Methyl 3-[3-(2H-benzotriazol-2-yl)-4-hydroxy-5-(2-methyl-2-propanyl)phenyl]propanoate

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1984 - 1985

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP and OECD testing guideline compliant study (Outdated OECD 407 version)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report date:

1985

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Deviations:

yes

Remarks:

No analytical verification of doses (but daily preparation of dosing suspension)

GLP compliance:

yes

Remarks:

Experimental Toxicology, CIBA-GEIGY Ltd., Basle, Switzerland

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

400-820-2

EC Name:

-

Cas Number:

84268-33-7

Molecular formula:

C20 H23 N3 O3

IUPAC Name:

methyl 3-[3-(2H-1,2,3-benzotriazol-2-yl)-5-tert-butyl-4-hydroxyphenyl]propanoate

Details on test material:

- Physical state: solid
- Analytical purity: purified
- Lot/batch No.: Z 288/1U
- Storage condition of test material: room temperature

Test animals

Species:

rat

Strain:

other: RAIf (SPF)

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Animal Production, CIBA-GEIGY LTD., 4332 Stein / Switzerland
- Age at study initiation: approx. 6 weeks
- Weight at study initiation: 197-205 g in males, 185-195 g in females
- Housing: 5/cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 8 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2
- Humidity (%): 55 ± 10
- Air changes (per hr): 16 - 20
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: Carboxymethyl-cellulose 0.5% with 0.1% Tween 80

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: The suspensions were freshly prepared every day immediately prior to the dosing of the animals and administered within 4 hours.

Analytical verification of doses or concentrations:

no

Details on analytical verification of doses or concentrations:

Analytical verification was not performed, but the suspension for gavage application was prepared daily prio to dosing.
As the substance is insoluble in water, no stability test for hydrolysis was performed.

Duration of treatment / exposure:

29 days

Frequency of treatment:

daily

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 50, 200, 1000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

- Post-exposure recovery period in satellite groups: no

Positive control:

not applicable

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

BODY WEIGHT: Yes
- Time schedule for examinations: weekly (midweek)

FOOD CONSUMPTION: weekly


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: Yes

WATER CONSUMPTION: Yes
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: before the beginning (day -6) and towards the end (day 20) of the treatment period
- Dose groups that were examined: highest dosage group and control group

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of treatment period
- Anaesthetic used for blood collection: Yes: ether
- Animals fasted: Yes
- How many animals: all animals
- Parameters checked: Erythrocyte Count, Hemoglobin, Hematocrit, Leucocyte Count, Differential Leucocyte Count, Prothrombin Time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of treatment period
- Animals fasted: Yes
- How many animals: all
- Parameters checked: Urea, Creatinine, Total bilirubin, Total protein, Albumin, Globulins, A/G Ratio, Aspartate aminotransferase, Alanine aminotransferase

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

OTHER: Hearing test: Animals of the highest dosage group and of the control group were examined before the beginning (day -6) and towards the end (day 20) of the treatment period.

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

For each time point and parameter a uni-variate statistical analysis was conducted. Due to the routine manner of the analysis system, parameter free methods were applied. Each treated group was compared to the control group in respect of dispersion and displacement. In addition a trend test<2> was applied considering all groups.

Results and discussion

Results of examinations

Details on results:

CLINICAL SIGNS AND MORTALITY:No death occurred during the course of the study.
Besides emaciation in the treated males of group 4 (1000 mg/kg bw.) and one female animal (group 4, 1000 mg/kg bw.) with straub tail no clinical symptoms were observed and no signs of systemic toxicity were found.

BODY WEIGHT AND WEIGHT GAIN: The mean body weight gain of male group 4 (1000 mg/kg bw.) was depressed, while the body weight development of the other treated male groups and all female groups was similar to that of the respective control groups during the course of this study.

FOOD CONSUMPTION: The food consumption in treated male and female groups 3 and 4 (200 and 1000 mg/kg bw.) were subject to large variation. In male group 4 (1000 mg/kg bw.) food consumption increased at week 3 after a transient decrease during the first two weeks of treatment and showed a second decrease at the last week of the study. Male group 3 (200 mg/kg bw.) showed a marked increase of feed intake at the last week of treatment,
due to spillage of diet. In females food consumption was increased in group 4 (1000 mg/kg bw.) beginning at the second week of treatment; spillage
of diet was observed in this group at week 3 and 4 of the study. Animals of group 3 (200 mg/kg bw.) showed only a transient increase at the first week of the study.

FOOD CONVERSION: A tendency to lower food conversion was observed in treated male group 4 (1000 mg/kg bw.), except at week 3 of treatment where it was markedly increased. In treated female group 4 (1000 mg/kg bw.) the food conversion was increased, while a transient increase was observed
only at the first week of treatment in female group 3 (200 mg/kg bw.).

WATER CONSUMPTION: The mean water consumption of male group 4 (1000 mg/kg bw.) was depressed at week 2 and 4 of the study, while the water
consumption in male group 3 (200 mg/kg bw.) was slightly elevated throughout the treatment period. The mean water consumption in female group 4 (1000 mg/kg bw.) showed a tendency to higher intake during the whole duration of the treatment.

OPHTHALMOSCOPIC EXAMINATION: Ophthalmic inspections and hearing examinations performed before and towards the end of the application period revealed no evidence of a reaction to the treatment.

HAEMATOLOGY: No changes were observed throughout the treated groups which could be related to the administration of the test compound.

CLINICAL CHEMISTRY: The activity of plasma alanine aminotransferase was increased in both males and females from the high dose group (1000
mg/kg) and increased activity of aspartate aminotransferase was noted in males and females of group 4 (1000 mg/kg) and in the females of group 3 (200 mg/kg). Increased levels of plasma urea and bilirubin occurred in the males of group 4 (1000 mg/kg). A minimal decrease in plasma globulin concentration in the male group 4 (1000 mg/kg) was noted. Total protein and albumin levels remained within the normal physiological limits and the albumin to globulin ratio was increased accordingly.

ORGAN WEIGHTS: The mean weight of the exsanguinated body was significantly lower than that of the controls in male group 4 (1000 mg/kg bw.). Both absolute and relative mean liver weights of all treated males were increased when compared to the control values. A statistically significant increase of liver weight was also observed in treated female groups 3 and 4 (200 and 1000 mg/kg bw.), whereas in females of group 2 (50 mg/kg bw.) this
increase was less marked and not significant. Both absolute and relative kidney weight was significantly increased in treated females of group 3 and 4 (200 and 1000 mg/kg bw.), while in treated males the weight of kidneys showed no marked alteration.

GROSS PATHOLOGY: All treated male rats of groups 2 and 3 (50 and 200 mg/kg bw.) showed enlarged livers. In males of treated group 4 (1000 mg/kg bw.) 3/5 rats had small thymuses and 2/5 small seminal vesicles. No gross anatomial changes were noted in treated female animals.

HISTOPATHOLOGY: There was extensive necrosis in the liver of one male rat (M20) of group 4 (1000 mg/kg bw.). Recent necrosis was found in 1/5 males of group 3 (200 mg/kg bw.) and in 2/5 males of group 2 (50 mg/kg bw.). Diffuse hypertrophy of hepatocytes was present in all treated male and female animals except for 4/5 female rats of group 2 (50 mg/kg bw.). All other microscopical changes noted in some control and test animals were only incidental in nature and not due to the application of the tested compound.

Effect levels

Dose descriptor:

NOAEL

Effect level:

< 50 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: liver weights, liver enzymes and the macroscopical and microscopical findings in the livers

Target system / organ toxicity

Critical effects observed:

not specified