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EC number: - | CAS number: -
- 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
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- 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
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 05 May 2008 to 29 May 2008.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 008
- Report date:
- 2008
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
- Qualifier:
- according to guideline
- Guideline:
- JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Remarks:
- Date of Signature: 15/10/2007; Date of Inspection: 21/08/2007
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Reaction mass of disodium N-(1-oxooctadecyl)-L-glutamate and disodium N-(1-oxohexadecyl)-L-glutamate
- EC Number:
- 700-014-3
- Molecular formula:
- See remarks
- IUPAC Name:
- Reaction mass of disodium N-(1-oxooctadecyl)-L-glutamate and disodium N-(1-oxohexadecyl)-L-glutamate
Constituent 1
Method
- Target gene:
- Histidine operon for Salmonella triphimurium
Tryptophan operon for Escherichia coli
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not applicable
- Species / strain / cell type:
- E. coli WP2 uvr A
- Details on mammalian cell type (if applicable):
- Not applicable
- Additional strain / cell type characteristics:
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- phenobarbitone/betanaphthoflavone
- Test concentrations with justification for top dose:
- Preliminary toxicity test:
0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate
Experiment 1 (Range finding test)
Salmonella strains, with and without S9: 5, 15, 50, 150, 500, 1500, 5000 µg/plate.
E. coli strain WP2uvrA-, with and without S9: 50, 150, 500, 1500, 5000 µg/plate.
Experiment 2 (Main test)
Salmonella strains, with and without S9: 5, 15, 50, 150, 500, 1500, 5000 µg/plate.
E. coli strain WP2uvrA-, with and without S9: 50, 150, 500, 1500, 5000 µg/plate. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: sterile distilled water
- Justification for choice of solvent/vehicle: The test material was fully soluble in sterile distilled water at 12.5 mg/mL in solubility checks performed in-house. Sterile distilled water was therefore selected as the vehicle.
Controlsopen allclose all
- Untreated negative controls:
- yes
- Remarks:
- Spontaneous mutation rates
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Sterile distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: N-ethyl-N'-Nitrosoguanidine 2 µg/plate for WP2uvrA-, 3 µg/plate for TA100 and 5µg/plate for TA1535.
- Remarks:
- Without S9
- Untreated negative controls:
- yes
- Remarks:
- Spontaneous mutation rates
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Sterile distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- Remarks:
- Without S9 Migrated to IUCLID6: 80 µg/plate for TA1537
- Untreated negative controls:
- yes
- Remarks:
- Spontaneous mutation rates
- Negative solvent / vehicle controls:
- yes
- Remarks:
- sterile distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitroquinoline-1-oxide 0.2 µg/plate for TA98
- Remarks:
- Without S9
- Untreated negative controls:
- yes
- Remarks:
- Spontaneous mutation rates
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Sterile distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2 Aminoanthracene (2AA) at 1 µg/plate for TA100, 2 µg/plate for TA1535 and TA1537 and 10 µg/plate for WP2uvrA-
- Remarks:
- With S9
- Untreated negative controls:
- yes
- Remarks:
- Spontaneous mutation rates
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Sterile distilled water
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- Remarks:
- With S9 Migrated to IUCLID6: 5 µg/plate for TA98
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 to 72 hours
- Fixation time (start of exposure up to fixation or harvest of cells): 48 to 72 hours
NUMBER OF REPLICATIONS: triplicate plating
DETERMINATION OF CYTOTOXICITY
- Method: lawn deficiency and colony reduction - Evaluation criteria:
- Acceptance Criteria
The reverse mutation assay may be considered valid if the following criteria are met:
All tester strain cultures exhibit a characteristic number of spontaneous revertants per plate in the vehicle and untreated controls. Acceptable ranges are presented in the standard test method with historical control ranges from 2006 and 2007 were used.
The appropriate characteristics for each tester strain have been confirmed, e.g. rfa cell-wall mutation and pKM101 plasmid R-factor etc.
All tester strain cultures should be in the approximate range of 1 to 9.9 x 10^9 bacteria per mL.
Each mean positive control value should be at least twice the respective vehicle control value for each strain, thus demonstrating both the intrinsic sensitivity of the tester strains to mutagenic exposure and the integrity of the S9-mix. The positive control historical ranges were used from 2006 and 2007.
There should be a minimum of four non-toxic test material dose levels.
There should not be an excessive loss of plates due to contamination.
