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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP and guideline study: restriction: E. coli not tested

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. certificate)
Remarks:
testing lab.
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
no data about impurities

Method

Target gene:
S. tyhpimurium
Species / strain
Species / strain / cell type:
other: S. typhimurium TA 98, TA 100, TA 102, TA 1535, TA 1537
Metabolic activation:
with and without
Metabolic activation system:
rat liver S-9 mix
Test concentrations with justification for top dose:
0, 125, 250, 1000, 2000 µg/plate
Vehicle / solvent:
distilled water
Controls
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
yes
Positive controls:
yes
Positive control substance:
other: without S-9 mix: sodium azide, 9-aminoacridine, 2-nitrofluorene and mitomycin C; with S-9 mix: 2-anthramine and danthron
Details on test system and experimental conditions:
Preincubation method : the test substance solution (maximum volume: 0.1 ml), 0.5 ml of S9 mix and 0 .1 ml of the strain were incubated for 60 minutes at 37°C prior adding the overlay agar and pouring onto the surface of a minimum agar plate.
Direct plate incorporation method: the test substance solution (maximum volume: 0.1 ml), 0 .5 ml of S9 mix (when required) and 0 .1 ml of the strain were added to 2 ml molten agar containing traces of histidine and biotin and maintained at +45°C. After rapid homogenization, the mixture was spread out on a Petri plate containing minimum medium.
The methods used were the direct plate incorporation method (both tests without S9 mix, first test with S9 mix) or the preincubation method (second test with S9 mix) described by Maron and Ames.
Evaluation criteria:
The following criteria were used as an aid for determining a positive response: a reproducible and significant dose relationship using a linear regression analysis, considered as significant if p<0 .05 (for n = 18 values, the correlation coefficient must be r >0 .47) and/or a reproducible and significant increase (i.e. a doubling in the number of revertants for at least one of the tested strains when compared to that of the negative and/or solvent controls) for at least one of the tested concentrations.
A test substance is considered as non-mutagenic in this test system if the above two criteria are not fully met. Biological and statistical significances were considered during the evaluation.

Results and discussion

Test results
Species / strain:
other: S. typhimurium TA98, TA100, TA102, TA1535, TA1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
other: see additional information on results
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
CYTOTOXICITY IN PRELIMINARY TEST
- Without metabolic activation moderate effects on bacterial lawn at 2260 ug/plate, slight effects at 1130 ug/plate; number of revertants decreased to ca. 50% of solvent control at 1130 ug/plate and ca. 7% at 2260 ug/plate
- With metabolic activation no effects on bacterial lawn but decrease in the number of revertants at 452 and 1130 μg/plate (ca. 50% of solvent control), at 2260 μg/plate ca. 7%.
GENOTOXIC EFFECTS IN THE MAIN STUDY
- The test substance 2-Mercaptoethanol, sodium salt did not induce any significant increase in the number of revertants, with or without S9 mix, in any of the 5 strains tested.
- Negative and positive controls were valid.
- The high dose level decreased the number of revertants to ca. 50% of the solvent control in TA100 and TA102 (with and without MA), to ca. 70% in TA98 and TA1537 (with and without MA) in at least one out of 2 trials.
Remarks on result:
other: strain/cell type:
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Under our experimental conditions, the test substance 2-MERCAPTOETHANOL, SODIUM SALT did not show mutagenic activity in the Ames test .