reaction products of oxidation of 2-aminoanthracene-9,10-dione and chlorine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

other: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Remarks:

The test was conducted by means of Read Across approach. Further information was attached at section 13

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

7,16-dichloro-6,15-dihydroanthrazine-5,9,14,18-tetrone

EC Number:

204-980-2

EC Name:

7,16-dichloro-6,15-dihydroanthrazine-5,9,14,18-tetrone

Cas Number:

130-20-1

Molecular formula:

C28H12Cl2N2O4

IUPAC Name:

7,16-dichloro-6,15-dihydrodinaphtho[2,3-a:2',3'-h]phenazine-5,9,14,18-tetrone

Test material form:

solid: particulate/powder

Details on test material:

Vat Blue 6

Method

Target gene:

Salmonella typhimurium histidine (his) reversion system

Species / strain

Species / strain / cell type:

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

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix (Rat liver enzymes)

Test concentrations with justification for top dose:

2, 20, 100, 300 µg per plate
The test substance is insoluble in water and organic solvents

Vehicle / solvent:

anhydrous dimethylsulphoxide

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

Remarks:

In addition, strains were periodically checked for deoxycholate sensitivity, penicillin sensitivity or resistance to the following mutagens TA1535 - methylnitrosourea TA1537 - 9 -aminoacridine TA98 - N-hydroxyaminofluorene TA100 - methylnitrosourea

Details on test system and experimental conditions:

A sample of the test item was tested in Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 at four dose levels: 2, 20, 100, 300 µg per plate. The compound, just prior to testing was dissolved in anhydrous dimethylsulphoxide. For assay, various amounts of the solution was added to give the appropriate concentration in 3.5 ml of molten soft-agar (0.9% Difco agar and 0.5% w/v of NaCl) containing 0.1 micromol biotin, 0.1 micromol histidine, 20 micromol glucose-6-phosphate, 3 micromol NADP+ , 25 micromol MgCl2, 300 micromol sodium phosphate buffer, pH 7.4 and 0.1 mL of nutrient broth culture of bacteria. Where liver activation was required, 0.1 mL of liver PMS was added just prior to overlaying onto plates containing 25 mL of minimal Davis agar. A positive control compound, acetylaminofluorene, was also tested using TA98. Negative controls contained all the above ingredients except the test chemical. Assays were carried out in duplicate.
After the agar had hardened, the plates were inverted and left to incubate at 37°C for 48 hrs. Counting was carried out using a New Brunswick Biotran II colony counter set at maximum sensitivity and the size setting left off. The counter had been previously calibrated using a test plate. Each experiment was repeated on two separate occasions.

Evaluation criteria:

All results were meaned using a DEC-10 computer and the standard deviations calculated. Each test plate result was compared with the appropriate control using the SPSS programme for Students [-test for equal or unequal variances. Results in which the values statistically differ, i.e. p <=.0.05 are considered to be mutagenic

Statistics:

SPSS: Students [-test for equal or unequal variances

Results and discussion

Test results

Key result

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA100, TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

The test item was not mutagenic in the bacteria reverse mutation assay

Executive summary:

The test item was tested with the Salmonella typhimurium strains TA 1535, TA 1537, TA 98 and TA 100 at concentrations from 2 to 300 µg per plate both in the presence and absence of metabolic activation. No mutagenic effect was observed