Disodium benzene-1,3-disulphonate

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

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 peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Mutagenic activity of 27 dyes and related chemicals in the Salmonella/microsome and mouse lymphoma TK +/- assays

Author:

T.P. Cameron, T.J. Hughes, P.E. Kirby, V.A. Fung and V.C. Dunkel

Year:

1987

Bibliographic source:

Mutation Research, 189 (1987) 223-261

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Gene mutation toxicity study was performed to evaluate mutagenicity of C.I. Acid blue 74 by the standard plate-incorporation assay

GLP compliance:

no

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 5,5'-(2-(1,3-dihydro-3-oxo-2H-indazol-2-ylidene)-1,2-dihydro-3H-indol-3-one)disulphonate

EC Number:

212-728-8

EC Name:

Disodium 5,5'-(2-(1,3-dihydro-3-oxo-2H-indazol-2-ylidene)-1,2-dihydro-3H-indol-3-one)disulphonate

Cas Number:

860-22-0

Molecular formula:

C16H10N2O8S2.2Na

IUPAC Name:

disodium 3,3'-dioxo-1,1',3,3'-tetrahydro-2,2'-biindole-5,5'-disulfonate

Constituent 2

Reference substance name:

Indigo tin disulfonate sodium

IUPAC Name:

Indigo tin disulfonate sodium

Details on test material:

Details on test material
- Name of test material (as cited in study report): C.I. Acid blue 74
- Molecular formula (if other than submission substance):
C16-H10-N2-O8-S2.2Na
C16-H8-N2-Na2-O8-S2
C16-H8-N2-O8-S2.2Na
- Molecular weight (if other than submission substance): 466.3572 g/mol
- Substance type: Organic
- Physical state: Solid
Purity: 85%

Specific details on test material used for the study:

- Name of test material: C.I. Acid blue 74
- Molecular formula: C16-H10-N2-O8-S2.2Na
C16-H8-N2-Na2-O8-S2
C16-H8-N2-O8-S2.2Na
- Molecular weight: 466.3572 g/mol
- Substance type: Organic
- Physical state: Solid
- Purity: 85%
- Impurity: < 10% H2O, < 7% NaCl, Na2SO4

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537, TA1538, TA98 and TA100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

S9 mix from Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters

Test concentrations with justification for top dose:

0, 3333, 1000, 6666 and 10000 µg/plate

Vehicle / solvent:

Vehicle
- Vehicle(s)/solvent(s) used: Yes
- Justification for choice of solvent/vehicle: No data available

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

2-nitrofluorene

sodium azide

other: 2-aminoanthracene (All strains, +S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: No data available
- Exposure duration: No data available
- Expression time (cells in growth medium): No data available
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

SELECTION AGENT (mutation assays): No data available
SPINDLE INHIBITOR (cytogenetic assays): No data available
STAIN (for cytogenetic assays): No data available

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: No data available

DETERMINATION OF CYTOTOXICITY
- Method: No data available

OTHER EXAMINATIONS:
- Determination of polyploidy: No data available
- Determination of endoreplication: No data available
- Other: No data available

Rationale for test conditions:

No data available

Evaluation criteria:

Dose-related increase in the number of revertants above spontaneous solvent controls, with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537.

Statistics:

Mean and standard deviation.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA1538, TA98 and TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No data available

Any other information on results incl. tables

MUTAGENICITY OF MISCELLANEOUS DYE IN THE S. typhimurium ASSAY:

Revertants per plate:

µg/plate	TA1535			TA1537
	(-)S9	(R)S9	(H)S9	(-)S9	(R)S9	(H)S9
Solvent control	23±3	9±5	15±5	9±2	10±3	9±4
Positive control	814±42	156±17	171±14	269±28	294±14	157±11
333	23±2	13±4	11±3	7±1	7±4	Toxic
1000	17±3	10±4	13±5	4±1	8±4	6±3
3333	21±2	11±2	9±3	3±2	7±3	3±1
6666	19±7	12±0	14±3	5±1	5±2	6±1
10000	19±3	22±4	10±4	3±1	Toxic	4±2

 

µg/plate	TA1538			TA98			TA100
	(-)S9	(R)S9	(H)S9	(-)S9	(R)S9	(H)S9	(-)S9	(R)S9	(H)S9
Solvent control	20±6	19±6	22±7	20±4	33±7	30±6	161±26	160±17	183±4
Positive control	677±27	750±33	449±23	269±12	1725±66	775±60	1104±36	2818±14	1469±35
333	10±2	17±3	27±7	18±2	23±6	32±4	160±7	147±20	171±17
1000	6±5	15±5	19±6	17±1	22±6	28±3	158±10	161±7	187±2
3333	5±3	13±3	13±3	13±6	16±7	22±3	158±16	169±11	205±48
6666	7±3	9±2	18±2	13±3	20±5	15±4	193±28	182±15	246±46
10000	7±1	7±5	13±3	23±11	17±2	15±7	167±31	198±17	201±11

Applicant's summary and conclusion

Conclusions:

The substance C.I. Acid blue 74 is considered to be not mutagenic in Salmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.

Executive summary:

Gene mutation toxicity study was performed to evaluate mutagenicity of C.I. Acid blue 74 by the standard plate-incorporation assay.The study was performed using S.typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100. All strain were tested at dose levels of 0, 3333, 1000, 6666 and 10000 µg/plate both with and without metabolic activation (Aroclor 1254-induced male Fischer). Positive control and solvent control as used. There was no dose-related increase in the number of revertants above spontaneous solvent controls, with a 2-fold increase for strains TA1535, TA1538, TA98 and TA100, and a 3-fold increase for TA1537. Hence, the substance C.I. Acid blue 74 is considered to be not mutagenic in Salmonella typhimurium strain TA1535, TA1537, TA1538, TA98 and TA100 with and without metabolic activation.