyte count,
platelet count, hematocrit, hemoglobin, and red blood cell indices (mean
corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular
hemoglobin concentration). Serum was analyzed for albumin, globulin, total
protein, blood urea nitrogen, total bilirubin, glucose, glutamic pyruvic
transaminase (SGPT), alkaline phosphatase, glutamic oxaloacetic
transaminase (SGOT), T3, T4, thiocyanate and lactate dehydrogenase.
Urine was analyzed for the presence of thiocyanates.
Detailed necropsies were conducted on all rats that died during the course
of the study, those that were killed moribund, and those that survived to
25 / 25
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5. Toxicity
Date 02.10.2003
study termination. The adrenal glands (both together), testes (with
epididymides, heart, kidneys, liver, pituitary and spleen were weighed. The
aforementioned organs and the following tissues were fixed in 10% neutral
formalin: abdominal aorta, bone and bone marrow (femur), brain,
esophagus, ovaries, colon, ileum, lung, lymph nodes (mesenteric),
mammary gland, nasal turbinates, pancreas, thyroid/parathyroid, prostate,
salivary gland, sciatic nerve, skeletal muscle, skin, spinal cord, stomach,
thymus, trachea, urinary bladder, uterus (with cervix) and gross lesions.
Eyes (with optic nerve) were fixed in a solution of 2% glutaraldehyde and
10% neutral buffered formalin. Tissues were processed, embedded in
paraffin, cut at five microns, stained with hematoxylin and examined
microscopically.
Statistical analyses: In life and terminal body weights and organ weight
data were analyzed using Dunnett's test. Organ to body weight ratios were
analyzed using the Mann-Whitney test, with the Bonferroni inequality. Data
for frequencies of microscopic lesions were evaluated with the Fisher's
exact test with the Bonferroni inequality. Hematological and serum and
urine chemistry variables were examined using Dunnett's test.
Test material was propionitrile (CAS No. 107-12-0). The purity of the test
Test substance :
material was 96%. Impurities were not listed.
(1) valid without restriction
Reliability :
The study is comparable to a guideline study; however, a NOAEL was not
established.
Critical study for SIDS endpoint
Flag :
10.08.2003 (27)
5.5 GENETIC TOXICITY `IN VITRO`
Bacterial reverse mutation assay
Type :
Salmonella typhimurium/TA98, 100, 1535, 1537, and Escherichia
System of testing :
coli/WP2uvrA(pKM101)
100, 333, 1000, 3330 and 5000 micrograms/plate
Test concentration :
> 5000 micrograms/plate
Cytotoxic concentr. :
with and without
Metabolic activation :
negative
Result :
other: EEC Annex V Guideline number B.14, "Other Effects-Mutagenicity
Method :
Salmonella typhimurium-Reverse Mutation Assay", and Guideline number
B.13, Other Effects-Mutagenicity, Escherichia coli-Reverse Mutation
Assay"
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
This is the critical study for the mutagenesis endpoint.
Remark :
No positive responses were induced in any of the tester
Result :
strains. None of the concentrations tested caused toxicity. No precipitate
was observed at the maximum concentration tested. All criteria for a valid
test were met.
Test strains: The S. typhimurium and E. coli strains were obtained from Dr.
Test condition :
Bruce Ames, University of California Berkeley and the National Collection
of Industrial Bacteria, Torry Research Station, Scotland, respectively.
Frozen permanent stocks were prepared by growing fresh overnight
cultures, adding DMSO (0.09 ml/ml culture) and freezing small aliquots at <
= -70 degrees C. Master plates were prepared by streaking each test
strain from a frozen permanent stock onto minimal agar supplemented with
histidine, biotin, ampicillin and/or tryptophan (depending on the strain).
Tester strain master plates were stored at 5 +/- 3 degrees C. Overnight
cultures were inoculated by transferring colonies from the master plates to
a flask containing culture medium. Inoculated flasks were placed in
26 / 26
Id 109-74-0
5. Toxicity
Date 02.10.2003
a shaker/incubator (125 +/- 25 rpm, 37 +/- 2 degrees C). Cultures were
harvested once a predetermined turbidity was reached (at least 0.5 x 10E9
cells/ml). Test stains were checked for rfa wall mutation (all Salmonella
strains), pKM101 plasmid R-factor (Salmonella TA98 and TA100 and E.
coli only), and characteristic number of spontaneous revertants (all strains)
on the day the mutagenesis test was conducted.
Test medium: The broth used to grow overnight cultures of the tester
strains was Vogel-Bonner salt solution supplemented with 2.5% Oxoid
Nutrient Broth No. 2. Bottom agar was Vogel-Bonner minimal medium E
supplemented with 1.5% (w/v) agar and 0.2% (w/v) glucose. Overlay agar
contained 0.7% agar (w/v) and 0.5% NaCl (w/v) supplemented with either
10 ml of 0.5 mM histidine/biotin solution or 0.5 mM tryptophan solution.
S-9 mix: S9 homogenate was purchased from Molecular Toxicology Inc.
This was prepared from male Sprague-Dawley rats that had been injected
i.p. with 500 mg/kg Aroclor 1254. S-9 mix was prepared immediately prior
to use.
Concentrations of test material: The concentrations tested (100, 333, 1000,
3330 and 5000 micrograms/plate) were selected based on the results of a
dose range-finding study using test strains TA100 and WP2uvrA(pKM101)
and 10 doses of test material ranging from 6.67 to 5000 micrograms/plate
(both in the presence and absence of S-9 mix).
Positive, negative and sterility controls: Positive controls [2
aminoanthracene (2.5 and 5.0 micrograms/plate), 2-nitrofluorene (1.0
micrograms/plate), sodium azide (2.0 micrograms/plate), ICR-191 (2.0
micrograms/plate), and 4-nitroquinoline-N-oxide (2 micrograms/plate)] were
run concurrently. DMSO (50 microliters) was used as a vehicle and vehicle
control. The most concentrated test material dilution and S-9 mix were
tested for sterility by plating a 50 microliter aliquot on selective agar.
Test conduct: A plate incorporation methodology was used. Test material
or positive control (50 microliters), test strains (100 microliters) and S-9 mix
or vehicle (500 microliters) were combined in 2.0 ml of molten, selective
top agar. This was overlaid onto 25 ml of minimal agar that had been
plated into 15 x 100 mm Petri dishes. All concentrations of test material,
vehicle controls and positive controls were plated in triplicate. Revertant
colonies were counted after 48 +/- 8 hours of inverted incubation at 37 +/- 2
degrees C. The condition of the background lawn was evaluated for
evidence of cytotoxicity and precipitate.
Evaluation: The mean number of revertants and standard deviation were
calculated. Various criteria were established to constitute a valid assay
(test strain integrity, characteristic number of spontaneous revertants, cell
density > = 0.5 x 10E9, at least a 3-fold increase in revertants in positive
controls, and a minimum of 3 non-toxic doses). A positive response was
indicated by a 2-3 fold increase in mean revertant number depending on
the bacterial tester strain.
Purity was 99.9%
Test substance :
Material was not genotoxic under conditions of this assay.
Conclusion :
(1) valid without restriction
Reliability :
This was a well-documented EEC Annex guideline study conducted under
GLP assurances.
10.08.2003 (3)
Chromosomal aberration test
Type :
Chinese Hamster Ovary (CHO) Cells
System of testing :
up to 701 micrograms/ml
Test concentration :
>701 microgams/ml
Cytotoxic concentr. :
with and without
Metabolic activation :
27 / 27
Id 109-74-0
5. Toxicity
Date 02.10.2003
negative
Result :
OECD Guide-line 473
Method :
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
This is the critical study for the chromosomal aberration endpoint.
Remark :
No significant increases in cells with chromosomal aberrations, polyploidy,
Result :
or endoreduplication were observed in analyzed cultures. No precipitate
was observed at the maximum concentration tested. None of the
concentrations caused a significant reduction in the mitotic index. All
criteria for validity were met.
Cells: The Chinese hamster ovary cells used in the assay (CHO-WBL)
Test condition :
were from a permanent cell line originally obtained from Dr. S. Wolff,
University of California, San Francisco. Stock cultures were maintained for
up to 8 weeks after thawing. Mycoplasma testing was performed twice
during this period. Cells were grown at 37 +/- 2 degrees C (in 5% +/- 1.5%
Co2 in air) in McCoy's 5a culture medium which was supplemented with
10% fetal bovine serum, 2 mM L-glutamine, 100 units/ml penicillin G and
100 micrograms/ml streptomycin.
S-9 mix: S-9 was isolated from the liver of rats (sex not stated) 5 days after
i.p. treatment with 500 mg/kg Aroclor 1254. S-9 was stored frozen at < =
70 degrees C until use. S-9 mix was prepared by adding an energy-
producing system (NADP plus isocitric acid) to S-9.
Test material and negative and positive controls: The test material was
dissolved in DMSO. The top concentration tested (approximately 700
micrograms/ml or 10 mM) was the recommended high dose for the assay.
The negative control was 10 microliters/ml DMSO. The positive controls
were mitomycin C (without activation) and cyclophosphamide (with
activation).
Initial test: Cultures were initiated by seeding approximately 1.2 x 10E6
cells per 75 cm2 flask into 10 ml of complete McCoy's 5a medium. For the
test without metabolic activation, cultures were incubated with test
material for 3.0 hrs at 37 degrees C. For the test with metabolic activation,
cells were incubated for approximately 3.0 hours with test material and S-9
mix in McCoy's 5a medium that did not contain fetal bovine serum.
Replicate cultures for each concentration of test material (4.77, 6.81, 9.73,
13.9, 19.9, 28.4, 40.5, 57.8, 82.6, 118, 169, 241, 344, 491 and 701
micrograms/ml), positive control (0.75 and 1.5 micrograms/ml mitomycin C
and 5.0 and 10.0 micrograms/ml cyclophosphamide), vehicle and untreated
controls were prepared. Cultures with or without S-9 were then washed
with buffered saline, and incubated with complete McCoy's 5a medium for
17 hours. Colcemid (0.1 micrograms/ml) was present during the last 2
hours of incubation. Cells were visually inspected for cytotoxicity prior to
harvest. Cells were then trypsinized and spun in a centrifuge. The
supernatant was discarded and the cells swollen with 75 mM KCl hypotonic
solution. The cells were then fixed with an absolute methanol: glacial
acetic acid (3:1, v:v) fixative. They were then placed on glass slides and
air-dried. Cells were stained with 5% Giemsa and analyzed for mitotic
index and chromosomal aberrations.
Confirmatory assay: As in the previous test, cells were harvested after 20
total hours of incubation. In this test, the test with metabolic activation was
conducted the same as in the initial test, but with different concentrations of
test material (217, 289, 385, 513 and 684 micrograms/ml). In the test
without metabolic activation, the test material (27.3, 54.5, 109, 217, 289,
385, 513 and 684 micrograms/ml), positive control (0.20 or 0.40
micrograms/ml mitomycin C or 5.0 or 10.0 micrograms/ml
cyclophosphamide) and negative controls were incubated with the cells for
28 / 28
Id 109-74-0
5. Toxicity
Date 02.10.2003
17.8 hours (instead of 3), and colcemid was present for the last 2.2 hours
of harvest (instead of 2). The slides were prepared as described for the
previous test.
Evaluation: Cells were selected for good morphology and
only cells with the number of centromeres equal to the modal number 21
+/-2 were analyzed. One hundred cells (if possible) were analyzed from
each replicate of the vehicle control, 4 concentrations of the test material
(the highest 4 concentrations tested), and one concentration of positive
control for the different types of chromosomal aberrations. At least 25 cells
were analyzed from those cultures that had greater than 25% of cells with
one or more aberrations. The number of mitotic cells in 1000 cells was
determined and the ratio expressed as percentage of mitotic cells. Percent
polyploidy and endoreduplication were analyzed by evaluating 100
metaphases (if possible). Chromatid and isochromatid gaps were noted but
were not used in calculating the total number of aberrations.
Acceptance criteria: The assay was considered valid if the negative
(untreated) and vehicle controls contained < 5% cells with aberrations, the
positive control result was significantly higher than that of the vehicle
control, a high dose of 10 mM or the highest soluble concentration was
used if the material did not cause a reduction of the mitotic index at the
tested concentrations, and at least 3 concentrations were analyzed.
Data analysis: The statistical analysis employed a Cochran-Armitage test
for linear trends and Fisher's Exact Test to compare the percentage of cells
with aberrations. Data for polyploidy and/or endoreduplication were also
analyzed separately. A test was considered positive if a significant
increase in the number of cells with aberrations (p < = 0.01) was observed
at one or more concentrations.
Purity was 99.9%
Test substance :
Material was not genotoxic under conditions of this assay.
Conclusion :
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted under GLP
assurances.
06.08.2003 (2)
5.6 GENETIC TOXICITY `IN VIVO`
5.7 CARCINOGENICITY
5.8.1 TOXICITY TO FERTILITY
Fertility
Type :
rat
Species :
female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
21 to 33 days (depending on day of mating)
Exposure period :
6 hr/day, 7 days/week
Frequency of treatm. :
Premating exposure period
0 days
Male :
21 days
Female :
to gestation days 13-15
Duration of test :
No. of generation :
studies
60, 120 and 210 ppm
Doses :
29 / 29
Id 109-74-0
5. Toxicity
Date 02.10.2003
yes
Control group :
= 60 ppm
NOAEL parental :
= 210 ppm
other: NOAEL :
Reproductive Toxicity
other
Method :
1984
Year :
yes
GLP :
other TS
Test substance :
Exposure concentrations: The average mean daily analytical exposure
Result :
concentrations (60.1, 120.2 and 209.2 ppm) were within 1% of the target
levels (60, 120 and 210 ppm, respectively). Mean temperatures and
humidities ranged from 22-25 degrees C and 26-29%, respectively.
Signs of toxicity: None of the animals died. There was no effect of test
material on body weight. Animals exposed to 210 ppm exhibited arched
back (N = 4 on days 1-10 and N=2 on days 11-20), lacrimation (N = 2 on
days 1-10 and N = 1 on days 21-30), salivation (N= 15 on days 1-10, N =
22 on days 11-20 and N = 21 on days 21-30) hypoactivity (N = 13 on days
1-10, N = 5 on days 11-20 and N = 3 on days 21-30), staining of facial fur
(N = 2 on days 1-10, N = 4 on days 11-20 and N = 4 on days 21-30) and
red nasal encrustation (N = 1 on days 1-10, N = 5 on days 11-20 and N = 5
on days 21-20) after exposure. Animals exposed to 120 ppm also exhibited
salivation (N = 6 on days 11-20 and N = 4 on days 21-30, staining of facial
fur (N = 7 on days 1-10, N = 5 on days 11-10 and N = 2 on days 21-30)
and red nasal encrustation (N = 2 on days 1 1-0, N = 8 on days 11-20 and
N = 6 on days 21-30). A few animals in the 60 ppm group also exhibited
red nasal encrustation (N = 1 on days 1-10, N = 3 on days 11-20 and N = 1
on days 21-30) and staining of facial fur (N = 1 on days 1-10, N = 3 on days
11-20 and N = 1 on days 21-20). One control animal had stained facial fur
on days 21-30 and another had red nasal encrustation on days 1-10. Signs
of toxicity generally abated by the morning after exposure. Alopecia was
observed in animals (N = 2 controls, N = 3 low dose, N = 5 mid dose, N =
9) at one or more of their weekly physical examinations.
The only remarkable findings at gross necropsy were bilateral uterine
hydrometra in one animal exposed to 210 ppm and hydrometra in the left
uterine horn of one animal exposed to 120 ppm.
Fertility: There was no effect of treatment on fertility. Efficiency of mating
(32.0%, 32.0%, 30.7% and 25.0% in the control, low, mid and high dose
groups) and pregnancy rate (100%, 95.8%, 100% and 91.3% in the
respective groups) were comparable between groups. There was no
difference in the numbers of live implants (ranged from 13.4 - 13.9),
resorptions (ranged from 0.6 - 0.8), nidations (ranged from 14.1 - 14.5),
corpora lutea (ranged from 13.0 - 15.2), preimplantation loss (4-8%) and
postimplantation loss (4-6%). Evaluation of the vaginal smears of 2 females
that did not copulate showed one that did not cycle (but was pregnant at
necropsy), and another that only went through the cycling stage of
proestrus.
