N-acetylsulphanilyl chloride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from an authoritative database.

Data source

Reference

Reference Type:

other: Authoritative database

Title:

Bacterial reverse mutation study (BRM) using the test chemical

Author:

J-Check

Year:

2001

Bibliographic source:

J-Check_121608

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

According to OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N-acetylsulphanilyl chloride

EC Number:

204-485-1

EC Name:

N-acetylsulphanilyl chloride

Cas Number:

121-60-8

Molecular formula:

C8H8ClNO3S

IUPAC Name:

4-acetamidobenzenesulfonyl chloride

Test material form:

solid: crystalline

Details on test material:

- Name of test material (as cited in study report): N-acetylsulphanilyl chloride
- Substance type: Organic
- Physical state: solid

Specific details on test material used for the study:

Purity:99.7%

Method

Target gene:

His and Trp

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Liver S9 microsomal fraction was prepared from phenobarbital and 5,6-bezoflavone-induced male Sprague Dawley rats.

Test concentrations with justification for top dose:

i) Plate Incorporation Method:
Without S9: 0, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate in the strains of TA 100, TA 1535 and E.coli WP2.
: 0, 31.3, 62.5, 125, 250, 500 and 1000 ug/plate in the strains of TA 98 and TA 1537
With S9: 0, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate (For all strains)

ii) Pre-incubation Method
Without S9: 0, 78.1, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate in the strains of TA 100, TA 1535 and E.coli WP2.
: 0, 31.3, 62.5, 125, 250, 500 and 1000 ug/plate in the strains of TA 98 and TA 1537

With S9: 0, 78.1, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate (For all strains)

Vehicle / solvent:

N,N-dimethylformamide
- Justification for choice of solvent/vehicle: The test chemical was soluble in N,N-dimethylformamide

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

N,N-dimethylformamide

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

sodium azide

furylfuramide

other: 2- aminoanthracene

Details on test system and experimental conditions:

NUMBER OF REPLICATIONS:
- Number of cultures per concentration (single, duplicate, triplicate) : Triplicate per test
- Number of independent experiments : Two

METHOD OF TREATMENT/ EXPOSURE:
- Cell density at seeding (if applicable): NA
- Test substance added in medium; in agar (plate incorporation)

Rationale for test conditions:

A confirmatory test was carried out in pre-incubation method, as negative mutagenic effects was observed in plate incorporation method.

Evaluation criteria:

The test chemical was adjudged as a postive mutagen when the number of the revertant colonies in the test chemical treated increased dose dependently and became two-fold or more than that of the negative control and significant reproducible increase was noted at one or more concentration and at least in one strain with or without metabolic activation system and is considered as negative when any of the above criteria was not fulfilled.

Statistics:

No statistical analysis was performed.

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At 1000 ug/plate concentration without S9, above 2500 ug/plate with S9.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At and above 2500 ug/ml concentration with and without S9.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At 1000 ug/plate concentration without S9 and at 2500 ug/plate with S9.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At 1000 ug/plate concentration without S9, at 2500 ug/plate with S9.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Test results 5

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

At 2500 ug/ml concentration with and without S9.

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

True negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: No mutagenic potential of the test chemical was observed.

Applicant's summary and conclusion

Conclusions:

The registered chemical, 4-Acetamidobenzenesulfonyl chloride (CAS 120-60-8) was tested negative in bacterial reverse mutation test (according to OECD TG 471) in S. typhimurium TA 98, TA100, TA 1535 and TA 1537 and E. coli WP2 uvrA strains, both in the presence and absence of S9 metabolic activation system.

Executive summary:

A GLP-compliant bacterial reverse mutation study (OECD TG 471) was performed to assess the mutagenic potential of the registered substance in S. typhimurium and E.coli strains. The following strains were used: Salmonella typhimurium TA 98, TA100, TA 1535 and TA 1537 and E. coli WP2 uvrA. 

The following test concentrations were used:

i) Plate Incorporation Method:

Without S9: 0, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate in the strains of TA 100, TA 1535 and E.coli WP2.

                : 0, 31.3, 62.5, 125, 250, 500 and 1000 ug/plate in the strains of TA 98 and TA 1537

With S9: 0, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate (For all strains) 

ii) Pre-incubation Method

Without S9: 0, 78.1, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate in the strains of TA 100, TA 1535 and E.coli WP2.

                : 0, 31.3, 62.5, 125, 250, 500 and 1000 ug/plate in the strains of TA 98 and TA 1537

 

With S9: 0, 78.1, 156.3, 312.5, 625, 1250, 2500 and 5000 ug/plate (For all strains)

N,N-dimethylformamide was used as a vehicle. A confirmatory test according to the pre-incubation method was carried out, as the test chemical was found non-mutagenic in the first experiment performed according to the plate incorporation method. The test chemical was considered positive (mutagenic) when the number of the revertant colonies in treated cultures increased dose-dependently and became two-fold or more than that of the vehicle negative control, and a significant reproducible increase was noted at one or more concentration and at least in one strain with or without metabolic activation system. The test chemical was considered negative (non-mutagenic) when any of the above criteria were not fulfilled. No statistical analysis was performed. Results: The test substance did not induce a two-fold and/or biologically relevant increase in revertant colony counts compared with the vehicle control when it was tested up to 5000µg/plate neither in the presence nor the absence of S9 metabolic activation in S. typhimurium and E. coli strains used. Conclusion:The test substance was considered non-mutagenic (negative) in the bacterial reverse mutation test (OECD TG 471).