Registration Dossier

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 from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study conducted comparable to OECD guideline tested with the source substance CAS 102-71-6. Based on the structural similarities and the fact that the target substance is an adduct of the source substance, this study is considered valid for read-across.
Cross-reference
Reason / purpose for cross-reference:
reference to same study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Deviations:
yes
Remarks:
treatmant of animals for 13 prior to blood sampling
Principles of method if other than guideline:
According to standard NTP protocol
GLP compliance:
yes
Type of assay:
micronucleus assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid - liquid: suspension
Details on test material:
Name of test material (as cited in study report): Triethanolamine
- Physical state: clear, colorless, viscous liquid
- Analytical purity: 99%
- Impurities (identity and concentrations): <0.5% water; <.4% primary or secondary amines
- Lot/batch No.: Lot 3B-1-84
- Storage condition of test material: Neat chemical stored at room temperature; triethanolamine:acetone mixtures stored at 5°C, protected from light


Test animals

Species:
mouse
Strain:
B6C3F1
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Simonsen Laboratories (Gilroy, CA)
- Age at study initiation: 6 weeks old
- Weight at study initiation: mean 23 g (males), 19 g (females)
- Housing: individually in Polycarbonate (Lab Products, Inc., Garfield, NJ), changed weekly, with bedding of Beta-Chips® hardwood chips (Northeastern Products, Inc., Warrensburg, NY), changed weekly
- Diet (e.g. ad libitum): NIH-07 open formula pelleted diet (Zeigler Brothers, Inc.,Gardners, PA), available ad libitum, changed weekly
- Water (e.g. ad libitum): Tap water (City of Columbus municipal supply) via automatic watering system (Edstrom Industries, Waterford, WI), available
ad libitum
- Acclimatization: 11 to 14 da

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.4 - 23.9
- Humidity (%): 35 - 65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES: From: 30 June 1986 To:14 November 1986

Administration / exposure

Route of administration:
dermal
Vehicle:
- Vehicle(s)/solvent(s) used: Acetone
Details on exposure:
TEST SITE
- Area of exposure: area extending from the animal’s mid-back to the dorsal intrascapular region
- Time intervals for shavings or clipplings: clipped weekly


TEST MATERIAL
- Amount(s) applied (volume or weight with unit): if the dose volume exceeded 320 μL, half the total volume was administered in the morning and the
remainder was administered in the afternoon
- Concentration (if solution): 0, 125, 250, 500, 1,000, or 2,000 mg/kg bw
- Constant volume or concentration used: yes

VEHICLE
- Justification for use and choice of vehicle (if other than water): Acetone is miscible with triethanolamine and because acetone rapidly evaporates.

USE OF RESTRAINERS FOR PREVENTING INGESTION: no
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
daily, 5 days/week
Post exposure period:
none
Doses / concentrations
Remarks:
Doses / Concentrations:
500, 1,000, 2,000, 4000 mg/kg bw
Basis:
other: nominal per unit body weight
No. of animals per sex per dose:
10
Control animals:
yes, concurrent no treatment

Examinations

Tissues and cell types examined:
Peripheral blood samples
Details of tissue and slide preparation:
CRITERIA FOR DOSE SELECTION
Doses were based on a 14-a nd 16-day study, where skin irritation was observed with higher doses.

DETAILS OF SLIDE PREPARATION:
Smears were immediately prepared and fixed in absolute methanol and were later stained with a chromatin-specific fluorescent dye mixture of Hoechst 33258/pyronin Y (MacGregor et al., 1983)

METHOD OF ANALYSIS:
Slides were scanned to determine the frequency of micronuclei in 2000 polychromatic erythrocytes (PCEs) and 10000 normochromatic erythrocytes (NCEs) in each animal per dose group.

Statistics:
Log transformation of the NCE data, testing for normality by the Shapiro-Wilk test, and testing for heterogeneity of variance by Cochran’s test were performed before statistical analyses. The frequency of micronucleated cells among NCEs was analyzed by analysis of variance using the SAS GLM procedure. The NCE data for each dose group were compared with the concurrent solvent control with a Student’s t-test. The frequency of micronucleated cells among PCEs was analyzed by the Cochran-Armitage trend test, and individual dose groups were compared to the concurrent solvent control by Kastenbaum-Bowman’s binomial test.

