Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
supporting study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP-Study conducted according to guideline protocol tested with the hydrolysis product CAS 102-71-6.

Data source

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

Materials and methods

Test guidelineopen allclose all
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Qualifier:
according to guideline
Guideline:
other: Specifications for the Conduct of Studies to Evaluate the Toxic and Carcinogenic Potential of Chemical, Biological, and Physical Agents in Laboratory Animals for the National Toxicology Program (NTP) October 2006
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Reference substance name:
2,2',2''-nitrilotriethanol
EC Number:
203-049-8
EC Name:
2,2',2''-nitrilotriethanol
Cas Number:
102-71-6
IUPAC Name:
2,2',2''-nitrilotriethanol
Test material form:
other: liquid
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 5E C, protected from light

Test animals

Species:
rat
Strain:
Fischer 344
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 98 g (males), 84 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

Type of coverage:
open
Vehicle:
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
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
The dose formulations were prepared once every 2 weeks and were stored at 5°C in amber glass bottles under a nitrogen head space, protected from light, for up to 3 weeks. Stability studies of the dermal dose formulations were performed by the analytical chemistry laboratory; stability was confirmed for at least 3 weeks at room temperature in sealed glass vials, under a nitrogen head space, in the dark and for at least 3 hours under animal room conditions (open to air and light).
The dose formulations were analyzed at the beginning, midpoint, and end of the studies with gas chromatography.
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
daily, 5 days/week
Doses / concentrations
Remarks:
Doses / Concentrations:
125, 250, 500, 1,000, or 2,000 mg/kg bw
Basis:
nominal per unit body weight
No. of animals per sex per dose:
10
Control animals:
yes, concurrent no treatment
Details on study design:
- Dose selection rationale: Based on a 14-a nd 16-day study, where skin irritation was observed with higher doses.

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily;
- Cage side observations checked in table were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly

DERMAL IRRITATION (if dermal study): Yes
- Time schedule for examinations: weekly

BODY WEIGHT: Yes
- Time schedule for examinations: initially, weekly, and at the end of the study

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION: No
- Time schedule for examinations:

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at days 4 ± 1 and 21 ± 2
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All
- Parameters examined: hematocrit; hemoglobin concentration; erythrocyte, reticulocyte, and nucleated erythrocyte counts; mean cell volume; mean cell hemoglobin; mean cell hemoglobin concentration; platelet count; and total leukocyte count and differentials

CLINICAL CHEMISTRY: Yesa
- Time schedule for collection of blood: at days 4 ± 1 and 21 ± 2
- Animals fasted: No data
- How many animals: All
- Parameters examined: urea nitrogen, creatinine, glucose, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, and sorbitol dehydrogenase

URINALYSIS: Yes
- Time schedule for collection of urine: overnight (16 hours) from clinical pathology group rats during weeks 1, 3, 7, and 13 and from clinical pathology group mice during weeks 7 and 12.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters examined: glucose, protein, volume, and specific gravity

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY AND HISTOPATHOLOGY: Yes
Complete histopathology was performed on core study rats in the vehicle control and 2,000 mg/kg groups and mice in the vehicle control and 4,000 mg/kg groups. In addition to gross lesions and tissue masses, the tissues examined included: adrenal gland, bone and marrow, brain, clitoral gland, epididymis, esophagus, gallbladder (mice), heart, kidney, large intestine (cecum, colon, and rectum), liver, lung, lymph nodes (mandibular and mesenteric), mammary gland, nose, ovary, pancreas, parathyroid gland, pituitary gland, preputial gland, prostate gland, salivary gland, seminal vesicle, skin (lesions and unaffected skin from site of application; inguinal skin), small intestine (duodenum, jejunum, and ileum), spinal cord and sciatic nerve (if neurologic signs were present), spleen, stomach (forestomach and glandular stomach), testis, thymus, thyroid gland, trachea, urinary bladder, uterus, and vagina (females in vaginal cytology studies only). Additionally, the kidney of female rats, pituitary gland of male and female rats, and skin (site of application) of male and female rats and mice in the lower exposure groups were examined until a no-effect level was reached.

