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EC number: 209-060-4 | CAS number: 554-12-1
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
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- Nanomaterial pour density
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- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
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- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 4 November 2003 to 9 December 2003
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 003
- Report date:
- 2003
Materials and methods
Test guidelineopen allclose all
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Methyl propionate
- EC Number:
- 209-060-4
- EC Name:
- Methyl propionate
- Cas Number:
- 554-12-1
- Molecular formula:
- C4H8O2
- IUPAC Name:
- methyl propanoate
- Test material form:
- liquid
- Details on test material:
- - Appearance: colourless liquid
- Storage conditions: room temperature, in the dark
Constituent 1
Method
- Target gene:
- - Histidine requirement in the Salmonella typhimurium strains (Histidine operon).
- Tryptophan requirement in the Escherichia coli strain (Tryptophan operon).
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix
- Test concentrations with justification for top dose:
- - Toxicity Test
A toxicity test, using strain TA 100 only, was performed to establish suitable dose levels for the mutation tests. One plate of each of the following concentrations of test material was prepared:
17, 50, 167, 500, 1667 and 5000 μg per plate (with and without S9 mix)
- Mutation Tests
The dose levels for the mutation assays, based on the results of the toxicity test, were:
17, 50, 167, 500, 1667 and 5000 μg per plate (with and without S9 mix) - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Sterile ultra-pure water
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Sterile ultra-pure water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- 2-nitrofluorene
- sodium azide
- N-ethyl-N-nitro-N-nitrosoguanidine
- other: 2-aminoanthracene
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation) and pre-incubation
EXPERIMENT 1- PLATE INCORPORATION METHOD
Volumes of soft agar (2 mL) were dispensed into small sterile tubes. To this, 0.5 mL of S9 mix or 0.05 M phosphate buffer, pH 7.4 were added, followed by 0.1 mL of bacteria. The solvent or test solution (0.1 mL) was added last. The tube contents, which were continually cooling, were mixed and poured onto minimal medium plates, prepared in-house. These plates contained 20 mL of 1.5% purified agar, in Vogel-Bonner Medium E (Vogel et al (1956)) with 2% glucose. When the soft agar had set, the plates were inverted and incubated at 37°C for 2 or 3 days. All testing for this experiment was performed in triplicate.
EXPERIMENT 2- PRE-INCUBATION METHOD
Volumes of S9 mix or 0.05 M phosphate buffer, pH 7.4 (0.5 mL) were dispensed into small sterile tubes. This was followed by 0.1 ml of bacteria per tube and, finally, the solvent or test solution (0.1 mL per tube). The tube tops were then screwed on tightly and the tubes placed in a shaking incubator at 37°C for 20 min. After this, the tube tops were removed and 2 mL of soft agar added to each tube. The tube contents, which were continually cooling, were mixed and then poured onto agar plates, as above. When the soft agar had set, the plates were inverted and incubated at 37°C for 2 or 3 days. All testing for this experiment was performed in triplicate.
INCUBATION AND SCORING
After incubation, the colonies were counted using a Cardinal Colony Counter set at maximum sensitivity, i.e. colonies of 0.1 mm or more in diameter were counted. The data was captured electronically using a validated software system. The plates were also examined for precipitates and microscopically, for microcolony growth.
NUMBER OF REPLICATIONS: 3
ACCEPTABILITY CRITERIA
A test was considered acceptable if, for each strain:
i) the bacteria demonstrated their typical responses to crystal violet, ampicillin and u.v. light.
ii) at least 2 of the vehicle control plates were within the following ranges: TA 1535, 4-30; TA 1537, 1-20; TA 98, 10-60; TA 100, 60-200 and E. coli WP2uvrA 1-60.
iii) on at least 2 of the positive control plates there were at least x 2 the mean vehicle control mutant numbers per plate, or in the case of TA 100, at least x 1.5 the mean vehicle control mutant numbers per plate.
iv) no toxicity or contamination was observed in at least 4 dose levels.
