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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1993-04-20 to 1993-07-23 (administration period)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Cross-reference
Reason / purpose:
reference to same study
Reference
Endpoint:
toxicity to reproduction
Remarks:
other: subchronic oral toxicity study
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1993-04-20 to 1993-07-23 (administration period)
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study.
Reason / purpose:
reference to same study
Qualifier:
according to
Guideline:
other: OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents) adopted May 12, 1981
Deviations:
no
Qualifier:
according to
Guideline:
other: EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents) 1988
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
Male and female Wistar rats (Chbb : THOM (SPF)
- Source: Dr. Karl THOMAE , Biberach / Riss , FRG
- Age at study initiation: 42 days
- Weight at study initiation: 179 (168 - 188) g for the males, 137 (125 - 147) g for the females .
- Fasting period before study: no
- Housing: individually in type DK III stainless steel wire cages (floor area about 800 cm² )
- Diet (ad libitum): ground Kliba maintenance diet rat/mouse/hamster, 343 meal, KLINGENTALMÜHLE AG, Kaiseraugst, Switzerland
- Water (ad libitum): tap water
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1993-04-13 To: 1993-07-23
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on mating procedure:
no mating
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
3 months
Frequency of treatment:
continuously in the diet
Remarks:
Doses / Concentrations:
ca. 58, 175, 340, 1085 mg/kg bw/d
Basis:
actual ingested
Remarks:
Doses / Concentrations:
750, 2250, 4500, 15000 ppm in the diet
Basis:
nominal in diet
No. of animals per sex per dose:
10 males and 10 females per group
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale:
The doses were selected on two previous studies with the test substance.

In a gavage study, Uvinul N 539 was administered to 5 male and 5 female Wistar rats (Chbb :THOM SPF) in olive oil at a dose of 2,000 mg/kg body weight for 15 consecutive days. As a control, 5 male and 5 female rats were treated with vehicle only. Following findings were obtained:
Mortality and body weight was not affected. In the second week of the study, salivation was observed each time after administration of the test substance. The blood sampling at the end of the study revealed in treated males and females statistically significantly increased serum-gamma-glutamyltransferase (SGGT) values and cholesterole values; in treated males, creatinine, total protein, and globulins were increased, in females red blood cells, hemoglobin, and hematocrit were decreased. However, from the above mentioned changes in clinical chemistry and hematology only the changes in SGGT and cholesterole were assessed as being treatment-related. Urine was discolored (slightly red). In both sexes, liver weights were increased.

In a palatability study (BASF 10S0227/92057), Uvinul N 539 was administered to 3 Wistar rats (CHbb:THOM SPF) per sex and dose at doses of 0; 4500 and 15000 ppm in the diet for 2 weeks. No substance-related impairment of food consumption was observed. At the end of the study, the body weights of the animals of the 15000 ppm group were about 97/93% (males/females) of the control values. However, due to the slightness of the effect and the low number of animals, it cannot be ruled out that this was an incidental finding.
Positive control:
no
Parental animals: Observations and examinations:
The first day of dosing was designated as day 0.

CAGE SIDE OBSERVATIONS and DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: The animals were examined for evident signs of toxicity twice a day (Mondays to Fridays) and once a day (Saturdays, Sundays and on public holidays). Once a week an additional comprehensive clinical examination was carried out.

BODY WEIGHT: Yes
- Time schedule for examinations: once weekly
The body weight was determined before the start of the administration period in order to randomize the animals. During the conduct of the study the body weight was determined on day 0 and thereafter once a week. The difference between the body weight on the respective day of weighing and the body weight on day 0 was calculated as the body weight change.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Food consumption over a period of 7 days was determined weekly and calculated as mean food consumption in grams per animal and day.
Oestrous cyclicity (parental animals):
no
Sperm parameters (parental animals):
no
Postmortem examinations (parental animals):
SACRIFICE
- Male animals: All surviving animals
- Maternal animals: All surviving animals

GROSS NECROPSY: Yes

HISTOPATHOLOGY / ORGAN WEIGHTS: Yes
Statistics:
The statistical evaluation and calculation of the data were carried out on the computer systems of the Department of Toxicology of BASF Aktiengesellschaft.
CLINICAL SIGNS AND MORTALITY
The administration of Uvinul N 539 in the diet in different dosages did not result in any substance-related findings. All findings obtained were incidental and not attributable to the test substance administration.
During the entire study period no animal died prematurely.

BODY WEIGHT AND WEIGHT GAIN
Body weight was impaired in high dose males and females, resulting in reduced values of 10/ 8% (males / females) at the end of the study.
Body weight change was impaired in high dose males and females , resulting in reduced values of 16 / 15 % (males / females) at the end of the study.
Although the difference was not always statistically significantly, the effect was assessed as being substance-related.