Other examinations:
Sperm Morphology and Vaginal Cytology Evaluations:
Rats in the 0, 500, 1,000, and 2,000 mg/kg groups and mice in the 0, 1,000, 2,000, and 4,000 mg/kg groups were evaluated.
Sperm samples were collected at the end of the studies and evaluated for sperm count, motility, and morphology. The right cauda, epididymis, and testis were weighed.
Vaginal samples were collected for 7 consecutive days before the end of the studies and evaluated for the relative frequency of estrous stages and for
estrous cycle length.
Statistics:
The probability of survival was estimated by the product-limit procedure of Kaplan and Meier (1958) and is presented in the form of graphs. Animals found dead of other than natural causes or missexed were censored from the survival analyses; animals dying from natural causes were not censored. Statistical analyses for possible dose-related effects on survival used Cox’s (1972) method for testing two groups for equality and Tarone’s (1975) life table test to identify dose-related trends. All reported P values for the survival analyses are two sided.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Dermal irritation:
effects observed, treatment-related
Description (incidence and severity):
signs of irritation, scaliness, crustiness and ulceration
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Reduced final mean body weights and weight gains of males and females in the 2,000 mg/kg groups
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
only minimal to mild changes in the high dose group with correlation to aobserved skin inflammation
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
minimal changes in males and females of the high dose groups
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
minimal changes in males and females of the high dose groups
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
increased kidney weights in males and females from 500/kg bw/d onwards: other weight changes are attributed to changed body weights
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
chronic inflammation at application sites
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Dosed females had greater incidences of nephropathy than did the vehicle controls
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
No mortality occured during the study.

BODY WEIGHT AND WEIGHT GAIN
Final mean body weights and weight gains of males and females in the 2000 mg/kg groups were significantly less than those of the vehicle controls; the mean body weight gain of females in the 1,000 mg/kg group was also significantly less than that of the vehicle controls.

HAEMATOLOGY
Mild increases in segmented neutrphil counts occurred in male and female rats in the 2000 mg/kg groups; in males, this change was accompanied by increased leukocyte and eosinophil counts. Minimal decreases occurred in mean red cell volume in male and female rats administered 2000 mg/kg and in hematocrit in females administered 2,000 mg/kg.


CLINICAL CHEMISTRY
Minimal increases in albumin and urea nitrogen concentrations occurred in females that received 2000 mg/kg. Serum aspartate aminotransferase
activities were mildly increased in male rats receiving 250 mg/kg or greater and in females receiving
2000 mg/kg. Serum alanine aminotransferase activity was minimally increased in males in the 1000 and 2000 mg/kg groups. The activity of sorbitol dehydrogenase, which is also liver specific, decreased minimally in females administered 500 or 1000 mg/kg.


URINALYSIS
Increased urine specific gravity in females in the 1,000 and 2,000 mg/kg groups at weeks 7 (day 44) and 13 and in male rats in the 2000 mg/kg group at week 13. Urine protein excretion was decreased in males in the 2,000 mg/kg group on day 16, in males administered 500 mg/kg or greater at week 7, and in males in the 1,000 and 2,000 mg/kg groups at week 13.

ORGAN WEIGHTS
No change in absolute liver weights. Kidney weights were generally greater in males and females administered 500, 1000, or 2000 mg/kg than in the vehicle controls.

GROSS PATHOLOGY
Lesions attributed to triethanolamine application included crust at the site of application for males and females administered 1000 or 2000 mg/kg. The compound-related skin lesions were chronic-active inflammation and acanthosis, which contained epidermal and dermal components. The epidermal component was characterized by acanthotic, hyperkeratotic, focally parakeratotic epidermis that occasionally contained rete pegs and was graded as acanthosis. Severely affected epidermis contained focal hemorrhage, fibrin and/or mineral deposits, bacterial colonies, serum pockets, pustules, erosions and/or ulcers. The dermal component in severely affected rats was characterized bydermal fibrosis, neocapillarization, minimally distorted adnexal organs, and variably severe mixed inflammatory infiltrates consisting of histiocytes, lymphocytes, neutrophils and eosinophils


HISTOPATHOLOGY:
No microscopic evidence of hepatic injury. Dosed females had greater incidences of mineralization and nephropathy than did the vehicle controls. The incidences of hypertrophy of the pituitary gland pars intermedia were significantly greater in males and females in the 2000 mg/kg group than in the vehicle controls. However,as the severity of this lesion did not vary between dose groups, it was regarded as incidental.


