Reaction mass of sodium sulphate and disodium dodecanedioate and Dodecanoic acid, 12-amino-, sodium salt (1:1)

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:

from 15 May 2009 to 30 October 2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Compliant to GLP and testing guideline; adequate coherence between data, comments and conclusions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

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

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate

IUPAC Name:

Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate

Details on test material:

- Name of test material (as cited in study report): Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate
- Substance type: UVCB
- Physical state: brown liquid
- Composition of test material, percentage of components: sodium hydroxyde: 1 - 8 % ; sodium sulfate: 7 - 20 % ; sodium amino-12-dodecanoate: 2 - 8 ; sodium dodecanoedioate: 0.5 - 5 % ; sodium chlorure: 0.05 - 0.3 % ; lactame 12: 0.2 - 1 % ; organic fraction : about 10 - 18 %
- Purity test date: 26 March 2009
- Lot/batch No.: T710/712
- Expiration date of the lot/batch: 20 March 2010
- Storage condition of test material: at room temperature

Method

Target gene:

Histidine operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

312.5 ; 625 ; 1250 ; 2500 and 5000 µg/plate, for all mutagenicity experiments, with and without S9 mix.

Vehicle / solvent:

- Vehicle used: water for injections

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

Migrated to IUCLID6: and 9-Aminoacridine; 2-Nitrofluorene; Mitomycin C; 2-Anthramine; Benzo(a)pyrene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar
The preliminary test, all experiments without S9 mix and the first and third experiments with S9 mix were performed according to the direct plate incorporation method. The second experiment with S9 mix was performed according to the preincubation method.


DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 or 72 hours

Evaluation criteria:

A reproducible 2-fold increase (for the TA 98, TA 100 and TA 102 strains) or 3-fold increase (for the TA 1535 and TA 1537 strains) in the number of revertants compared with the vehicle controls, in any strain at any dose-level and/or evidence of a dose-relationship was considered as a positive result. Reference to historical data, or other considerations of biological relevance may also be taken into account in the evaluation of the data obtained.

Statistics:

Not applicable.

Results and discussion

Test results

Test results 1

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:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

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

The test item did not show any mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.

Executive summary:

A preliminary toxicity test was performed to define the dose-levels of Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate to be used for the mutagenicity study. The test item was then 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.

 

Both experiments were performed according to the direct plate incorporation method except for the second test with S9 mix, which was performed according to the preincubation method (60 minutes,).

Five strains of bacteriaSalmonella typhimurium: TA 1535, TA 1537, TA 98, TA 100 and TA 102 were used. Each strain was exposed to at least five dose-levels of the test item (three plates/dose‑level). After 48 to 72 hours of incubation at, the revertant colonies were scored.

 

The evaluation of the toxicity was performed on the basis of the observation of the decrease in the number of revertant colonies and/or a thinning of the bacterial lawn.

 

The test item Reaction mass of sodium sulfate, sodium amino-12-dodecanoate and sodium dodecanoedioate was dissolved in water for injections.

The dose-levels of the positive controls were as follows:

without S9 mix

·          1 µg/plate of sodium azide (NaN₃): TA 1535 and TA 100 strains,

·          50 µg/plate of 9-Aminoacridine (9AA): TA 1537 strain,

·          0.5 µg/plate of 2-Nitrofluorene (2NF): TA 98 strain,

·          0.5 µg/plate of Mitomycin C (MMC): TA 102 strain.

with S9 mix

·          2 µg/plate of 2-Anthramine (2AM): TA 1535, TA 1537 and TA 98 strains,

·          5 µg/plate of Benzo(a)pyrene (BAP): TA 100 strain,

·          10 µg/plate of 2-Anthramine (2AM): TA 102 strain.

Results

 

The number of revertants for the vehicle and positive controls was as specified in the acceptance criteria. The study was therefore considered as valid.

Since the test item was freely soluble and non-toxic in the preliminary assay, the highest dose‑level was 5000 µg/plate, according to the criteria specified in the international guidelines.

 

The selected treatment-levels were: 312.5, 625, 1250, 2500 and 5000 µg/plate for all mutagenicity experiments, with and without S9 mix.

No precipitate was observed in the Petri plates when scoring the revertants at any of the tested dose-levels.

No noteworthy toxicity was noted towards all the strains used, either with or without S9 mix.

A noteworthy increase (up to 2.0-fold the vehicle control value) in the number of revertants was noted in the TA 100 strain, in the first experiment with S9 mix (direct plate incorporation method). This increase exceeded the threshold of 2‑fold the vehicle control value, however it was not observed in the second experiment, using the preincubation method. In the third experiment (performed under the same experimental conditions as for the first one), no noteworthy increase in the number of revertant was observed. Consequently, since it was not reproducible, the increase was not considered as biologically relevant.

The test item did not induce any noteworthy increase in the number of revertants, in any of the four other strains, either with or without S9 mix.

 

Conclusion

 

Under the experimental conditions of the study, the test item Reaction mass of sodium sulfate, sodium amino‑12-dodecanoate and sodium dodecanoedioate did not show any mutagenic activity in the bacterial reverse mutation test withSalmonella typhimurium.