Evaluation Criteria
There are several criteria for determining a positive result, such as a dose-related increase in revertant frequency over the dose range tested and/or a reproducible increase at one or more concentrations in at least one bacterial strain with or without metabolic activation. Biological relevance of the results will be considered first, statistical methods, as recommended by the UKEMS (5) can also be used as an aid to evaluation, however, statistical significance will not be the only determining factor for a positive response.
A test material will be considered non-mutagenic (negative) in the test system if the above criteria are not met.
Although most experiments will give clear positive or negative results, in some instances the data generated will prohibit a definitive judgement about the test material activity. Results of this type will be reported as equivocal. - Statistics:
- Standard deviation
Results and discussion
Test resultsopen allclose all
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- 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:
- valid
- Positive controls validity:
- valid
- Species / strain:
- E. coli WP2 uvr A
- 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:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none reported
- Effects of osmolality: none reported
- Evaporation from medium: none reported
- Water solubility: The test material was fully soluble in sterile distilled water at 12.5 mg/mL in solubility checks performed in-house.
- Precipitation: A pale flakey precipitate was observed at and above 1500 µg/plate, this did not prevent the scoring of revertant colonies.
- Other confounding effects: none
RANGE-FINDING/SCREENING STUDIES:
Preliminary toxicity test:
The test material caused a reduction in the frequency of TA100 revertant colonies and/or a weakening of the background lawn, initially from 500 µg/plate. Substantial reductions in the frequency of WP2uvrA- revertant colonies were also noted at 5000 µg/plate. The test material formulation and S9-mix used in this experiment were both shown to be sterile.
COMPARISON WITH HISTORICAL CONTROL DATA:
Prior to use, the master strains were checked for characteristics, viability and spontaneous reversion rate (all were found to be satisfactory). These data are not given in the report. The amino acid supplemented top agar and the S9-mix used in both experiments was shown to be sterile.
ADDITIONAL INFORMATION ON CYTOTOXICITY:
None
CONTROLS
The vehicle (sterile distilled water) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated.
The test material caused a visible reduction in the growth of the bacterial background lawn and/or a substantial decrease in the frequency of revertant colonies for all of the Salmonella strains in both the presence and absence of S9, initially from 500 µg/plate. Small decreases in revertant colony frequency were also noted for E.coli strain WP2uvrA- at 5000 µg/plate in both the presence and absence of S9. The sensitivity of the tester strains to the toxicity of the test material varied slightly between strain type, exposure with or without S9 and experiment number.
A pale flakey precipitate was observed at and above 1500 µg/plate, this did not prevent the scoring of revertant colonies.
No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation. - Remarks on result:
- other: all strains/cell types tested
- Remarks:
- Migrated from field 'Test system'.
Any other information on results incl. tables
The test material caused a visible reduction in the growth of the bacterial background lawn and/or a substantial decrease in the frequency of revertant colonies for all of the Salmonella strains in both the presence and absence of S9, initially from 500 µg/plate. Small decreases in revertant colony frequency were also noted for E .coli strain WP2uvrA- at 5000 µg/plate in both the presence and absence of S9. The sensitivity of the tester strains to the toxicity of the test material varied slightly between strain type, exposure with or without S9 and experiment number.
A pale flakey precipitate was observed at and above 1500 µg/plate, this did not prevent the scoring of revertant colonies.
No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.
All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies thus confirming the activity of the S9-mix and the sensitivity of the bacterial strains.
