4',5'-dibromo-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

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 reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Mutagenicity of Some Lipsticks and Their Dyes

Author:

Marie R. Green and Jullia V. Pastewka

Year:

1980

Bibliographic source:

Journal of the National Cancer Institute (1980) 64 (3): 665-669.

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as below

Principles of method if other than guideline:

Twenty-four lipsticks of various shades and colors were tested for mutagenicity with the histidine-requiring tester strain Salmonella typhlmurlum TA98 of which the test compound of interest was D & C Orange no. 5.

GLP compliance:

not specified

Type of assay:

bacterial gene mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

4',5'-dibromo-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

EC Number:

209-876-0

EC Name:

4',5'-dibromo-3',6'-dihydroxyspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

Cas Number:

596-03-2

Molecular formula:

C20H10Br2O5

IUPAC Name:

4',5'-dibromo-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one

Details on test material:

- Name of test material: D & C Orange no. 5- Molecular formula: C20H10Br2O5- Molecular weight: 490.10 g/mol- Substance type: Organic

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

rat liver S9

Test concentrations with justification for top dose:

1, 10, 100, 250, 500 µg/plate

Vehicle / solvent:

DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

7,12-dimethylbenzanthracene

2-nitrofluorene

Remarks:

For without S9: 2-Nitrofluorene For with S9: 7,12-dimethylbenz[a]anthracene (DMBA)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar DURATION- Preincubation period: no data available- Exposure duration: 15 hours- 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 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: mitotic index; cloning efficiency; relative total growth; other: no data availableOTHER EXAMINATIONS:- Determination of polyploidy: no data available- Determination of endo replication: no data available- Other:

Evaluation criteria:

Dose responses in terms of the numbers of His+/ revertants were observed.

Statistics:

Mean and standard deviation

Results and discussion

Test results

Test results 1

Species / strain:

S. typhimurium TA 98

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

positive

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

Remarks on result:

other: strain/cell type:

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Compound	Amount/plate, µg	No of revertants/plate
		-S9	+S9
DMSO	0	26±5	47±7
D & C orange no. 5	1	26±2	ND
	10	29±1	ND
	100	58±14	43±7
	250	139±10	44±9
	500	226±20	42±5

·        Lipsticks containing D&C Orange No. 5 would probably not exhibit mutagenicity at the concentrations tested, inasmuch as the response with this dye was seen only at and above the 100-µg level.

·        Lipstick E, containing D&C Orange No.5, was not mutagenic in the absence of S9 mix.

·        They observed mutagenicity for D&C Orange No.5. This fluorescent dye may also contain other fluorescent dyes that have been shown to damage DNA of other microbial systems as a result of photodynamic action. It is possible, therefore, that these or other impurities may account their results.

·        DMSO solvent control (-S-9): 26±5 revertant/plate (30 plates); DMSO solvent control (+S-9): 47±7 revertants/plate (18 plates).

Positive controls: 20µg DMBA (+S-9), 344±62 revertants/plate (18 plates); 5µg 2-nitrofluorene (-S-9), 1,123±169 revertants/plate (3 plates);

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):negative without metabolic activationpositive with metabolic activationThe test material D & C Orange no. 5 is mutagenic in the tester strain Salmonella typhimuriumTA98 in the absence of S9 metabolic activation system. However, the mutagenic response with this dye was observed only at and above the 100µg level.D & C Orange no. 5 is non-mutagenic in the tester strain Salmonella typhimuriumTA98 in the presence of S9 metabolic activation system.

Executive summary:

Some lipsticks and their dyes were tested for mutagenicity using S. typhimurium strain TA98. One of these dyes was the test chemical D & C Orange no. 5.

 

The standard quantitative mutagenicity assay in which the bacteria are incorporated into an agar overlay with the test agent was performed.

 

2-Nitrofluorene was used as a positive control in the absence of S-9. DMBA was used as a positive control in the presence of S-9. DMSO was used as a solvent control.

 

D&C Orange Nos. 5 was mutagenic without S-9. Dose responses in terms of the numbers of His+ revertants were observed for this dye in three experiments. The mutagenic effect was diminished or lost in the presence of S-9.

 

The test material D & C Orange no. 5 is mutagenic in the tester strain Salmonella typhimurium TA98 in the absence of S9 metabolic activation system. However, the mutagenic response with this dye was observed only at and above the 100µg level. However, this fluorescent dye may also contain other fluorescent dyes that have been shown to damage DNA of other microbial systems as a result of photodynamic action. It is possible, therefore, that these or other impurities may account their results.

 

D & C Orange no. 5 is non-mutagenic in the tester strain Salmonella typhimurium TA98 in the presence of S9 metabolic activation system.