FOOD CONSUMPTION and COMPOUND INTAKE
The overall food consumption was reduced in high dose males and females, the values being about 13 and 10% below the control values, respectively. The food consumption of all other treated animals was within the range of biological variation.
For calculated compound intake, see table below.

ORGAN WEIGHTS, PATHOLOGY, HISTOPATHOLOGY
The following substance-related findings concerning the reproductive system were recorded:

15000 ppm group:
- Increased number of hypertrophic cells in the pars distalis of the pituitary gland of males. The cells had
abundant pale eosinophilic cytoplasm and often contained large vacuoles. They tended to occur
in the middle of the pars distalis rather than in the lateral regions.
- Slight or moderate hypertrophy of the thyroid foilicular epithelium (all animals) and associated pale staining colloid in both sexes

4500 ppm group:
- Minimal or slight hypertrophy of the thyroid follicular epithelium and associated pale staining colloid in both sexes

2250/750 ppm group:
Few incidences of minimal changes in the thyroid gland (not considered as treatment-related)

No treatment related gross lesions were observed.
No changes in absolute and relative testes weights in males and adrenal weigths in males/females were observed.
No treatment related incidences of microscopic findings were observed in adrenal glands, epididymides, prostate, testes of males.
No treatment related incidences of microscopic findings were observed in adrenal glands, mammary gland, ovaries, uterus, vagina of females.

For further details see chapter 7.5.1.
Reproductive effects observed:
not specified

Approximate food intake

Test

group

Concentration

[ppm]

Mean daily test substance intake

[mg/kg bw/d]

males

females

all animals

1

750

53

63

58

2

2250

163

187

175

3

4500

315

365

340

4

15000

1027

1143

1085

For results other than those to reproductive organs, see section 7.5.1, Repeated dose toxicity: oral.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1999

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Version / remarks:
adopted May 12, 1981
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
1988
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Uvinul N 539
- Analytical purity: 98.3%
- Lot/batch No.: 505396-70182
- Stability under test conditions: confirmed
- Storage condition of test material: room temperature

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
Male and female Wistar rats (Chbb : THOM (SPF)
- Source: Dr. Karl THOMAE , Biberach / Riss , FRG
- Age at study initiation: 42 days
- Weight at study initiation: 179 (168 - 188) g for the males, 137 (125 - 147) g for the females .
- Fasting period before study: no
- Housing: individually in type DK III stainless steel wire cages (floor area about 800 cm² )
- Diet (ad libitum): ground Kliba maintenance diet rat/mouse/hamster, 343 meal, KLINGENTALMÜHLE AG, Kaiseraugst, Switzerland
- Water (ad libitum): tap water
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24°C
- Humidity (%): 30-70%
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1993-04-13 To: 1993-07-23

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
3 months
Frequency of treatment:
continuously in the diet
Doses / concentrationsopen allclose all
Dose / conc.:
750 ppm
Remarks:
53 (males); 63 (females); 58 (all animals) mg/kg bw/d
Dose / conc.:
2 250 ppm
Remarks:
163 (males); 187 (females); 175 (all animals) mg/kg bw/d
Dose / conc.:
4 500 ppm
Remarks:
315 (males); 365 (females); 340 (all animals) mg/kg bw/d
Dose / conc.:
15 000 ppm
Remarks:
1027 (males); 1143 (females); 1085 (all animals) mg/kg bw/d
No. of animals per sex per dose:
10 males and 10 females per group
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale:
The doses were selected on two previous studies with the test substance.

In a gavage study, Uvinul N 539 was administered to 5 male and 5 female Wistar rats (Chbb :THOM SPF) in olive oil at a dose of 2,000 mg/kg body weight for 15 consecutive days. As a control, 5 male and 5 female rats were treated with vehicle only. Following findings were obtained:
Mortality and body weight was not affected. In the second week of the study, salivation was observed each time after administration of the test substance. The blood sampling at the end of the study revealed in treated males and females statistically significantly increased serum-gamma-glutamyltransferase (SGGT) values and cholesterole values; in treated males, creatinine, total protein, and globulins were increased, in females red blood cells, hemoglobin, and hematocrit were decreased. However, from the above mentioned changes in clinical chemistry and hematology only the changes in SGGT and cholesterole were assessed as being treatment-related. Urine was discolored (slightly red). In both sexes, liver weights were increased.

In a palatability study (BASF 10S0227/92057), Uvinul N 539 was administered to 3 Wistar rats (CHbb:THOM SPF) per sex and dose at doses of 0; 4500 and 15000 ppm in the diet for 2 weeks. No substance-related impairment of food consumption was observed. At the end of the study, the body weights of the animals of the 15000 ppm group were about 97/93% (males/females) of the control values. However, due to the slightness of the effect and the low number of animals, it cannot be ruled out that this was an incidental finding.
Positive control:
no

Examinations

Observations and examinations performed and frequency:
The first day of dosing was designated as day 0.

