Registration Dossier

Administrative data

Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented study which meets scientific principles

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1992
Reference Type:
publication
Title:
Micronucleated erythrocyte frequency in peripheral blood of B6C3F1 mice from short-term, prechronic and chronic studies of the NTP carcinogenesis bioassay program
Author:
Witt KL, et al.
Year:
2000
Bibliographic source:
Environmental Molecular Mutagenesis, 36, 163-194

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Principles of method if other than guideline:
Micronucleus assay on peripheral blood lymphocytes
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Diethanolamine
- Physical state: liquid, colorless to pale yellowish
- Analytical purity: >99% (GC)
- Storage condition of test material: room temperature, protected from light
- Source: Kodak Laboratory and Specialty Chemicals (Rochester, NY, USA)

Test animals

Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals and environmental conditions:
- Source: Taconic Farms, Germantown, NY,  USA
- Age at study initiation: about 5-6 weeks
- Housing: individually
- Diet ad libitum
- Water ad libitum
- Acclimation period: 12-13 day

Administration / exposure

Route of administration:
dermal
Vehicle:
- Vehicle(s)/solvent(s) used: ethanol
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
daily on working days (5 days/week)
Post exposure period:
none
Doses / concentrationsopen allclose all
Remarks:
Doses / Concentrations:
37.5, 75, 150, 300, and 600 mg/ml
Basis:
nominal conc.
Remarks:
Doses / Concentrations:
80, 160, 320, 630, 1250 mg/kg bw
Basis:
nominal conc.
No. of animals per sex per dose:
10 males and 10 females
Control animals:
yes, concurrent vehicle
Positive control(s):
urethane
- Route of administration: drinking water
- Doses / concentrations: 0.2%

Examinations

Details of tissue and slide preparation:
Smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13-week  
study. Smears were immediately prepared and fixed in absolute methanol.  The methanol-fixed slides were stained with Hoechst 33258/pyronin Y  
(MacGregor et al., 1983) and coded. Slides were scanned to determine the  frequency of micronuclei in 10,000 normochromatic erythrocytes (NCEs) 
in  up to 10 male and 10 female mice per dose group.
Statistics:
Log transformation of NCE data, testing for normality by the Shapiro-Wilk  test, and testing for heterogeneity of variance by Cochran's test. The frequency of micronucleated cells among NCEs was analyzed by analysis  of variance with the SAS GLM procedure. The NCE data for each dosed group  were compared with the concurrent solvent control group by Student's  t-test.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Remarks:
local and systemic signs of toxicity down to the lowest dose level tested
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
The test substance did not induce micronuclei in peripheral blood erythrocytes of mice exposed dermally to concentrations of up to 1250 mg/kg bw for 13 weeks.
The sensitivity of the test system was verified since the positive control substance showed the expected positive reactions.

Any other information on results incl. tables

Results summary:

The test substance did not induce micronuclei in peripheral blood erythrocytes of mice exposed dermally to concentrations of up to 1250 mg/kg bw for 13 weeks.

The sensitivity of the test system was verified since the positive control substance showed the expected positive reactions.

Detailed results:

Peripheral Blood

Study ID

Result

Route

Male

Female

A08796

Negative

Negative

Skin Paint


A08796 - 1

Start Date

Sample Collection Time

Sex

Cell

Dosing Regimen

Trend Test P-Value

01/02/1990

24 Hours

Male

NCE

SKIN x 65, 90 Days

1.000

 

Dose (mg/kg)

No. of Animals Scored

Mean MN-NCE/1000 NCE ± SEM

Pairwise P

Vehicle Control:

Ethanol

0         

10

1.36 ± 0.15

Test Chemical:

Diethanolamine

80         

10

1.38 ± 0.17

0.465

160         

9

1.44 ± 0.13

0.370

320         

9

1.09 ± 0.13

0.901

630         

10

1.06 ± 0.16

0.933

1250         

8

0.81 ± 0.10

0.997

Positive Control:

Urethane

0.2       

3

18.62 ± 2.43

0.0001

A08796 - 2

Start Date

Sample Collection Time

Sex

Cell

Dosing Regimen

Trend Test P-Value

01/03/1990

24 Hours

Female

NCE

SKIN x 65, 90 Days

0.847

 

Dose (mg/kg)

No. of Animals Scored

Mean MN-NCE/1000 NCE ± SEM

Pairwise P

Vehicle Control:

Ethanol

0         

10

0.82 ± 0.11

Test Chemical:

Diethanolamine

80         

10

0.79 ± 0.08

0.600

160         

10

0.81 ± 0.07

0.554

320         

10

0.73 ± 0.06

0.766

630         

10

0.71 ± 0.12

0.822

1250         

5

0.71 ± 0.14

0.776

Applicant's summary and conclusion