Animals: Virgin female Sprague Dawley rats (43 days old upon receipt) and
Test condition :
virgin male Sprague Dawley rats (50 days old upon receipt) were
acclimated for one week and examined for general health and the
presence of pinworms and ectoparasites. Body weights of ten females and
ten males that were taken upon receipt were 128-144 g and 178-233 g,
respectively. No significant health problems were noted during the
acclimation period, and the animals were released for study. Males and
females were individually caged (except during mating). Food and water
were available ad libitum (except food was not available to females during
exposure). Animals were housed in a room maintained at 72 +/- 2 degrees
F and 40-60% relative humidity, under a 12 hr light/dark cycle.
30 / 30
Id 109-74-0
5. Toxicity
Date 02.10.2003
Exposures: Exposures (6 hr/day, 7 days/week) occurred in 10 m3
Rochester-style stainless steel and glass inhalation chambers. Due to
inclement weather and building equipment failures, 2 exposures (days 2
and 16) were only for 4 hours and one exposure (day 1) was for 5 hours.
Atmospheres of propionitrile vapor were generated using bubbler systems.
The concentrations of material in the chambers were controlled by either
adjusting the nitrogen flow though the propionitrile in the bubblers or by
changing the amount of test material in the bubblers. Concentrations of
test material in the chambers were measured 4 times daily (except during
day 16) using a Miran 1A General Purpose Gas Analyzer. The nominal
concentration of material, temperature and humidity also were measured
(at intervals that were not listed).
Study conduct: Twenty four females per group were assigned to be
exposed to target concentrations of 0, 60, 120 or 210 ppm test material.
Exposure began when females were 63 days old. Animals were observed
during exposure for signs of toxicity. After 21 days of exposure (which was
sufficient to cover 3-4 estrus cycles), females were randomly mated (1:1) to
an untreated male that had been assigned to the corresponding treatment
group (30 males were assigned per group). At night, after exposure,
females were caged with their assigned male until copulation was
confirmed (by presence of a copulatory plug or sperm in vaginal smear) or
5 nights without confirmed copulation. Females that failed to mate with the
assigned male were mated with another male that had copulated with
another female in the same group. Nightly co-housing with the second
male occurred until copulation was confirmed (or for a maximum of 7
nights). The day on which copulation was confirmed was considered
gestation day 0. Exposure of females continued until copulation was
confirmed or a maximum of 12 nights of cohabitation with males without
signs of copulation. Vaginal smears were taken on 5 consecutive days for
females that did not exhibit copulation.
After assignment to the study, males and females were weighed once per
week. Mated females were weighed on gestation days 0 and 13. Females
were given a thorough physical examination once per week and were
observed for clinical signs of toxicity before and after exposures. All
animals were checked twice daily for mortality and gross abnormalities.
Females were killed on gestation day 13 (or the nearest working day after
gestation day 13, up to gestation day 15). Females without confirmed
copulation ware euthanized in the second week after the last day of co-
housing. Each female was given an external examination and weighed.
The tissues and organs of the thoracic and abdominal cavities were
examined for gross lesions. Pregnancy status was determined, nidation
sites and were classified, and corpora lutea were counted. The ovaries
and uteri of females were preserved in 10% neutral buffered formalin.
Males were killed after mating and were not examined.
Statistical analyses: Body weight data were analyzed using Dunnett's test.
Mating and pregnancy rate data were analyzed by a Fisher's exact test and
an uncorrected chi-square test. Other data were analyzed by the Mann-
Whitney U test. The Bonferroni inequality was assumed when comparing
multiple treatments to control values for all tests except Dunnett's test. The
critical level for significance was p < 0.05.
The test material was propionitrile (CAS No. 107-12-0). Purity of the test
Test substance :
material was 96.1%. Impurities included acrylonitrile (0.1%), adiponitrile
(0.7%), p-nitrosodiphenylamine (0.08%) and water (< 0.1%). Analyses
indicated no significant decomposition of the test material over the course
of the study.
The authors concluded that the incidences of red nasal encrustation in the
Conclusion :
low dose animals, alopecia in the mid and high dose animals and staining
of facial fur in all treated groups were too low to be definitely related to
31 / 31
Id 109-74-0
5. Toxicity
Date 02.10.2003
administration of test material. There was no effect of treatment on fertility
of females.
(1) valid without restriction
Reliability :
Study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (20)
Fertility
Type :
rat
Species :
male
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
57 days
Exposure period :
6 hours/day, 5 days/week
Frequency of treatm. :
Premating exposure period
46 days
Male :
0 days
Female :
to gestation day 13-15
Duration of test :
No. of generation :
studies
60, 120 and 210 ppm
Doses :
yes
Control group :
= 60 ppm
NOAEL parental :
= 210 ppm
other: NOAEL :
Reproductive Toxicity
other
Method :
1985
Year :
yes
GLP :
other TS
Test substance :
Exposure concentrations: The average mean daily analytical exposure
Result :
concentrations (60.2, 120.4 and 208.9 ppm) were within 1% of the target
levels (60, 120 and 210 ppm, respectively). Mean temperatures and
humidities ranged from 22-25.5 degrees C and 24-27%, respectively.
Signs of toxicity: One of animals exposed to 210 ppm died after 2 days of
exposure. On the previous day, this animal exhibited labored breathing,
hypoactivity, poor control of the hind limbs, difficulty in standing, body
tremors and involuntary movements. No unusual findings were observed
at necropsy.
Body weights of males exposed to 210 ppm were approximately 6-9%
lower than those of the control group during most of the exposure period,
and remained lower than control (but were not significantly different) until
the end of the study.
Animals exposed to 210 ppm exhibited signs of toxicity such as arched
back (N = 8 on days 1-10, N = 3 on days 11-20 and 51-57, and N = 5 on
days 41-50), hypoactivity (N = 12-15 at each 10-day interval up to day 50,
and N = 4 from days 51-57), labored breathing (N = 10 on days 1-10, N = 3
on days 11-20 and 31-40, N = 5 on days 21-30 and N = 1 on days 51-57),
and salivation (N = 3 on days 1-10, and N = 10 - 12 at all other intervals). A
few high dose animals (individual numbers were not stated) also exhibited
abnormal behavior such as grinding of teeth, head bobbing, body tremors,
involuntary movements, and pawing at the cage. A few of the animals
exposed to 120 ppm exhibited salivation (N = 3-8 at all intervals) and
hypoactivity (N = 3 at days 11-20). Signs of toxicity generally abated by the
morning after exposure. Alopecia was observed in animals (N = 1 control,
N = 2 low dose, N = 1 mid dose, N = 5 high dose) at one or more of their
weekly physical examinations. No unusual treatment-related signs were
observed in rats exposed to 60 ppm. The only remarkable finding at gross
necropsy was a small right testis in one animal exposed to 120 ppm.
32 / 32
Id 109-74-0
5. Toxicity
Date 02.10.2003
Fertility: There was no effect of treatment on male fertility. Efficiency of
mating (34.4%, 30.6%, 29.8% and 27.1% in the control, low, mid and high
dose groups) and pregnancy rate (90.5%, 97.6%, 90.0% and 97.4% in the
respective groups) were comparable between groups. There was no
difference in the numbers of live implants (ranged from 12.7 - 13.9),
resorptions (ranged from 0.7 - 1.1), nidations (ranged from 13.8 - 14.9),
corpora lutea (ranged from 13.1 - 15.2), preimplantation loss (4-8%) and
postimplantation loss (5-10%).
Animals: Virgin female Sprague Dawley rats (28 days old upon receipt) and
Test condition :
virgin male Sprague Dawley rats (50 days old upon receipt) were
acclimated for one week and examined for general health and the
presence of pinworms and ectoparasites. Body weights of fifteen females
and ten males that were taken upon receipt were 155-181 g and 80-103 g,
respectively. No significant health problems were noted during the
acclimation period, and the animals were released for study. Males and
females were individually caged (except during mating). Food and water
were available ad libitum (except food was not available to males during
exposure). Animals were housed in a room maintained at 72 +/- 2 degrees
F and 40-60% relative humidity, under a 12 hr light/dark cycle.
Exposures: Exposures (6 hr/day, 5 days/week) occurred in 10 m3
Rochester-style stainless steel and glass inhalation chambers. A
scheduled exposure day was cancelled due in clement weather. A new
exposure day (exposure day 41) was used in its place. Due to inclement
weather and building equipment failures, 2 exposures (days 33 and 43)
were only for 4 hours and one exposure (day 32) was for 5 hours.
Atmospheres of propionitrile vapor were generated using bubbler systems.
The concentrations of material in the chambers were controlled by either
adjusting the nitrogen flow though the propionitrile in the bubblers or by
changing the amount of test material in the bubblers. Concentrations of
test material in the chambers were measured 4 times daily (except during
day 43) using a Miran 1A General Purpose Gas Analyzer. The nominal
concentration of material, temperature and humidity also were measured
(at intervals that were not listed).
Study conduct: Fifteen males per group were assigned to be exposed to
target concentrations of 0, 60, 120 or 210 ppm test material. Exposure
began when males were 43 days old. Mating was initiated when males and
females were 16 and 12 weeks old, respectively. At this time, males had
been 69 days on the study (which was sufficient to cover the
spermatogenesis cycle of the rat), and had 46 days of exposure. Males
were randomly mated (1: 1) with three untreated females (consecutively)
that had been assigned to the corresponding treatment group (45 females
were assigned per group). Exposure of males continued until the day after
the last mating opportunity (57 exposure days). At night, after exposure,
males were caged with their assigned female until copulation was
confirmed (by presence of a copulatory plug or sperm in vaginal smear) or
5 nights without confirmed copulation. The day on which copulation was
confirmed was considered gestation day 0.
After assignment to the study, males and females were weighed once per
week. Mated females were weighed on gestation days 0 and 13. Males
were given a thorough physical examination once per week and were
observed for clinical signs of toxicity before and after exposures. All
animals were checked twice daily for mortality and gross abnormalities
(except for one day prior to mating when inclement weather permitted
observations).
One half of the males of each group were euthanized on each of the 2
consecutive days at the end of the study. They had not been exposed to
33 / 33
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5. Toxicity
Date 02.10.2003
propionitrile for about 2 weeks prior to termination. Each male was given
an external examination and weighed. The tissues and organs of the
thoracic, scrotal and abdominal cavities were examined for gross lesions
and the testes, epididymides, prostate glands and seminal vesicles were
preserved in 10% neutral buffered formalin. Females that were not mated
with males were euthanized and were not examined.
Mated females were euthanized on gestation day 13 (or the nearest
workday up to gestation day 15). Females that were co-housed with males
without confirmed copulation were euthanized during the second week
after the last day of co-housing. Gross necropsies were performed on
females that had copulated and those that had not. The tissues and organs
of the thoracic and abdominal cavities were examined. Pregnancy status
was determined, nidation sites and were classified, and corpora lutea were
counted.
Statistical analyses: Body weight data were analyzed using Dunnett's test.
Mating and pregnancy rate data were analyzed by a Fisher's exact test and
an uncorrected chi-square test. Other data were analyzed by the Mann-
Whitney U test. The Bonferroni inequality was assumed when comparing
multiple treatments to control values for all tests except Dunnett's test. The
critical level for significance was p < 0.05.
Test material was propionitrile (CAS No. 107-12-0). Purity of the test
Test substance :
material was 96.1%. Impurities included acrylonitrile (0.1%), adiponitrile
(0.7%), p-nitrosodiphenylamine (0.08%) and water (< 0.1%). Analyses
indicated no significant decomposition of the test material over the course
of the study.
There was no effect of treatment on fertility of males.
Conclusion :
(1) valid without restriction
Reliability :
Study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (21)
5.8.2 DEVELOPMENTAL TOXICITY/TERATOGENICITY
rat
Species :
female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
Days 6-20 of gestation
Exposure period :
6 hr/day
Frequency of treatm. :
to day 21 of gestation
Duration of test :
50, 100, 150, and 200 ppm
Doses :
yes, concurrent no treatment
Control group :
= 200 ppm
NOAEL maternal tox. :
= 200 ppm
NOAEL teratogen. :
= 150 ppm
NOAEL Fetotoxicity :
= 200
NOAEL Embryotoxicity :
butyronitrile was not teratogenic
Result :
other: similar to OECD 414
Method :
1992
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Four different concentrations of acetonitrile, priopionitrile, isobutylnitrile,
Remark :
acrylonitrile, allylnitrile, methacrylonitrile and 2-chloroacrylonitrile also were
tested in this study.
The methodology described in the manuscript is essentially identical to that
under OECD TG-414. However, information is lacking in the report in
34 / 34
Id 109-74-0
5. Toxicity
Date 02.10.2003
regard to food intake. Therefore, it is unknown if reduced fetal body
weights were a consequence of reduced material feed intake. In addition,
it is unknown as to whether the study was conducted under GLP
assurances. However, based on the date in which this study was
completed it is likely to have been a GLP study. All other parameters noted
in the guideline appear in the manuscript.
In a preliminary study, 300 ppm induced mortality in 6 of 6 pregnant rats.
Since a repeated dose toxicity study has not been conducted, and whether
inhalation of 200 ppm caused toxicity to dams was not assessed in the
developmental study (other than effect on body weight), the actual
maternal NOAEL may be less than 200 ppm.
Maternal: All animals survived and there was no effect on weight gain.
Result :
Indices of pregnancy were comparable among groups. There was no
significant effect of treatment on the mean numbers of implantations, or
incidences of nonsurviving implants and resorptions.
Fetal: There was no effect of treatment on the mean numbers of live
fetuses or sex ratio. There was a concentration-related trend towards a
decrease in fetal body weights, with weights of females from the 200 ppm
group significantly less than control (5.08 g/litter in treated vs. 5.41/litter in
control). A single case of skeletal malformation (fused ribs) was observed
at 100 ppm. The incidences of visceral and skeletal variations in treated
fetuses were similar to controls.
Animals: Male (350 g) and primiparous female (200-220 g) were
Test condition :
acclimated for 1-2 weeks prior to breeding. Females were then placed with
males (one male: 3 females) overnight and examined by vaginal smear for
the presence of sperm the following morning. Sperm-positive females were
considered to be at Day 0 of gestation. These animals were randomly
assigned to groups of 20-23 rats each.
Exposure conditions: Exposures were conducted in 200 -liter stainless-
steel inhalation chambers at an air flow of 10-20 m3/hr. Chambers were
maintained at a negative pressure of < = 3 mm water. Chamber
temperatures and humidities were 23 +/- 2 degrees and 50 +/- 5%,
respectively. Vapor was generated by bubbling an additional air flow
through a flask containing test material. The vapor was mixed with filtered
room air to achieve the desired concentration. Analytical concentrations
were determined by analyzing the atmosphere once/hour (by gas-liquid
chromatography) during each 6 hour exposure. The nominal
concentrations of n-butyronitrile were 50, 100, 150 and 200 ppm.
Corresponding analytical concentrations were 53 +\- 2.3, 104 +/- 4.5, 154
+/- 11.5, and 208 +/- 98.7 ppm.
Test conduct: Animals were exposed 6 hours/day on Days 6 through 20 of
gestation. Control animals were exposed concurrently to filtered room air in
an adjacent chamber with flow characteristics identical to those of the
treated groups. Food and water were available ad libitum (except
during exposure). All rats were observed daily and maternal body weights
were recorded on Days 0, 6 and 21 of gestation. Females were euthanized
on Day 21 of gestation and the uterus was removed and weighed. The
uterus horns were then opened and the numbers of implantation and
absorption sites and live and dead fetuses were recorded. Live fetuses
were removed and weighed, examined for external anomalies (including
those of the oral cavity) and sexed. The numbers of fetuses (and litters)
examined for external anomalies in the 0, 50, 100, 150 and 200 ppm
groups were 227 (17), 239 (17), 219 (18), 280 (21) and 187 (15),
respectively. Half of the fetuses from each litter were fixed in Bouin's
solution and examined microscopically for visceral abnormalities. The
remaining half were fixed in 70% ethanol, eviscerated, macerated in 1%
KOH , stained in alizarin red S, and examined microscopically for skeletal
anomalies.
35 / 35
Id 109-74-0
5. Toxicity
Date 02.10.2003
Statistical analysis: Depending on the parameter evaluated: one-way
analysis of variance followed by Dunnett's test, Wilcoxon test after arc-sine-
square root transformation, Fisher's test, or least squares analysis. The
litter was used as the basis for analysis of fetal variables.
Purity was > 99%.
Test substance :
(1) valid without restriction
Reliability :
The study was comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
10.08.2003 (24)
5.8.3 TOXICITY TO REPRODUCTION, OTHER STUDIES
5.9 SPECIFIC INVESTIGATIONS
Mechanistic Studies
Endpoint :
Study descr. in chapter :
Reference :
Type :
mouse
Species :
male
Sex :
CD-1
Strain :
i.p.
Route of admin. :
No. of animals :
other
Method :
1981
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Propionitrile also was tested in this study. The results with this material
Remark :
were similar to those of n-Butyronitrile.
This study is considered to be valid without restriction. The study was
conducted and documented in a thorough manner.
In the first study, the mortality rate for animals treated only with 55 mg/kg n
Result :
butyronitrile was 8/10. Co-treatment with sodium nitrite or sodium
thiosulfate reduced the rate to 2/10 and 1/10, respectively.
In the carbon tetrachloride study, the mortality rate for animals treated only
with 55 mg/kg n-butyronitrile was 10/10. None of the animals co-treated
with carbon tetrachloride died.
Cyanide concentrations in liver and brain of mice co-treated with sodium
thiosulfate or carbon tetrachloride were significantly less than those of mice
treated with n-butyronitrile alone. In mice receiving n-butyronitrile only,
21.4 +/- 16.1 (mean +/- SD) and 16.8 +/- 12.6 nmol/g cyanide were found
in liver and brain, respectively. In mice receiving n-butyronitrile plus sodium
thiosulfate, 2.4 +/- 0.8 and 2.4 +/- 2.6 nmol/g cyanide were found in liver
and brain, respectively. In mice receiving propionitrile plus carbon
tetrachloride, 1.3 +/- 1.5 and 1.6 +/- 2.0 nmol/g cyanide were found in liver
and brain, respectively.
Male CD mice (30 g) were divided into 3 groups of ten animals each. One
Test condition :
group received 55 mg/kg i.p. n-butyronitrile only, another received i.p.
injections of 75 mg/kg sodium nitrite (a cyanide antagonist) 20 minutes
before and 100 minutes after i.p. injection of 55 mg/kg n-butyronitrile, and
another received i.p. injections of 1 g/kg sodium thiosulfate (a cyanide
antagonist) 20 minutes before and 80 and 180 minutes after i.p. injection of
55 mg/kg n-butyronitrile.
36 / 36
Id 109-74-0
5. Toxicity
Date 02.10.2003
Two other groups of 10 mice received either 0.2 ml of vegetable oil or 0.2
ml of 20% carbon tetrachloride (a hepatotoxic dose) in vegetable oil
subcutaneously, 24 hours before i.p. treatment with 55 mg/kg n
butyronitrile .
In both experiments, animals were observed for 7 days. Mortality data were
analyzed statistically by the chi-square test. The criterion for significance
was p < 0.05.
In an additional study, the concentrations of cyanide in liver and brain were
determined in a) 5 mice treated only with 38 mg/kg n-butyronitrile (i.p.), b)
5 mice given 1 g/kg sodium thiosulfate 20 minutes before and 80 minutes
after 38 mg/kg n-butyronitrile (i.p.), and c) 5 mice given 0.2 ml of 20%
carbon tetrachloride subcutaneously 24 hours before i.p. treatment with 38
mg/kg n-butyronitrile. All mice were killed 2.5 hours after n-butyronitrile
injection (if still alive at this time). The livers and brains were excised as
soon as possible after death, quick-frozen and weighed. Cyanide
concentrations were determined by the method of Bruce et al. (Anal Chem
27: 1346-1347, 1955). Results were analyzed using an unpaired t-test.
The purity of the test material was 98%. No free cyanide was found in
Test substance :
solutions made in distilled, deionized water.
n-Butyronitrile is activated by the liver to release cyanide, which is
Conclusion :
responsible for acute toxicity.
06.08.2003 (28)
5.10 EXPOSURE EXPERIENCE
5.11 ADDITIONAL REMARKS
37 / 37
Id 109-74-0
6. Analyt. Meth. for Detection and Identification
Date 02.10.2003
6.1 ANALYTICAL METHODS
6.2 DETECTION AND IDENTIFICATION
38 / 38
Id 109-74-0
7. Eff. Against Target Org. and Intended Uses
Date 02.10.2003
7.1 FUNCTION
7.2 EFFECTS ON ORGANISMS TO BE CONTROLLED
7.3 ORGANISMS TO BE PROTECTED
7.4 USER
7.5 RESISTANCE
39 / 39
Id 109-74-0
8. Meas. Nec. to Prot. Man, Animals, Environment
Date 02.10.2003
8.1 METHODS HANDLING AND STORING
8.2 FIRE GUIDANCE
8.3 EMERGENCY MEASURES
8.4 POSSIB. OF RENDERING SUBST. HARMLESS
8.5 WASTE MANAGEMENT
8.6 SIDE-EFFECTS DETECTION
8.7 SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER
8.8 REACTIVITY TOWARDS CONTAINER MATERIAL
40 / 40
Id 109-74-0
9. References
Date 02.10.2003
(1) Chapatwala KD, Babu GRV, Nawaz MS. 1992. Degradation of acetonitrile and biphenyl
compounds by a mixed microbial culture. Environ Toxicol and Chem 11: 1145-1151.
(2) Covance Laboratories Inc. Chromosomal Aberrations in Chinese Hamster Ovary (CHO)
Cells with EC98-0254, NBN (unpublished study). Study Number 20877-0-437OECD,
December 28, 1999.
(3) Covance Laboratories Inc. Mutagenicity Test with EC98-0254 NBN in the Salmonella-
Escherichia coli/mammalian-microsome reverse mutation assay with a confirmatory assay
(unpublished study). Covance study number 20877-0-409R, December 8, 1999.
(4) Daubert TE and Danner RP. Physical & Thermodynamic Properties of Pure Chemicals:
Data Compilation; NY: Hemisphere Publishing Corporation Co., 1989.
(5) Eastman Chemical Company. Material Safety Data Sheet for "EASTMAN" n-Butyronitrile,
dated August 8, 2000.
(6) Eastman Kodak Company, Environmental Analytical Services,
Chemicals Quality Services Division. n-Butyronitrile:
Biochemical Oxygen Demand Determination (unpublished study). Report No. L8092-BOD,
March 29, 1999.
(7) Eastman Kodak Company, Environmental Analytical Services, Chemicals Quality Services
Division. n-Butyronitrile: Chemical Oxygen Demand Determination (unpublished study).
Report No. L8092-COD, March 29, 1999.
(8) Eastman Kodak Company, Environmental Sciences Section, Health and Environment
Laboratories. n-Butyronitrile: A Growth Inhibition Test with the Alga, Selenastrum
capricornutum (unpublished study). Study No. EN-512-900741-A, January 28, 2000.
(9) Eastman Kodak Company, Environmental Sciences Section, Health and Environment
Laboratories. n-Butyronitrile: An Acute Aquatic Effects Test with the Daphnid (unpublished
study). Study No. EN-431-900741-A, March 30, 1999.
(10) Eastman Kodak Company, Environmental Sciences Section, Health and Environment
Laboratories. n-Butyronitrile: An Acute Aquatic Effects Test with the Fathead Minnow
(unpublished study). Study No. EN-430-900741-A, March 30, 1999.
(11) Eastman Kodak Company, Health and Environment Laboratories, Toxicological Sciences
Section. Acute inhalation toxicity of n-Butyronitrile in the rat (unpublished study).
Document Number 215515L, September 4, 1986.
(12) Eastman Kodak Company, Laboratory of Industrial Medicine. Notebook No. 59, p. 476,
February 15, 1960 (unpublished study).
(13) Eastman Kodak Company, Toxicological Sciences Section, Health and Environment
Laboratories. Acute inhalation toxicity and 1-hour LC10 value of n-butyronitrile in the
rat (unpublished study). Document Number 236009R, April 6, 1987.
(14) EPIWIN Aop Program (v1.90).
(15) EPIWIN ECOSAR Program (0.99).
(16) EPIWIN Hydrowin Program (v1.67).
(17) EPIWIN Kowwin Program (v1.66).
(18) EPIWIN Level III Fugacity model program.
(19) EPIWIN Wskow (v1.40)
41 / 41
Id 109-74-0
9. References
Date 02.10.2003
(20) Kier LD. 1984. Female fertility study of Sprague-Dawley rats exposed by the inhalation
route to propionitrile. Unpublished Monsanto Report No MSL-4438, dated December 31,
1984.
(21) Kier LD. 1984. Male fertility study of Sprague-Dawley rats exposed by the inhalation route
to propionitrile. Unpublished Monsanto Report No MSL-4422, dated December 17, 1984
(22) Lutin PA. 1970. Removal of organic nitriles from wastewater systems. J Water Pollut
Control Fed 42: 1632-42.
(23) Riddick JA, Bunger WB, Sackano TK. 1986. Organic Solvents: Physical Properties &
Methods of Purification. In: Techniques of Chemistry, Vol. II (4th Ed). NY:Wiley
Interscience, p. 583-7.
(24) Saillenfait AM, Bonnet P, Guenier JP, and DeCeaurriz J. Relative Developmental
Toxicities of Inhaled Aliphatic Mononitriles in Rats. Fund Appl Toxicol 20: 365-375, 1993.
(25) Sangster J. 1989. Octanol-water partition coefficients of simple organic compounds. J Phys
Chem Ref Data 18:1111-1230.
(26) Smyth HF et al. Range-finding toxicity data: list VI. Amer Ind Hyg Ass J. 23: 95-107, March
- April 1962.
(27) Velasquez DJ and Thake DC. 1984. Three-month toxicity study of propionitrile vapor
administered to male and female Sprague-Dawley rats by inhalation. Unpublished
Monsanto Report No MSL-4113, dated October 1, 1984.
(28) Willhite CC and Smith RP. 1981. The role of cyanide liberation in the acute toxicity of
aliphatic nitriles. Toxicol Appl Pharmacol 59: 589-602.
(29) Windholz M, et al. The Merck Index - An Encyclopedia of Chemicals, Drugs and
Biologicals. 10th Edition. Rahway, NJ: Merck & Co., Inc., 1983.
42 / 42
Id 109-74-0
10. Summary and Evaluation
Date 02.10.2003
10.1 END POINT SUMMARY
10.2 HAZARD SUMMARY
10.3 RISK ASSESSMENT
43 / 43
201-14860B1
IUCLID
Data Set
Existing Chemical : ID: 78-82-O
: 78-82-O
CAS No.
EINECS Name : lsobutyronitrile
TSCA Name : Propanenitrile, 2-methyl-
Molecular Formula . : C4H7N
Producer related part
Company : Eastman Chemical Company
Creation date : 02.07.2003
Substance related part
: Eastman Chemical Company
Company
Creation date : 02.07.2003
Status
Memo
Printing date : 06.10.2003
Revision date : 13.11.2003
Date of last update : 06.10.2003
Number of pages : 43
Chapter (profile) : Chapter: 1,2, 3,4, 5, 6, 7, 8, 10
Reliability (profile) : Reliability: without reliability, 1, 2, 3,4
Flags (profile) : Flags: without flag, confidential, non confidential, WGK (DE), TA-Luft (DE),
Material Safety Dataset, Risk Assessment, Directive 67/548/EEC, SIDS
l/l
Id 78-82-0
1. General Information
Date 02.10.2003
1.0.1 APPLICANT AND COMPANY INFORMATION
1.0.2 LOCATION OF PRODUCTION SITE, IMPORTER OR FORMULATOR
1.0.3 IDENTITY OF RECIPIENTS
1.0.4 DETAILS ON CATEGORY/TEMPLATE
1.1.0 SUBSTANCE IDENTIFICATION
IUPAC Name :
C(#N)C(C)C
Smiles Code :
C4H7N
Molecular formula :
69.11
Molecular weight :
Petrol class :
15.08.2003
1.1.1 GENERAL SUBSTANCE INFORMATION
1.1.2 SPECTRA
1.2 SYNONYMS AND TRADENAMES
-Methylpropanenitrile
-Methylpropionitrile
1-Cyano-1-methylethane
2-Cyanopropane
2-Methylpropanenitrile
2-Methylpropionitrile
Dimethylacetonitrile
Isobutanenitrile
2/2
Id 78-82-0
1. General Information
Date 02.10.2003
Isopropyl cyanide
Isopropyl nitrile
Propanenitrile, 2-methyl
Propanoic acid, 2-methyl-, nitrile
1.3 IMPURITIES
1.4 ADDITIVES
1.5 TOTAL QUANTITY
1.6.1 LABELLING
1.6.2 CLASSIFICATION
1.6.3 PACKAGING
1.7 USE PATTERN
industrial
Type of use :
Chemical industry: used in synthesis
Category :
Chemical intermediate
Remark :
(2) valid with restrictions
Reliability :
(5)
1.7.1 DETAILED USE PATTERN
1.7.2 METHODS OF MANUFACTURE
1.8 REGULATORY MEASURES
1.8.1 OCCUPATIONAL EXPOSURE LIMIT VALUES
1.8.2 ACCEPTABLE RESIDUES LEVELS
3/3
Id 78-82-0
1. General Information
Date 02.10.2003
1.8.3 WATER POLLUTION
1.8.4 MAJOR ACCIDENT HAZARDS
1.8.5 AIR POLLUTION
1.8.6 LISTINGS E.G. CHEMICAL INVENTORIES
1.9.1 DEGRADATION/TRANSFORMATION PRODUCTS
1.9.2 COMPONENTS
1.10 SOURCE OF EXPOSURE
1.11 ADDITIONAL REMARKS
1.12 LAST LITERATURE SEARCH
1.13 REVIEWS
4/4
Id 78-82-0
2. Physico-Chemical Data
Date 02.10.2003
2.1 MELTING POINT
= -71.5 癈
Value :
Sublimation :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of the material is unknown. Data obtained from Hazardous
Remark :
Substances Data Bank Number: 5221. Last revision date: 9/21/1999.
(2) valid with restrictions
Reliability :
Primary source is peer-reviewed published data.
Critical study for SIDS endpoint
Flag :
(31)
2.2 BOILING POINT
= 103.8 癈 at 1016 hPa
Value :
Decomposition :
other: not specified
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Purity of the material is unknown. Data obtained from Hazardous
Remark :
Substances Data Bank Number: 5221. Last revision date: 9/21/1999
(2) valid with restrictions
Reliability :
Primary source is peer-reviewed published data.
Critical study for SIDS endpoint
Flag :
06.08.2003 (31)
2.3 DENSITY
relative density
Type :
= .77 at 20 癈
Value :
other
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
(2) valid with restrictions
Reliability :
Source of data is a Material Safety Data Sheet.
(5)
2.3.1 GRANULOMETRY
2.4 VAPOUR PRESSURE
= 55.2 hPa at 20 癈
Value :
Decomposition :
other (measured)
Method :
1986
Year :
5/5
Id 78-82-0
2. Physico-Chemical Data
Date 02.10.2003
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The study results were reported in Appendix 2 of a GLP acute inhalation
Remark :
toxicity study discussed below under acute toxicity effects.
Vapor pressures at 40, 23, and 20 degrees C were 92.1, 47.0, 41.4 and
Result :
mmHg, respectively.
The vapor pressures at 40 and 23 degrees C were determined by head
Test condition :
space gas chromatography with flame ionization detection. The vapor
pressure at 20 degrees C was calculated by linear extrapolation.
Purity was 99.7%
Test substance :
(2) valid with restrictions
Reliability :
Basic data are given.
Critical study for SIDS endpoint
Flag :
06.08.2003 (6)
2.5 PARTITION COEFFICIENT
Partition coefficient :
= .46 at 癈
Log pow :
pH value :
other (measured)
Method :
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The value obtained from the experiment was 0.46. The confidence limit
Result :
was 0.25.
The test was performed at ambient temperature (20-25 degrees C). The
Test condition :
value was obtained using the Shake-Flask method. The aqueous phase
was octanol-saturated water. The concentration of material in the aqueous
phase was measured using gas-liquid chromatography.
Purity of the test material was not mentioned.
Test substance :
(2) valid with restrictions
Reliability :
Data were from a peer reviewed, published source.
Critical study for SIDS endpoint
Flag :
13.08.2003 (27)
octanol-water
Partition coefficient :
= .76 at 25 癈
Log pow :
=7
pH value :
other (calculated)
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program were the CAS No., melting point = - 71.5
Remark :
degrees C, boiling point = 103.8 degrees C, water solubility = 39,000 mg/l,
and vapor pressure = 41.4 mm Hg.
(2) valid with restrictions
Reliability :
Approved program for estimating Log Kow.
13.08.2003 (20)
2.6.1 SOLUBILITY IN DIFFERENT MEDIA
water
Solubility in :
= 39000 mg/l at 25 癈
Value :
pH value :
6/6
Id 78-82-0
2. Physico-Chemical Data
Date 02.10.2003
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
Deg. product :
other
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Seventeen ml of isobutyronitrile (IBN) was added to a beaker containing
Remark :
500 ml of distilled water that was being stirred with a stir bar at moderate
speed. A thermometer was placed in the beaker to monitor temperature
(25 degrees C). IBN was added in 2 ml increments under constant stirring,
and the water was inspected for saturation after each addition. After 25 ml
IBN had been added, IBN was added in 1 ml increments because of
noticeable signs of reaching the saturation point. A total of 27 ml IBN was
added.
The solubility was determined by analyzing 20 ml of the final solution with a
nitrogen analyzer. This instrument measured the amount of nitrogen
dissolved in the water.
The concentration of IBN = 1.049 g N/100 g solution x 69 g IBN/14g N x
0.755 (instrument response factor derived from nitrogen standard) = 3.90
wt % IBN.
Purity of the test material was not listed. According to a MSDS from the
Test substance :
supplier (dated 1/9/2002), the purity of the material is 100%.
(2) valid with restrictions
Reliability :
The study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
23.09.2003 (4)
water
Solubility in :
= 32,500 mg/l at 25 癈
Value :
pH value :
at 癈
concentration :
Temperature effects :
Examine different pol. :
at 25 癈
pKa :
Description :
Stable :
Deg. product :
other: calculated
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program were the CAS No., melting point = - 71.5
Remark :
degrees C, boiling point = 103.8 degrees C, water solubility = 39,000 mg/l,
and vapor pressure = 41.4 mm Hg.
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
11.09.2003 (22)
2.6.2 SURFACE TENSION
7/7
Id 78-82-0
2. Physico-Chemical Data
Date 02.10.2003
2.7 FLASH POINT
2.8 AUTO FLAMMABILITY
2.9 FLAMMABILITY
2.10 EXPLOSIVE PROPERTIES
2.11 OXIDIZING PROPERTIES
2.12 DISSOCIATION CONSTANT
2.13 VISCOSITY
2.14 ADDITIONAL REMARKS
8/8
Id 78-82-0
3. Environmental Fate and Pathways
Date 02.10.2003
3.1.1 PHOTODEGRADATION
air
Type :
Sun light
Light source :
nm
Light spectrum :
based on intensity of sunlight
Relative intensity :
INDIRECT PHOTOLYSIS
OH
Sensitizer :
Conc. of sensitizer :
= .0000000000007032 cm?(molecule*sec)
Rate constant :
= 50 % after 15.2 day(s)
Degradation :
Deg. product :
other (calculated)
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program were the CAS No., melting point = - 71.5
Remark :
degrees C, boiling point 103.8 degrees C, water solubility = 39,000 mg/l,
and vapor pressure = 41.4 mm Hg.
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
Critical study for SIDS endpoint
Flag :
19.08.2003 (17)
3.1.2 STABILITY IN WATER
abiotic
Type :
other
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
: EPIWIN Hydrowin cannot calculate hydrolysis rate constants for nitriles.
Remark
The theoretical hydrolysis of the related material propionitrile and several
other chemicals has been examined by Dr. Lee Wolfe at the USEPA
Environmental Research Laboratory in Athens, Georgia. The results of
these analyses were published in a report by Dr. Wolfe that could not be
located. In a personal communication, Dr. Wolfe stated that propionitrile
can hydrolyze (albeit slowly). According to a study cited in the Hazardous
Substances Data Bank, the chemical hydrolysis of the related material
acetonitrile in water is base-catalyzed (the rate constant for base catalyzed
hydrolysis is 5.8X10-3/M-hr), but the half-life at pH 7 is more than 150,000
yrs (Ellington et al., 1988). Acetonitrile (CH3CN, CAS No. 75-05-8) is the
2-carbon analog of the category members, possessing the same
functionality, but having one less carbon than propionitrile. Taken together,
these data suggest that hydrolysis of butyronitrile at environmentally
relevant pHs will occur too slowly to be a significant means of degradation.
(2) valid with restrictions
Reliability :
Experimental results for the test material could not be located. Results are
for a related material.
19.08.2003 (19)
9/9
Id 78-82-0
3. Environmental Fate and Pathways
Date 02.10.2003
3.1.3 STABILITY IN SOIL
3.2.1 MONITORING DATA
3.2.2 FIELD STUDIES
3.3.1 TRANSPORT BETWEEN ENVIRONMENTAL COMPARTMENTS
fugacity model level III
Type :
other: air, water, soil and sediment
Media :
17.8 % (Fugacity Model Level I)
Air :
47.3 % (Fugacity Model Level I)
Water :
.0809 % (Fugacity Model Level II/III)
Biota :
34.7 % (Fugacity Model Level II/III)
Soil :
other: calculated
Method :
2003
Year :
Measured inputs to the program were the CAS No., melting point = - 71.5
Remark :
degrees C, boiling point = 103.8 degrees C, water solubility = 39,000 mg/l,
and vapor pressure = 41.4 mm Hg.
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
Critical study for SIDS endpoint
Flag :
19.08.2003 (21)
3.3.2 DISTRIBUTION
3.4 MODE OF DEGRADATION IN ACTUAL USE
3.5 BIODEGRADATION
aerobic
Type :
Inoculum :
Deg. product :
other
Method :
1998
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Refer to Section 3.6 below.
Remark :
(1) valid without restriction
Reliability :
Critical study for SIDS endpoint
Flag :
06.08.2003
aerobic
Type :
Inoculum :
14 day(s)
Contact time :
readily biodegradable
Result :
Deg. product :
other: MITI protocol
Method :
1978
Year :
10 / 10
Id 78-82-0
3. Environmental Fate and Pathways
Date 02.10.2003
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Isobutylnitrile was shown to biodegrade readily using the Japanese MITI
Remark :
protocol (2 week incubation, 100 ppm concentration), with BODs of 53.9 -
66.3%.
Hazardous Substances Data Bank for 2-methylpropanenitrile.
Source :
(4) not assignable
Reliability :
The primary references were not consulted.
10.08.2003 (25) (28)
aerobic
Type :
other: mixed microbial culture
Inoculum :
1000 mg/l
Concentration :
48 hour(s)
Contact time :
other: biodegradable
Result :
Deg. product :
other
Method :
1992
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The final protein and ammonia concentrations and pH were 6.76 mg/l, 51.6
Result :
micromoles/ml and 8.21, respectively, indicating that the mixed culture
could use this material as a growth substrate.
A mixed microbial culture (protein concentration of 0.085 mg/l) was isolated
Test condition :
from an environment contaminated with organic cyanides and
polychlorinated biphenyls. This was grown for 48 hours on phosphate
buffer (pH 7.0, 30 degrees C) containing propionitrile (1 g/l) as the sole
source of carbon and nitrogen. The final concentration of protein, ammonia
and pH were determined.
Test material was obtained from Aldrich Chemical Co. It is presumed that
Test substance :
the material has high purity.
(4) not assignable
Reliability :
The study shows that the test material was used as a substrate (and
therefore was metabolized); however, the extent to which the test material
biodegraded is difficult to determine from the study.
07.08.2003 (1)
3.6 BOD5, COD OR BOD5/COD RATIO
BOD5
other: Method C.5., "Degradation, Biochemical Oxygen Demand", Official
Method :
Journal of the European Communities, No. L251/212, 19.9.84
1998
Year :
mg/l
BOD5 :
yes
GLP :
COD
other:Method C.6., "Degradation, Chemical Oxygen Demand", Official
Method :
Journal of the European Communities, No. L383A/227, 29 December 1992.
1998
Year :
= 1910 mg/g substance
COD :
yes
GLP :
RATIO BOD5 / COD
= .28
BOD5/COD :
Method is similar to OECD: TG-301C: Modified MITI Test. The study is the
Remark :
critical study for the biodegradation endpoint. One of the two 5-day dilution
water bottles had a dissolved oxygen drop greater than 0.2 ppm. This was
a protocol deviation. It was not considered significant enough to negate
11 / 11
Id 78-82-0
3. Environmental Fate and Pathways
Date 02.10.2003
the test.
BOD analysis: The values obtained for the 2 different concentrations were
Result :
0.567 and 0.494 g BOD/g test material at 5 days and 2.78 and 2.51 g
BOD/g test material at 20 days. The average BOD5 and BOD20 values
were 0.53 grams BOD/gram of test substance and 2.6 grams BOD/gram
of test substance, respectively.
COD analysis: The values obtained for the 3 replicates were 2.126, 1.625
and 1.964 g COD/g test material. The average value was 1.91 g COD/g
test material. The percent recovery of the reference sample was 93.5%.
BOD5/COD: The BOD5/COD ratio is 0.28 (0.53/1.91)
Test conditions: COD Determination: A 0.50 N potassium dichromate
Test condition :
solution was used to standardize the ferrous ammonium sulfate titrant.
Mercuric sulfate was added to minimize chloride interference (if any). Three
separate replicates were tested. A potassium phthalate standard was
analyzed as a positive control. The test was considered valid if the
recovery of the standard fell between the limits of 83.47 - 116.06%. The
COD was calculated by subtracting the amount of titrant needed for the
sample from the amount of titrant needed for a blank. This result was
multiplied by the normality of the titrant and the equivalent weight of
oxygen. The product was then divided by the sample weight.
BOD Determination: The test was performed according to an Eastman
Kodak Company protocol [Need to have protocol for details]. Two separate
concentrations (0.00033%, 0.00050%) were tested. The 5-day BOD was
calculated by subtracting the final dissolved oxygen reading and the 5-day
seeded dilution water drop from the initial dissolved oxygen reading. The
20-day BOD was calculated by subtracting the average 20-day seeded
dilution water drop from the total dissolved oxygen drop over 20 days. The
results were multiplied by 100. The products were divided by the product
of the percent concentration of the stock solution in the BOD bottle and the
concentration of test chemical in the stock solution. The products were
divided by 1,000,000. The results were in units of grams of BOD per gram
of test substance for the 5-day (or 20-day) incubation.
BOD5/COD ratio: This ratio was calculated by dividing the average 5-day
BOD value by the average COD value.
Purity was 99.8%.
Test substance :
The test material is not considered to be "Readily Biodegradable" based on
Conclusion :
a BOD5/COD ratio of <0.5 (0.53/1.91 = 0.28)
(1) valid without restriction
Reliability :
This was a well-documented guideline study conducted under GLP
assurances.
10.08.2003 (7) (8)
3.7 BIOACCUMULATION
3.8 ADDITIONAL REMARKS
12 / 12
Id 78-82-0
4. Ecotoxicity
Date 02.10.2003
4.1 ACUTE/PROLONGED TOXICITY TO FISH
static
Type :
Pimephales promelas (Fish, fresh water)
Species :
96 hour(s)
Exposure period :
mg/l
Unit :
= 102.1 measured/nominal
NOEC :
> 102.1 measured/nominal
LC50 :
yes
Limit test :
yes
Analytical monitoring :
other: OECG:TG-203 and EEC/Annex V C.1.
Method :
1998
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The dissolved oxygen concentration was slightly less than 60% of the initial
Remark :
values at the end of the test. This did not adversely affect the outcome.
Through 48 hours, solutions containing test material were clear and
colorless. At 72 hours through the end of the test, exposure solutions were
slightly cloudy; however no slicks or precipitates were observed.
The LC50 value indicates that the test substance would not be classified
according to the European Union's labeling directive and would correspond
to a "low concern level" according to the U.S. EPA's assessment criteria.
No mortality occurred and all fish exhibited normal behavior and
Result :
appearance. The mean concentration of test material was 102.1 mg/l. The
analyzed percent loss of the test material ranged from 12.3 - 21.5%. The
temperatures of all solutions ranged from 20 - 21 degrees throughout the
test. The pH and dissolved oxygen values ranged from 7.8 - 8.4 and 4.7 -
8.5 mg/l, respectively. The temperature, pH and dissolved oxygen values
were considered to be acceptable for the organisms used in the test. The
test was considered to be valid.
Organisms: Juvenile fathead minnows were acclimated to test water for at
Test condition :
least two weeks prior to testing. They were randomized to 6 sets of 7 fish
each. Two sets of minnows (7/set) were killed before the start of the test to
determine average wet weight (0.17 and 0.16 g/set) and mean standard
length (2.91 and 2.90 cm/set).
Test water: The water was pumped from Lake Ontario into a large
underground storage vessel. Water from this vessel was subsequently
pumped into the laboratory where it passed through polypropylene filter
tubes, activated carbon filter tubes, and another set of polypropylene filter
tubes. The filtered water stream was treated with sodium thiosulfate to
further reduce trace levels of residual chlorine. The water was then heated
to 20 +/- 2 degrees C and distributed to an open basin for seasoning prior
to use. Representative values for hardness and total alkalinity (both as
CaCO3) were 120.0 and 89.6 mg/l, respectively.
Test material: A stock solution of test material (12 mg/l) was prepared in a
500 ml volumetric flask containing test water. The exposure solutions were
prepared at a nominal concentration of 120 mg/l by adding the appropriate
amount of stock solution to glass vessels (30.5 cm Pyrex seamless,
cuboidal chromatography jars) containing 20 liters of test water. The
solutions in each test vessel were stirred with a stir rod prior to adding fish.
Duplicate test and dilution water control vessels were prepared.
Test conduct: Immediately after stirring, fish were placed into each of the
replicate test and control vessels (7 per vessel). Glass lids were placed on
13 / 13
Id 78-82-0
4. Ecotoxicity
Date 02.10.2003
top of each test vessel and sealed with Parafilm. Biological loading within
test vessels was kept below 1.0 g wet weight/l test solution. The vessels
were placed in a certified hood under 8 hours of fluorescent lighting/day.
Animals were observed for mortality and signs of stress at 0, 4, 24, 48, 72
and 96 hours. Temperature, dissolved oxygen concentration and pH of
each solution also were measured at 24 hour intervals. Concentrations of
test material in the test vessels at 0 and 96 hours were analyzed by
GC/MS. The geometric mean of the concentrations was calculated. Since
no mortality was observed, statistical analyses were not performed.
The test was considered valid if control mortality was <= 10%, dissolved
oxygen did not fall below 60% of the initial oxygen level, the temperature
was 20 +/- 1 degrees C and there were no abnormal occurrences that
could influence the outcome.
Purity was 99.8%.
Test substance :
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
07.08.2003 (11)
4.2 ACUTE TOXICITY TO AQUATIC INVERTEBRATES
static
Type :
Daphnia magna (Crustacea)
Species :
48 hour(s)
Exposure period :
mg/l
Unit :
= 94.3 measured/nominal
NOEC :
> 94.3 measured/nominal
EC50 :
yes
Limit Test :
yes
Analytical monitoring :
other: OECD: TG-202 and EEC/Annex V C.2
Method :
1998
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
No protocol deviations were noted.
Remark :
All daphnids exposed to test-article exhibited behavior comparable to
Result :
controls. No immobility was observed. No precipitation of test material was
observed. The concentrations of test material at time 0 were 108.7 and
115.5 mg/l and the concentrations at 48 hours were 76.9 and 81.9 mg/l.
The geometric mean of the test concentrations at 0 and 48 hours was 94.3
mg/l. The analyzed percent loss of the test material over 48 hours was
29.2%. The temperatures of all solutions were maintained at 21 degrees C
throughout the test. The pH and dissolved oxygen values ranged from
8.2 - 8.5 and 8.3 - 8.5 mg/l, respectively. The temperature, pH and
dissolved oxygen values were considered to be acceptable for the
organisms used in the test. The test was considered to be valid.
Organisms: Adult Daphnia magna were reared within the testing facility in
Test condition :
100 l culturing flasks. Gravid daphnids used to produce test animals were
obtained from rearing tanks that had been established for at least two
weeks. Prior to the study, approximately 100 gravid daphnids were
transferred by net into two glass bowls containing test water and food. After
18 hours in the bowls, all adult daphnids were removed. Neonates were
collected by pipette and transferred directly into exposure vessels. A total
of 10 daphnids were placed into each of the replicate test and control
vessels.
