Sodium N-chlorobenzenesulphonamide

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:

2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to GLP and valid methods, therefore it is considered adequate, reliable and relevant for classification.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2006

Report date:

2006

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

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

CAS 127-52-6 trihydrate

IUPAC Name:

CAS 127-52-6 trihydrate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Chloramin B; sodium N-chlorobenzenesulphonamide trihydrate
- Substance type: Biocide
- Physical state: White to light yellow powder
- Analytical purity: 98%
- Impurities (identity and concentrations): 0.6% NaOH.
- Composition of test material, percentage of components: 77% CAS 127-52-6 (98% pure), 22.4% Water incl. trihydrate, 0.6% NaOH.
- Isomers composition: Not applicable
- Purity test date: No data
- Lot/batch No.: 54/MA/20/06.
- Expiration date of the lot/batch: 03/2011.
- Stability under test conditions: Stable, c.f. Chapter 4.18.
- Storage condition of test material: Stable, c.f. Chapter 4.18.

Method

Target gene:

Histidine; Tryptophan

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media: Nutrient broth Merck; Agar-agar Fluka
- Properly maintained: yes

- Periodically "cleansed" against high spontaneous background: yes/no

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

First toxicity test: 0, 10, 100, 500, 1000, 2500 and 5000µg/plate
Second toxicity test: 0, 1, 5, 10, 25, 50 and100 µg/plate
First mutagenicity test: 0, 1, 3, 10, 30 and 100 µg/plate
Second mutagenicity test: 0, 0.3, 1, 3, 10 and 30 µg/plate (due to the substance´s toxicity lower doses were chosen for the repeated test)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: demineralized water
- Justification for choice of solvent/vehicle:

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

without solvent

Negative solvent / vehicle controls:

yes

Remarks:

0.1mL demineralized water

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

1.5 µg/plate TA 100 and TA 1535

Controls 2

Untreated negative controls:

yes

Remarks:

without solvent

Negative solvent / vehicle controls:

yes

Remarks:

0.1mL demineralized water

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine

Remarks:

20 µg/plate TA 98

Controls 3

Untreated negative controls:

yes

Remarks:

without solvent

Negative solvent / vehicle controls:

yes

Remarks:

0.1mL demineralized water

Positive controls:

yes

Positive control substance:

other: 9-aminoacridine hydrochloride monohydrate

Remarks:

100 µg/plate TA 1537

Controls 4

Untreated negative controls:

yes

Remarks:

without solvent

Negative solvent / vehicle controls:

yes

Remarks:

0.1mL demineralized water

Positive controls:

yes

Positive control substance:

other: 2-aminofluorene

Remarks:

10µg/plate TA100 and TA98

Controls 5

Untreated negative controls:

yes

Remarks:

without solvent

Negative solvent / vehicle controls:

yes

Remarks:

0.1mL demineralized water

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Remarks:

1.0µg/plate –TA 1535, 2.5µg/plate –TA 1537, 25 µg/plate – E.coli

Controls 6

Untreated negative controls:

yes

Remarks:

without solvent

Negative solvent / vehicle controls:

yes

Remarks:

0.1mL demineralized water

Positive controls:

yes

Positive control substance:

other: N-methyl-N’-nitro-N-nitrosoguanidine

Remarks:

20µg/plate E. Coli

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48-72h (exposure time in selection medium)
- Selection time (if incubation with a selection agent): 48-72h

SELECTION AGENT (mutation assays): histidine; tryptophan

NUMBER OF REPLICATIONS: Triplicate

DETERMINATION OF CYTOTOXICITY
- Method: decrease in number of revertants, changes in background

Evaluation criteria:

The main criterion for the evaluation of results was the modified two-fold increase rule, its using is comparable with using of statistical methods. After this rule the result is positive, when reproducible dose-effect and/or doubling of ratio of number of revertants at tested dose to number of revertants in negative control (Rt/Rc ) is reached.

Results and discussion

Test results

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:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Cytotoxicity at 100 µg/plate

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 with metabolic activation
negative without metabolic activation

In the arrangement given above, the test substance Chloramin B trihydrate was nonmutagenic for all used bacterial strains with as well as without metabolic activation.

Executive summary:

The test Substance Chloramin B trihydrate was assayed for the mutagenicity by the Bacterial Reverse Mutation Test. The test was performed according to OECD Test Guideline 471 Bacterial Reverse Mutation Test which is analogous to the EU method B.13/14. Four indicator Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537 and one indicator Escherichia coli WP2uvrA strain were used. The test substance was dissolved in demineralized water and assayed in doses of 0.3-100 µg which were applied to the plates in emulsion in amount of 0.05 mL. Two series of experiments were performed with each strain- without metabolic activation and with supernatant of rat liver and a mixture of cofactors. In the arrangement given above, the test substance Chloramin B trihydrate was nonmutagenic for all used bacterial strains with as well as without metabolic activation.