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Toxicological information

Genetic toxicity: in vitro

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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:
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

Reference
Name:
Unnamed
Type:
Constituent
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 resultsopen allclose all
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
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.