Test water: The water was pumped from Lake Ontario into a large
underground storage vessel. Water from this vessel was subsequently
14 / 14
Id 78-82-0
4. Ecotoxicity
Date 02.10.2003
pumped into the laboratory where it passed through polypropylene filter
tubes, activated carbon filter tubes, and another set of polypropylene filter
tubes. The filtered water stream was treated with sodium thiosulfate to
further reduce trace levels of residual chlorine. The water was then heated
to 20 +/- 2 degrees C and distributed to an open basin for seasoning prior
to use. Representative values for hardness and total alkalinity (both as
CaCO3) were 120.0 and 89.6 mg/l, respectively.
Test material: A stock solution of test material (12 mg/l) was prepared in a
500 ml volumetric flask containing test water. The exposure solutions were
prepared at a nominal concentration of 120 mg/l by adding the appropriate
amount of stock solution to glass vessels (300 ml Kimax glass, lipless
beakers) containing 20 liters of test water. The solutions in each test vessel
were stirred with a stir rod prior to adding daphnids. Duplicate test and
control vessels were prepared.
Test conduct: Immediately after stirring, daphnids were placed into each of
the replicate test and control vessels (10 per vessel). Watch glasses were
placed on top of each test vessel and sealed with Parafilm. The vessels
were placed in a certified hood under 8 hours of fluorescent lighting/day.
Animals were observed for mortality and signs of stress at 0, 24 and 48
hours. Temperature, dissolved oxygen concentration, and pH of each
solution were measured at 0 (prior to adding organisms) and 48 hours.
Concentrations of test material in the test vessels at 0 and 48 hours were
analyzed by GC/MS. The geometric mean of the concentrations was
calculated. Since no mortality was observed, statistical analyses were not
performed.
The test was considered valid if control mortality was <= 10%, dissolved
oxygen did not fall below 60% of the initial oxygen level, the temperature
was 20 +/- 2 degrees C, test daphnids in the control groups were not
trapped at the surface of the water and there were no abnormal
occurrences that could influence the outcome.
Purity was 99.8%.
Test substance :
The 48-hour EC50 value indicates that the test substance would not be
Conclusion :
classified according to the European Union's labeling directive and would
correspond to a "low concern level" according to the U.S. EPA's
assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
31.07.2003 (10)
4.3 TOXICITY TO AQUATIC PLANTS E.G. ALGAE
Selenastrum capricornutum (Algae)
Species :
other: biomass and growth rate
Endpoint :
72 hour(s)
Exposure period :
mg/l
Unit :
= 87.8 measured/nominal
NOEC :
> 87.8 measured/nominal
EC50 :
yes
Limit test :
yes
Analytical monitoring :
other: OECD: TG-201 and EEC/Annex V C.3
Method :
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
No protocol deviations were noted. The EbC50 (0-72 hr) and the ErC50 (0
Remark :
15 / 15
Id 78-82-0
4. Ecotoxicity
Date 02.10.2003
72 hr) were inestimable as greater than 50% inhibition in growth and/or
biomass was not achieved. The significant loss (up to 80.7% over the
course of the study) in test material was attributed to volatilization.
Algae exposed to test material exhibited normal growth with respect to
Result :
control. No deformed cells were noted. At the end of the test, the mean
cell density in treated cultures was 1.365 x 10E6 cells /ml (compared to
1.356 x 10E6 cells in control).
The average concentrations of material in the test flasks at the beginning of
the test and after 72 hours were 200.68 and 38.65 mg/l, respectively.
Approximately 80.74% of the material was lost over the course of the
experiment. The mean concentration was 87.82 mg/l. This concentration
was listed as the NOEC.
Results of the photostability tests were similar to those in flasks containing
test material and algae. The control solutions that were exposed to light or
were in the dark exhibited 78.11% and 69.35% losses of test material.
The mean temperature and illumination were 24 degrees C and 746 foot
candles (range 744 - 748 foot-candles) throughout the test. The pH ranged
from 7.42 - 7.88. The shaker speed was maintained at 100 rpm.
The test was considered to be valid since the mean cell concentration in
control cultures increased by a factor of 136-fold within 72 hours.
Test Organisms: A 4-day culture of Selenastrum capricornutum SF-3148
Test condition :
(passage 5 in liquid algal medium) was used as the test algae. Several
passages were performed prior to the test to confirm exponential growth.
Test medium: Sterile growth medium was prepared using high quality
distilled water. The pH of the medium was measured and adjusted to 7.5
(+/- 0.1) using 0.01N NaOH.
Test material stock solution: Test material (0.156 ml) was added to 600.0 g
of algal growth medium (to produce a nominal concentration of 200 mg/l).
The solution was immediately capped and stirred for 1-2 minutes. An
aliquot of the solution was removed for analysis of concentration at time 0.
Test conduct: All steps were carried out aseptically in a hood to prevent
contamination. Test vessels were sterile 250 ml Erlenmeyer flasks. Test
material stock solution (100 ml) was added to 5 flasks and test medium that
did not contain test material was added to 3 flasks. Algae (515 microliters
of algal stock culture to achieve an initial cell density of 1 x 10E4 cells/ml)
were added to 3/5 flasks that contained test material and the three that did
not. The two flasks that contained test material but were not inoculated
served as photostability controls. One of the flasks was exposed to light
and one was wrapped in foil to shield it from light. All flasks were secured
with foam stoppers and transferred to a shaking incubator (24 degrees C,
100 rpm). They were illuminated at an average of 746.2 footcandles
throughout the study.
Temperature, light intensity, and shaker speed (rpm) were assessed at the
0, 24, 48, and 72 hours. Concentrations of test material in the flasks that
contained algae also were assessed at these times. The pH and was
assessed at time 0 and after 72 hours. Concentrations of test material in
the photostability controls also were measured at 0 and 72 hours.
Concentrations of test material were analyzed using gas chromatography
with flame ionization detection (GC/FID).
The exposure concentration was calculated as the geometric mean of the
test concentrations analyzed at the 4 time points. Cell counts were
performed after 24, 48 and 72 hours of exposure using a calibrated Coulter
Counter. The mean algal cell count for the test and control curves was
16 / 16
Id 78-82-0
4. Ecotoxicity
Date 02.10.2003
calculated. Two measures of growth [biomass (area under the growth
curve) and growth rate] were used to determine the effect of the material
on algae. The concentrations that produced a 50% inhibition of biomass
(EbC50) and growth rate (ErC50) relative to control were to be calculated
by fitting linear regression models to the data.
The test was considered valid if the mean cell concentration in the control
cultures increased by a factor of at least 16 within 72 hours.
Purity was 99.9%.
Test substance :
The results of this study indicate that the test substance would not be
Conclusion :
classified according to the European Union's labeling directive and would
correspond to a "low concern level" according to the U.S. EPA's
assessment criteria.
(1) valid without restriction
Reliability :
This was a well-documented OECD-study conducted under GLP
assurances.
Critical study for SIDS endpoint
Flag :
07.08.2003 (9)
other algae: green algae
Species :
biomass
Endpoint :
96 hour(s)
Exposure period :
mg/l
Unit :
= 429.46 calculated
EC50 :
other: model calculation
Method :
2003
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Measured inputs to the program were melting point = - 71.5 degrees C,
Remark :
boiling point = 103.8 degrees C, water solubility = 39,000 mg/l and vapor
pressure = 41.4 mm Hg.
Model compound class is neutral organic.
(2) valid with restrictions
Reliability :
Data were obtained by modeling.
19.08.2003 (18)
4.4 TOXICITY TO MICROORGANISMS E.G. BACTERIA
4.5.1 CHRONIC TOXICITY TO FISH
4.5.2 CHRONIC TOXICITY TO AQUATIC INVERTEBRATES
4.6.1 TOXICITY TO SEDIMENT DWELLING ORGANISMS
4.6.2 TOXICITY TO TERRESTRIAL PLANTS
4.6.3 TOXICITY TO SOIL DWELLING ORGANISMS
17 / 17
Id 78-82-0
4. Ecotoxicity
Date 02.10.2003
4.6.4 TOX. TO OTHER NON MAMM. TERR. SPECIES
4.7 BIOLOGICAL EFFECTS MONITORING
4.8 BIOTRANSFORMATION AND KINETICS
4.9 ADDITIONAL REMARKS
18 / 18
Id 78-82-0
5. Toxicity
Date 02.10.2003
5.0 TOXICOKINETICS, METABOLISM AND DISTRIBUTION
5.1.1 ACUTE ORAL TOXICITY
LD50
Type :
= .1 ml/kg bw
Value :
rat
Species :
other: Carworth-Wistar
Strain :
male
Sex :
5
Number of animals :
Vehicle :
Doses :
other
Method :
1962
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The LD50 value (with confidence interval) was 0.10 ml/kg (77 - 130 mg/kg).
Result :
Based on a relative density of 0.77 (at 20 degrees C), the LD50 value in
mg/kg is 77 mg/kg.
Test material was given by gavage to groups of 5 nonfasted rats (4-5
Test condition :
weeks old, 90-120 g). Dosages (not listed) were arranged in a logarithmic
series differing by a factor of 2. The material was administered in a suitable
vehicle (water, corn oil, 1% Tergitol Penetrant 7, or semi-solid agar). The
number of deaths was monitored over 14 days. The LD50 value and its
confidence interval was estimated by the method of Thompson (Bacteriol
Rev 11:115, 1947) using the tables of Weil (Biometrics 8:249, 1952).
(2) valid with restrictions
Reliability :
The number of deaths at each concentration and the purity of the test
material were not mentioned.
08.07.2003 (29)
LD50
Type :
= 50 mg/kg bw
Value :
rat
Species :
other: unknown
Strain :
no data
Sex :
20
Number of animals :
water
Vehicle :
10 - 3200 mg/kg bw
Doses :
other
Method :
1961
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The LD50 value was 50 mg/kg. The numbers of deaths at each dose were
Result :
not reported. However, death was noted to have occurred between 15
minutes and one day. Clinical signs included moderate to very weak, rapid
and labored respiration, prostration, and very severe vasodilation
(especially at lower does). Survivors gained weight over the observation
period.
A total of 20 rats were administered oral doses of isobutyronitrile ranging
Test condition :
from 10-3200 mg/kg (either undiluted or 10% in water). Animals were
monitored for clinical observations and weight change for 14 days after
exposure.
Material is considered highly toxic
Conclusion :
(2) valid with restrictions
Reliability :
19 / 19
Id 78-82-0
5. Toxicity
Date 02.10.2003
Basic data are given. Purity of the material is unknown.
10.08.2003 (13)
LD50
Type :
= 50 - 100 mg/kg bw
Value :
rat
Species :
no data
Strain :
no data
Sex :
5
Number of animals :
other: 1/2% sodium cellulose sulfate in water
Vehicle :
25 - 400 mg/kg bw
Doses :
other
Method :
1957
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The LD50 value was 50-100 mg/kg. The numbers of deaths at each dose
Result :
were not reported. However, death was noted to have occurred between 1
to 4 hours. Clinical signs included weakness, ataxia, pink feet and ears,
and occasional kicking. Survivors gained weight over the 2 week
observation period.
A total of 5 rats were administered oral doses of isobutyronitrile ranging
Test condition :
from 25-400 mg/kg (as a 10% emulsion in 1/2% sodium cellulose sulfate in
water). Animals were monitored for clinical observations and weight
change for 14 days after exposure.
The material is moderately toxic in rats.
Conclusion :
(2) valid with restrictions
Reliability :
Basic data are given. Purity of the material is unknown.
10.08.2003 (14)
LD50
Type :
> 50 mg/kg bw
Value :
rat
Species :
no data
Strain :
no data
Sex :
2
Number of animals :
other: 1/2% sodium cellulose sulfate in water
Vehicle :
50 and 100 mg/kg bw
Doses :
other
Method :
1957
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Neither of the animals died. Both animals gained weight during the
Result :
observation period. Urine from rats given 50 mg/kg bw orally contained 0.1
mg thiocyanate per cc, indicating the material was converted to cyanide in
the body. The amount of thiocyanate in urine from rats given 25 mg/kg bw
was not listed.
A total of 2 rats were administered 25 and 50 mg/kg bw isobutyronitrile (as
Test condition :
a 10% emulsion in 1/2% sodium cellulose sulfate in water). Animals were
monitored for clinical observations and weight change for 14 days after
exposure. Urine was collected for 24 hours after exposure.
(4) not assignable
Reliability :
Not enough animals were tested for an accurate LD50 determination.
31.07.2003 (14)
5.1.2 ACUTE INHALATION TOXICITY
other: LC10
Type :
= 1173 ppm
Value :
20 / 20
Id 78-82-0
5. Toxicity
Date 02.10.2003
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
30
Number of animals :
Vehicle :
target vapor concentrations were 1200, 1800, and 2700 ppm
Doses :
1 hour(s)
Exposure time :
OECD Guide-line 403 "Acute Inhalation Toxicity"
Method :
1986
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
A pulmonary function test on 12 male rats (4/treatment group) was
Remark :
conducted in conjunction with this test. All four animals exposed to 2709
ppm and two exposed to 1778 ppm died on day 4. All animals exposed to
1248 ppm lived. The most consistent finding in the survivors (5/6) was a
decrease in dynamic compliance (up to 76% compared to pre-exposure
values). The forced expiratory flow at 10% vital capacity also was
decreased (up to 67%) in the 2 survivors exposed to 1778 ppm. The
changes observed were associated with pulmonary edema or congestion.
LC10 value: LC10 (+/- 95% confidence interval) = 1143 ppm (males), 1630
Result :
ppm (females), 1173 ppm (combined).
Exposure concentrations: Actual concentrations were 1248 +\- 62, 1778 +\
16, and 2709 +\- 34 ppm. No aerosol was present. Overall mean chamber
temperature and relative humidity varied from 22 - 23 degrees C and 51 -
59%, respectively.
Deaths at each dose:
1248 ppm: 1/10; 1 male on Day 1
1778 ppm: 5/10; 4 males and 1 female on Day 1
2709 ppm: 8/10; 5 males on Day 1 and 3 females (2 on Day 1 and one on
Day 2)
Remarks: Lethargy was noted at 1248 ppm (4/4 males and 1/5 females).
The lethargic female also exhibited sialorrhea, which resolved after Day 2.
At 1778 ppm, all animals exhibited lethargy. At 2709 ppm, all animals
exhibited lethargy, gait disturbances and narcosis. These clinical signs
were seen during or just after exposure cessation, the severity was dose-
related, and they resolved after 24-hours. The mean body weight increased
in all animals during the 14-day observation period. No compound-related
gross pathology was noted in animals that died spontaneously or in
animals terminated on Day 14.
Rats [CRL:CD(SD)BR] were exposed to target vapor concentrations of
Test condition :
1200, 1800, and 2700 ppm (5/sex/concentration). Males weighed 232-263
g and females weighed 210-237 g at the start of the study. Food and water
were available ad libitum (except during exposure).
Exposures were conducted in 420 l stainless steel and glass inhalation
chambers. Chambers were maintained under negative pressure (-0.5"
water) and at 13 air changes per hour. Vapors were generated by metering
the test material dropwise into a heated glass bead-packed column
supplied with metered, dried, oil-free compressed air. Chamber vapor
samples were continuously collected and analyzed with an infrared
analyzer equipped for automated sampling and analyses. Temperature
and humidity were measured twice per hour. Twice during exposure, a
particle counter sampled the chamber atmosphere for nongaseous
airborne material. Particles greater than 0.3 microns were counted. The
results were compared to those of the air control chamber. Distribution of
test material was determined by measuring the concentrations in 27
different positions throughout the chamber and comparing them to a fixed
21 / 21
Id 78-82-0
5. Toxicity
Date 02.10.2003
reference position. Subsequently, the chamber vapor concentration was
measured from the fixed reference position.
Exposure was for 1-hour in duration followed by a 14-day observation
period. Rats visible through the chamber windows were observed for signs
of toxicity during exposure. Each rat was removed from its cage before and
after exposure and twice daily thereafter and examined. Animals that died
during the study were necropsied as soon as possible after discovery of
death. Survivors were fasted for 16 hours prior to euthanization and
necropsy on Day 15. The following tissues were examined: trachea, lungs,
heart, liver, esophagus, stomach, duodenum, jejunum, ileum, cecum,
colon, pancreas, liver, salivary glands, kidneys, urinary bladder, pituitary
gland, adrenal, thyroid, parathyroid, thymus, spleen, mesenteric lymph
nodes, bone marrow (femoral), brain, testes, epididiymides, male
accessory sex glands, ovaries, vagina, uterus, and Fallopian tubes.
