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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1976
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was conducted non-GLP, however it was conducted state of the art at the time of conduct. Although the animals were young at start of dosing, the study is considered relevant, reliable and adequate.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1976
Report Date:
1976

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
no
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other: liquid

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, Mass.
- Age at study initiation: weanlings
- Weight at study initiation: 50-79 g
- Fasting period before study: No data
- Housing: individual drawer-type wire mesh cages
- Diet : Charles River, Rat, Mouse and Hamster Meal.
- Water: ad libitum
- Acclimation period: Not provided

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24.4°C
- Humidity (%):45-65%
- Air changes (per hr):10
- Photoperiod (hrs dark / hrs light):12/12

Administration / exposure

Route of administration:
oral: feed
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: 0; 0.50; 2.00; 8.00 (four weeks) and 4.00 g/kg body weight/day for the remaining nine weeks
The diet was prepared by mixing, on a weigth basis, the compound with the basic diet in a Patterson-Kelley twin shell blender, equiped with a high speed mixing bar having slanted blades. Calculations were based on the weight of the animals for the second preceding week. The dosages express active compound, based on the 35% solid concentration of the sample.

DIET PREPARATION
- Rate of preparation of diet (frequency):Fresh diets were prepared each week
- Mixing appropriate amounts with (Type of food): Charles River, Rat, Mouse and Hamster Meal.
- Storage temperature of food: Not provided

VEHICLE No Vehicle
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days
Frequency of treatment:
daily
Doses / concentrations
Remarks:
Doses / Concentrations:
0; 0.50; 2.00; 8.00 (four weeks) and 4.00 g/kg body weight/day for the remaining nine weeks
Basis:
nominal in diet
No. of animals per sex per dose:
20
Control animals:
yes, plain diet

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS:
- Time schedule:daily observations were made and any abnormal conditions recorded.

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined : Yes, weekly
and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at six weeks and at the termination of the study
- Anesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 5 rats of each sex (=10) from each group (4 groups)>40 rats
- Following parameters were examined:
Hematocrit Percentage
Hemoglobin Concentration
Total Leukocyte Count
Differential Leukocyte Count
RBC Morphology


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the conclusion of the experiment when possible
- Animals fasted: No data
- How many animals: the same rats as those used for hematology at the conclusion of the experiment= 40
- Following parameters were examined:
Serum Glucose Concentration
Blood Urea Nitrogen (BUN) Concentration
Serum Glutamic Oxaloacetic Transaminase (SGOT)
Serum Glutamic-Pyruvic Transaminase (SGPT)
Gamma Glutamyl Transpeptidase (GGTP)

URINALYSIS: Yes
- Time schedule for collection of urine: Twenty-four hour urine specimens were collected from 10 rats/group at six weeks and prior to sacrifice.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Following parameters were examined:
Color
Appearance
Reaction(pH)
Specific Gravity
Protein
Sugar
WBC/h.p.f.
RBC/h.p.f.

NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes , all rats underwent a thorough gross necropsy. Rats which had positive gross findings had either complete or partial (major organ) histology examination performed.
HISTOPATHOLOGY: Yes
Statistics:
Numerical data were treated statistically (student t test)

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
Two high dose rats died during the course of the experiment; male rat #5105 of acute hemorraghic prostatitis on the 27th day of the test, and female rat #5090 of hydrophilic degeneration of the liver and kidney on the 89th day.
Mortality:
mortality observed, treatment-related
Description (incidence):
Two high dose rats died during the course of the experiment; male rat #5105 of acute hemorraghic prostatitis on the 27th day of the test, and female rat #5090 of hydrophilic degeneration of the liver and kidney on the 89th day.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
[Trade name] depressed significantly the rate of body weight gain in the mid dose and high dose rats under the conditions of this experiment. This depression appeared more pronounced in the male than in the female rats.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
During weeks four through nine the feed consumption of the high dose male animals was significantly depressed. The other groups showed only an occasional significant depression.
Food efficiency:
effects observed, treatment-related
Description (incidence and severity):
The feed efficiency of the mid dose and high dose test groups is decidedly different from that of the control group, more so in the male than in the female rats.
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Some rats in all levels exhibited slightly elevated SGOT values and 4 high dose rats exhibited elevated SGPT levels. Histologically, however, there was no basis for these elevations.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Some of the rats in the mid dose and high dose group displayed hematuria to a greater or lesser degree. Their urinary system was not remarkably changed histologically. Only one rat, #5103, high dose group, displayed hematuria and moderate acute cystitis.
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Decrease in thyroid weight in males of high dose group; decrease in adrenal and gonadal weight in high dose groups; decrease in pituitary weight in females of high dose group; decrease in brain weight in males of mid dose and high dose group.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
Rats of various groups had chronic respiratory disease. Two male rats had lower urinary tract pathology, 1 control rat had a pulmonary granuloma, 1 high dose rat hydrophilic degeneration of liver and kidneys, 1 high dose rat hepatic lipidosis.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Rats of various groups had chronic respiratory disease. Two male rats had lower urinary tract pathology, 1 control rat had a pulmonary granuloma, 1 high dose rat hydrophilic degeneration of liver and kidneys, 1 high dose rat hepatic lipidosis.
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL SIGNS AND MORTALITY
Due to decrease in body weight and feed consumption in the high dose group (8.00 g/kg/day) the level was lowered to 4.00 g/kg/day for weeks five through the termination of the study. Two high dose rats died during the course of the experiment; male rat #5105 of acute hemorraghic prostatitis on the 27th day of the test, and female rat #5090 of hydrophilic degeneration of the liver and kidney on the 89th day.