Table1: Spontaneous Mutation Rates (Concurrent Negative Controls)
Range-finding Test
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
|||||
84 |
|
14 |
|
29 |
|
19 |
|
7 |
|
95 |
(94) |
18 |
(16) |
22 |
(26) |
20 |
(19) |
5 |
(6) |
102 |
|
16 |
|
26 |
|
19 |
|
7 |
|
Main Test
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
TA100 |
TA1535 |
WP2uvrA- |
TA98 |
TA1537 |
|||||
137 |
|
21 |
|
34 |
|
15 |
|
12 |
|
140 |
(136) |
18 |
(19) |
26 |
(35) |
8 |
(11) |
11 |
(12) |
131 |
|
19 |
|
45 |
|
11 |
|
13 |
|
Table2: Test Results: Range-Finding Test– Without Metabolic Activation
Test Period |
From: 19 May 2008 |
To: 22 May 2008 |
||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||||||||
Base-pair substitution type |
Frameshift type |
|||||||||||
TA100 |
TA1535 |
WP2uvrA‑ |
TA98 |
TA1537 |
||||||||
- |
0 |
108 91 97 |
(99) 8.6# |
12 14 13 |
(13) 1.0 |
37 20 15 |
(24) 11.5 |
12 11 16 |
(13) 2.6 |
3 5 8 |
(5) 2.5 |
|
- |
5 |
86 106 92 |
(95) 10.3 |
13 15 13 |
(14) 1.2 |
N/T |
11 11 12 |
(11) 0.6 |
4 5 4 |
(4) 0.6 |
||
- |
15 |
97 90 111 |
(99) 10.7 |
11 9 18 |
(13) 4.7 |
N/T |
15 18 15 |
(16) 1.7 |
3 7 8 |
(6) 2.6 |
||
- |
50 |
118 112 88 |
(106) 15.9 |
19 10 10 |
(13) 5.2 |
14 32 19 |
(22) 9.3 |
16 15 14 |
(15) 1.0 |
5 2 3 |
(3) 1.5 |
|
- |
150 |
87 106 89 |
(94) 10.4 |
7 7 11 |
(8) 2.3 |
26 24 15 |
(22) 5.9 |
13 11 10 |
(11) 1.5 |
1 5 4 |
(3) 2.1 |
|
- |
500 |
38 41 41 |
(40) 1.7 |
2 10 2 |
(5) 4.6 |
20 22 15 |
(19) 3.6 |
8 2 2 |
(4) 3.5 |
1 1 5 |
(2) 2.3 |
|
- |
1500 |
10 P* 9 P* 14 P* |
(11) 2.6 |
1 P* 1 P* 1 P* |
(1) 0.0 |
15 P 13 P 13 P |
(14) 1.2 |
5 P 8 P 0 P |
(4) 4.0 |
0 P 0 P 1 P |
(0) 0.6 |
|
- |
5000 |
0 P* 0 P* 0 P* |
(0) 0.0 |
0 P* 0 P* 0 P* |
(0) 0.0 |
9 P 5 P 10 P |
(8) 2.6 |
0 P* 0 P* 0 P* |
(0) 0.0 |
0 P* 0 P* 0 P* |
(0) 0.0 |
|
Positive controls
S9-Mix
- |
Name Concentration (μg/plate) No. colonies per plate |
ENNG |
ENNG |
ENNG |
4NQO |
9AA |
||||||
3 |
5 |
2 |
0.2 |
80 |
||||||||
682 696 685 |
(688) 7.4 |
354 375 305 |
(345) 35.9 |
881 841 654 |
(792) 121.2 |
588 129 155 |
(291) 257.8 |
1168 1144 1107 |
(1140) 30.7 |
ENNG = N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO = 4-Nitroquinoline-1-oxide
9AA = 9-Aminoacridine
N/T = Not tested at this dose level
P = Precipitate
* = Partial or complete absence of bacterial background lawn
# = Standard deviation
Table3: Test Results: Range-Finding Test– With Metabolic Activation
Test Period |
From: 19 May 2008 |
To: 22 May 2008 |
|||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
|||||||||||
Base-pair substitution type |
Frameshift type |
||||||||||||
TA100 |
TA1535 |
WP2uvrA‑ |
TA98 |
TA1537 |
|||||||||
+ |
0 |
92 97 77 |
(89) 10.4# |
15 15 12 |
(14) 1.7 |
22 29 18 |
(23) 5.6 |
26 30 32 |
(29) 3.1 |
7 7 7 |
(7) 0.0 |
||
+ |
5 |
71 90 90 |
(84) 11.0 |
9 7 8 |
(8) 1.0 |
N/T |
31 22 24 |
(26) 4.7 |
13 9 3 |
(8) 5.0 |
|||
+ |
15 |
75 99 96 |
(90) 13.1 |
3 9 10 |
(7) 3.8 |
N/T |
16 21 15 |
(17) 3.2 |
10 4 4 |
(6) 3.5 |
|||
+ |
50 |
80 81 77 |
(79) 2.1 |
8 4 8 |
(7) 2.3 |
31 21 22 |
(25) 5.5 |
21 23 12 |
(19) 5.9 |
7 7 5 |
(6) 1.2 |
||
+ |
150 |
80 95 80 |
(85) 8.7 |
9 7 2 |
(6) 3.6 |
21 22 14 |
(19) 4.4 |
20 14 19 |
(18) 3.2 |
9 8 3 |
(7) 3.2 |
||
+ |
500 |
97 89 75 |
(87) 11.1 |
3 2 4 |
(3) 1.0 |
18 14 18 |
(17) 2.3 |
16 27 16 |
(20) 6.4 |
4 3 4 |
(4) 0.6 |
||
+ |
1500 |
26 P 30 P 43 P |
(33) 8.9 |
4 P* 2 P* 4 P* |
(3) 1.2 |
14 P 12 P 7 P |