Results and discussion

Test results
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Remarks:
animals of high dose groups showed acanthosis and inflammation at the site of application were observed in both males and females at 4000 mg/kg bw per day, and liver and kidney weights were increased at that dose.
Vehicle controls validity:
valid
Negative controls validity:
not examined
Positive controls validity:
not examined

Any other information on results incl. tables

Results:

Sex Sample Collection Time Dose (g/kg) Animal Number Normochromatic Erythrocytes
Trend P: 0.42
Male 24 hour No. Examined Total MN Cells Percent NCE MN Cells
per 1000
Vehicle Control Acetone 0 11 20   1.4  
0 12 23   2  
0 13 29   2.2  
0 14 38   3.2  
0 15 14   1.1  
0 16 17   1.5  
0 17 20   1.7  
0 18 27   2  
0 19 15   1.2  
0 20 13   1.2  
Average ± SEM   ± 1.75 ± 0.20
 
Test Chemical Triethanolamine 1 71 22   2.2  
1 72 18   1.8  
1 73 21   1.8  
1 74 21   1.6  
1 75 36   2.9  
1 76 22   1.6  
1 77 25   2.4  
1 78 21   1.8  
1 79 20   1.9  
1 80 8   0.8  
Average ± SEM   ± 1.88 ± 0.17
Pairwise P   0.3138
 
Test Chemical Triethanolamine 2 100 15   1.2  
2 91 18   1.5  
2 92 15   1.5  
2 93 18   1.8  
2 94 11   1.1  
2 95 15   1.2  
2 96 11   1  
2 97 18   1.6  
2 98 22   1.7  
2 99 14   1  
Average ± SEM   ± 1.35 ± 0.10
Pairwise P   0.935
 
Test Chemical Triethanolamine 4 111 18   1.5  
4 112 30   2.2  
4 113 18   1.7  
4 114 11   1  
4 115 19   1.7  
4 116 25   1.9  
4 117 14   1.4  
4 118 17   1.4  
4 119 19   1.7  
4 120 46   4.5  
Average ± SEM   ± 1.89 ± 0.30
Pairwise P   0.3047

Sex Sample Collection Time Dose (g/kg) Animal Number Normochromatic Erythrocytes
Trend P: 0.925
Female 24 hour No. Examined Total MN Cells Percent NCE MN Cells
per 1000
Vehicle Control Acetone 0 131 10   0.8  
0 132 25   2  
0 133 10   1  
0 134 13   1.1  
0 135 11   0.9  
0 136 10   0.8  
0 137 12   1.1  
0 138 13   1.1  
0 139 12   1.1  
0 140 18   1.7  
Average ± SEM   ± 1.16 ± 0.12
 
Test Chemical Triethanolamine 1 191 17   1.5  
1 192 20   1.5  
1 193 16   1.6  
1 194 12   1.2  
1 195 15   1.2  
1 196 9   0.9  
1 197 13   1  
1 198 15   1.1  
1 199 13   1.2  
1 200 10   0.9  
Average ± SEM   ± 1.20 ± 0.08
Pairwise P   0.389
 
Test Chemical Triethanolamine 2 211 18   1.8  
2 212 10   0.7  
2 213 9   0.9  
2 214 13   1.1  
2 215 17   1.2  
2 216 16   1.4  
2 217 8   0.8  
2 218 16   1.5  
2 219 9   0.7  
2 220 7   0.7  
Average ± SEM   ± 1.07 ± 0.12
Pairwise P   0.7393
 
Test Chemical Triethanolamine 4 231 8   0.8  
4 232 14   1.3  
4 233 11   1  
4 234 12   1  
4 235 4   0.4  
4 236 11   0.9  
4 237 21   1.7  
4 238 9   0.8  
4 239 12   1.1  
4 240 13   1.1  
Average ± SEM   ± 1.00 ± 0.11
Pairwise P   0.8813

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information): negative
Treatment of male and female mice with triethanolamineup to 4000 mg/kg bw/d for 13 weeks by dermal application did not result in any change in the frequency of micronuclei in their blood cells.
Executive summary:

Male and female B6C3F mice were treated dermally with triethanolamine up to 4000 mg/kg bw/d for 13 weeks. Analysis of periperhal blood cells after the application period did don reveal any change in the frequency of micronuclei when compared with untreated control animals. Thus, no in vivo mutagenicity could be shown for triethanolamine.