Lithium amide

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

publication

Title:

Micronucleus Tests in Mice on 39 Food Additives and Eight Miscellaneous Chemicals

Author:

Hayashi, M. et al.

Year:

1988

Bibliographic source:

Food and Chemical Toxicology, 26(6): 487-500

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

not specified

Type of assay:

other: micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Ammonium chloride

EC Number:

235-186-4

EC Name:

Ammonium chloride

Cas Number:

12125-02-9

Molecular formula:

ClH4N

IUPAC Name:

ammonium chloride

Test material form:

solid

Test animals

Species:

mouse

Strain:

other: ddY strain

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Shizuoka Agricultural Cooperative Association for Laboratory Animals, Japan
- Age at study initiation: 8 weeks
- Diet: ad libitum, CE-2 pellets (Japan Clea, Tokyo)
- Water: ad libitum

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle used: no data

Duration of treatment / exposure:

24 hours

Frequency of treatment:

singly within 24 hours; and 4 injections within 24 hours

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

31.3 mg/kg bw (total dose)

Doses / concentrations 2

Dose / conc.:

62.5 mg/kg bw (total dose)

Doses / concentrations 3

Dose / conc.:

125 mg/kg bw (total dose)

Doses / concentrations 4

Dose / conc.:

250 mg/kg bw (total dose)

Doses / concentrations 5

Dose / conc.:

500 mg/kg bw (total dose)

No. of animals per sex per dose:

5

Control animals:

yes

Examinations

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
The doses were determined via pre-tests. Up to the maximum tolerated dose was used.

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Two experimental set ups were performed. A single dose was applied in the first experiment at dose levels of 0, 62.5, 125, 250 or 500 mg/kg bw. In the second experiment four injections were given within 24 hours using dose leves of 31.3, 62.5, 125 or 250 mg/kg bw.

DETAILS OF SLIDE PREPARATION: The bone marrrow cells were rinsed with foetal bovine serum. Afterwards the cells were fixed with methanol and stained with Giemsa.

Results and discussion

Test results

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

not examined

Additional information on results:

RESULTS OF DEFINITIVE STUDY
Micronucleated erythrocytes occurred to a degree of 0.12 % after single injection. 0.17 % were observed after four injections.

Applicant's summary and conclusion

Conclusions:

No genotoxicity was observed during the micronucleus test with ammonium sulfate.

Executive summary:

In a mouse bone marrow micronucleus assay equivalent or similar to OECD guideline 474, 5 male ddY mice per dose were treated intraperitoneal (ip) with ammonium chloride at doses of 0, 62.5, 125, 250 or 500 mg/kg bw or as four injections within 24 hours at dose levels of 31.3, 62.5, 125 or 250 mg/kg bw. Bone marrow cells were harvested at 24 hours post treatment. No vehicle was indicated. The maximum tolerated dose was tested. 1000 PCEs were evaluated per animal and the number of micronucleated erythrocytes was recorded. Based on the findings no evidence of genotoxicity has been observed under the experimental conditions described.