Sodium 4-[[4-[(4-hydroxy-2-methylphenyl)azo]phenyl]amino]-3-nitrobenzenesulphonate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian germ cell study: gene mutation

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

19th May to 2nd July 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1983

Report date:

1983

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: Salamone, Heddle and Katz (1981) Mutagenic activity of 41 compounds in the in vitro micronucleus assay. Prog.Mut.Res.Vol 1, p.686-697.

Deviations:

no

Principles of method if other than guideline:

The test is based on the following principles:
-After chromosomal damage has been induced by test compound or its metabolites, acentric fragments of chromosomal material lag behind at anaphase. After telophase a large portion of the fragments is not included in the main nucleus of the cell, hence micronuclei are formed.
Micronuclei can be formed in a wide variety of cell types, but in this test system erythrocytes are observed because micronuclei can easily be detected in this cell type since the nucleus proper is extruded during maturation.

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Sodium 4-[[4-[(4-hydroxy-2-methylphenyl)azo]phenyl]amino]-3-nitrobenzenesulphonate

EC Number:

274-950-1

EC Name:

Sodium 4-[[4-[(4-hydroxy-2-methylphenyl)azo]phenyl]amino]-3-nitrobenzenesulphonate

Cas Number:

70865-20-2

Molecular formula:

C19H16N4O6S.Na

IUPAC Name:

sodium 4-[[4-[(4-hydroxy-2-methylphenyl)azo]phenyl]amino]-3-nitrobenzenesulphonate

Test material form:

solid: particulate/powder

Details on test material:

Acid Yellow 199

Test animals

Species:

mouse

Strain:

other: C57BL/6J

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal Breeding Unit, Alderley Park, Macclesfield, Cheshire
- Age at study initiation: 6-8 weeks
- Assigned to test groups randomly: [no/yes, under following basis: ]
- Fasting period before study:
- Housing: 5 per cage
- Diet (ad libitum): Porton Combined Diet [PCD] supplied by BP Nutrition Ltd, Stepfield, Witham, Essex, UK
- Water (ad libitum): tap water
- Acclimation period: yes

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21-23
- Humidity (%): 39-76
- Air changes (per hr): 22
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 19th May to 2nd July 1982

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: Kraft Wesson Corn Oil (Kraft Foods Ltd, Liverpool, UK)
- Justification for choice of solvent/vehicle: limited solubility in water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The est substance was suspended in corn oil

Duration of treatment / exposure:

72 hours

Frequency of treatment:

Animals were dosed with 2 consecutive Intraperitoneal Injections, administered 24 hours apart

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day

Doses / concentrations 2

Dose / conc.:

625 mg/kg bw/day

No. of animals per sex per dose:

15 males and 15 females per dose group
5 animals/sex each were sacrificed 24, 48, 72 hours after the last treatment

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide
- Justification for choice of positive control(s): according to the used method
- Route of administration: intraperitoneal

- Doses / concentrations: 75 mg/kg

Examinations

Tissues and cell types examined:

500 mg polychromatic erythrocytes were examined and the number containing micronuclei scored. The sample was also examined for any evidence of cytotoxicity.

Details of tissue and slide preparation:

The animals were killed by cervical dislocation after the last dose of compound.
Femurs were removed and stripped clean of muscle.
The iliac end of the femur was removed and a fine paint brush wetted with a solution of albumen (6% v/v in saline) was dipped into marrow canal.
3 to 4 streaks of marrow suspension were then applied to appropriately labelled clean, dry microscope slides.
The slides were allowed to air dry.
The slides were then stained in Wright's stain using an Ames Hema-Tek staining machine (Hema-Tek, Miles Laboratory Limited, Stoke Court, Stoke Poges, Slough, UK)
Slides were coded and scored blind.

Evaluation criteria:

One of the evaluation criteria is that, the test is considered positive if: The test item induces any statically significant increase in the frequency of the micronuclei when compare to the control animals.

Statistics:

The results were analysed for significant differences from the control using a one slide Student's "t" test.

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Acid Yellow 199 gave negative results at all three sampling times, even though there were a number of deaths which occurred in the higher dose level animals. These deaths were unexpected based on the LD50/7T, data and were probably due to administration of the compound as a split dose rather than a single treatment. However, the 625mg/kg dose group would qualify as a maximum tolerated dose at the 48 and 72hr sampling times.

Applicant's summary and conclusion

Conclusions:

The results indicate that Acid Yellow 199 has no clastogenic activity in the mouse micronucleus test.

Executive summary:

Acid Yellow 199 was tested for clastogenic activity in the mouse micronucleus test using C57BL/6J mice. The test item did not induce any statistically significant increase in the frequency of the micronuclei when compared to control animals. Throughout the study the positive control substance, cyclophosphamide gave the expected responses. The results indicate that Acid Yellow 199 has no clastogenic activity in the mouse micronucleus test.