BODY WEIGHT AND WEIGHT GAIN
Aerosol 22 depressed significantly the rate of body weight gain in the mid dose and high dose rats under the conditions of this experiment. This depression appeared more pronounced in the male than in the female rats.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
During weeks four through nine the feed consumption of the high dose male animals was significantly depressed. The other groups showed only an occasional significant depression.

FOOD EFFICIENCY
The feed efficiency of the mid dose and high dose test groups was decidedly different from that of the control group, more so in the male than in the female rats.

HAEMATOLOGY
All rats had normal hematocrit concentrations for the duration of the experiment
All rats had normal hemoglobin concentrations for the duration of the experiment
Rat #5097 (4.00 g/kg/day group) had a WBC of 35000 cells/mL at six weeks but by three months its WBC was normal. The rest of the rats had normal white cell counts at all times examined.
All rats examined at six and 13 weeks displayed lymphocytosis.
The RBC’s of all animals were morphologically normal at all times.

CLINICAL CHEMISTRY
All rats had normal blood glucose values when examined.
All rats examined had normal BUN values.
Some rats in all levels exhibited slightly elevated SGOT values. Histologically, however, there was no basis for these elevations.
Four high dose rats exhibited elevated SGPT levels. Histologically, however, there was no basis for these elevations.
All rats examined had normal GGPT values.

URINALYSIS
Some of the rats in the mid dose and high dose group displayed hematuria to a greater or lesser degree. Their urinary system, however, was not remarkable histologically. Only one rat, #5103, high dose group, displayed hematuria and moderate acute cystitis

ORGAN WEIGHTS
Cfr. Table 3 and 4 (absolute & relative organ weights)

GROSS PATHOLOGY
Rats of various groups had chronic respiratory disease. Two male rats had lower urinary tract pathology, 1 control rat had a pulmonary granuloma, 1 high dose rat hydrophilic degeneration of liver and kidneys, 1 high dose rat hepatic lipidosis.

Effect levels

Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (nominal)
Based on:
act. ingr.
Sex:
male/female
Basis for effect level:
other: overall effects

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Table 1. Body weight data for rats receiving [Trade name] for 13 weeks

Dose levels (g/kg).

Mean male Body Weight (g)

Weight Gain (g)

Mean female Body Weight (g)

Weight Gain (g)

Initial

Terminal

Initial

Terminal

0

68.4

509.0

440.6

60.0

289.3

229.3

0.50

68.3

505.6

437.3

60.1

276.7

216.6

2.00

68.4

434.5*

366.1

60.1

249.9*

189.8

8.00 (4.00)

68.2

269.8*

192.6

59.8

189.1*

129.3

* Statistically significant from control at P=0.05.

 

Table 2. Feed consumption for rats receiving [Trade name] for 13 weeks (g/rat/week)

Week

on

diet

Males

Females

Dose levels (g/kg)

Dose levels (g/kg)

0

0.50

2.00

8.00 (4.00)

0

0.50

2.00

8.00 (4.00)

1

103.8

97.2

112.5

101.5

83.3

90.3

86.8

84.0

2

124.8

119.0

133.5

137.0

103.5

116.5

106.3

115.3

3

157.3

157.8

162.8

163.0

134.8

144.3

150.5*

156.5*

4

157.3

135.0*

132.5*

186.1*

137.5

123.8

125.5

122.3

5

199.0

189.0

195.3

152.1*

158.3

138.0*

140.0*

150.8

6

185.7

185.7

190.1

162.8

145.0

159.2

168.8*

138.0

7

184.8

183.0

188.0

206.6*

130.8

131.0

137.1

170.8*

8

185.0

186.0

183.0

196.1*

150.3

138.5

152.8

164.3

9

175.3

178.5

184.3

194.2*

149.5

139.8

136.5*

166.0

10

181.0

179.5

184.0

176.3

134.8

129.3

138.8

150.8

11

182.5

191.0

183.7

177.1

147.5

153.2

143.2

164.5

12

179.0

178.4

176.3

182.4

160.0

145.0

158.3

172.8

13

190.0

188.9

177.0

160.5*

142.5

126.8

131.0

167.1

Mean

169.7

166.8

169.5

168.9

136.8

133.5

136.6

147.9

* Statistically significant from control at P=0.05.

