9-[2-(ethoxycarbonyl)phenyl]-3,6-bis(ethylamino)-2,7-dimethylxanthylium, salt with 4,5-dihydro-5-oxo-1-(4-sulphophenyl)-4-[(4-sulphophenyl)azo]-1H-pyrazole-3-carboxylic acid (3:1)

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from publication.

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity Testing of Di (2- ethylhexyl)phthalate and Related Chemicals in Salmonella

Author:

Errol Zeiger, Steve Haworth, Kristien Mortelmans, and William Speck

Year:

1985

Bibliographic source:

Environmental Mutagenesis 7:213-232 (1985)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: AS mention below

Principles of method if other than guideline:

To evaluate the mutagenic potential of Dioctyl phthalate in Salmonella Typhimurium strains TA1535, TA1537, TA98, and TAl00 by Ames test.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Dioctyl phthalate

EC Number:

204-214-7

EC Name:

Dioctyl phthalate

Cas Number:

117-84-0

Molecular formula:

C24H38O4

IUPAC Name:

1, 2-dioctyl benzene-1, 2-dicarboxylate

Specific details on test material used for the study:

Details on test material
- Name of test material (as cited in study report): Dioctyl phthalate
- Substance type: Organic

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537, TA98, and TAl00

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

Liver homogenates (S-9 fraction) from Aroclor 1254-induced male Sprague-Dawley rats (RLI) or male Syrian hamsters (HLI).

Test concentrations with justification for top dose:

100-10000µg/plate

Vehicle / solvent:

Vehicle
- Vehicle(s)/solvent(s) used: DMSO

Controls

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: TA1535 and TA100, sodium azide; TA98, 4-nitro-o-phenylenediamine;a nd TA1537, 9-aminoacridine. 2-Aminoanthracene was used as the positive control for metabolic activation in all strains.

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: Preincubation method

DURATION
- Preincubation period: 20 min
- Exposure duration: 3Days

Rationale for test conditions:

Not specified.

Evaluation criteria:

The strains were evaluated for dose dependent increase in the number of reveretants\plate.

Statistics:

Mean ±Standard deviations for strains were observed.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: strains TA1535, TA1537, TA98, and TAl00

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:The final dose level selection was based on the results of a preliminary range-finding study conducted with TAl00 in the presence and absence of S-9.

Remarks on result:

other: No mutagenic effect were observed.

Any other information on results incl. tables

Table 17:Di-Octyl Phthalate (Lab:SRI solvent : DMSO)

TA lOO

Dose	NA		10% HLI (-)		10%RLI (-)
µg/PLATE	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.000	114	4.3	118	5.0	103	5.4
100.000	124	10.9	120	4.7	109	11.7
333.000	103	3.8	105	6.9	105	10.7
1000.000	105	4.0	109	5.7	110	0.6
3333.000	97	7.0	105	2.6	101	7.2
10000.000	111	7.9	100	3.2	107	12.4
POS	331	13.6	1290	45.3	551	3.5

 

 

 

 

 

 

TA1535

Dose	NA		10% HLI (-)		10%RLI (-)
µg/PLATE	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.000	24	2.3	8	3.0	12	1.3
100.000	23	4.0	13	2.0	11	1.2
333.000	21	5.7	6	1.2	7	2.5
1000.000	22	4.0	12	2.2	8	2.4
3333.000	19	2.9	10	1.8	12	2.3
10000.000	17	1.5	10	2.5	7	0.7
POS	209	16.2	359	24.8	279	18.0

 

 

 

 

 

 

TA1537

Dose	NA		10% HLI (-)		10%RLI (-)
µg/PLATE	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.000	7	0.0	4	0.7	10	2.5
100.000	6	1.5	6	0.3	11	1.8
333.000	5	2.0	7	0.6	8	2.4
1000.000	7	0.9	9	2.0	5	0.6
3333.000	7	0.9	6	2.8	10	2.2
10000.000	4	1.2	5	0.7	9	3.7
POS	269	14.7	345	15.0	244	17.8

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

TA98

Dose	NA		10% HLI (-)		10%RLI (-)
µg/PLATE	MEAN	SEM	MEAN	SEM	MEAN	SEM
0.000	13	0.9	31	3.0	25	0.9
100.000	14	1.7	33	1.8	30	3.3
333.000	13	2.6	29	5.3	29	1.9
1000.000	18	1.8	29	1.9	26	1.2
3333.000	18	2.1	30	4.7	17	2.7
10000.000	18	1.5	26	3.8	20	2.4
POS	621	56.6	1027	53.1	382	27.0

 

Applicant's summary and conclusion

Conclusions:

Dioctyl phthalate (117-84-0) was evaluated for its mutagenic potential in Salmonella Typhimurium strains TA1535, TA1537, TA98, and TAl00 by Ames test. The test result was considered to be negative in the presence and absence of S9.

Executive summary:

Dioctyl phthalate was assessed for its possible mutagenic potential. For this purpose bacterial reverse mutation assay was performed on Salmonella Typhimurium strains TA1535, TA1537, TA98, and TAl00. The test substance was exposed at the test concentration of 100-10000µg/plate byPreincubation method. The test was conducted in the presence and absences of S9.No mutagenic effect were observed in all strain both in the presence and absence of metabolic activation. ThereforeDioctyl phthalate was considered to be non mutagenic in bacterial reverse mutation assay. Hence the test substance cannot be classified as gene mutant in vitro.