Sodium 3-(p-anilinophenylazo)benzenesulphonate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Data source

Reference

Reference Type:

publication

Title:

Effects Of Dyes (Indigo Carmine, Metanil Yellow, Fast Green FCF) And Nitrite In Vivo In Bone Marrow Chromosomes Of Mice

Author:

Ashok Kumar Giri, Tara Shankar Banerjee, Geeta Talukder And Archana Sharma

Year:

1986

Bibliographic source:

Cancer Letters, 30 (1986) 315-320

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

In vivo mammalian chromosome aberration assay was performed to determine the clastogenic nature of Metanil yellow

GLP compliance:

not specified

Type of assay:

other: In vivo mammalian bone marrow chromosome aberration assay

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 3-(p-anilinophenylazo)benzenesulphonate

EC Number:

209-608-2

EC Name:

Sodium 3-(p-anilinophenylazo)benzenesulphonate

Cas Number:

587-98-4

Molecular formula:

C18H14N3O3S.Na

IUPAC Name:

sodium 3-(p-anilinophenylazo)benzenesulphonate

Test material form:

solid

Details on test material:

- Name of test material: Ext. D&C Yellow No. 1
- IUPAC name: sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
- Molecular formula: C18H15N3O3SNa
- Molecular weight: 375.383 g/mol
- Substance type: organic
- Physical state: solid
- Purity: No data available
- Impurities (identity and concentrations): no data available

Specific details on test material used for the study:

Name of test material: Metanil yellow
- IUPAC name: sodium 3-[(4-anilinophenyl)diazenyl]benzenesulfonate
- Molecular formula: C18H15N3O3SNa
- Molecular weight: 375.383 g/mol
- Substance type: organic
- Physical state: solid
- Purity: No data available
- Impurities (identity and concentrations): no data available

Test animals

Species:

mouse

Strain:

Swiss

Remarks:

Albino

Details on species / strain selection:

No data

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Laboratory bred animals were used for the study
- Age at study initiation: Between 90-100 days
- Weight at study initiation: 30 g approx..
- Assigned to test groups randomly: No data
- Fasting period before study: No data
- Housing: No data
- Diet (e.g. ad libitum): No data
- Water (e.g. ad libitum): No data
- Acclimation period: No data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data
- Humidity (%):No data
- Air changes (per hr): No data
- Photoperiod (hrs dark / hrs light): No data

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: Distilled water
- Justification for choice of solvent/vehicle: The chemical was soluble in distilled water
- Concentration of test material in vehicle: 0 or 2 mg/Kg bw
- Amount of vehicle (if gavage or dermal): 0.05mL
- Type and concentration of dispersant aid (if powder): No data
- Lot/batch no. (if required): No data
- Purity: No data

Details on exposure:

For oral route
PREPARATION OF DOSING SOLUTIONS: Metanil yellow was dissolved in distilled water at dose level of 2 mg/Kg bw

DIET PREPARATION
- Rate of preparation of diet (frequency): No data
- Mixing appropriate amounts with (Type of food): No data
- Storage temperature of food: No data

Duration of treatment / exposure:

30 days

Frequency of treatment:

Once, daily

Post exposure period:

No data

Doses / concentrations

Remarks:

0 or 2 mg/Kg bw

No. of animals per sex per dose:

Total: 20
0 mg/Kg bw: 10 mice
2 mg/Kg bw: 10 mice

Control animals:

yes, concurrent vehicle

Positive control(s):

No data

Examinations

Tissues and cell types examined:

Bone marrow chromosomes

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: The dose in this experiment was calculated based on the idea that 200 ppm of the dye is allowed in food in India. The dose selected for dye depends on the assumption that a person can consume a maximum of about 100 mg of dye from foods every day. If a person weighing 50 kg consumes 100 mg of dye or nitrite per day then automatically the value comes to 2 mg/kg. These doses may appear a bit higher than normal human consumption, but
since the investigation was limited to only 30 days, it is conjectured that these doses were justified if the cumulative effects of these dyes on human beings for years was considered.

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Afetr 30 days treatment with metanil yellow

DETAILS OF SLIDE PREPARATION: Colchicine (0.04%) was injected intraperitoneally at the rate of 1 ml/100 g body wt, 90 min before the animal was killed. For chromosome studies, flame dried preparations of bone marrow chromosomes were stained in Giemsa by the usual schedule

METHOD OF ANALYSIS: No data

OTHER: Chromosomal aberrations were scanned at the metaphase stage.

Evaluation criteria:

Chromatid and chromosomal breaks, chromatid and chromosomal exchanges and cells with more than 10 aberrations were considered basically abnormal and hence scored accordingly.

The number of breaks per metaphase analysed Z/B, represents the fundamental index for comparison. In the Z/B category were the chromatid and chromosomal breaks, exchanges, dicentric chromosomes and cells with more than 10 aberrations. In qualitative evaluation the number of breaks were added in the following way: chromatid break, 1 break; chromosomal break, 1 break; chromosomal exchange, 2 breaks; dicentric chromosomes, 2 breaks; cells with more than 10 aberrations, 10 breaks. Gaps were not included

Statistics:

No data

Results and discussion

Test results

Sex:

male

Genotoxicity:

positive

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

No data

Any other information on results incl. tables

TABLE 1

Chromosomal Aberrations Induced by Metanil yellow on Bone Marrow Cells of Mice

Substance	Dose (mg/Kg bw)	No. of animals	n^a	Aberrant cells		Breaks	Cells with 10 aberrations	Z/B^b
				No.	%``
Distilled water (control)	0	10	600	11	1.83	12	2	0.053
Metanil yellow	2	10	600	89	14.83	44	55	0.990

^an = number of cells analysed.

^bZ/B = number of aberrations per cell

Applicant's summary and conclusion

Conclusions:

Metanil yellow induced chromosome aberrations in the bone marrow of Swiss albino male mice at a dose level of 2 mg/Kg bw and hence it is likely to be mutagenic in vivo.

Executive summary:

In vivo mammalian chromosome aberration assay was performed to determine the clastogenic nature of Metanil yellow. Laboratory bred 10 Swiss albino male mice were given metanil yellow in distilled water by gavage at dose level of 0 or 2 mg/Kg bw once daily for 30 days.Controls were fed 0.05 ml of distilled water daily for 30 days. The animals were killed on the completion of 30 days treatment, starting from 0 day. Colchicine (0.04%) was injected intraperitoneally at the rate of 1 ml/100 g body wt, 90 min before the animal was killed.

 

For chromosome studies, flame dried preparations of bone marrow chromosomes were stained in Giemsa by the usual schedule. A total of 600 metaphase plates were scanned from the 10 animals killed in each set of experiments and the slides were observed for chromatid and chromosomal breaks, chromatid and chromosomal exchanges and number of breaks per metaphase was analysed by the Z/B ratio.

 

A significant increase in the frequency of the chromosomal aberrations (index Z/B) was found with metanil yellow treated series when compared with the controls.

 

Metanil yellow induced chromosome aberrations in the bone marrow of Swiss albino male mice at a dose level of 2 mg/Kg bw and hence it is likely to be mutagenic in vivo.