CAGE SIDE OBSERVATIONS and DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: The animals were examined for evident signs of toxicity twice a day (Mondays to Fridays) and once a day (Saturdays, Sundays and on public holidays). Once a week an additional comprehensive clinical examination was carried out.

BODY WEIGHT: Yes
- Time schedule for examinations: once weekly
The body weight was determined before the start of the administration period in order to randomize the animals. During the conduct of the study the body weight was determined on day 0 and thereafter once a week. The difference between the body weight on the respective day of weighing and the body weight on day 0 was calculated as the body weight change.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
Food consumption over a period of 7 days was determined weekly and calculated as mean food consumption in grams per animal and day.

FOOD EFFICIENCY: No

WATER CONSUMPTION: No

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: 1 days prior to the first dose and 84 days after beginning of treatment
- Dose groups that were examined: control and 15000 ppm group

HAEMATOLOGY: Yes
- Time schedule for collection of blood: 85 days after beginning of treatment
- Anaesthetic used for blood collection: No
- Animals fasted: No
- How many animals: all animals
- Parameters examined: leukocytes, erythrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets, differential blood count, reticulocytes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: 85 days after beginning of treatment
- Animals fasted: No
- How many animals: all animals
- Parameters examined: prothrombin time (Hepato Quick's test), alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, serum gamma-glutamyltransferase, sodium, potassium, chloride, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol, magnesium

URINALYSIS: Yes
- Time schedule for collection of urine: 79 days after beginning of treatment
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters examined: volume, color, turbidity, nitrite, pH, protein, glucose, ketones, urobilinogen, bilirubin, blood, specific gravity, sediment

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes
Statistics:
The statistical evaluation and calculation of the data were carried out on the computer systems of the Department of Toxicology of BASF Aktiengesellschaft.

Results and discussion

Results of examinations

Details on results:
CLINICAL SIGNS AND MORTALITY
The administration of Uvinul N 539 in the diet in different dosages did not result in any substance-related findings. All findings obtained were incidental and not attributable to the test substance administration.
During the entire study period no animal died prematurely.

BODY WEIGHT AND WEIGHT GAIN
Body weight was impaired in high dose males and females, resulting in reduced values of 10/ 8% (males / females) at the end of the study.
Body weight change was impaired in high dose males and females , resulting in reduced values of 16 / 15 % (males / females) at the end of the study.
Although the difference was not always statistically significantly, the effect was assessed as being substance-related.

FOOD CONSUMPTION and COMPOUND INTAKE
The overall food consumption was reduced in high dose males and females, the values being about 13 and 10% below the control values, respectively. The food consumption of all other treated animals was within the range of biological variation.
For calculated compound intake, see table below.

OPHTHALMOSCOPIC EXAMINATION
The ophthalmological examinations revealed no substance-related findings.

HAEMATOLOGY, CLINICAL CHEMISTRY, URINALYSIS
The following substance-related findings were recorded:

15000 ppm group
- decrease in hemoglobin mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), alanine aminotransferase and aspartate aminotransferase in the females
- increase in platelets in both sexes and shortened prothrombin time in the females
- increase in gamma-glutamyltransferase activity in both sexes
- increase in alkaline phosphatase, total protein, globulins and total cholesterol in the females
- dark yellow discoloration of the urine specimens in both sexes
- decrease in total bilirubin in both sexes

4500 ppm group
- increase in platelets, total protein and globulins in the females
- decrease in alanine aminotransferase and aspartate aminotransferase in the females
- decrease in total bilirubin in both sexes

ORGAN WEIGHTS, PATHOLOGY, HISTOPATHOLOGY
The following substance-related findings were recorded:

15000 ppm group:
- increased absolute and relative liver weights in male and female rats
- prominent acinar pattern of the liver in 7 out of 10 male rats
- hypertrophy of periacinar hepatocytes in both sexes
- hypertrophy of centriacinar hepatocytes in both sexes
- slight or moderate hypertrophy of the thyroid, follicular epithelium and associated pale staining colloid in both sexes
- increased number of hypertrophic cells in the pituitary gland of males

4500 ppm group:
- increased absolute and relative liver weights in female rats
- hypertrophy of periacinar hepatocytes in both sexes
- hypertrophy of centriacinar hepatocytes in males
- minimal or slight hypertrophy of the thyroid follicular epithelium and associated pale staining colloid in both sexes

2250/750 ppm group:
Few incidences of minimal changes in the thyroid gland (not considered as treatment-related)

Effect levels

Dose descriptor:
NOAEL
Effect level:
2 250 ppm
Sex:
male/female
Basis for effect level:
other: overall effects
Remarks on result:
other: approx. 175 mg/kg bw/d

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Approximate mean food intake

Test

group

Concentration

[ppm]

Mean daily test substance intake

[mg/kg bw/d]

males

females

all animals

1

750

53

63

58

2

2250

163

187

175

3

4500

315

365

340

4

15000

1027

1143

1085

Applicant's summary and conclusion