v) in cases where a mutagenic response was observed, no more than one dose level was discarded before the dose that gave the highest significant mean colony number. - Evaluation criteria:
- Where the acceptibility criteria were met, a significant mutagenic response was recorded if there was:
i) for S. typhimurium strains TA 1535, TA 1537, and TA 98 and for E. coli, at least a doubling of the mean concurrent vehicle control values at some concentration of the test material. For S. typhimurium strain TA 100, a 1.5-fold increase over the control value was considered significant. If the mean colony count on the vehicle control plates was less than 10, then a value of 10 was assumed for assessment purposes. In such cases, a minimum count of 20 was required before a significant mutagenic response was registered.
ii) a dose related response, although at high dose levels this relationship could be inverted because of, for example, (1) toxicity to the bacteria generally, (2) specific toxicity to the mutants and (3) inhibition of foreign compound metabolising enzymes where mutagens require metabolic activation by the liver.
iii) a reproducible effect in independent tests.
Results and discussion
Test results
- Key result
- Species / strain:
- other: S. typhimurium TA1535, TA1537, TA98, TA100; E. coli WP2uvrA
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- > Toxicity Test
No toxicity to the bacteria was observed and no precipitation of the test material occurred in either the presence or the absence of S9 mix.
> Mutation Tests
- Vehicle Control
The vehicle control values were generally within the normal ranges experienced at the Test Facility and reported in the literature with these strains of S. typhimurium and E. coli, (Ames et al, 1975; Gatehouse et al, 1994). The exception was TA 98 in the first mutation assay, in the absence of S9 mix. On this occasion the mean vehicle control counts for TA 98 were lower than the acceptance criterion (see test rejection)
- Positive Control
The results obtained in the positive control groups were generally within the normal ranges expected for each bacterial strain and activation condition. In the first mutation assay retest (see test rejection), a poor response was obtained for TA 1537 with 2-aminoanthracene. Poor responses were also obtained in the second mutation assay for all strains with 2-aminoanthracene. These parts of both assays were repeated successfully.
- Test material
The test material did not induce mutagenic activity in any of the 5 bacterial strains used, in either activation condition.
There was no toxicity to the bacteria and no precipitation of the test item was observed in either mutation assay, in either the presence or the absence of S9 mix.
- Test Rejection
The first mutation assay in the presence of S9 mix was rejected due to an operator error in the preparation of the S9 mix. In the retest, TA 1537 was rejected due to the poor response obtained with 2-aminoanthracene. In the absence of S9 mix, TA 98 was repeated due to a dosing error at 17 μg per plate and also the vehicle control counts were outwith the acceptance criterion. WP2uvrA was also repeated due to a dosing error at 500 μg per plate.
In the second mutation assay, all strains were repeated in the presence of S9 mix due to poor responses obtained with 2-aminoanthracene. Also in this test TA 100 was rejected in the absence of S9 mix due to a dosing error at 500 μg per plate. These parts of the tests were repeated successfully.
Any other information on results incl. tables
Table 1: Summary of Results from Experiment 1
Mean Number of Revertant Colonies Per Plate in the Absence of S9 Mix |
||||||
Test item |
Dose level (µg per plate) |
TA1535 |
TA1537 |
TA98* |
TA100 |
WP2uvrA* |
Water |
100 µL |
22 |
5 |
22 |
153 |
36 |
Test material |
17 |
22 |
5 |
21 |
141 |
43 |
50 |
25 |
7 |
16 |
150 |
44 |
|
167 |
23 |
7 |
21 |
131 |
40 |
|
500 |
20 |
5 |
15 |
145 |
41 |
|
1667 |
26 |
8 |
18 |
141 |
