Glycerides, C16-18 mono- and di-

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

key study

Study period:

November 27th, 1984 - December 20th, 1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP - guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

: only 1000 instead of 2000 erythrocytes per animal counted, but 7 instead of 5 requested animals per sex tested; lack of details on test substance

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Glycerides, C16-18 and C18-hydroxy mono- and di-

EC Number:

295-191-2

EC Name:

Glycerides, C16-18 and C18-hydroxy mono- and di-

Cas Number:

91845-19-1

IUPAC Name:

91845-19-1

Details on test material:

- Name of test material (as cited in study report): Glycerides, C16-18 and C18 mono- and dihydroxy fatty acids
- Physical state: white, solid matter
- Analytical purity: no data

Test animals

Species:

mouse

Strain:

other: CFW 1 (Winkelmann)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 8 weeks
- Weight at study initiation: 21-29 g
- Assigned to test groups randomly: yes, using a table of random numbers
- Fasting period before study: yes, at least 4 hours
- Housing: males individually in Makrolon cages type I, up to 4 females in Makrolon cages type II
- Diet (e.g. ad libitum): Altromin 1324
- Water (e.g. ad libitum): tap water
- Acclimation period:


ENVIRONMENTAL CONDITIONS
- Temperature (°C): no data
- Humidity (%): no data
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: peanut oil;
- Purity: DAB 8
- Source: Caesar and Loretz, Hilden, Germany.

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
An aliquot of the test substance was molten at 70ºC and mixed with 70ºC warm peanut oil. After cooling the solution was applicated by oral gavage.

Dosing volume: 20 mL/kg bw

Duration of treatment / exposure:

single application

Frequency of treatment:

single application

Post exposure period:

up to 72 h

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
10000 mg/kg bw
Basis:
actual ingested
for analysis after 24, 48 and 72 h

Doses / concentrations 2

Remarks:

Doses / Concentrations:
1000 and 5000 mg/kg bw
Basis:
actual ingested
for analysis after 24 h

No. of animals per sex per dose:

7

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide: 7 male and 7 female animals received a single intraperitoneal dose of 10 mg/kg bw Endoxan (R) dissolved in water.
Bone marrow was analyzed 24 hours after application.

Examinations

Tissues and cell types examined:

femoral bone marrow

Details of tissue and slide preparation:

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
24, 48 and 72 h after administration animals were euthanized by carbon dioxide asphyxie. Bone marrow was taken from both femurs.

DETAILS OF SLIDE PREPARATION:
3 slides were prepared from each animal and stained with Giemsa.

METHOD OF ANALYSIS:
The number of micronuclei was counted in 1000 polychromatic erythrocytes per animal.
The ratio of polychromatic and normochromatic erythrocytes was recorded.

OTHER:

Statistics:

Kastenbaum, M.A. and Bowman, K.O., 1970
Tables for Determining the Statistical Significance of Mutation Frequencies

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Remarks:

at doses up to 10000 mg/kg bw

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 5000, 7500 and 10000 mg/kg bw was given to 1 male and 1 female animal each. At the middle and highest dose ruffled fur was observed up to 20 hours after application. No clinical signs of systemic toxicity were observed after 20 and 48 hours, respectively.

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no significant increase of micronuclei number was observed
- Ratio of PCE/NCE (for Micronucleus assay): comparable between all test groups
- Appropriateness of dose levels and route: given
- Statistical evaluation: performed according to Kastenbaum and Bowman

Any other information on results incl. tables

As there were no indications for genotoxicity in animals dosed with 10000 mg/kg bw, analysis of micronuclei was not performed in animals dosed with 1000 and 5000 mg/kg bw. No deaths occured during the study.

Table 1: Results of the in vivo micronucleus assay in male animals

Exp group	Number of animals	Sampling time (hours after administration)	Dose	Ratio polychromatic/ normochromatic erythrocytes (average)	micronuclei per 1000 polychromatic erythrocytes
					mean values	range
Vehicle control (peanut oil)	7	24	20 mL/kg	1.11	1.57	0-2
Positive control (Cyclophosphamide)	7	24	10 mg/kg	1.10	7.71	4-11
Test substance	7	24	10000 mg/kg	1.20	2.14	1-4
Test substance	7	48	10000 mg/kg	1.00	1.00	0-2
Test substance	7	72	10000 mg/kg	1.25	1.43	0-3

 

 

Table 2: Results of the in vivo micronucleus assay in female animals

Exp group	Number of animals	Sampling time (hours after administration)	Dose	Ratio polychromatic/ normochromatic erythrocytes (average)	micronuclei per 1000 polychromatic erythrocytes
					mean values	range
Vehicle control (peanut oil)	7	24	20 mL/kg	1.26	1.14	0-3
Positive control (Cyclophosphamide)	7	24	10 mg/kg	1.15	6.86	4-10
Test substance	7	24	10000 mg/kg	1.07	2.00	1-4
Test substance	7	48	10000 mg/kg	1.06	2.29	0-6
Test substance	7	72	10000 mg/kg	1.15	1.86	0-3

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative The test substance was not genotoxic in the micronucleus assay in vivo.