(11) 3.6 |
8 P 13 P 10 P |
(10) 2.5 |
8 P 4 P 4 P |
(5) 2.3 |
||
+ |
5000 |
0 P* 0 P* 0 P* |
(0) 0.0 |
0 P* 0 P* 0 P* |
(0) 0.0 |
4 P 16 P 14 P |
(11) 6.4 |
0 P* 0 P* 0 P* |
(0) 0.0 |
0 P* 0 P* 0 P* |
(0) 0.0 |
||
Positive controls
S9-Mix
+ |
Name Concentration (μg/plate) No. colonies per plate |
2AA |
2AA |
2AA |
BP |
2AA |
|||||||
1 |
2 |
10 |
5 |
2 |
|||||||||
2094 1811 1780 |
(1895) 173.0 |
310 198 288 |
(265) 59.3 |
508 475 435 |
(473) 36.6 |
216 222 224 |
(221) 4.2 |
275 365 197 |
(279) 84.1 |
||||
BP = Benzo(a)pyrene
2AA = 2-Aminoanthracene
N/T = Not tested at this dose level
P = Precipitate
* = Partial absence of bacterial background lawn
# = Standard deviation
Table4: Test Results: Main Test– Without Metabolic Activation
Test Period |
From: 26 May 2008 |
To: 29 May 2008 |
|||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
|||||||||||
Base-pair substitution type |
Frameshift type |
||||||||||||
TA100 |
TA1535 |
WP2uvrA‑ |
TA98 |
TA1537 |
|||||||||
- |
0 |
135 172 134 |
(147) 21.7# |
20 20 22 |
(21) 1.2 |
33 31 31 |
(32) 1.2 |
12 21 13 |
(15) 4.9 |
9 22 11 |
(14) 7.0 |
||
- |
5 |
153 142 123 |
(139) 15.2 |
16 20 15 |
(17) 2.6 |
N/T |
19 10 14 |
(14) 4.5 |
9 16 13 |
(13) 3.5 |
|||
- |
15 |
129 112 140 |
(127) 14.1 |
25 20 15 |
(20) 5.0 |
N/T |
11 14 10 |
(12) 2.1 |
14 14 7 |
(12) 4.0 |
|||
- |
50 |
121 136 119 |
(125) 9.3 |
25 18 24 |
(22) 3.8 |
31 36 20 |
(29) 8.2 |
19 12 12 |
(14) 4.0 |
15 9 9 |
(11) 3.5 |
||
- |
150 |
119 111 146 |
(125) 18.3 |
20 10 18 |
(16) 5.3 |
27 29 32 |
(29) 2.5 |
9 19 15 |
(14) 5.0 |
11 11 9 |
(10) 1.2 |
||
- |
500 |
56 74 68 |
(66) 9.2 |
2 3 7 |
(4) 2.6 |
38 48 36 |
(41) 6.4 |
13 9 9 |
(10) 2.3 |
0 0 0 |
(0) 0.0 |
||
- |
1500 |
27*P 18*P 25*P |
(23) 4.7 |
0*P 0*P 0*P |
(0) 0.0 |
23 P 25 P 30 P |
(26) 3.6 |
7 P 10 P 2 P |
(6) 4.0 |
0*P 0*P 0*P |
(0) 0.0 |
||
- |
5000 |
0*P 0*P 0*P |
(0) 0.0 |
0*P 0*P 0*P |
(0) 0.0 |
25P 25P 13P |
(21) 6.9 |
0*P 0*P 0*P |
(0) 0.0 |
0*P 0*P 0*P |
(0) 0.0 |
||
Positive controls
S9-Mix
- |
Name Concentration (μg/plate) No. colonies per plate |
ENNG |
ENNG |
ENNG |
4NQO |
9AA |
|||||||
3 |
5 |
2 |
0.2 |
80 |
|||||||||
472 498 578 |
(516) 55.2 |
255 183 196 |
(211) 38.4 |
715 713 730 |
(719) 9.3 |
112 111 121 |
(115) 5.5 |
2294 2350 1477 |
(2040) 488.7 |
ENNG = N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO = 4-Nitroquinoline-1-oxide
9AA = 9-Aminoacridine
N/T = Not tested at this dose level
P = Precipitate
* = Partial or complete absence of bacterial background lawn
# = Standard deviation
Table5: Test Results: Main Test– With Metabolic Activation
Test Period |
From: 26 May 2008 |
To: 29 May 2008 |
|||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
|||||||||||
Base-pair substitution type |
Frameshift type |
||||||||||||
TA100 |
TA1535 |
WP2uvrA‑ |
TA98 |
TA1537 |
|||||||||
+ |
0 |
99 126 128 |
(118) 16.2# |
8 16 16 |
(13) 4.6 |
40 42 41 |
(41) 1.0 |
34 29 23 |
(29) 5.5 |
13 21 18 |
(17) 4.0 |
||
+ |
5 |
128 117 117 |
(121) 6.4 |
9 10 14 |
(11) 2.6 |
N/T |
27 26 23 |
(25) 2.1 |
16 18 21 |
(18) 2.5 |
|||
+ |
15 |
125 114 118 |
(119) 5.6 |
10 11 18 |
(13) 4.4 |
N/T |
23 21 23 |
(22) 1.2 |
14 10 15 |
(13) 2.6 |
|||
+ |
50 |
154 115 120 |
(130) 21.2 |
13 8 11 |
(11) 2.5 |
34 37 31 |
(34) 3.0 |
22 22 16 |
(20) 3.5 |
12 12 14 |
(13) 1.2 |
||
+ |
150 |
109 109 113 |
(110) 2.3 |
19 10 9 |
(13) 5.5 |
38 40 32 |
(37) 4.2 |
27 21 24 |
(24) 3.0 |
