1,10-Decanediamine, 4-methylbenzenesulfonate (1:2)

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

This study was performed to investigate the potential of Clevios K Primer W8 dry to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and TA 102.

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations:

Pre-Experiment/Experiment I:	3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate
Experiment II:	33; 100; 333; 1000; 2500; and 5000 μg/plate

No precipitation of the test item occurred up to the highest investigated dose.

Reduced background growth was observed on the plates incubated with the test item in strains TA 98 and TA 100 in experiment I and in strains TA 1537 and TA 100 in experiment II.

Additional minor toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in experiment II in strain TA 100 with and without S9 mix at 5000 μg/plate. In experiment I and the remaining test groups of experiment II no further toxic effects were observed.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Clevios K Primer W8 dry at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

Therefore, Clevios K Primer W8 dry is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.

Link to relevant study records

Reference

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed between 15 May 2019 and 27 May 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

histidine (Salmonella typhimurium)

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/β-naphthoflavone induced rat liver

Test concentrations with justification for top dose:

Experiment I (pre-experiment): 3; 10; 33; 100; 333; 1000; 2500; 5000 μg/plate
Experiment II: 33; 100; 333; 1000; 2500; and 5000 μg/plate

Vehicle / solvent:

Deionised water. The solvent was chosen because of its solubility properties and its relative nontoxicity to the bacteria (Maron et al.; 1981).

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: slight toxicity

Remarks:

a reduction in the number of revertants below the indication factor of 0.5 at 5000 μg/plate in one of two experiments

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Additional information on results:

Reduced background growth was observed on the plates incubated with the test item in strains TA 98 and TA 100 in experiment I and in strains TA 1537 and TA 100 in experiment II.

Conclusions:

During the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. Therefore, Clevios K Primer W8 dry is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.

Executive summary:

This study was performed to investigate the potential of Clevios K Primer W8 dry to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, TA 100, and TA 102.

The assay was performed in two independent experiments both with and without liver microsomal activation. Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations:

Pre-Experiment/Experiment I:	3; 10; 33; 100; 333; 1000; 2500; and 5000 μg/plate
Experiment II:	33; 100; 333; 1000; 2500; and 5000 μg/plate

No precipitation of the test item occurred up to the highest investigated dose.

Reduced background growth was observed on the plates incubated with the test item in strains TA 98 and TA 100 in experiment I and in strains TA 1537 and TA 100 in experiment II.

Additional minor toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in experiment II in strain TA 100 with and without S9 mix at 5000 μg/plate. In experiment I and the remaining test groups of experiment II no further toxic effects were observed.

No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with Clevios K Primer W8 dry at any dose level, neither in the presence nor absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentrations in the range below the generally acknowledged border of biological relevance.

Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies.

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used.

Therefore, Clevios K Primer W8 dry is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Justification for classification or non-classification

During the described mutagenicity test and under the experimental conditions reported, the test item did not induce gene mutations by base pair changes or frameshifts in the genome of the strains used. Therefore, Clevios K Primer W8 dry is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.