Mortality data were evaluated by Probit analysis.
Purity was 99.7%
Test substance :
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted
under GLP assurances.
Critical study for SIDS endpoint
Flag :
10.08.2003 (15) (16)
LC50
Type :
> 1000 ppm
Value :
rat
Species :
Sprague-Dawley
Strain :
male/female
Sex :
10
Number of animals :
Vehicle :
1200 ppm (nominal)
Doses :
1 hour(s)
Exposure time :
other: DOT Final Rule 49 CFR (parts 172 and 173
Method :
1986
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Isolation and housing room low temperatures were 69 and 68 degrees F,
Remark :
respectively, exceeding tolerances specified in the protocol (73 +/- 3
degrees F). This deviation did not affect the outcome of the study.
All animals survived to the end of the study. Mean vapor concentrations of
Result :
the test material were 1233 +/- 15 and 1177 +/- 53 ppm for male and
female rats, respectively. The saturated vapor concentration at 20 degrees
C was 54, 474 ppm. An aerosol was not present. The temperature and
relative humidity of the room ranged from 68-70 degrees F and 55-60%,
respectively.
Groups of 5 male (243-260 g) and 5 female (220-233 g) rats
Test condition :
[CRL:CD(SD)BR] were exposed to a target vapor concentration of 1200
ppm for 1 hour. Feed and water were available ad libitum (except during
exposure). Rats were observed for mortality during exposure and twice
daily on subsequent workdays for 14 days.
Chamber atmospheres were produced by passing dried, oil-free
compressed air through a glass bead-packed distilling column into which
the test material was pumped dropwise. The exposure began when the
vapor concentration reached 1000 ppm. Males and females were exposed
in separate 20 l bell jars. Four samples of the atmosphere were analyzed
for iso-butyronitrile concentration with an infrared analyzer. Two samples
of atmosphere were analyzed with a particle analyzer to insure that an
aerosol was not present. The temperature was monitored 4 times during
exposure.
Purity of the test material was 99.7% (GC-MS). No attempt was made to
Test substance :
22 / 22
Id 78-82-0
5. Toxicity
Date 02.10.2003
identify trace components.
The material is not subject to regulation under 49 CFR Parts 172 and 173,
Conclusion :
since the LC50 value was greater than 1000 ppm.
(1) valid without restriction
Reliability :
The study is comparable to an OECD guideline, GLP study.
07.08.2003 (12)
LC50
Type :
= 500 - 1000 ppm
Value :
rat
Species :
other: albino
Strain :
no data
Sex :
18
Number of animals :
Vehicle :
Doses :
4 hour(s)
Exposure time :
other
Method :
1962
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
All six animals died after inhalation exposure to 1000 ppm, and 0/6 died
Result :
after exposure to 500 ppm test material for 4 hours. All six rats exposed to
a concentrated vapor for 10 minutes died.
Purity of the test material was not listed.
Test substance :
(4) not assignable
Reliability :
Study documentation is poor. Purity of the material is not known.
08.07.2003 (29)
5.1.3 ACUTE DERMAL TOXICITY
LD50
Type :
= .31 ml/kg bw
Value :
rabbit
Species :
New Zealand white
Strain :
male
Sex :
Number of animals :
Vehicle :
Doses :
other: Draize test
Method :
1962
Year :
no
GLP :
as prescribed by 1.1 - 1.4
Test substance :
The LD50 value (with confidence interval) was 0.31 (0.21 - 0.46) ml/kg.
Result :
Based on a density of 0.77 (at 20 degrees C), the LD50 value in mg/kg is
239.
Fur was clipped from the entire trunk of rabbits (2.5 to 3.5 kg) and the
Test condition :
material was applied beneath an impervious plastic film. Groups of 4
animals were exposed to various concentrations (not listed). The animals
were immobilized during the 24-hour contact period. The film was then
removed and the animals were caged for the subsequent 14 day
observation period. The LD50 value and its confidence interval was
estimated by the method of Thompson (Bacteriol Rev 11:115, 1947) using
the tables of Weil (Biometrics 8:249, 1952).
(2) valid with restrictions
Reliability :
Purity of the material, test concentrations and the number of deaths at each
concentration were not mentioned.
08.07.2003 (29)
23 / 23
Id 78-82-0
5. Toxicity
Date 02.10.2003
5.1.4 ACUTE TOXICITY, OTHER ROUTES
5.2.1 SKIN IRRITATION
5.2.2 EYE IRRITATION
5.3 SENSITIZATION
5.4 REPEATED DOSE TOXICITY
Sub-chronic
Type :
rat
Species :
male/female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
14 weeks (total of 63 exposure days)
Exposure period :
6 hours/day, 5 days per week
Frequency of treatm. :
Post exposure period :
60, 120, 209 ppm
Doses :
yes
Control group :
< 60 ppm
NOAEL :
= 60 ppm
LOAEL :
other
Method :
1984
Year :
no data
GLP :
other TS
Test substance :
Test material concentrations: The nominal concentrations (+/- SD) were
Result :
56.0 +/- 8.8, 119.1 +/- 16.9 and 203.0 +/- 19.6. Corresponding analytical
concentrations were 60.2 +/- 1.0, 120.3 +/- 1.1 and 209.0 +/- 1.3 ppm.
Since the nominal analytical ratios were close to 1, the material was a true
vapor. There was no indication that the test material was unstable.
Distribution was uniform (> 96%) for each of the exposure concentrations.
Airflow, temperature and relative humidity ranged from 1719-1765 l/min,
21.0 - 26.1 degrees C and 17-50%, respectively.
Effects at all exposure concentrations: Signs of toxicity (labored breathing,
nasal discharge, salivation, discharge from the eyes, hypoactivity and/or
alopecia) were observed in all exposed groups. Incidences of these signs
increased in a dose-dependent manner. Males and/or females in all
exposed groups had significant decreases in red blood cells and
hemoglobin values. Urine thiocyanate concentrations increased in all
exposed groups, with concentrations from animals exposed to 120 ppm
similar to or higher than those exposed to 210 ppm. However, since a
dose-dependent diuresis occurred, the total amount of urine thiocyanate
present (concentration x urine volume) increased with increasing
concentrations.
Effects at 209 ppm: Three males died or were killed in extremis (2
between exposures 2 and 3 and one between exposures 18 and 19).
Arched back, ataxia, tremors or convulsions, biting, pawing or rubbing chin
against cage, irritation of the conjunctiva and breathing difficulties were
noted in a few animals during exposure. Males and females exhibited
significant decreases in body weight throughout the study (P <= 0.01).
24 / 24
Id 78-82-0
5. Toxicity
Date 02.10.2003
Average final body weights of males and females were lower than controls
(377.5 g in exposed males vs. 435.4 g in controls and 255.0 g in exposed
females vs. 276.6 g in controls). Absolute and/or relative heart, liver, spleen
and kidney weights were increased in males and/or females. Absolute
testes weights were decreased in males. Mean corpuscular hemoglobin
concentrations (males only) were lower than control (all P <= 0.05). Serum
alkaline phosphatase (males and females), SGOT (males only) and SGPT
(males only) were increased, and BUN concentrations were decreased. No
gross lesions were observed at termination. There was mildly increased
hemosiderin deposition in the spleens of 10/15 females. Other microscopic
changes were considered to be spontaneous.
Effects at 120 ppm: Ataxia was observed during exposure in 2 females.
Males had significant body weight decreases (P < = 0.05) at two time
points. Absolute and/or relative liver weights were increased in males and
females and absolute and/or relative spleen weights were increased in
males. Mean corpuscular hemoglobin concentrations (males only, P < =
0.05) were lower than control. No gross lesions were observed at
termination. There was mildly increased hemosiderin deposition in the
spleens of 11/15 females. Other microscopic changes were considered to
be spontaneous.
Effects at 60 ppm: Absolute and relative spleen weights were increased in
males. Serum thiocyanate concentrations were marginally increased in
males and females.
Animals: Rats were acclimated for at least 10 days prior to use. Two days
Test condition :
prior to the start of the study, males weighed 174-200 g and females
weighed 132-145 g. On the first day of the study, the animals were 43
days old. Animals were randomly allocated by body weight into 4 groups of
15 animals/sex/group. Animals were individually housed in suspended
mesh cages and given food and water ad libitum (except during exposure).
Animal rooms were maintained at 70-74 degrees C and 35-60% relative
humidity, with a 12 hour light/dark cycle.
Exposure conditions: Exposures (6 hr/day, 5 days/week) occurred in 10
m3 Rochester-style stainless steel and glass inhalation chambers. Rats
were placed individually in wire mesh cages that were suspended in the
chambers by 3-tiered racks. Males were placed on one side and females
on the other. The concentrations of material in the chambers (20, 120 or
210 ppm) were controlled by either adjusting the nitrogen flow though the
propionitrile in the bubblers or by changing the amount of test material in
the bubblers. The bubbler was connected to a side port in the vertical
particle-size separator, which was in turn connected to the air inlet at the
top of the inhalation chamber. One bubbler was used in each of the
generation systems. Airflow was maintained at a constant flow of 1727
liters/min. Nominal concentration measurements were determined daily for
each chamber following exposure, by dividing the amount of test material
delivered to the chamber (the difference between the pre- and post-
exposure weights) over the 6-hr exposure period by the total air volume
during the same period. Concentrations of test material in the chambers
were measured 4 times daily using a Miran 1A General Purpose Gas
Analyzer. Additional samples of atmosphere from 9 specified locations in
each chamber were also taken at 3 different times to determine if the vapor
was distributed uniformly.
Test conduct: Animals were observed for clinical signs between the second
and fifth hour of each exposure. Estimations of the percentages of animals
exhibiting hypoactivity, eye irritation and breathing difficulties were made.
All animals were individually examined for gross signs of toxicity preceding
and following each exposure and checked for mortality. Each animal was
weighed and given a thorough examination for gross signs of toxicity on a
weekly basis.
25 / 25
Id 78-82-0
5. Toxicity
Date 02.10.2003
Animals were euthanized after 14 total weeks on the study. Terminal body
weights were obtained (following an overnight fast). Blood and urine were
collected. Whole blood was treated with an anticoagulant and was
analyzed for total and differential erythrocyte count, total leukocyte count,
platelet count, hematocrit, hemoglobin, and red blood cell indices (mean
corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular
hemoglobin concentration). Serum was analyzed for albumin, globulin, total
protein, blood urea nitrogen, total bilirubin, glucose, glutamic pyruvic
transaminase (SGPT), alkaline phosphatase, glutamic oxaloacetic
transaminase (SGOT), T3, T4, thiocyanate and lactate dehydrogenase.
Urine was analyzed for the presence of thiocyanates.
Detailed necropsies were conducted on all rats that died during the course
of the study, those that were killed moribund, and those that survived to
study termination. The adrenal glands (both together), testes (with
epididymides, heart, kidneys, liver, pituitary and spleen were weighed. The
aforementioned organs and the following tissues were fixed in 10% neutral
formalin: abdominal aorta, bone and bone marrow (femur), brain,
esophagus, ovaries, colon, ileum, lung, lymph nodes (mesenteric),
mammary gland, nasal turbinates, pancreas, thyroid/parathyroid, prostate,
salivary gland, sciatic nerve, skeletal muscle, skin, spinal cord, stomach,
thymus, trachea, urinary bladder, uterus (with cervix) and gross lesions.
Eyes (with optic nerve) were fixed in a solution of 2% glutaraldehyde and
10% neutral buffered formalin. Tissues were processed, embedded in
paraffin, cut at five microns, stained with hematoxylin and examined
microscopically.
Statistical analyses: In life and terminal body weights and organ weight
data were analyzed using Dunnett's test. Organ to body weight ratios were
analyzed using the Mann-Whitney test, with the Bonferroni inequality. Data
for frequencies of microscopic lesions were evaluated with the Fisher's
exact test with the Bonferroni inequality. Hematological and serum and
urine chemistry variables were examined using Dunnett's test.
Test material was propionitrile (CAS No. 107-12-0). The purity of the test
Test substance :
material was 96%. Impurities were not listed.
(1) valid without restriction
Reliability :
The study is comparable to a guideline study; however, a NOAEL was not
established
Critical study for SIDS endpoint
Flag :
10.08.2003 (30)
5.5 GENETIC TOXICITY `IN VITRO`
Bacterial reverse mutation assay
Type :
Salmonella typhimurium/TA98, 100, 1535, 1537, and Escherichia
System of testing :
coli/WP2uvrA(pKM101)
100, 333, 1000, 3330 and 5000 micrograms/plate
Test concentration :
> 5000 micrograms/plate
Cytotoxic concentr. :
with and without
Metabolic activation :
negative
Result :
other: EEC Annex V Guideline number B.14, "Other Effects-Mutagenicity
Method :
Salmonella typhimurium-Reverse Mutation Assay", and Guideline number
B.13, Other Effects-Mutagenicity, Escherichia coli-Reverse Mutation
Assay"
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
This is the critical study for the mutagenesis endpoint.
Remark :
26 / 26
Id 78-82-0
5. Toxicity
Date 02.10.2003
No positive responses were induced in any of the tester strains. None of
Result :
the concentrations tested caused toxicity. No precipitate was observed at
the maximum concentration tested. All criteria for a valid test were met.
Test strains: The S. typhimurium and E. coli strains were obtained from Dr.
Test condition :
Bruce Ames, University of California Berkeley and the National Collection
of Industrial Bacteria,Torry Research Station, Scotland, respectively.
Frozen permanent stocks were prepared by growing fresh overnight
cultures, adding DMSO (0.09 ml/ml culture) and freezing small aliquots at <
= -70 degrees C. Master plates were prepared by streaking each test
strain from a frozen permanent stock onto minimal agar supplemented with
histidine, biotin, ampicillin and/or tryptophan (depending on the strain).
Tester strain master plates were stored at 5 +/- 3 degrees C. Overnight
cultures were inoculated by transferring colonies from the master plates to
a flask containing culture medium. Inoculated flasks were placed in
a shaker/incubator (125 +/- 25 rpm, 37 +/- 2 degrees C). Cultures were
harvested once a predetermined turbidity was reached (at least 0.5 x 10E9
cells/ml). Test stains were checked for rfa wall mutation (all Salmonella
strains), pKM101 plasmid R-factor (Salmonella TA98 and TA100 and E.
coli only), and characteristic number of spontaneous revertants (all strains)
on the day the mutagenesis test was conducted.
Test medium: The broth used to grow overnight cultures of the tester
strains was Vogel-Bonner salt solution supplemented with 2.5% Oxoid
Nutrient Broth No. 2. Bottom agar was Vogel-Bonner minimal medium E
supplemented with 1.5% (w/v) agar and 0.2% (w/v) glucose. Overlay agar
contained 0.7% agar (w/v) and 0.5% NaCl (w/v) supplemented
with either 10 ml of 0.5 mM histidine/biotin solution or 0.5 mM tryptophan
solution.
S-9 mix: S9 homogenate was purchased from Molecular Toxicology Inc.
This was prepared from male Sprague-Dawley rats that had been injected
i.p. with 500 mg/kg Aroclor 1254. S-9 mix was prepared immediately prior
to use.
Concentrations of test material: The concentrations tested (100, 333, 1000,
3330 and 5000 micrograms/plate) were selected based on the results of a
dose range-finding study using test strains TA100 and WP2uvrA(pKM101)
and 10 doses of test material ranging from 6.67 to 5000 micrograms/plate
(both in the presence and absence of S-9 mix).
Positive, negative and sterility controls: Positive controls [2
aminoanthracene (2.5 and 5.0 micrograms/plate), 2-nitrofluorene (1.0
micrograms/plate), sodium azide (2.0 micrograms/plate), ICR-191 (2.0
micrograms/plate), and 4-nitroquinoline-N-oxide (2.0 micrograms/plate)]
were run concurrently. DMSO (50 microliters) was used as a vehicle
and vehicle control. The most concentrated test material dilution and S-9
mix were tested for sterility by plating a 50 microliter aliquot on selective
agar.
