4,4'-[1,2-ethenediylbis[(3-sulfo-4,1-phenylene)imino[6-[bis(2-hydroxyethyl)amino]-1,3,5-triazine-4,2-diyl]imino]]bisarenoate sodium salt

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

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

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented study report which meets basic scientific principles

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1983

Deviations:

yes

Remarks:

(no E.coli or TA 102 strain tested, only 2-amino anthracene as positive control with metabolic activation, only one experiment)

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Details on test material:

- Name of test material (as cited in study report): FAT 66031/B
- Analytical purity: 88.1%
- Lot/batch No.: Mg. 636 vom 21.7.87, BSG ex Pa 34/34.
- Storage condition of test material: 4°C, light protected

Method

Target gene:

his-operon

Species / strain

Species / strain / cell type:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix) prepared from the livers of rats treated with Aroclor 1254

Test concentrations with justification for top dose:

Pre-Experiment: 1, 3, 10, 33, 100, 333, 1000 and 5000 µg/plate
Experiment: 10.0, 100.0, 333.3, 1000.0 and 5000.0 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: H20 bidest
- Justification for choice of solvent/vehicle: The solvent was chosen because of its solubility properties and its relative non-toxicity to the bacteria.

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

bidest. water

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

other: 4-nitro-o-phenylene-diamine (NOPD); 2-aminoanthracene (2AA)

Remarks:

(-S9) sodium azide (TA 1535, 100), 4-NOPD (TA 1537, 1538, 98); (+S9) 2-aminoanthracene (all strains)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 72 hours at 37°C

NUMBER OF REPLICATIONS: 3 plates per condition; only one experiment

Evaluation criteria:

A test substance is considered as mutagen if in strain TA 100 the number of reversions is at least twice as high with two concentrations of the substance, and in strains TA 1535, TA 1537, TA 1538, and TA 98 it is at least three times higher with two concentrations of the test substance as compared to the spontaneous reversion rate. If in a repeated experiment there is a clear increase in the reversion rate by factors as indicated even in only one strain, then this also leads to the substance being classified as a mutagen.
Also, a continuous dose-dependent increase in the number of revertants is regarded as an indication of possibly existing mutagenic properties of the test substance, regardless whether the highest dose induces the above described enhancement factors or not. Such findings would perhaps not be sufficient to declare the substance mutagenic. However, it recommends further testing to exclude mutagenicity.

Statistics:

Revertant colony numbers were calculated as average and standard deviation. No statistical analysis test was performed.

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

The test article precipitated weakly at 5000.0 µg/plate. 

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

No toxic effects were observed in all strains. No reduction of revertant numbers for more than 70% was found in any strain used.The plates incubated with the test substance showed normal background growth up to 5000.0 µg/plate in the strains used with and without S9 mix. The test article did not induce an increase in revertant colony numbers in the absence and in the presence of S9 mix in all the strains used.

Results:

Dose µg/plate	metabolic activation	TA 1535	TA 1537	TA 1538	TA 98	TA 100
0	-	16	16	14	27	154
10	-	12	6	11	26	192
100	-	13	19	10	22	205
333.3	-	20	13	16	21	182
1000	-	18	21	10	25	187
5000	-	14	18	13	25	169
Sodium azide	-	1542				1599
4-NOPD	-		323	1963	1761
0	+	16	19	23	51	139
10	+	12	21	23	36	150
100	+	11	19	22	46	150
333.3	+	14	22	23	51	173
1000	+	10	22	26	41	179
5000	+	11	12	25	43	154
2-Aminoanthracene	+	1025	373	2144	1928	1550

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test item did not induce point mutations by base pair changes or frameshifts in the genome of the strains used. Therefore, the test substance is considered to be non-mutagenic in this Salmonella typhimurium reverse mutation assay.