26 13 16 |
(18) 6.8 |
||
+ |
500 |
122 123 141 |
(129) 10.7 |
8 9 11 |
(9) 1.5 |
29 43 38 |
(37) 7.1 |
C 26 33 |
(30) 4.9 |
9 9 10 |
(9) 0.6 |
||
+ |
1500 |
95 P 102 P 76 P |
(91) 13.5 |
7*P 12*P 4*P |
(8) 4.0 |
24 P 24 P 23 P |
(24) 0.6 |
22 P 24 P 30 P |
(25) 4.2 |
12*P 11*P 9*P |
(11) 1.5 |
||
+ |
5000 |
0*P 0*P 0*P |
(0) 0.0 |
0*P 0*P 0*P |
(0) 0.0 |
17P 26P 28P |
(24) 5.9 |
0*P 0*P 0*P |
(0) 0.0 |
0*P 0*P 0*P |
(0) 0.0 |
||
Positive controls
S9-Mix
+ |
Name Concentration (μg/plate) No. colonies per plate |
2AA |
2AA |
2AA |
BP |
2AA |
|||||||
1 |
2 |
10 |
5 |
2 |
|||||||||
2319 4009 2002 |
(2777) 1078.9 |
397 379 340 |
(372) 29.1 |
451 494 472 |
(472) 21.5 |
148 179 177 |
(168) 17.3 |
330 419 653 |
(467) 166.8 |
||||
BP = Benzo(a)pyrene
2AA = 2-Aminoanthracene
C = Contaminated
N/T = Not tested at this dose level
P = Precipitate
* = Partial absence of bacterial background lawn
# = Standard deviation
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative with and without metabolic activation
The test material was considered to be non-mutagenic under the conditions of this test. - Executive summary:
The method was designed to conform to the guidelines for bacterial mutagenicity testing published by the major Japanese Regulatory Authorities including METI, MHLW and MAFF. It also meets the requirements of the OECD Guidelines for Testing of Chemicals No. 471 "Bacterial Reverse Mutation Test", Method B13/14 of Commission Directive 2000/32/EC and the, EPA (TSCA) OPPTS harmonised guidelines.
Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA- were treated with the test material using the Ames plate incorporation method at up to seven dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system (10 % liver S9 in standard co-factors). The dose range for the range-finding test was determined in a preliminary toxicity assay and ranged between 5 and 5000 µg/plate, depending on bacterial strain type. The experiment was repeated on a separate day using the same dose range as the range-finding test, fresh cultures of the bacterial strains and fresh test material formulations.
Additional dose levels (5 and 15 µg/plate) were included (where applicable) to allow for test material induced toxicity, in Salmonella typhimurium strains only, ensuring that at least four non‑toxic doses were achieved.
The vehicle (sterile distilled water) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated.
The test material caused a visible reduction in the growth of the bacterial background lawn and/or a substantial decrease in the frequency of revertant colonies for all of the Salmonella strains in both the presence and absence of S9, initially from 500 µg/plate. Small decreases in revertant colony frequency were also noted for E.coli strain WP2uvrA-at 5000 µg/plate in both the presence and absence of S9. The sensitivity of the tester strains to the toxicity of the test material varied slightly between strain type, exposure with or without S9 and experiment number.
A pale flakey precipitate was observed at and above 1500 µg/plate, this did not prevent the scoring of revertant colonies.
No significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test material, either with or without metabolic activation.
The test material was considered to be non-mutagenic under the conditions of this test.
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