Test conduct: A plate incorporation methodology was used. Test material
or positive control (50 microliters), test strains (100 microliters) and S-9 mix
or vehicle (500 microliters) were combined in 2.0 ml of molten, selective
top agar. This was overlaid onto 25 ml of minimal agar that had been
plated into 15 x 100 mm Petri dishes. All concentrations of test material,
vehicle controls and positive controls were plated in triplicate. Revertant
colonies were counted after 48 +/- 8 hours of inverted incubation at 37 +/- 2
degrees C. The condition of the background lawn was evaluated for
evidence of cytotoxicity and precipitate.
Evaluation: The mean number of revertants and standard deviation were
calculated. Various criteria were established to constitute a valid assay
(test strain integrity, characteristic number of spontaneous revertants, cell
27 / 27
Id 78-82-0
5. Toxicity
Date 02.10.2003
density > = 0.5 x 10E9, at least a 3-fold increase in revertants in positive
controls, and a minimum of 3 non-toxic doses). A positive response was
indicated by a 2-3 fold increase in mean revertant number depending on
the bacterial tester strain.
Purity of the test material was not confirmed in this study. However, the lot
Test substance :
of test material used (D-7) was the same as that used in the aquatic toxicity
studies, where the purity was analyzed to be 99.8 - 99.9 %.
Material was not genotoxic under conditions of this assay
Conclusion :
(1) valid without restriction
Reliability :
This was a well-documented EEC Annex guideline study conducted under
GLP assurances.
10.08.2003 (3)
Chromosomal aberration test
Type :
Chinese Hamster Ovary (CHO) Cells
System of testing :
up to 700 micrograms/ml
Test concentration :
> 700 micrograms/ml
Cytotoxic concentr. :
with and without
Metabolic activation :
negative
Result :
OECD Guide-line 473
Method :
1999
Year :
yes
GLP :
as prescribed by 1.1 - 1.4
Test substance :
This is the critical study for the chromosomal aberration
Remark :
endpoint.
Without activation: In the initial study without metabolic activation,
Result :
reductions of 30%, 31%, 6% and 6% were observed in the mitotic indices
of the cultures treated with 239, 342, 489 and 669 micrograms/ml,
respectively. Chromosomal aberrations were analyzed from cultures
treated with these concentrations. In the confirmatory study without
activation, reductions of 44%, 24%, 23%, 16% and 34% were observed in
the mitotic indices of cultures treated with 222, 296, 394, 525 and 700
micrograms/ml, respectively. Chromosomal aberrations were analyzed
from cultures treated with 296, 394, 525 and 700 micrograms/ml. No
significant increases in cells with chromosomal aberrations, polyploidy,
or endoreduplication were observed in analyzed cultures.
With activation: In the initial study with metabolic activation, reductions of
25% and 46% were observed in the mitotic indices of cultures treated with
342 and 489 micrograms/ml, respectively. Chromosomal aberrations were
analyzed from cultures treated with 239, 342, 489 and 699 micrograms/ml.
No reductions in mitotic index were noted in any of the cultures in the
confirmatory study. In this study, chromosomal aberrations were analyzed
from cultures treated with 296, 394, 525 and 700 micrograms/ml. No
significant increases in cells with chromosomal aberrations, polyploidy, or
endoreduplication were observed in analyzed cultures.
No precipitate was observed at the maximum concentration tested in any of
the studies.
All criteria for validity were met in each study.
Cells: The Chinese hamster ovary cells used in the assay (CHO-WBL)
Test condition :
were from a permanent cell line originally obtained from Dr. S. Wolff,
University of California, San Francisco. Stock cultures were maintained for
up to 8 weeks after thawing. Mycoplasma testing was performed twice
during this period. Cells were grown at 37 +/- 2 degrees C (in 5% +/- 1.5%
CO2 in air) in McCoy's 5a culture medium which was supplemented with
10% fetal bovine serum, 2 mM L-glutamine, 100 units/ml penicillin G and
100 micrograms/ml streptomycin.
S-9 mix: S-9 was isolated from the liver of rats (sex not stated) 5 days after
28 / 28
Id 78-82-0
5. Toxicity
Date 02.10.2003
i.p. treatment with 500 mg/kg Aroclor 1254. S-9 was stored frozen at < =
70 degrees C until use. S-9 mix was prepared by adding an energy-
producing system (NADP plus isocitric acid) to S-9.
Test material and negative and positive controls: The test material was
dissolved in DMSO. The top concentration tested (approximately 700
micrograms/ml or 10 mM) was the recommended high dose for the assay.
The negative control was 10 microliters/ml DMSO. The positive controls
were mitomycin C (without activation) and cyclophosphamide (with
activation).
Initial test: Cultures were initiated by seeding approximately 1.2 x 10E6
cells per 75 cm2 flask into 10 ml of complete McCoy's 5a medium. For the
test without metabolic activation, cultures were incubated with test
material for 3.0 hrs at 37 degrees C. For the test with metabolic activation,
cells were incubated for approximately 3.0 hours with test material and S-9
mix in McCoy's 5a medium that did not contain fetal bovine serum.
Replicate cultures for each concentration of test material (4.73, 6.76, 9.66,
13.8, 19.7, 28.1, 40.1, 57.3, 81.9, 117, 167, 239, 342, 489 and 699
micrograms/ml), positive control (0.75 and 1.5 micrograms/ml mitomycin C
and 5.0 and 10.0 micrograms/ml cyclophosphamide), vehicle and untreated
controls were prepared. Cultures with or without S-9 were then washed
with buffered saline, and incubated with complete McCoy's 5a medium for
17 hours. Colcemid (0.1 micrograms/ml) was present during the last 2
hours of incubation. Cells were visually inspected for cytotoxicity
prior to harvest. Cells were then trypsinized and spun in a centrifuge. The
supernatant was discarded and the cells swollen with 75 mM KCl hypotonic
solution. The cells were then fixed with an absolute methanol: glacial
acetic acid (3:1, v:v) fixative. They were then placed on glass slides and
air-dried. Cells were stained with 5% Giemsa and analyzed for mitotic
index and chromosomal aberrations.
Confirmatory assay: The test with metabolic activation was conducted the
same as in the initial test, but with different concentrations of test material
(222, 296, 394, 525 and 700 micrograms/ml). In the test without metabolic
activation, the test material (27.8, 55.5, 111, 222, 296, 394, 525 and 700
micrograms/ml), positive control (0.20 or 0.40 micrograms/ml mitomycin C
or 5.0 or 10.0 micrograms/ml cyclophosphamide) and negative controls
were incubated with the cells for 18.3 hours (instead of 3). For both tests,
Colcemid was present for the last 1.9 hours of incubation and cells were
harvested after 20.2 total hours of incubation. The slides were prepared as
described for the previous test.
Evaluation: Cells were selected for good morphology and only cells with
the number of centromeres equal to the modal number 21 +/-2 were
analyzed. One hundred cells (if possible) were analyzed from each
replicate of the vehicle control, 4 concentrations of the test material, and
one concentration of positive control for the different types of chromosomal
aberrations. At least 25 cells were analyzed from those cultures that had
greater than 25% of cells with one or more aberrations. The number of
mitotic cells in 1000 cells was determined and the ratio expressed as
percentage of mitotic cells. Percent polyploidy and endoreduplication
were analyzed by evaluating 100 metaphases (if possible). Chromatid and
isochromatid gaps were noted but were not used in calculating the total
number of aberrations.
Acceptance criteria: The assay was considered valid if the negative
(untreated) and vehicle controls contained < 5% cells with aberrations, the
positive control result was significantly higher (p < = 0.01) than that of the
vehicle control, a high dose of 10 mM or the highest soluble concentration
was used if the material did not cause at least a 50% reduction of the
mitotic index at the tested concentrations, and at least 3 concentrations
29 / 29
Id 78-82-0
5. Toxicity
Date 02.10.2003
were analyzed.
Data analysis: The statistical analysis employed a Cochran-Armitage test
for linear trends and Fisher's Exact Test to compare the percentage of cells
with aberrations. Data for polyploidy and/or endoreduplication were also
analyzed separately. A test was considered positive if a significant
increase in the number of cells with aberrations (p < = 0.01) was observed
at one or more concentrations.
Purity of the test material was not confirmed in this study. However, the lot
Test substance :
of test material used (D-7) was the same as that used in the aquatic toxicity
studies, where the purity was analyzed to be 99.8 - 99.9 %.
Material was not genotoxic under conditions of this assay.
Conclusion :
(1) valid without restriction
Reliability :
This was a well-documented OECD guideline study conducted
under GLP assurances.
10.08.2003 (2)
5.6 GENETIC TOXICITY `IN VIVO`
5.7 CARCINOGENICITY
5.8.1 TOXICITY TO FERTILITY
Fertility
Type :
rat
Species :
female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
21 to 33 days (depending on day of mating)
Exposure period :
6 hr/day, 7 days/week
Frequency of treatm. :
Premating exposure period
0 days
Male :
21 days
Female :
to gestation days 13-15
Duration of test :
No. of generation :
studies
60, 120 and 210 ppm
Doses :
yes
Control group :
= 60 ppm
NOAEL parental :
= 210 ppm
other: NOAEL :
Reproductive Toxicity
other
Method :
1984
Year :
yes
GLP :
other TS
Test substance :
Exposure concentrations: The average mean daily analytical exposure
Result :
concentrations (60.1, 120.2 and 209.2 ppm) were within 1% of the target
levels (60, 120 and 210 ppm, respectively). Mean temperatures and
humidities ranged from 22-25 degrees C and 26-29%, respectively.
Signs of toxicity: None of the animals died. There was no effect of test
material on body weight. Animals exposed to 210 ppm exhibited arched
back (N = 4 on days 1-10 and N=2 on days 11-20), lacrimation (N = 2 on
days 1-10 and N = 1 on days 21-30), salivation (N= 15 on days 1-10, N =
22 on days 11-20 and N = 21 on days 21-30) hypoactivity (N = 13 on days
30 / 30
Id 78-82-0
5. Toxicity
Date 02.10.2003
1-10, N = 5 on days 11-20 and N = 3 on days 21-30), staining of facial fur
(N = 2 on days 1-10, N = 4 on days 11-20 and N = 4 on days 21-30) and
red nasal encrustation (N = 1 on days 1-10, N = 5 on days 11-20 and N = 5
on days 21-20) after exposure. Animals exposed to 120 ppm also exhibited
salivation (N = 6 on days 11-20 and N = 4 on days 21-30, staining of facial
fur (N = 7 on days 1-10, N = 5 on days 11-10 and N = 2 on days 21-30)
and red nasal encrustation (N = 2 on days 1 1-0, N = 8 on days 11-20 and
N = 6 on days 21-30). A few animals in the 60 ppm group also exhibited
red nasal encrustation (N = 1 on days 1-10, N = 3 on days 11-20 and N = 1
on days 21-30) and staining of facial fur (N = 1 on days 1-10, N = 3 on days
11-20 and N = 1 on days 21-20). One control animal had stained facial fur
on days 21-30 and another had red nasal encrustation on days 1-10. Signs
of toxicity generally abated by the morning after exposure. Alopecia was
observed in animals (N = 2 controls, N = 3 low dose, N = 5 mid dose, N =
9) at one or more of their weekly physical examinations.
The only remarkable findings at gross necropsy were bilateral uterine
hydrometra in one animal exposed to 210 ppm and hydrometra in the left
uterine horn of one animal exposed to 120 ppm.
Fertility: There was no effect of treatment on fertility. Efficiency of mating
(32.0%, 32.0%, 30.7% and 25.0% in the control, low, mid and high dose
groups) and pregnancy rate (100%, 95.8%, 100% and 91.3% in the
respective groups) were comparable between groups. There was no
difference in the numbers of live implants (ranged from 13.4 - 13.9),
resorptions (ranged from 0.6 - 0.8), nidations (ranged from 14.1 - 14.5),
corpora lutea (ranged from 13.0 - 15.2), preimplantation loss (4-8%) and
postimplantation loss (4-6%). Evaluation of the vaginal smears of 2 females
that did not copulate showed one that did not cycle (but was pregnant at
necropsy), and another that only went through the cycling stage of
proestrus.
Animals: Virgin female Sprague Dawley rats (43 days old upon receipt) and
Test condition :
virgin male Sprague Dawley rats (50 days old upon receipt) were
acclimated for one week and examined for general health and the
presence of pinworms and ectoparasites. Body weights of ten females and
ten males that were taken upon receipt were 128-144 g and 178-233 g,
respectively. No significant health problems were noted during the
acclimation period, and the animals were released for study. Males and
females were individually caged (except during mating). Food and water
were available ad libitum (except food was not available to females during
exposure). Animals were housed in a room maintained at 72 +/- 2 degrees
F and 40-60% relative humidity, under a 12 hr light/dark cycle.
Exposures: Exposures (6 hr/day, 7 days/week) occurred in 10 m3
Rochester-style stainless steel and glass inhalation chambers. Due to
inclement weather and building equipment failures, 2 exposures (days 2
and 16) were only for 4 hours and one exposure (day 1) was for 5 hours.
Atmospheres of propionitrile vapor were generated using bubbler systems.
The concentrations of material in the chambers were controlled by either
adjusting the nitrogen flow though the propionitrile in the bubblers or by
changing the amount of test material in the bubblers. Concentrations of
test material in the chambers were measured 4 times daily (except during
day 16) using a Miran 1A General Purpose Gas Analyzer. The nominal
concentration of material, temperature and humidity also were measured
(at intervals that were not listed).
Study conduct: Twenty four females per group were assigned to be
exposed to target concentrations of 0, 60, 120 or 210 ppm test material.
Exposure began when females were 63 days old. Animals were observed
during exposure for signs of toxicity. After 21 days of exposure (which was
sufficient to cover 3-4 estrus cycles), females were randomly mated (1:1) to
an untreated male that had been assigned to the corresponding treatment
31 / 31
Id 78-82-0
5. Toxicity
Date 02.10.2003
group (30 males were assigned per group). At night, after exposure,
females were caged with their assigned male until copulation was
confirmed (by presence of a copulatory plug or sperm in vaginal smear) or
5 nights without confirmed copulation. Females that failed to mate with the
assigned male were mated with another male that had copulated with
another female in the same group. Nightly co-housing with the second
male occurred until copulation was confirmed (or for a maximum of 7
nights). The day on which copulation was confirmed was considered
gestation day 0. Exposure of females continued until copulation was
confirmed or a maximum of 12 nights of cohabitation with males without
signs of copulation. Vaginal smears were taken on 5 consecutive days for
females that did not exhibit copulation.
After assignment to the study, males and females were weighed once per
week. Mated females were weighed on gestation days 0 and 13. Females
were given a thorough physical examination once per week and were
observed for clinical signs of toxicity before and after exposures. All
animals were checked twice daily for mortality and gross abnormalities.
Females were killed on gestation day 13 (or the nearest working day after
gestation day 13, up to gestation day 15). Females without confirmed
copulation ware euthanized in the second week after the last day of co-
housing. Each female was given an external examination and weighed.
The tissues and organs of the thoracic and abdominal cavities were
examined for gross lesions. Pregnancy status was determined, nidation
sites and were classified, and corpora lutea were counted. The ovaries
and uteri of females were preserved in 10% neutral buffered formalin.
Males were killed after mating and were not examined.
Statistical analyses: Body weight data were analyzed using Dunnett's test.
Mating and pregnancy rate data were analyzed by a Fisher's exact test and
an uncorrected chi-square test. Other data were analyzed by the Mann-
Whitney U test. The Bonferroni inequality was assumed when comparing
multiple treatments to control values for all tests except Dunnett's test. The
critical level for significance was p < 0.05.
The test material was propionitrile (CAS No. 107-12-0). Purity of the test
Test substance :
material was 96.1%. Impurities included acrylonitrile (0.1%), adiponitrile
(0.7%), p-nitrosodiphenylamine (0.08%) and water (< 0.1%). Analyses
indicated no significant decomposition of the test material over the course
of the study.
The authors concluded that the incidences of red nasal encrustation in the
Conclusion :
low dose animals, alopecia in the mid and high dose animals and staining
of facial fur in all treated groups were too low to be definitely related to
administration of test material. There was no effect of treatment on fertility
of females.
