Ammonium sulphate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

no data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Limited documentation

Data source

Reference

Reference Type:

publication

Title:

Micronucleus test in mice on 39 food additives and eight miscellaneous chemicals.

Author:

Hayashi M et al.

Year:

1988

Bibliographic source:

Fd Chem Toxic 26: 487-500.

Materials and methods

Principles of method if other than guideline:

The maximum doses of the test compounds were determined by pilot experiments using the multisampling at multi-dose levels method according to Hayashi M et al (1984). A pilot experiment for the micronucleus test. The multi-sampling at multi-dose levels method. Mutat Res 141:, 165.

GLP compliance:

not specified

Type of assay:

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 animals

Species:

mouse

Strain:

other: ddY

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
male ddY mice
- Source: Shizuoka Agricultural Cooperative Association for Laboratory Animals, Shizuoka
- Age at study initiation: 8 weeks
- Diet (ad libitum): food pellets CE-2 (Japan Clea, Tokyo)
- Water: ad libitum
No further data.

ENVIRONMENTAL CONDITIONS: no data

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: physiol. saline

Duration of treatment / exposure:

single dose (pilot study) or 4 doses, divided by 24 hour intervals (see table below, Remarks on results)

Post exposure period:

24 hours after dosing

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

62.5 mg/kg bw/day

Remarks:

single dosing

Doses / concentrations 2

Dose / conc.:

125 mg/kg bw/day

Remarks:

single dosing

Doses / concentrations 3

Dose / conc.:

250 mg/kg bw/day

Remarks:

single dosing

Doses / concentrations 4

Dose / conc.:

500 mg/kg bw/day

Remarks:

single dosing

Doses / concentrations 5

Dose / conc.:

31.3 mg/kg bw/day

Remarks:

multiple dosing

Doses / concentrations 6

Dose / conc.:

62.5 mg/kg bw/day

Remarks:

multiple dosing

Doses / concentrations 7

Dose / conc.:

125 mg/kg bw/day

Remarks:

multiple dosing

Doses / concentrations 8

Dose / conc.:

250 mg/kg bw/day

Remarks:

multiple dosing

No. of animals per sex per dose:

6 mice per group

Control animals:

yes, concurrent vehicle

Positive control(s):

mitomycin C, 2.0 mg/kg bw

Examinations

Tissues and cell types examined:

bone marrow (femur)

Details of tissue and slide preparation:

Mice were killed by cervical dislocation at the appropriate time after an administration. Femoral marrow cells were flushed out with fetal bovine serum and smeared on clean glass slides. Cells were fixed with methanol for 5 min, and stained with Acridine Orange for the pilot experiment and with Giemsa for the full-scale test.
The preparations were coded and analysed without any knowledge of the treatment. One thousand polychromatic erythrocytes per mouse were scored using a light microscope, with a high power objective (x 100), and the number of micronucleated polychromatic erythrocytes (MNPCEs) was recorded. The proportion of polychromatic erythrocytes (PCEs) among the total erythrocytes was also evaluated by observing 1000 erythrocytes on the same slide.

Statistics:

The dose-response relationships were tested using the Cochran-Armitage trend test. A positive dose-response was considered significant at P < 0.05.

Results and discussion

Test results

Key result

Sex:

male

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

There was no statistically significant increase in the frequency of micronucleated polychromatic erythrocytes.

Compound	Route	No. of doses	Time between doses (hr)	Sampling Time (hr)	Dose level (mg/kg)	NMPCE (%)	PCE (%)	Mortality
Single dosing
Ammonium chloride	ip	1		24	0 62.5 125 250 500	0.18 +/- 0.18 0.12 +/- 0.12 0.15 +/- 0.14 0.13 +/- 0.05 0.12 + -/ 0.08	56.8 +/- 4.7 60.9 +/- 4.2 91.7 +/- 3.8 64.3 +/- 2.5 64.3 +/- 2.5	0/6 0/6 0/6 0/6 0/6
Mitomycin (positive control)	ip	1		24	2.0	4.18 +/- 1.30 ^*	52.3 +/- 4.6	0/6
Repeated dosing
Ammonium chloride	ip	4	24	24	0 31.3 62.5 125 250	0.20 +/- 0.09 0.25 +/- 0.19 0.19 +/- 0.10 0.20 +/- 0.08 0.17 +/- 0.08	59.9 +/- 8.3 67.2 +/- 13.5 63.7 +/- 4.5 64.0 +/- 9.2 61.6 +/- 6.9	0/6 0/6 0/6 0/6 0/6
Mitomycin (positive control)	ip	1		24	2.0	7.15 +/- 3.92 ^*	32.2 +/- 11.0	0/6

(*) p<0.01