Alanine, N,N-bis(carboxymethyl)-, sodium salt (1:3)

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

yes (incl. QA statement)

Remarks:

BASF

Type of assay:

other: mammalian micronucleus assay in vivo

Test material

Test material information

Constituent 1

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 3043/V9Z

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River GmbH, WIGA, Sulzfeld, FRG
- Assigned to test groups randomly: yes
- Fasting period before study: no
- Housing: small groups for acclimatization, individually after administration
- Identifications of animals: cage cards
- Diet: (Kliba Haltungsdiät, Klingentalmühle AG, Kaiseraugst, Switzerland; ad libitum
- Water: from bottles, ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature: 20 – 24°C
- Humidity: 30-70 %
- Air changes (per hr): fully air-conditioned
- Photoperiod (hrs dark / hrs light): 12 h / 12 h

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: immediately before administration; all concentrations were administered at 10 mL volume.

Frequency of treatment:

single treatment

Post exposure period:

24 h (all concentrations and controls); 48 h (negative control and 2000 mg/kg)

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (actual dose received)

Doses / concentrations 2

Dose / conc.:

500 mg/kg bw/day (actual dose received)

Doses / concentrations 3

Dose / conc.:

1 000 mg/kg bw/day (actual dose received)

Doses / concentrations 4

Dose / conc.:

2 000 mg/kg bw/day (actual dose received)

No. of animals per sex per dose:

5/sex (vehicle ctrl.; all dose groups) 5 animals (2 males and 3 females or 3 males and 2 females) for pos. ctrl.

Control animals:

yes, concurrent vehicle

Positive control(s):

As a positive control, 20 mg of cyclophosphamide (CPP)/kg body weight or 0.15 mg of vincristine sulphate (VCR)/kg body weight, both, dissolved in purified water, were administered to male and female animals once orally or intraperitoneally each in a volume of 10 mL/kg body weight.

Examinations

Tissues and cell types examined:

erythrocytes of the femural bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: The respective OECD guideline suggests 2000 mg/kg bw as the highest dose. In a pretest, all animals survived this dosage.

DETAILS OF SLIDE PREPARATION: Bone marrow was prepared from the femura and transferred onto microscopic slides using a standard protocol. The resulting slides were stained with eosin-methylene blue and Giemsa and finally embedded in Corbit-Balsam.

METHOD OF ANALYSIS: 2000 polychromatic erythrocytes were counted for each animal. The number of nor-chromatic erythrocytes was scored, too.

Evaluation criteria:

The test chemical is to be considered positive in this assay if the following criteria are met: A dose-related and significant increase in the number of micronucleated polychromatic erythrocytes at any of the intervals. The proportion of cells containing micronuclei exceeded both the values of the concurrent negative control range and the negative historical control range.
A test substance is generally considered negative in this test system if: There was no significant increase in the number of micronucleated polychromatic erythrocytes at any dose above concurrent control frequencies and at any time. The frequencies of cells containing micronuclei were within the historical control range.

Statistics:

Statistical evaluation of data: MUKERN (BASF AG).
Comparison of dose groups: One-sided Wilcoxon test for the hypothesis of equal medians.

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

No inhibition of erythropoiesis was observed.