OTHER FINDINGS
There were no biologically significant differences in sperm morphology or vaginal cytology parameters between dosed and vehicle control rats.

Effect levels

open allclose all
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
125 mg/kg bw/day
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Increased relative liver weight
Dose descriptor:
NOAEL
Remarks:
systemic
Effect level:
500 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Increased relative liver weight
Dose descriptor:
NOAEL
Remarks:
local
Effect level:
125 mg/kg bw/day
Based on:
test mat.
Sex:
male
Basis for effect level:
other: Acanthosis and chronic inflammation
Dose descriptor:
NOAEL
Remarks:
local
Effect level:
250 mg/kg bw/day
Based on:
test mat.
Sex:
female
Basis for effect level:
other: Acanthosis and chronic inflammation

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Results:

1. Body weights:

Mean Body Weights (g) Finale Weights Relative to Controls (%)
Dose (mg/kg) Initial Final Change
Male 0 98 ± 2 296 ± 6 198 ± 6
125 94 ± 2 279 ± 5 185 ± 5 94
250 95 ± 2 279 ± 6 184 ± 5 94
500 96 ± 2 288 ± 5 193 ± 5 97
1000 98 ± 3 290 ± 10 192 ± 9 98
2000 95 ± 3 252 ± 9** 156 ± 8** 85
Female 0 84 ± 2 176 ± 3 92 ± 3
125 84 ± 2 171 ± 3 87 ± 2 97
250 86 ± 2 175 ± 3 89 ± 2 100
500 85 ± 2 173 ± 3 88 ± 3 98
1000 87 ± 2 170 ± 3 83 ± 3* 97
2000 82 ± 1 156 ± 2** 74 ± 2** 89

* Significantly different (P≤0.05) from the vehicle control group by Williams’ or Dunnett’s test

** P≤0.01

2. Changes in Organ Weights and Organ-Weight-to-Body-Weight Ratios

0 mg/kg 125 mg/kg 250 mg/kg 500 mg/kg 1000 mg/kg 2000 mg/kg
Female Necropsy body weight  174 ± 4 170 ± 4 174 ± 4 174 ± 3 170 ± 3 158 ± 4**
Brain Absolute 1.748 ± 0.020 1.712 ± 0.023 1.754 ± 0.014 1.159 ± 0.024 1.761 ± 0.022 1.770 ± 0.013
Relative 10.14 ± 0.35 10.17 ± 0.33 10.14 ± 0.23 10.11 ± 0.12 10.35 ± 0.14 11.27 ± 0.19**
Kidney Absolute 0.744 ± 0.017 0.745 ± 0.017 0.758 ± 0.019 0.810 ± 0.015* 0.847 ± 0.019** 0.891 ± 0.019**
Relative 4.31 ± 0.14 4.41 ± 0.11 4.37 ± 0.08 4.66 ± 0.08* 4.98 ± 0.12** 5.67 ± 0.15**
Liver Absolute 6.982 ± 0.290 6.860 ± 0.188 7.154 ± 0.222 7.340 ± 0.289 7.483 ± 0.342 7.067 ± 0.272
Relative 40.28 ± 1.52 40.52 ± 0.71 41.15 ± 0.68 42.10 ± 1.37 43.88 ± 1.76 44.96 ± 1.64*
Spleen Absolute 0.468 ± 0.010 0.461 ± 0.010 0.462 ± 0.009  0.456 ± 0.011 0.461 ± 0.013 0.427 ± 0.009*
Relative 2.71 ± 0.08 2.73 ± 0.08 2.67 ± 0.03 2.62 ± 0.04 2.71 ± 0.09 2.72 ± 0.07
Male Necropsy body weight  299 ± 6 284 ± 7 286 ± 6 287 ± 7 295 ± 11 248 ± 11**
Brain Absolute 1.911 ± 0.021 1.867 ± 0.021 1.868 ± 0.019 1.900 ± 0.011 1.919 ± 0.018 1.880 ± 0.030
Relative 6.41 ± 0.11 6.60 ± 0.15 6.55 ± 0.12 6.65 ± 0.18 6.58 ± 0.19 7.74 ± 0.39**
Kidney Absolute 1.187 ± 0.025 1.134 ± 0.026 1.188 ± 0.031 1.264 ± 0.028 1.366 ± 0.039** 1.366 ± 0.051**
Relative 3.97 ± 0.05 4.00 ± 0.06 4.16 ± 0.06 4.41 ± 0.06* 4.65 ± 0.09** 5.58 ± 0.24**
Liver Absolute 13.949 ± 0.446 12.404 ± 0.4463 13.418 ± 0.693 15.174 ± 0.382 15.516 ± 0.421 12.517 ± 0.405
Relative 46.58 ± 0.86 43.68 ± 1.14 46.73 ± 1.61 53.12 ± 1.90* 52.92 ± 1.05* 51.18 ± 2.14*
Spleen Absolute 0.666 ± 0.016 0.646 ± 0.013 0.626 ± 0.009 0.666 ± 0.013 0.690 ± 0.028 0.612 ± 0.024
Relative 2.23 ± 0.03 2.28 ± 0.05 2.19 ± 0.04 2.33 ± 0.05 2.34 ± 0.03 2.50 ± 0.11*

