Disodium 5-amino-4-hydroxy-3-(phenylazo)naphthalene-2,7-disulphonate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

4 (not assignable)

Data source

Reference

Reference Type:

secondary source

Title:

Unnamed

Year:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

D&C Red No. 33

IUPAC Name:

D&C Red No. 33

Constituent 2

Reference substance name:

Disodium 5-amino-4-hydroxy-3-(phenylazo)naphthalene-2,7-disulphonate

EC Number:

222-656-9

EC Name:

Disodium 5-amino-4-hydroxy-3-(phenylazo)naphthalene-2,7-disulphonate

Cas Number:

3567-66-6

Molecular formula:

C16H13N3O7S2.2Na

IUPAC Name:

disodium 5-amino-4-hydroxy-3-(phenylazo)naphthalene-2,7-disulphonate

Test material form:

not specified

Details on test material:

Name of test material (as cited in study report): - D&C Red No. 33- Molecular formula (if other than submission substance): C16-H13-N3-O7-S2.2Na- Molecular weight (if other than submission substance): 467.3889 g/mol- Substance type: Organic - Physical state: No data available.Purity- 93 %- Impurities (identity and concentrations): No data available.

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

No data available.

Administration / exposure

Route of administration:

oral: unspecified

Vehicle:

- Vehicle(s)/solvent(s) used: Deionised water

Details on exposure:

No data available.

Duration of treatment / exposure:

48 hour

Frequency of treatment:

once

Post exposure period:

No data available.

Doses / concentrations

Remarks:

Doses / Concentrations:500, 1000 and 2000 mg/kg bwBasis:

No. of animals per sex per dose:

Total no of animals-30 500 mg/kg bw- 5 male and 5 female1000 mg/kg bw-5 male and 5 female2000 mg/kg bw- 5 male and 5 female

Control animals:

yes, concurrent vehicle

Positive control(s):

No data available.

Examinations

Tissues and cell types examined:

Erythrocytes were examined.

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: The test concentrations were based on the result of a pre-experiment for toxicity in which 2 mice were orally exposed to 2000 mg/kg bw D&C Red 33. The animals were examined for acute toxic symptoms at intervals of around 1, 2-4, 6, 24, 30, and 48h after administration of D&C Red 33. 2000 mg/kg was selected as the maximum tolerated dose level.TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): The bone marrow cells were collected 24h and 48h (highest dose only) after dosing.DETAILS OF SLIDE PREPARATION: Bone marrow preparations were stained and examined microscopically for the NCE/PCE ratio and micronuclei.METHOD OF ANALYSIS: Toxicity and thus exposure of the target cells was determined by measuring the ratio between normo-chromatic to polychromatic erythrocytes (PCE/NCE ratio).OTHER:

Evaluation criteria:

The number of micro nucleated PCEs compared to the concurrent vehicle.

Statistics:

No data available.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDYThe test concentrations were based on the result of a pre-experiment for toxicity in which 2 mice were orally exposed to 2000 mg/kg bw D&C Red 33. The animals were examined for acute toxic symptoms at intervals of around 1, 2-4, 6, 24, 30, and 48h after administration of D&C Red 33. 2000 mg/kg was selected as the maximum tolerated dose level. In the pre-experiment for toxicity, all animals expressed toxic effects like reduction ofspontaneous activity, abdominal position, eyelid closure and ruffled fur 1 and 2-4 h after treatment. From 6h after treatment these toxic effects decreased and were lost at 30 h.RESULTS OF DEFINITIVE STUDY- Types of structural aberrations for significant dose levels (for Cytogenetic or SCE assay):- Induction of micronuclei (for Micronucleus assay): D&C Red 33 did not induce micronuclei in bonemarrow cells of treated mice.- Ratio of PCE/NCE (for Micronucleus assay): yes - Appropriateness of dose levels and route:- Statistical evaluation:

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negativeThe result was found to be negative for D&C Red 33 in male and female NMRI mice for Mammalian Erythrocyte Micronucleus Test.

Executive summary:

In gene toxicity study for D&C Red 33was observed in male and female NMRI mice by Mammalian Erythrocyte Micronucleus Test. Mice were exposed at the concentration of500, 1000 and 2000 mg/kg bw. Bone marrow cells were collected 24h and 48h (highest dose only) after dosing. Toxicity and thus exposure of the target cells was determined by measuring the ratio between normo-chromatic to polychromatic erythrocytes (PCE/NCE ratio). The animals of the highest dose group were examined for acute toxic symptoms at intervals around 1, 2-4, 6 and 24 h after treatment. In order to quantify the concentration of D&C Red 33 in blood serum 2 animals per sex were treated with 2000 mg/kg bw D&C Red 33. 1 and 4 h after treatment the animals were sacrificed, their blood was collected and analyzed by HPLC. Clinical signs like reduction of spontaneous activity, abdominal position, eyelid closure and ruffled fur were observed after 1 and 2-4 h treatment. From 6h after treatment these toxic effects decreased and were lost at 30 h. The urine colour of the treated animals was red which together with the observed toxic effects indicate the systemic distribution and thus bioavailability in the bone marrow of D&C Red 33. This was confirmed by the serum analysis showing substantial amounts of D&C Red 33 in the serum 1 h after treatment; after 4 h the levels dropped below the detection limit. Biological relevant increases in the number of micro nucleated PCEs compared to the concurrent vehicle controls were not found following treatment with D&C Red 33 at any time point. Therefore it was considered that D&C Red 33 did not induce micronuclei in bone marrow cells of treated mice. The result was found to be negative for D&C Red 33for Mammalian Erythrocyte Micronucleus Test.