3-chloroperbenzoic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

6 November 2017 - 14 December 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

An incorrect amount of the positive control was added to the plates. Since all positive control values were within the laboratory historical control data ranges, this deviation has no influence on the results of the study.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

3-chloroperbenzoic acid

EC Number:

213-322-3

EC Name:

3-chloroperbenzoic acid

Cas Number:

937-14-4

Molecular formula:

C7H5ClO3

IUPAC Name:

3-chlorobenzene-1-carboperoxoic acid

Test material form:

liquid - solid: mixture of

Remarks:

The substance in its marketed form is not in solid or granular form. The marketed product contains 18% water and is thus a wet, paste-like consistency.

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch No.of test material: A0378021

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: In refrigerator (2-8°C)
- Stability under test conditions: 31 October 2019 (retest date)

TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: The test item was soluble in ethanol.

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

Rat liver microsomal enzymes (S9 homogenate) from animals treated with Aroclor 1254.

Test concentrations with justification for top dose:

Dose-range finding test - eight concentrations, 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate were tested in triplicate. The highest concentration of the test item used in the first mutation assay was the level at which the test item inhibited bacterial growth.
First Experiment: Direct Plate Assay - 0.18, 0.55, 1.7, 5.4, 17 and 52 μg/plate
Second Experiment: Pre-Incubation Assay - 0.17, 0.55, 1.7, 5.4, 17, 52 and 164 µg/plate using TA100 and WP2uvrA. Based on the results of this test the following concentrations were used for TA1535, TA1537 and TA98:
Absence of S9-mix: 0.028, 0.088, 0.28, 0.86, 2.7 and 8.4 μg/plate.
Presence of S9-mix: 0.28, 0.86, 2.7, 8.4, 26 and 82 μg/plate.

Vehicle / solvent:

Ethanol.

Controls

Untreated negative controls:

yes

Remarks:

Ethanol

Negative solvent / vehicle controls:

no

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: ICR-191; 2-aminoanthracene

Details on test system and experimental conditions:

All incubations were carried out in a controlled environment at a temperature of 37.0 ± 1.0°C.

Evaluation criteria:

A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is considered acceptable if it meets the following criteria:
a) The vehicle control and positive control plates from each tester strain (with or without S9-mix) must exhibit a characteristic number of revertant colonies when compared against relevant historical control data generated at Charles River Den Bosch.
b) The selected dose-range should include a clearly toxic concentration or should exhibit limited solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 5 mg/plate.
c) No more than 5% of the plates are lost through contamination or some other unforeseen event. If the results are considered invalid due to contamination, the experiment will be repeated.

A test item is considered negative (not mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 or WP2uvrA is not greater than two (2) times the concurrent control, and the total number of revertants in tester strains TA1535, TA1537 or TA98 is not greater than three (3) times the concurrent control.
b) The negative response should be reproducible in at least one follow up experiment.

A test item is considered positive (mutagenic) in the test if:
a) The total number of revertants in tester strain TA100 or WP2uvrA is greater than two (2) times the concurrent control, or the total number of revertants in tester strains TA1535, TA1537 or TA98 is greater than three (3) times the concurrent control.
b) In case a repeat experiment is performed when a positive response is observed in one of the tester strains, the positive response should be reproducible in at least one follow up experiment.

Statistics:

Not applicable.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

In conclusion, the substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Executive summary:

In the in vitro genotoxicity study (Ames test) the substance was tested for mutagenicity in Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100, and in Escherichia coli strain WP2 uvrA. A dose-range finding test was conducted using eight concentrations of the substance (1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate) were tested in triplicate.  The highest concentration of the substance used in the first mutation assay was the level at which the substance inhibited bacterial growth. This was followed with the first experiment (Direct Plate Assay) employing concentrations of 0.18, 0.55, 1.7, 5.4, 17 and 52 μg/plate. The second experiment (Pre-Incubation Assay) used concentrations of 0.17, 0.55, 1.7, 5.4, 17, 52 and 164 µg/plate with TA100 and WP2uvrA. Based on the results of this test the following concentrations were used for TA1535, TA1537 and TA98:

in the absence of S9-mix: 0.028, 0.088, 0.28, 0.86, 2.7 and 8.4 μg/plate and in the presence of S9-mix: 0.28, 0.86, 2.7, 8.4, 26 and 82 μg/plate. No evidence of mutagenic activity was seen at any concentration of the substance in either mutation test. It was concluded that the substance showed no evidence of mutagenic activity in these bacterial systems under the test conditions employed.