Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda

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:

no data available

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is sufficiently documented. Comparable to guideline study with acceptable restrictions (no E. coli or TA102 strain, only one positive control substance for test with metabolic activation)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no E.coli or TA102 strain, only one positive control substance for test with metabolic activation

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

sodium diisobutylnaphtalene sulfonate

Cas Number:

91078-64-7

IUPAC Name:

sodium diisobutylnaphtalene sulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

See confidential details on test material section

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

Standard plate test: 0, 60, 120, 100, 250, 500, 1000, 2000, 2500, 5000, 7500 µg/plate
Preincubation test: 0, 20, 100, 500, 1000, 2000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: water

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (with S-9 mix for all strains); N-methyl-N-nitro-N-nitrosoguanidine (without S-9 mix for TA100 and TA1535); 4-nitro-o-phenyldiamine (without S-9 mix for TA98); 9-aminoacridine chloride monohydrate (without S-9 mix for TA1537)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION
- Preincubation period: 20 minutes
- Exposure duration: 48 hours

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth

Evaluation criteria:

In general, a substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results.

Statistics:

no data available

Results and discussion

Test results

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:

cytotoxicity

Remarks:

A bacteriotoxic effect was observed depending on the strain and experimental conditions from about 1000 µg - 2500 µg/plate onward.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: Complete solubility of test substance in aqua dest.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

Under these experimental conditions, the test item Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.

Executive summary:

This study was performed to investigate the potential of the test item, Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda, to induce reverse mutation in Salmonella typhimurium. The test item was tested in two independent experiments, with and without a metabolic activation system, the S9 mix, prepared from a liver post‑mitochondrial fraction (S9 fraction) of rats induced with Aroclor 1254. The first experiment was performed according to the direct plate incorporation method and the second was performed according to the pre-incubation method (20 minutes). 

Four strains of bacteria Salmonella typhimurium TA 1535, TA 1537, TA 98 and TA 100 were used. Each strain was exposed to dose-levels of the test item. The selected dose-levels were: 60, 120, 100, 250, 500, 1000, 2000, 2500, 5000, 7500 µg/plate in the first test and 20, 100, 500, 1000, 2000 µg/plate in the second test. The number of mutations were scored.

A bacteriotoxic effect was observed depending on the strain and test conditions from about 1000 µg – 2500 µg/plate onward. An increase in the number of his revertants was not observed both in the standard plate test and in the pre-incubation test either without S-9 mix or after the addition of the metabolizing system.

In conclusion, the test item did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium, in the absence or in the presence of a rat metabolising system therefore Reaction product of naphthalene, butanol, sulfonated and neutralized by caustic soda is not classified according to Annex VI of the Directive 67/548/CEE and according to EU Regulation 1272/2008 (CLP).