Organ weights (absolute weights) and body weights are given in grams; organ-weight-to-body-weight ratios (relative weights) are given as mg organ weight/

g body weight (mean ± standard error).

* Significantly different (P≤0.05) from the vehicle control group by Williams’ or Dunnett’s test

** P≤0.01

3. Incidences of Selected Nonneoplastic Lesions

0 mg/kg 125 mg/kg 250 mg/kg 500 mg/kg 1000 mg/kg 2000 mg/kg
Male Skin, Site of Application (a) 10 10 10 10 10 10
Acanthosis (b) 0 0 6** (1.2)c 9**(1.2) 10**(2.0) 10**(3.5)
Inflammation, Chronic Active 0 0 2 (1.5) 2 (2.0) 10**(2.7) 10**(4.0)
Pituitary Gland, Pars Intermedia 9 6 7 8 9 8
Hypertrophy 0 0 0 0 0 5**(1.0)
Female Skin, Site of Application (a) 10 10 10 10 10 10
Acanthosis (b) 0 0 0 4*(1.3) 8**(1.6) 10**(2.8)
Inflammation, Chronic Active 0 0 0 1 (2.0) 5*(2.6) 10**(3.9)
Kidney, Renal Tubule 10 10 10 10 10 10
Regeneration (Nephropathy) 2(1.0) 3(1.0) 5(1.0) 7*(1.0) 10**(1.4) 8*(1.4)
Mineralization 3(1.0) 9**(1.1) 6(1.0) 7(1.6) 9**(1.9) 9**(1.4)
Pituitary Gland, Pars Intermedia 6 8 8 8 6 9
Hypertrophy 0 0 0 0 1(1.0) 9**(1.7)

* Significantly different (P≤0.05) from the vehicle control group by Williams’ or Dunnett’s test

** P≤0.01

(a) Number of animals with organ examined microscopically

(b) Number of animals with lesion

(c) Average severity of lesions in affected rats: 1=minimal; 2=mild; 3=moderate; 4=marked

Applicant's summary and conclusion

Executive summary:

Groups of 10 male and 10 female rats were topically administered 0, 125, 250, 500, or 1000 mg triethanolamine per kilogram body weight in acetone or 2000 mg/kg neat triethanolamine, 5 days per week, for 13 weeks. All rats survived to the end of the study. Final mean body weights and weight gains of males and females administered 2000 mg/kg and the mean body weight gain of females administered 1000 mg/kg were significantly less than those of the vehicle controls. Clinical observations included irritation, scaliness, and crustiness of the skin at the site of application for males and females. Males also had discoloration, and two males administered 2000 mg/kg had ulceration at the site of application. Changes in clinical pathology parameters were minor and consistent with inflammation at the site of application.

Kidney weights were generally greater in males and females administered 500, 1000, or 2000 mg/kg than in the vehicle controls. Microscopic lesions attributed to triethanolamine administration included acanthosis and inflammation at the site of application, nephropathy in females, and hypertrophy of the pituitary gland pars intermedia in males and females. The NOEL for systemic effects were found to be 500 mg/kg bw/d for females and 125 mg/kg bw/d for males, respectively.