(1) valid without restriction
Reliability :
Study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (23)
Fertility
Type :
rat
Species :
male
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
57 days
Exposure period :
6 hours/day, 5 days/week
Frequency of treatm. :
Premating exposure period
46 days
Male :
0 days
Female :
to gestation day 13-15
Duration of test :
32 / 32
Id 78-82-0
5. Toxicity
Date 02.10.2003
No. of generation :
studies
60, 120 and 210 ppm
Doses :
yes
Control group :
= 60 ppm
NOAEL parental :
= 210 ppm
other: NOAEL :
Reproductive Toxicity
other
Method :
1985
Year :
yes
GLP :
other TS
Test substance :
Exposure concentrations: The average mean daily analytical exposure
Result :
concentrations (60.2, 120.4 and 208.9 ppm) were within 1% of the target
levels (60, 120 and 210 ppm, respectively). Mean temperatures and
humidities ranged from 22-25.5 degrees C and 24-27%, respectively.
Signs of toxicity: One of animals exposed to 210 ppm died after 2 days of
exposure. On the previous day, this animal exhibited labored breathing,
hypoactivity, poor control of the hind limbs, difficulty in standing, body
tremors and involuntary movements. No unusual findings were observed
at necropsy.
Body weights of males exposed to 210 ppm were approximately 6-9%
lower than those of the control group during most of the exposure period,
and remained lower than control (but were not significantly different) until
the end of the study.
Animals exposed to 210 ppm exhibited signs of toxicity such as arched
back (N = 8 on days 1-10, N = 3 on days 11-20 and 51-57, and N = 5 on
days 41-50), hypoactivity (N = 12-15 at each 10-day interval up to day 50,
and N = 4 from days 51-57), labored breathing (N = 10 on days 1-10, N = 3
on days 11-20 and 31-40, N = 5 on days 21-30 and N = 1 on days 51-57),
and salivation (N = 3 on days 1-10, and N = 10 - 12 at all other intervals). A
few high dose animals (individual numbers were not stated) also exhibited
abnormal behavior such as grinding of teeth, head bobbing, body tremors,
involuntary movements, and pawing at the cage. A few of the animals
exposed to 120 ppm exhibited salivation (N = 3-8 at all intervals) and
hypoactivity (N = 3 at days 11-20). Signs of toxicity generally abated by the
morning after exposure. Alopecia was observed in animals (N = 1 control,
N = 2 low dose, N = 1 mid dose, N = 5 high dose) at one or more of their
weekly physical examinations. No unusual treatment-related signs were
observed in rats exposed to 60 ppm. The only remarkable finding at gross
necropsy was a small right testis in one animal exposed to 120 ppm.
Fertility: There was no effect of treatment on male fertility. Efficiency of
mating (34.4%, 30.6%, 29.8% and 27.1% in the control, low, mid and high
dose groups) and pregnancy rate (90.5%, 97.6%, 90.0% and 97.4% in the
respective groups) were comparable between groups. There was no
difference in the numbers of live implants (ranged from 12.7 - 13.9),
resorptions (ranged from 0.7 - 1.1), nidations (ranged from 13.8 - 14.9),
corpora lutea (ranged from 13.1 - 15.2), preimplantation loss (4-8%) and
postimplantation loss (5-10%).
Animals: Virgin female Sprague Dawley rats (28 days old upon receipt) and
Test condition :
virgin male Sprague Dawley rats (50 days old upon receipt) were
acclimated for one week and examined for general health and the
presence of pinworms and ectoparasites. Body weights of fifteen females
and ten males that were taken upon receipt were 155-181 g and 80-103 g,
respectively. No significant health problems were noted during the
acclimation period, and the animals were released for study. Males and
females were individually caged (except during mating). Food and water
were available ad libitum (except food was not available to males during
33 / 33
Id 78-82-0
5. Toxicity
Date 02.10.2003
exposure). Animals were housed in a room maintained at 72 +/- 2 degrees
F and 40-60% relative humidity, under a 12 hr light/dark cycle.
Exposures: Exposures (6 hr/day, 5 days/week) occurred in 10 m3
Rochester-style stainless steel and glass inhalation chambers. A
scheduled exposure day was cancelled due in clement weather. A new
exposure day (exposure day 41) was used in its place. Due to inclement
weather and building equipment failures, 2 exposures (days 33 and 43)
were only for 4 hours and one exposure (day 32) was for 5 hours.
Atmospheres of propionitrile vapor were generated using bubbler systems.
The concentrations of material in the chambers were controlled by either
adjusting the nitrogen flow though the propionitrile in the bubblers or by
changing the amount of test material in the bubblers. Concentrations of
test material in the chambers were measured 4 times daily (except during
day 43) using a Miran 1A General Purpose Gas Analyzer. The nominal
concentration of material, temperature and humidity also were measured
(at intervals that were not listed).
Study conduct: Fifteen males per group were assigned to be exposed to
target concentrations of 0, 60, 120 or 210 ppm test material. Exposure
began when males were 43 days old. Mating was initiated when males and
females were 16 and 12 weeks old, respectively. At this time, males had
been 69 days on the study (which was sufficient to cover the
spermatogenesis cycle of the rat), and had 46 days of exposure. Males
were randomly mated (1: 1) with three untreated females (consecutively)
that had been assigned to the corresponding treatment group (45 females
were assigned per group). Exposure of males continued until the day after
the last mating opportunity (57 exposure days). At night, after exposure,
males were caged with their assigned female until copulation was
confirmed (by presence of a copulatory plug or sperm in vaginal smear) or
5 nights without confirmed copulation. The day on which copulation was
confirmed was considered gestation day 0.
After assignment to the study, males and females were weighed once per
week. Mated females were weighed on gestation days 0 and 13. Males
were given a thorough physical examination once per week and were
observed for clinical signs of toxicity before and after exposures. All
animals were checked twice daily for mortality and gross abnormalities
(except for one day prior to mating when inclement weather permitted
observations).
One half of the males of each group were euthanized on each of the 2
consecutive days at the end of the study. They had not been exposed to
propionitrile for about 2 weeks prior to termination. Each male was given
an external examination and weighed. The tissues and organs of the
thoracic, scrotal and abdominal cavities were examined for gross lesions
and the testes, epididymides, prostate glands and seminal vesicles were
preserved in 10% neutral buffered formalin. Females that were not mated
with males were euthanized and were not examined.
Mated females were euthanized on gestation day 13 (or the nearest
workday up to gestation day 15). Females that were co-housed with males
without confirmed copulation were euthanized during the second week
after the last day of co-housing. Gross necropsies were performed on
females that had copulated and those that had not. The tissues and organs
of the thoracic and abdominal cavities were examined. Pregnancy status
was determined, nidation sites and were classified, and corpora lutea were
counted.
Statistical analyses: Body weight data were analyzed using Dunnett's test.
Mating and pregnancy rate data were analyzed by a Fisher's exact test and
an uncorrected chi-square test. Other data were analyzed by the Mann
34 / 34
Id 78-82-0
5. Toxicity
Date 02.10.2003
Whitney U test. The Bonferroni inequality was assumed when comparing
multiple treatments to control values for all tests except Dunnett's test. The
critical level for significance was p < 0.05.
Test material was propionitrile (CAS No. 107-12-0). Purity of the test
Test substance :
material was 96.1%. Impurities included acrylonitrile (0.1%), adiponitrile
(0.7%), p-nitrosodiphenylamine (0.08%) and water (< 0.1%). Analyses
indicated no significant decomposition of the test material over the course
of the study.
There was no effect of treatment on fertility of males.
Conclusion :
(1) valid without restriction
Reliability :
Study is comparable to a guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (24)
5.8.2 DEVELOPMENTAL TOXICITY/TERATOGENICITY
rat
Species :
female
Sex :
Sprague-Dawley
Strain :
inhalation
Route of admin. :
6 hours/day
Exposure period :
Days 6 to 20 of gestation
Frequency of treatm. :
to Gestation Day 21
Duration of test :
50, 100, 200, and 300 ppm
Doses :
yes
Control group :
= 100 ppm
NOAEL maternal tox. :
= 300 ppm
NOAEL teratogen. :
= 100 ppm
NOAEL Fetotoxicity :
= 200 ppm
NOAEL Embryotoxicity :
isobutyronitrile was not teratogenic
Result :
other: similar to OECD 414
Method :
1992
Year :
no data
GLP :
as prescribed by 1.1 - 1.4
Test substance :
Four different concentrations of acetonitrile, priopionitrile, n-butyronitrile,
Remark :
acrylonitrile, allylnitrile, methacrylonitrile and 2-chloroacrylonitrile
also were tested in this study.
The methodology described in the manuscript is essentially identical to that
under OECD TG-414. However, information is lacking in the report in
regard to food intake. Therefore, it is unknown if reduced fetal body
weights were a consequence of reduced material feed intake. In addition,
it is unknown as to whether the study was conducted under GLP
assurances. However, based on the date in which this study was
completed it is likely to have been a GLP study. All other parameters noted
in the guideline appear in the manuscript.
The NOAEL for fetoxicity is 100 ppm, since a reduction in fetal body weight
was observed in females from the 200 ppm group. The NOAEL for
embryotoxicity is 200 ppm, since 300 ppm caused an increase in
embryolethality.
Maternal: One out of 21 animals exposed to 200 ppm and 3/21 exposed to
Result :
300 ppm died before study termination. Maternal weight gains were similar
in all groups. Indices of pregnancy were comparable among groups. There
was no significant effect of treatment on the mean numbers of
implantations. The indices of nonsurviving implants and embryonic
resorptions (per litter) in rats treated with 300 ppm (11.07 +/- 9.29 per litter
for both indices) were significantly higher than control (2.02 +/- 3.63 per
litter for both indices).
35 / 35
Id 78-82-0
5. Toxicity
Date 02.10.2003
Fetal: There was no effect of treatment on the mean numbers of live
fetuses or sex ratio. There was a concentration- related decrease in fetal
body weights, with weights of females in the 200 ppm (5.11 +/- 0.41 g per
litter) and males and females in 300 ppm groups (4.96 +/- 0.37 and 4.68
+/- 0.34 g per litter, respectively) being significantly less than control (5.75
+/- 0.29 g per litter for males and 5.56 +/- 0.42 g per litter for females). A
single case of unilateral hydronephrosis was observed in one fetus from
the 300 ppm group. The incidences of visceral and skeletal variations in
treated fetuses were not significantly different from controls.
Animals: Male (350 g) and primiparous female (200-220 g) were
Test condition :
acclimated for 1-2 weeks prior to breeding. Females were then placed with
males (one male: 3 females) overnight and examined by vaginal smear for
the presence of sperm the following morning. Sperm-positive females were
considered to be at Day 0 of gestation. These animals were randomly
assigned to groups of 20-23 rats each.
Exposure conditions: Exposures were conducted in 200 璴iter stainless
steel inhalation chambers at an air flow of 10-20 m3/hr. Chambers were
maintained at a negative pressure of < = 3 mm water. Chamber
temperatures and humidities were 23 +/- 2 degrees and 50 +/- 5%,
respectively. Vapor was generated by bubbling an additional air flow
through a flask containing test material. The vapor was mixed with filtered
room air to achieve the desired concentration. Analytical concentrations
were determined by analyzing the atmosphere once/hour (by gas-liquid
chromatography) during each 6 hour exposure. The nominal
concentrations of isobutyronitrile were 50, 100, 200 and 300 ppm.
Corresponding analytical concentrations were 54 +\- 3.9, 98 +/- 10.0, 208
+/- 12.4, and 308 +/- 18.6 ppm.
Test conduct: Animals were exposed 6 hours/day on Days 6 through 20 of
gestation. Control animals were exposed concurrently to filtered room air in
an adjacent chamber with flow characteristics identical to those of the
treated groups. Food and water were available ad libitum (except
during exposure). All rats were observed daily and maternal body weights
were recorded on Days 0, 6 and 21 of gestation. Females were euthanized
on Day 21 of gestation and the uterus was removed and weighed. The
uterus horns were then opened and the numbers of implantation and
absorption sites and live and dead fetuses were recorded. Live fetuses
were removed and weighed, examined for external anomalies (including
those of the oral cavity) and sexed. The numbers of fetuses (and litters)
examined for external anomalies in the 0, 50, 100, 200 and 300 ppm
groups were 191 (16), 235 (19), 222 (19), 185 (14) and 193 (15),
respectively. Half of the fetuses from each litter were fixed in Bouin's
solution and examined microscopically for visceral abnormalities. The
remaining half were fixed in 70% ethanol, eviscerated, macerated in 1%
KOH , stained in alizarin red S, and examined microscopically for skeletal
anomalies.
Statistical analysis: Depending on the parameter evaluated: one-way
analysis of variance followed by Dunnett's test, Wilcoxon test after arc-sine-
square root transformation, Fisher's test, or least squares analysis. The
litter was used as the basis for analysis of fetal variables.
Purity was > 99%.
Test substance :
It was concluded that isobutyronitrile was not teratogenic. While evidence
Conclusion :
of embryolethality and fetotoxicity were noted, these effects occurred at
levels that also induced maternal toxicity (300 ppm).
(1) valid without restriction
Reliability :
This was a well-documented OECD-like guideline study.
Critical study for SIDS endpoint
Flag :
07.08.2003 (26)
36 / 36
Id 78-82-0
5. Toxicity
Date 02.10.2003
5.8.3 TOXICITY TO REPRODUCTION, OTHER STUDIES
5.9 SPECIFIC INVESTIGATIONS
5.10 EXPOSURE EXPERIENCE
5.11 ADDITIONAL REMARKS
37 / 37
Id 78-82-0
6. Analyt. Meth. for Detection and Identification
Date 02.10.2003
6.1 ANALYTICAL METHODS
6.2 DETECTION AND IDENTIFICATION
38 / 38
Id 78-82-0
7. Eff. Against Target Org. and Intended Uses
Date 02.10.2003
7.1 FUNCTION
7.2 EFFECTS ON ORGANISMS TO BE CONTROLLED
7.3 ORGANISMS TO BE PROTECTED
7.4 USER
7.5 RESISTANCE
39 / 39
Id 78-82-0
8. Meas. Nec. to Prot. Man, Animals, Environment
Date 02.10.2003
8.1 METHODS HANDLING AND STORING
8.2 FIRE GUIDANCE
8.3 EMERGENCY MEASURES
8.4 POSSIB. OF RENDERING SUBST. HARMLESS
8.5 WASTE MANAGEMENT
8.6 SIDE-EFFECTS DETECTION
8.7 SUBSTANCE REGISTERED AS DANGEROUS FOR GROUND WATER
8.8 REACTIVITY TOWARDS CONTAINER MATERIAL
40 / 40
Id 78-82-0
9. References
Date 02.10.2003
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(3) Covance Laboratories Inc. Mutagenicity Test with EC98-0256 IBN in the Salmonella-
Escherichia coli/mammalian-microsome reverse mutation assay with a confirmatory assay
(unpublished study). Covance study number 20878-0-409R, December 8, 1999.
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(6) Eastman Kodak Company, Chemical Quality Services Division, Report No. 215514K TX-
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(unpublished study). Study No. EN-430-907253-A, October 29, 1998.
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(13) Eastman Kodak Company, Laboratory of Industrial Medicine. Unpublished Study,
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(14) Eastman Kodak Company, Laboratory of Industrial Medicine. Unpublished Study,
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41 / 41
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9. References
Date 02.10.2003
(18) EPIWIN ECOSAR Program (v0.99).
(19) EPIWIN Hydrowin Program (v1.67).
(20) EPIWIN Kowwin Program (v1.66).
(21) EPIWIN Level III fugacity model.
(22) EPIWIN Wskow Program (v1.40).
(23) Kier LD. 1984. Female fertility study of Sprague-Dawley rats exposed by the inhalation
route to propionitrile. Unpublished Monsanto Report No MSL-4438, dated December 31,
1984.
(24) Kier LD. 1984. Male fertility study of Sprague-Dawley rats exposed by the inhalation route
to propionitrile. Unpublished Monsanto Report No MSL-4422, dated December 17, 1984.
(25) Kitano M. 1978. Biodegradation and bioaccumulation test on chemical substances. OECD
Tokyo Meeting. Reference Book TSU-No 3.
(26) Saillenfait AM, Bonnet P, Guenier JP, and DeCeaurriz J. Relative Developmental
Toxicities of Inhaled Aliphatic Mononitriles in Rats. Fund Appl Toxicol 20: 365-375, 1993.
(27) Sangster J. 1989. Octanol-water partition coefficients of simple organic compounds. J Phys
Chem Ref Data 18:1111-1230.
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Id 78-82-0
10. Summary and Evaluation
Date 02.10.2003
10.1 END POINT SUMMARY
10.2 HAZARD SUMMARY
10.3 RISK ASSESSMENT
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