2-(3,6-dihydroxy-2,4,5,7-tetraiodoxanthen-9-yl)benzoic acid, aluminium salt

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:

weight of evidence

Reliability:

2 (reliable with restrictions)

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:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

Mutagenicity testing was performed by the standard plate-incorporation assay for C I -ACID RED 51 in Salmonella typhimurium by using TA1535, TA1537, TA1538, TA98 and TA100 strains.

GLP compliance:

no

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

EC Number:

240-474-8

EC Name:

Disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

Cas Number:

16423-68-0

Molecular formula:

C20H8I4O5.2Na

IUPAC Name:

disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

Constituent 2

Reference substance name:

erythrosine BS

IUPAC Name:

erythrosine BS

Details on test material:

Details on test material
- Name of test material (as cited in study report): C I ACID RED 51
- Molecular formula (if other than submission substance): C20-H6-I4-O5.2Na
- Molecular weight (if other than submission substance): 879.8424 g/mol
- Substance type: Organic
- Physical state: Solid(Powder)
-Purity: 90.5% dye
- Impurities (identity and concentrations): 6.6%H20
2 0% NaCl
0.3% Na2SO4

Method

Target gene:

S.typhimurium mutagenicity assay

Species / strain

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

The test concentration were
33 .0µg/plate
100 .0µg/plate
333.0 µg/plate
1000.0µg/plate
3333.0µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: The solvents used were water, dimethyl sulfoxide, and acetone (exact solvent was not mention)

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: without metabolic activation the positive controls were as fallow:Sodium azide for strains TA1535 and TA100; 9-aminoacridine for TA1537; 2-nitrofluorene for TA1538 and TA98 with metabolic activationaminoanthracene for TA1535, TA1537, TA1538, TA98 TA100

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: 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 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: mitotic index; cloning efficiency; relative total growth; other: No data available

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

OTHER: The chemical was tested at 5 dose levels in triplicate.

Evaluation criteria:

A response was considered positive if there was a
dose-related increase in the number of revertants above spontaneous solvent controls

Statistics:

No data available

Results and discussion

Test results

Species / strain:

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

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:

RANGE-FINDING/SCREENING STUDIES: The range of
concentrations for testing was based on preliminary
toxicity tests in which the viability of the bacterial cells on complete medium was measured at concentrations up to 10 mg/plate or to the limit of solubility. When solubility and toxicity were not limiting factors, the maximum concentration tested was 10 mg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Other:

S. typhimuriumASSAY

Dose(µg/plate)	Revertants per plate
	TA1535
	(-S)S9                                   (R)S9                            (H)S9
Solvent Cont.	30±7	16±6	20±6
Positive Cont.^b	380±6	198±17^2	119±7
33.00	21±3	19±3	18±2
100.00	28±3	21±6	21±4
333.00	19±4	15±2	16±2
1000.00	15±4	9±1	10±1
3333.00	14±2	13±3	11±3

 

Dose(µg/plate)	Revertants per plate
	TA1537
	(-S)S9                                   (R)S9                            (H)S9
Solvent Cont.	5±2	9±5	9±3
Positive Cont.^b	199±30	36±28^2	79±6
33.00	5±4	7±2	9±8
100.00	8±3	10±3	5±2
333.00	6±3	6±3	9±1
1000.00	5±1	5±2	7±6
3333.00	5±1	4±4	7±2

 

 

Dose(µg/plate)	Revertants per plate
	TA1538
	(-S)S9                                   (R)S9                            (H)S9
Solvent Cont.	15±3	19±4	22±3
Positive Cont.^b	191±16^1	1040±33^2	525±15
33.00	16±4	22±5	14±9
100.00	12±5	16±4	14±9
333.00	12±2	21±7	15±4
1000.00	9±2	14±6	19±4
3333.00	11±1	14±1	10±5

 

 

Dose(µg/plate)	Revertants per plate
	TA98
	(-S)S9                                   (R)S9                            (H)S9
Solvent Cont.	21±3	34±5	34±3
Positive Cont.^b	196±34^2	1086±74^2	991±25
33.00	18±4	25±11	24±2
100.00	17±5	27±5	26±6
333.00	19±4	31±2	25±9
1000.00	15±5	23±6	20±6
3333.00	13±4	12±2	20±7

 

Dose(µg/plate)	Revertants per plate
	TA100
	(-S)S9                                   (R)S9                            (H)S9
Solvent Cont.	171±14	156±13	191±26
Positive Cont.^b	571±7	1630±63^2	1214±61
33.00	179±21	138±2	160±21
100.00	168±11	136±10	189±21
333.00	106±10	163±20	179±34
1000.00	117±35	164±14	175±7
3333.00	102±33	138±5	171±9
^aMean and standard deviation. ^bin assays without metabofic activation, the positive controls were sodium azide for TA1535 and TA100, 1.0 µg/plate, 9-aminoacridine for TA1537, 50.0 µg/plate; 2-nitrofluorene for TA1538, 1.0 or 5.0 µg/plate, and for TA98, 2.0 or 5.0 µg/plate. In those assays in which the lower dose of 2-nitrofluorene was used, the positive control value is followed by a superscript indicating the dose level used (either^1or^2). In assays with rat-liver S9, the positive control for all strains was 2-aminoanthracene, 2.5 µg/plate except for those positive control values, followed by a numerical superscript. This number is the dose used as a positive control. In assays with hamster-liver S9, the positive control for all strains was 2-aminoanthracene, 1.0 µg/plate, except for those positive controls followed by the superscript "c". In these cases 2.5µg/plate was used. 1 Dose level was 1.0 µg/plate. 2 Dose level was 2.0 µg/plate. 5 Dose level was 5.0 µg/plate. c Dose level was 2.5 µg/plate. d Only 2 plates

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative Negative (with and without S9 mix)

The dye C. I -ACID RED 51 was found to be negative ti induce gene mutation with and without S9 mix in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 by Plate-incorporation assay.

Executive summary:

Bacterial gene mutation assay was performed for the test material C. I -ACID RED 51 inSalmonella typhimuriumTA1535, TA1537, TA1538, TA100 and TA98 strains at a dose range of 33.0, 100.0, 333.0, 1000.0and3333.0µg/plate by Plate-incorporation method.

Chemical was tested without metabolic activation and with S9 mix from Aroclor 1254-induced male Fischer 344 rats and Syrian golden hamsters. Appropriate positive and solvent controls were also incorporated in the study.

C. I -ACID RED 51 failed to induce gene mutation in the bacterial strains TA1535, TA1537, TA1538, TA100 and TA98.

According to the CLP regulation, the test material C. I -ACID RED 51 does not classify as a gene mutant.