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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Jan - Feb 1992
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
E. coli WP2 strain or S. typhimurium TA 102 were not investigated.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
E. coli WP2 strain or S. typhimurium TA 102 were not investigated.
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Test material form:
solid
Specific details on test material used for the study:
Information from study report:
- Name of test material: Emulgator E30, Fest
- CAS number: 5896-54-8
The CAS number in report (5896-54-8) is misleading, since this CAS number does only represent 1-Penta-decanesulfonic acid, sodium salt (1:1), but it is confirmed that the substance used is correctly assigned to CAS 68188-18-1.
- Appearance/Further information: The test substance was supplied as white wax-like leaf-let and was received at the test institute on March 06, 1991.
- Batch No. of test material: 210291
- Purity: 95 %
- Analytical reference: TGL 39237
- Solubility and stability of the test substance in the solvent/vehicle: The substance is known to be stable in water.

Method

Target gene:
Histidine locus
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 preparation from the liver of Aroclor 1254 induced Sprague-Dawley rats.
Test concentrations with justification for top dose:
For initial test and independent repeat: 5, 1.5, 0.5, 0.15, 0.05, 0.015, 0.005 mg/plate with and without metabolic activation
Vehicle / solvent:
Water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
water
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
other: 2-aminoanthracene 10 µg/plate (all strains)
Remarks:
The positive controls 2-nitrofluorene, Na-azide and 9-aminoacridine were used without S9 mix; the positive control 2-aminoanthracene was used with S9 mix.
Details on test system and experimental conditions:
METHOD OF APPLICATION: Plate Incorporation Test
For each concentration and tester strain three parallel plates were used.

DETERMINATION OF CYTOTOXICITY
- Method: background growth
Evaluation criteria:
The threshold value given in the following tables is the number of revertants (average number of revertant colonies from test compound) for which X² is >/= 6.6 with the average number of spontaneous revertant colonies being the corresponding solvent control. If the number of revertants with the test compound would be greater than the calculated threshold value the number of revertants with the test substance would be significantly greater than the spontanous rate in the solvent control (error probability p
Statistics:
The statistical significance of the increase in the mutation frequency is examined in the X²-test with respect to its statistical significance. A statistically significant dose-related increase in the number of revertants occurs if several dilutions of the test compound are significant greater than the average number of spontaneous revertant colonies.

Results and discussion

Test results
Species / strain:
other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
Mutagenicity of the test material was not detected, neither in the initial experiment in concentrations up to and including 5 mg/plate, nor in the independent repetition with up to and including 0.5 mg/plate, both conducted with and without a metabolic activating system.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
In the first test toxic effects were exhibited when the highest concentration of 5 mg/plate was applied. The tester strains gave reduced numbers of revertants compared to the untreated control however, accompanied by a scattered background growth of the inoculated bacteria. Growth of strain TA 1538 was completely inhibited by a concentration of 0.5 mg/plate. Strain TA 1537 showed the same effect with a concentration of 1.5 mg/plate.
Therefore the highest concentration of the test substance was 0.5 mg/plate in the repetition experiments either with or without metabolic activation.

Applicant's summary and conclusion

Executive summary:

A bacterial reverse mutation assay (Ames test) according to OECD TG 471 was carried out with and without a metabolic activating system with the 5 tester strains S. thyphimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100, however no E. coli WP2 strain or S. typhimurium TA 102 was investigated. One independent repetition of the test was performed. Mutagenicity of the test material was not detected. Toxicity to S. typhimurium could be observed at the higher concentrations.