Disodium 3-[(2,4-dimethyl-5-sulphonatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate

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:

other: Data is from peer reviwed journal

Data source

Reference

Reference Type:

publication

Title:

Salmonella Mutagenicity Tests: V. Results from the Testing of 311 Chemicals

Author:

Errol Zeiger, Beth Anderson, Steve Haworth, Timothy Lawlor, and Kristien Mortelmans

Year:

1992

Bibliographic source:

Environmental and Molecular Mutagenesis Volume 19, Supplement 21:2-141 (1992)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: refer below principle

Principles of method if other than guideline:

The purpose of this study is to present the results and data from the test chemical for their ability to induce mutations in tester strains of Salmonella typhimurium.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 3-[(2,4-dimethyl-5-sulphonatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate

EC Number:

224-909-9

EC Name:

Disodium 3-[(2,4-dimethyl-5-sulphonatophenyl)azo]-4-hydroxynaphthalene-1-sulphonate

Cas Number:

4548-53-2

Molecular formula:

C18H16N2O7S2.2Na

IUPAC Name:

disodium 3-[(2,4-dimethyl-5-sulfonatophenyl)diazenyl]-4-hydroxynaphthalene-1-sulfonate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Ponceau SX (C.I. Food Red 1) - Molecular formula (if other than submission substance): C18H16N2O7S2.2Na- Molecular weight (if other than submission substance): 480.4276- Smiles notation (if other than submission substance): [Na+].[Na+].Cc1cc(C)c(cc1N=Nc2cc(c3ccccc3c2O)S(=O)(=O)[O-])S(=O)(=O)[O-]- InChl (if other than submission substance): 1S/C18H16N2O7S2.2Na/c1-10-7-11(2)16(28(22,23)24)8-14(10)19-20-15-9-17(29(25,26)27)12-5-3-4-6-13(12)18(15)21;;/h3-9,21H,1-2H3,(H,22,23,24)(H,25,26,27);;/q;2*+1/p-2/b20-19+;;- Substance type: Organic- Physical state: Solid

Method

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA100, TA98, TA97 and TA 1535

Details on mammalian cell type (if applicable):

No data available

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S-9

Test concentrations with justification for top dose:

0.00,1.000, 3.000, 10.00, 33.00, 100.00,333.00,1000.00,3333.00,10000.00 µg/Plate

Vehicle / solvent:

Solvent : DMSOJustification 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:

9-aminoacridine

sodium azide

methylmethanesulfonate

mitomycin C

other: 4-nitro-o phenylenediamine

Details on test system and experimental conditions:

METHOD OF APPLICATION: pre incubationDURATION- Pre incubation period: 20 min- Exposure duration: No data available- Expression time (cells in growth medium): 2 days- 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 availableSELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: No data availableNUMBER OF CELLS EVALUATED: No data availableDETERMINATION OF CYTOTOXICITY- Method: No data availableOTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication: No data available- Other: No data available

Evaluation criteria:

Histidine- independent (his+) colonies arising on the plates were counted

Statistics:

No data available

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA100, TA98, TA97 and TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data available- Effects of osmolality: No data available- Evaporation from medium: No data available- Water solubility: No data available- Precipitation: No data available- Other confounding effects: No data availableRANGE-FINDING/SCREENING STUDIES: yes,each chemical was initially tested in the preincubation test at half-log dose intervals up to a dose that elicited toxicity, or to a dose immediately below one that was toxic in the preliminary toxicity procedure.COMPARISON WITH HISTORICAL CONTROL DATA: No data availableADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Dose	TA100
	NA (-)                                    NA (-)
µg/plate	MEAN	SEM	MEAN	SEM
0.00	107	4.6	95	6.8
1.000
3.000
10.00
33.00
100.00	90	25.0	104	8.4
333.00	117	2.4	113	3.3
1000.00	133	3.7	100	3.2
3333.00	109	8.2	121	7.9
10000.00	116	9.0	96	4.6
POS	397	8.0	502	9.2

 

 

Dose	TA100
	10% HLI (-)		30% HLI (?)		30% HLI (-)
µg/plate	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.00	107	1.2	121	4.2	110	9.4
1.000					113	3.5
3.000					100	3.8
10.00					93	3.2
33.00					115	3.7
100.00	83	4.0	163	2.9	112	2.3
333.00	104	6.8	154	9.9	120	3.2
1000.00	106	6.2	154	10.8	122	1.9
3333.00	89	5.2	150	11.2	138	6.1
10000.00	117	8.4	162	3.0	114	8.2
POS	688	6.3	702	25.8	735	106.9

 

Dose	TA100
	10% RLI (-)                30% RLI (-)
µg/plate	MEAN	SEM	MEAN	SEM
0.00	106	12.0	144	5.0
1.000
3.000
10.00
33.00
100.00	97	5.7	131	2.2
333.00	94	8.5	155	5.2
1000.00	79	4.6	155	6.3
3333.00	102	13.9	146	1.9
10000.00	94	5.6	145	7.4
POS	691	152.3	967	21.5

 

 

Dose	TA1535
	NA (-)		10% HLI (-)		30% HLI (-)		10%RLI (-)		30%RLI (-)
µg/plate	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.00	27	2.6	23	3.3	14	2.9	18	3.5	16	2.0
1.000
3.000
10.00
33.00
100.00	30	1.2	17	1.3	14	3.4	25	1.8	13	3.2
333.00	24	5.5	15	2.2	12	1.2	17	4.4	18	3.2
1000.00	27	0.9	17	2.0	12	1.2	19	0.6	13	1.2
3333.00	30	5.4	22	1.0	18	1.2	17	3.0	13	1.9
10000.00	30	3.5	13	0.9	17	0.3	22	1.5	12	2.0
POS	327	20.3	145	2.8	188	12.2	1612x		183	4.0

 

 

Dose	TA97
	NA (-)		10% HLI (-)		30% HLI (-)		10%RLI (-)		30%RLI (-)
µg/plate	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.00	113	4.7	145	10.4	141	8.4	153	0.7	163	1.7
1.000
3.000
10.00
33.00
100.00	118	8.2	129	7.6	146	9.2	135	3.8	146	4.1
333.00	122	10.7	137	2.3	136	6.7	143	10.3	144	2.3
1000.00	104	6.1	140	4.7	160	2.1	136	1.2	139	7.7
3333.00	89	25.0	146	0.9	150	7.6	134	5.3	135	4.4
10000.00	116	1.2	159	4.6	131	10.8	130	4.7	119	3.8
POS	582	16.5	932	175.2	994	65.9	2046	86.0	699	14.4

 

 

Dose	TA98
	NA (-)				10% HLI (-)		30%HLI (-)		10%RLI (-)		30 % RLI(-)
µg/plate	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.00	20	2.5	18	3.0	29	0.9	25	4.6	29	3.2	22	0.7
1.000
3.000
10.00
33.00
100.00	13	4.1	14	1.5	34	3.5	40	7.5	27	2.7	23	3.4
333.00	20	2.0	19	1.7	32	1.5	36	4.9	33	7.3	24	1.2
1000.00	17	2.2	22	3.7	41	3.5	36	1.5	31	0.7	22	1.2
3333.00	19	0.0	20	3.3	39	3.2	39	3.3	33	1.5	23	2.0
10000.00	14	1.2	23	1.9	28	2.0	28	3.3	25	3.2	14	2.7
POS	355	21.4	425	11.6	664	42.2	372	8.4	460	164.9	374	18.8

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):negativeThe end point for the genetic toxicity test was found to be negative (with and without) when treated with Ponceau SX (4548-53-2).

Executive summary:

Genetic toxicity test was performed by Ponceau SX onS typhimurium(Strain TA100, TA98, TA97 and TA 1535).Initial testing was in strain without activation and with 30% rat and hamster S-9. If a positive response was obtained in one or both strains, only the positive test condition was repeated. Each chemical was initially tested in the preincubation test at half-log dose intervals up to a dose that elicited toxicity, or to a dose immediately below one that was toxic in the preliminary toxicity procedure.The test chemical (0.05 ml), overnight culture of Salmonella (0.10 ml at CWR and MIC, and 0.05 ml at SRI), and S-9 mix or buffer (0.50 ml), were incubated at 37◦C, without shaking, for 20 min.At least five doses of each chemical were tested in triplicate, and repeat experiments were performed at least one week following the initial trial.A maximum of 0.05 ml solvent was added to each plate.Different concentrations used are 0, 1, 3.10, 33, 100, 333, 1000, 3333, 10000, µg/plates.X

 

The positive controls in the absence of metabolic activation were sodium azide (TA1535 and TA100), 9-aminoacridine (TA97 and TA 1537), 4-nitro-o phenylenediamine (TA98 and TA1538), mitomycin C (TA102), and methyl methane sulfonate (TA 104) and the positive control for metabolic activation with all strains was 2-aminoanthracene, and either sterigmatocystin or 2 aminoanthracene were used.

 

 A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his⁺revertants did not meet the criteria for a “+W” response, or if only single doses produced increases in his⁺revertants in repeat trials. Chemicals were judged nonmutagenic (-) if they did not meet the criteria for a mutagenic or questionable response.

 

From the experiment onS typhimurium(Strain TA100, TA98, TA97 and TA 1535)it was concluded that thePonceau SX(CAS No 4548-53-2) was genetically non-toxic chemical with and without activation.