Disodium 8-hydroxynaphthalene-1,6-disulphonate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from peer reviwed journal

Justification for type of information:

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

Specific details on test material used for the study:

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): Not applicable
- Molecular weight (if other than submission substance): Not applicable
- Substance type: Organic

Method

Target gene:

Histidine

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 : DMSO
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:

9-aminoacridine

sodium azide

methylmethanesulfonate

mitomycin C

other: 4-nitro-o phenylenediamine

Details on test system and experimental conditions:

METHOD OF APPLICATION: pre incubation
DURATION
- 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 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

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 available

RANGE-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 available
ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: No mutagenic effect were observed

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:

The 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  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. No mutagenic effect were observed .From the experiment on Salmonella 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.