2-ethylhexyl oleate

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:

17 Jul - 26 Aug 1991

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-guideline study with acceptable restrictions (lack of details on test substance; no analytical purity given)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

analytical purity of test substance not specified

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Reference substance 001

Cas Number:

135800-37-2

Molecular formula:

C16H32O2 to C26H52O2

Details on test material:

- Name of test material (as cited in study report): Fatty acids, C8-16, 2-ethylhexyl esters
- Physical state: Yellow liquid
- Analytical purity: no data
- Storage condition of test material: RT
- Density: 0.86 g /cm³
- Stability: stable
- Solubility: insoluble in water

Test animals

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 7 - 12 weeks
- Assigned to test groups randomly: yes
- Housing: 5 animals of each sex in polycarbonate cages
- Acclimation period: 7 - 15 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18 - 26
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil

Details on exposure:

Dosing volume: 10 mL/kg

Duration of treatment / exposure:

Single dose

Frequency of treatment:

Single dose

Post exposure period:

Up to 72 h

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
0, 1250, 2500 and 5000 µL/kg bw
Basis:
nominal conc.

Doses / concentrations 2

Remarks:

Doses / Concentrations:
0, 1075, 2150 and 4300 mg/kg bw
Basis:
other: calculated based on a density of 860 mg/mL

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Route of administration: intreperitoneal
- Doses / concentrations: 30 mg/kg bw

Examinations

Tissues and cell types examined:

Erythrocytes of the bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Results of a pretest with 50, 160, 500, 1600 and 5000 µL/kg and the sponsor´s specification

DETAILS OF SLIDE PREPARATION: Immediately after sacrife, both femurs of the animal were removed and flushed with a few drops of fetal calf serum onto a clean microscope slide. A second slide was inverted and placed to the first slide. Using a circular motion, the two slides were rubbed together until the bone marrow was evenly dispersed. The two slides were gently pulled appart, air dried, fixed in methanol for approx. 10 min and again air dried.The slides were stained with Giemsa for 5 min and differentiated in distilled water afterwards.

METHOD OF ANALYSIS: at least 1000 polychromatic erythrocytes were counted for each category.

Evaluation criteria:

There should be a statistically significant increase in the number of micronucleated polychromatic erythrocytes in the treatment group over the negative control group.

Statistics:

The two-tailed Student´s t-test was applied to compare the frequency of micronucleated cells to the respective vehicle control.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Any other information on results incl. tables

Table 1: Mean numbers of micronucleated cells

Treatment	Micronucleated cells
	mean	SD
Vehicle control	2.26	1.41
5000 µL/kg	1.66	1.15
2500 µL/kg	1.93	1.17
1250 µL/kg	1.80	0.81
Positive control	17.20	2.62

There was no significant increase of micronucleated cells in the treatment groups compared to the negative control. The positve control was valid. No indications for mutagenicity in vivo were given.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative