Amides, C12-18 and C18-unsatd., N,N-bis(hydroxyethyl)

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:

Not available

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1999

Materials and methods

Principles of method if other than guideline:

The test substance was applied dermally to mice for 14 weeks. Peripheral blood samples were obtained from male and female mice at termination, and smears were immediately prepared and slides were scanned to determine the frequency of micronuclei.

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N-bis(2-hydroxyethyl)dodecanamide

EC Number:

204-393-1

EC Name:

N,N-bis(2-hydroxyethyl)dodecanamide

Cas Number:

120-40-1

Molecular formula:

C16H33NO3

IUPAC Name:

N,N-bis(2-hydroxyethyl)dodecanamide

Constituent 2

Reference substance name:

2,2'-iminodiethanol

EC Number:

203-868-0

EC Name:

2,2'-iminodiethanol

Cas Number:

111-42-2

Molecular formula:

C4H11NO2

IUPAC Name:

2,2'-iminodiethanol

Test material form:

other: Viscous light yellow liquid or a white to light yellow, waxy solid

Test animals

Species:

mouse

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Taconic Farms, Germantown, NY
- Age at study initiation: 7 wk
- Housing: Housed individually in Polycarbonate cages
- Bedding: Sani-Chip heat-treated hardwood chips (P.J. Murphy Forest Products Corp., Montville, NJ)
- Diet : NIH-07 open formula pelleted diet, ad libitum
- Water : Tap water, ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature : 21.1-23.9°C
- Humidity : 42- 57%
- Air changes : 10/hr
- Photoperiod : 12 h dark/12 h light

IN-LIFE DATES: From: 1992-01-15 To: 1992-04-17

Administration / exposure

Route of administration:

dermal

Vehicle:

- Vehicle(s)/solvent(s) used: Ethanol
- Concentration of test material in vehicle:
- Lot/batch no. (if required): 91D22U
- Purity: Purity of the bulk ethanol ranged from 97% to 103% relative to the reference standard

Duration of treatment / exposure:

14 wk

Frequency of treatment:

5 exposures/wk

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

50 mg/kg bw/day (nominal)

Remarks:

Corresponding to 20 mg/mL in ethanol

Doses / concentrations 3

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Corresponding to 40 mg/mL in ethanol

Doses / concentrations 4

Dose / conc.:

200 mg/kg bw/day (nominal)

Remarks:

Corresponding to 80 mg/mL in ethanol

Doses / concentrations 5

Dose / conc.:

400 mg/kg bw/day (nominal)

Remarks:

Corresponding to 160 mg/mL in ethanol

Doses / concentrations 6

Dose / conc.:

800 mg/kg bw/day (nominal)

Remarks:

Corresponding to 320 mg/mL in ethanol

No. of animals per sex per dose:

10

Control animals:

yes, concurrent vehicle

Examinations

Tissues and cell types examined:

Tissues - Peripheral blood.
Cells - Normochromatic erythrocytes.
Slides were scanned to determine the frequency of micronuclei in 2000 Normochromatic erythrocytes (NCEs) in each of five animals per dose group.

Details of tissue and slide preparation:

A detailed discussion of this assay is presented by MacGregor et al, 1990.

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): Blood samples collected at the end of 14 wk study period

DETAILS OF SLIDE PREPARATION: Peripheral blood samples smears were immediately prepared and fixed in absolute methanol. The methanol-fixed slides were stained with acridine orange and coded.

Evaluation criteria:

An individual trial is considered positive if the trend test P value is less than or equal to 0.025 or if the P value for any single dose group is less than or equal to 0.025 divided by the number of dose groups.

Statistics:

The frequency of micronucleated cells among NCEs was analyzed by a statistical software package that tested for increasing trend over dose groups with a one-tailed Cochran- Armitage trend test, followed by pairwise comparisons between each dosed group and the control group. In the presence of excess binomial variation, as detected by a binomial dispersion test, the binomial variance of the Cochran-Armitage test was adjusted upward in proportion to the excess variation.

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Any other information on results incl. tables

For detailed results table kindly refer to the attached background materials section of the IUCLID.

Applicant's summary and conclusion

Conclusions:

Under the study conditions, the test substance did not increase the frequency of micronuclei in peripheral blood cells of mice.

Executive summary:

A study was conducted to evaluate the in vivo genetic toxicity of the test substance, C12 DEA (90% active) in a peripheral blood micronucleus assay. The substance was applied dermally to mice for 14 weeks with a frequency of 5 exposures/week at doses of 50, 100, 200, 400 and 800 mg/kg bw. Peripheral blood samples were obtained from male and female mice, and smears were immediately prepared and fixed in absolute methanol. The methanol-fixed slides were stained with acridine orange and coded. Slides were scanned to determine the frequency of micronuclei in 2000 normochromatic erythrocytes (NCEs) in each of five animals per dose group. No increase in the frequency of micronucleated normochromatic erythrocytes was observed in the test at any dose level. Under the study conditions, the test substance did not increase the frequency of micronuclei in peripheral blood cells of mice (Irwin, 1999).