36 |
|
5000 |
22 |
8 |
13 |
131 |
41 |
|
Positive controls |
Compound |
NaN3 |
9AA |
2NF |
NaN3 |
ENNG |
Dose level (µg per plate) |
1 |
80 |
1 |
1 |
2 |
|
Mean |
412 |
4411 |
1310 |
1014 |
270 |
|
Mean Number of Revertant Colonies Per Plate in the Presence of S9 Mix |
||||||
Test item |
Dose level (µg per plate) |
TA1535 |
TA1537* |
TA98 |
TA100 |
WP2uvrA |
Water |
100 µL |
21 |
17 |
39 |
114 |
7 |
Test material |
17 |
29 |
15 |
32 |
124 |
8 |
50 |
17 |
18 |
38 |
125 |
14 |
|
167 |
26 |
15 |
36 |
114 |
11 |
|
500 |
27 |
22 |
31 |
124 |
10 |
|
1667 |
31 |
17 |
35 |
105 |
14 |
|
5000 |
31 |
15 |
36 |
128 |
12 |
|
Positive controls |
Compound |
2AAN |
2AAN |
2AAN |
2AAN |
2AAN |
Dose level (µg per plate) |
2 |
2 |
0.5 |
0.5 |
20 |
|
Mean |
602 |
526 |
2721 |
4449 |
246 |
* Results from re-tests
NaN3: sodium azide
9AA: 9-aminoacradene
2NF: 2-nitrofluorene
ENNG: N-ethyl-N-nitro-N-nitrosoguanidine
2AAN: 2-aminoanthracene
Table 2: Summary of results from Experiment 2
Mean Number of Revertant Colonies Per Plate in the Absence of S9 Mix |
||||||
Test item |
Dose level (µg per plate) |
TA1535 |
TA1537 |
TA98 |
TA100* |
WP2uvrA |
Water |
100 µL |
11 |
10 |
22 |
86 |
8 |
Test material |
17 |
14 |
9 |
17 |
97 |
13 |
50 |
17 |
12 |
21 |
89 |
6 |
|
167 |
10 |
8 |
18 |
89 |
8 |
|
500 |
11 |
7 |
19 |
86 |
9 |
|
1667 |
10 |
10 |
20 |
80 |
7 |
|
5000 |
12 |
9 |
20 |
85 |
7 |
|
Positive controls |
Compound |
NaN3 |
9AA |
2NF |
NaN3 |
ENNG |
Dose level (µg per plate) |
1 |
80 |
1 |
1 |
2 |
|
Mean |
283 |
3038 |
1610 |
1014 |
387 |
|
Mean Number of Revertant Colonies Per Plate in the Presence of S9 Mix |
||||||
Test item |
Dose level (µg per plate) |
TA1535 |
TA1537 |
TA98 |
TA100 |
WP2uvrA |
Water |
100 µL |
15 |
14 |
21 |
76 |
7 |
Test material |
17 |
13 |
12 |
26 |
79 |
6 |
50 |
11 |
13 |
22 |
75 |
9 |
|
167 |
15 |
7 |
24 |
72 |
13 |
|
500 |
14 |
11 |
22 |
76 |
8 |
|
1667 |
11 |
14 |
20 |
74 |
7 |
|
5000 |
9 |
5 |
18 |
74 |
7 |
|
Positive controls |
Compound |
2AAN |
2AAN |
2AAN |
2AAN |
2AAN |
Dose level (µg per plate) |
2 |
2 |
0.5 |
0.5 |
20 |
|
Mean |
65 |
36 |
79 |
190 |
11 |
* Results from re-test
NaN3: sodium azide
9AA: 9-aminoacradene
2NF: 2-nitrofluorene
ENNG: N-ethyl-N-nitro-N-nitrosoguanidine
2AAN: 2-aminoanthracene
Applicant's summary and conclusion
- Conclusions:
- It was concluded that the test material was not mutagenic to Salmonella typhimurium or Escherichia coli when tested in sterile, ultra-pure water up to a pre-determined maximum concentration of 5000 μg per plate.
- Executive summary:
The mutagenic potential of the test material was investigated in a study which was conducted in accordance with the standardised guidelines OECD 471 and EU Method B.13/14, under GLP conditions.
During the study the test material was tested for mutagenic activity in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 and Escherichia coli WP2uvrA at concentrations ranging from 17 to 5000 μg per plate. The solvent used was sterile, ultra-pure water.
Two independent tests (one direct plate and one pre-incubation) were conducted on agar plates in the presence and absence of an Aroclor-1254 induced rat liver preparation and the co-factors required for mixed-function oxidase activity (S9 mix). The first mutation assay was repeated in the presence of S9 mix only due to an operator error.
Concurrent positive controls demonstrated the sensitivity of the assay and the metabolising activity of the S9 mix.
No mutagenic activity was observed in any of the 5 bacterial strains, in either activation condition.
There was no toxicity to the bacteria and no precipitation of the test material was observed in either mutation assay, in either the presence or the absence of S9 mix.
It was concluded that the test material was not mutagenic to Salmonella typhimurium or Escherichia coli when tested in sterile, ultra-pure water up to a pre-determined maximum concentration of 5000 μg per plate.
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