Alkylated Naphthalene Sulphonic Acid

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

The test substance was tested in a plate incorporation and pre-incubation assay in Salmonella thypimurium strains TA100, TA102, TA97a, TA 98 and TA 1535 in presence and absence of metabolic activation. No mutagenic effects were seen in three experiments. The number of revertant colonies was not increased in comparison with the spontaneous revertants. Therefore the test substance is considered not mutagenic under the test conditions.

Link to relevant study records

Reference

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:

21-01-2014 to 07-02-2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: study according to the guideline under GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine

Species / strain / cell type:

S. typhimurium, other: LT2 TA 1535, TA 97a, TA 98, TA 100 and TA 102

Details on mammalian cell type (if applicable):

TA97a: hisD6610, uvrB, pKM 101, rfa
TA 98: hisD3052, uvrB, pKM 101, rfa
TA 100: hisG46, uvrB, pKM 101, rfa
TA102: hisG428, pKM 101, rfa
TA1535 : hisG46, uvrB, rfa.

Metabolic activation:

with and without

Metabolic activation system:

S9 produced from the livers of male Sprague-Dawley rats which were treated with 500 mg Aroclor 1254/kg body weight ip (Trinova Biochem. Gießen (Batch no. 3067)

Test concentrations with justification for top dose:

exp 1: 0.050, 0.150, 0.500, 1.500 and 5.000 mg/plate.
exp 2 (TA 100 and TA 102): 0.047, 0.094, 0.188, 0.375, 0.750 and 1.500 mg/plate
exp 3 (TA97a, TA 98 and TA 1535): 0.047, 0.094, 0.188, 0.375, 0.750 and 1.500 mg/plate

Vehicle / solvent:

- Vehicle used: ethanol
- Justification for choice of solvent/vehicle: the test substance was sufficiently soluble, and no effects on the viability of the bacteria or the number of spontaneous revertants are expected.

Untreated negative controls:

yes

Remarks:

DMSO (positive control solvent)

Negative solvent / vehicle controls:

yes

Remarks:

ethanol

True negative controls:

yes

Remarks:

H2O

Positive controls:

yes

Remarks:

without metabolic activarion TA 97a, TA98 and TA102 : 4-Nitro-1,2-phenylene diamine; TA 1535 and TA 100 : Sodium azide ; with metabolic activation TA 97a, TA 100, TA 102 and TA 1535 : Aminoanthracene ; TA98: Benzo-a-pyrene

Positive control substance:

sodium azide

benzo(a)pyrene

other: 4-Nitro-1,2-phenylene diamine, aminoanthracene

Details on test system and experimental conditions:

Cell cultures: Salmonella typhimurium (all strains used) were obtained from TRINOVA BioChem

METHOD OF APPLICATION: in medium; exp 1 preincubation; exp 2 and 3 in agar (plate incorporation)

DURATION
- Preincubation period (exp 1): 20 min
- Exposure duration: 48 h at 37 °C

SELECTION AGENT (mutation assays): histidine

CELL COUNT: visually

NUMBER OF REPLICATIONS: 4 per strain and concentration

NUMBER OF CELLS: titre determined to be 1.5E09 to 3.6E09 cells/mL

Evaluation criteria:

positive when: significant, reproducible increase of revertant colonies per plate (increase factor >2) in at least one strain. A concentration-related increase over the range tested can also be taken as a sign of mutagenic activity .
biological relevance is taken into account first

Statistics:

none performed

Species / strain:

S. typhimurium, other: LT2 TA 1535, TA 97a, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

exp 1: cytotoxicity at 1500 (slight) and 5000 ug/plate (turbidity at 500, 1500 and 5000 ug/plate ); exp 2 and 3 no cytotoxicity (turbidity at 375, 750 and 1500 ug/plate)

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results (migrated information):
negative

The test substance did not induce mutations in 5 strains of Salmonella typhimurium both in presence and absence of metabolic activation

Executive summary:

The test substance was tested in a plate incorporation and pre-incubation assay in Salmonella thypimurium strains TA100, TA102, TA97a, TA 98 and TA 1535 in presence and absence of metabolic activation. No mutagenic effects were seen in three experiments. The number of revertant colonies was not increased in comparison with the spontaneous revertants. Therefore the test substance is considered not mutagenic under the test conditions.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

An Ames test on the structural analogue DNNSA was negative with and without metabolic activation. The Ames test on DDNSA is one of the bridging studies used to substantiate the read-across. In view of structural similarities and the outcome of the bridging Ames test, the results of the tests on BaDNNSA are considered representative for DDNSA. A chromosome aberration test and a Mouse Lymphoma cell mutation assay on BaDNNSA were both negative. Therefore it is concluded that also DDNSA is not expected to exhibit mutagenic or clastigenic properties.

Justification for selection of genetic toxicity endpoint
Guideline study under GLP with the test substance

Justification for classification or non-classification

The test material, DDNSA, was negative in the Ames test and a structural analogue was negative in the two other in vitro genetic toxicity tests (mouse lymphoma cell mutation assay and chromosome aberration test in human lymphocytes). Therefore DNNSA is not classified for genetic toxicity according to CLP (Regulation EC No 1272/2008) or DSD (Directive 67/548/EEC).