 

Table 3. Summary of Mean Body Weights and Organ Weights on rats receiving [Trade name] in their diet for 13 weeks

Sex

Dose

Level.

g/kg

Terminal

Body Weight

   (g)

Organ Weights

Thyroids

Heart

Liver

Adrenals

Kidneys

Gonads

Pituitary

Brain

Male

0

503.9

0.034

1.262

20.390

0.060

3.527

3.361

0.013

2.216

 

0.50

503.7

0.031

1.274

22.540

0.060

3.566

3.491

0.013

2.107

 

2.00

443.3*

0.033

1.158

15.884*

0.064

2.984*

3.179

0.014

2.084*

 

8.00(4)

286.9*

0.025*

0.809*

9.890*

0.046*

1.926*

2.709*

0.010

1.906*

 

Female

0

301.6

0.033

0.879

11.326

0.085

1.966

0.152

0.017

1.969

 

0.50

280.9

0.034

0.799

9.759*

0.080

1.900

0.169

0.017

1.969

 

2.00

254.3*

0.031

0.755*

10.529

0.077

1.912

0.153

0.017

1.897

 

8.00(4)

200.2*

0.032

0.636*

8.316*

0.061*

1.582*

0.101*

0.012*

1.938

*Statistically significant from control at P=0.05

 

Table 4. Organ-to-body weight ratios for male and female beagle dogs receiving [Trade name] in their diet for 13 weeks

 

Dose

Levels

g/kg

Terminal

Body Weight

(g)

(Grams of organ weight/kilograms of body weight)

 

Sex

Thyroids

Heart

Liver

Adrenals

Kidneys

Gonads

Pituitary

Brain

Male

0

503.9

0.067

2.504

40.464

0.119

6.999

6.670

0.026

4.398

 

0.50

503.7

0.073

2.529

44.749

0.119

7.080

6.931

0.026

4.183

 

2.00

443.3*

0.074

2.612

35.831

0.144

6.731

7.171

0.032

4.701

 

8.00(4)

286.9*

0.087

2.820

34.472

0.160

6.713

9.442

0.035

6.643

 

Female

0

301.6

0.109

2.914

37.553

0.282

6.519

0.504

0.056

6.529

 

0.50

280.9

0.121

2.844

34.742

0.285

6.764

0.602

0.061

7.010

 

2.00

  254.3*

0.122

2.969

41.404

0.303

7.519

0.602

0.067

7.460

 

8.00(4)

 200.2*

0.160

3.177

41.538

0.305

7.902

0.504

0.060

9.680

*Statistically significant from control at P=0.05.

 

 

 

 

Applicant's summary and conclusion

Conclusions:
These studies have shown that the test item, when fed to random bred albino rats under the conditions of this experiment, decreased the rate of body weight gain, feed consumption and food efficiency at the mid dose and high dose levels, and increased SGOT and SGPT at the high doses. Further hematuria was seen in mid and high dosed rats, various organ weights were decreased and lower urinary tract pathology was seen in 2 high dosed rats.It thus appeared that the no effect level in the rat is below 0.50 g/kg/day, however 0.50g act.ingr./kg bw/day can be considered as NOAEL.
Executive summary:

Sprague-Dawley albino rats, 80 of each sex, were fed a control diet, or 0.50, 2.00 or 8.00 g/kg /day of test item, containing 35.8% active ingredient, mixed in the diet. The high dose was reduced to 4.00 g/kg/day of test item mixed in the diet for weeks five through to termination.
The rats were carefully observed for the duration of the experiment and pertinent hematological and biochemical parameters were conducted while the experiment was in progress. At the conclusion of the experiment all rats were necropsied and all organs and tissues were examined histologically from ten rats (five male and five female) from the control and high dose groups.
These studies have shown that the test item, when fed to random bred albino rats under the conditions of this experiment, decreased body weight gain, feed consumption and food efficiency at the mid dose and high dose levels and increased SGOT and SGPT at the high doses. Further hematuria was seen in the mid and high dose rats, various organ weights were decreased (e.g. decrease in adrenal and gonadal weight in high dose groups; decrease in pituitary weight in females of high dose group) and lower urinary tract pathology was seen in 2 high dosed rats. It thus appeared that the no effect level in the rat is below 0.50 g/kg/day, however 0.5g act.ingr./kg bw/day can be considered as NOAEL.