(carboxymethyl)dimethyl-3-[(1-oxotetradecyl)amino]propylammonium hydroxide

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:

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:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

adopted in 2016

Deviations:

yes

Remarks:

no historical controls given, immature and mature micronucleated erythrocytes counted in 400 erythrocytes/animal

GLP compliance:

yes

Type of assay:

mammalian erythrocyte micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

(carboxymethyl)dimethyl-3-[(1-oxotetradecyl)amino]propylammonium hydroxide

EC Number:

261-684-6

EC Name:

(carboxymethyl)dimethyl-3-[(1-oxotetradecyl)amino]propylammonium hydroxide

Cas Number:

59272-84-3

Molecular formula:

C21H42N2O3

IUPAC Name:

(carboxymethyl)dimethyl-3-[(1-oxotetradecyl)amino]propylammonium hydroxide

Test material form:

solid

Test animals

Species:

mouse

Strain:

Swiss

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Central Institute for the Breeding of Laboratory Animals, TNO, Zeist, The Netherlands
- Weight at study initiation: males: about 23 g; females: about 19 g
- Assigned to test groups randomly: yes
- Diet: ad libitum
- Water: ad libitum

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: water
- Concentration of test material in vehicle: 10%
- Amount of vehicle (if gavage or dermal): 10 mL/kg bw

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: The test substance was dissolved in water as a 10% solution. As the aqueous solution of W-1108 foamed rather strongly, a drop (appr. 50 mg) of an antifoam agent (Polymekor 1488, Goldschmidt, FRG) was added per 100 mL solution.

Duration of treatment / exposure:

not applicable

Frequency of treatment:

single treatment

Post exposure period:

24, 48, and 72 hours

Doses / concentrations

Dose / conc.:

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

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

mitomycin C
- Justification for choice of positive control(s): as described in OECD 474
- Route of administration: i.p.
- Doses / concentrations: 1.5 mg mitomycin C/20 mL saline/kg bw

Examinations

Tissues and cell types examined:

Tissue: bone marrow
Cell type: bone marrow cells

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: It is widely accepted that for a meaningful micronucleus test a very high dose should be administered to the test animals. Usually a proportion (20 - 80%) of the LD50 value is used. The LD50 value for the test substance is 1875 mg/kg bw.

DETAILS OF SLIDE PREPARATION: The femurs were dissected free, the bone marrow was flushed from the femurs into a centrifuge tube containing calf serum and the mixture was centrifuged. The excess serum was removed. The cells were then resuspended by mixing gently, a drop of the suspension was placed on a slide and spread with a haemocytometer cover glass. Five slides were prepared from each animal. The smears were air dried, fixed in methanol and stained according to May-Grünwald Giemsa.

METHOD OF ANALYSIS: The stained smears were examined by light microscopy to determine the incidence of micronucleated poly- and normochromatic erythrocytes and the ratio of polychromatic and normochromatic erythrocytes. The incidence of micronucleated poly- and normochromatic erythrocytes was determined in a total of 1000 erythrocytes per animal (200 erythrocytes per slide).
The counting procedure followed in the positive control animals was slightly different. Generally the determination of the incidence of micronucleated erythrocytes per 1000 erythrocytes was restricted to four animals (two males and two females) at each sacrifice time: 24 h, 48 h, 72 h. In the remaining animals the incidence of micronucleated cells was recorded per 400 erythrocytes.
For all animals (solvent control, positive control and test animals) the ratio of poly- and normochromatic erythrocytes was determined by counting the number of poly- and normochromatic erythrocytes in a total of 400 erythrocytes per animal.

Evaluation criteria:

not specified

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Toxicity: Two males treated with the test substance showed a rough coat on the first day after treatment and one female showed signs of slight diarrhoea.
Body weight was significantly reduced on Day 1 in males and females.

Any other information on results incl. tables

Table 1: results

Group	Number of animals	Dose [mg/kg bw]	% PCEs (= PCE x 100 / (PCE + NCE) at sampling time			Total micronuclei per 1000 erythrocytes at sampling time (range)
			24h	48h	72 h	24h	48h	72 h
Vehicle control (water)	5 males	0	51	44	62	2.4 (1-5)	2.6 (1-5)	1.8 (0-4)
	5 females	0	56	54	49	1.4 (0-5)	1 (0-2)	0.8 (0-2)
Positive control	5 males	1.5	50	27	36	29 (17-40)	28 (18-44)	19 (13-24)
	5 females	1.5	57	27	29	30 (18-43)	11 (3-20)	9 (3-13)
Test substance	5 males	1000	42	33	58	2.4 (1-4)	2 (1-4)	1.2 (0-3)
	5 females	1000	53	42	54	0.6 (0-2)	2.6 (0-5)	1.2 (0-3)
PCE = polychromatic erythrocytes NCE = normochromatic erythrocytes

Applicant's summary and conclusion

Conclusions:

The results of the present micronucleus test did not provide any evidence of chromosomal damage and/or damage to the mitotic apparatus caused by oral administration of high doses of the test substance.