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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:
From 25 July 1978 to 24 October 1978
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1980
Report Date:
1980

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
information on temperature and humidity were not recorded.
GLP compliance:
no
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Chlorendic anhydride- Physical state: White somewhat chunky powder.- Lot/batch No.: 8093-1

Test animals

Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Inc., Portage, Michigan
- Weight at study initiation: Male 75 to 106 g. Female 71 to 97 g.
- Fasting period before study: not applicable
- Housing: Suspended wire-mesh cages.
- Diet: ad libitum
- Water: ad libitum

ENVIRONMENTAL CONDITIONS
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS:

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Purine Laboratory Chow
Analytical verification of doses or concentrations:
no
Duration of treatment / exposure:
90 days
Frequency of treatment:
Daily
Doses / concentrationsopen allclose all
Dose / conc.:
100 ppm
Remarks:
Treatment level at 100 ppm gave a compound consumption of 8 mg/kg bw/day for males and females.
Dose / conc.:
500 ppm
Remarks:
Treatment level at 500 ppm gave a compound consumption of 39 mg/kg bw/day for males and 45 mg/kg bw/day for females.
Dose / conc.:
2 500 ppm
Remarks:
Treatment level at 2500 ppm gave a compound consumption of 202 mg/kg bw/day for males and 226 mg/kg bw/day for females.
No. of animals per sex per dose:
15
Control animals:
yes, concurrent no treatment
Positive control:
None

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice daily


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: weekly


BODY WEIGHT: Yes
- Time schedule for examinations: weekly


FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined 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: No


WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No


OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: pre-test period and at 3 months
- Dose groups that were examined: All groups


HAEMATOLOGY: Yes
- Time schedule for collection of blood: 1, 2 and 3 months.
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals : Five rats per sex per group


CLINICAL CHEMISTRY: Yes / No / No data
- Time schedule for collection of blood:1, 2 and 3 months.
- Animals fasted: No data
- How many animals: Five rats per sex per group


URINALYSIS: Yes
- Time schedule for collection of urine: 1, 2 and 3 months.
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data


NEUROBEHAVIOURAL EXAMINATION: No
Sacrifice and pathology:
GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes
Statistics:
All statistical analyses compared the treatment groups with the control group by sex Body weights (week 13), food consumption (week 13), hematological biochemical and urinalysis parameters (1, 2 and 3 months) and absolute and relative organ weights (terminal sacrifice) were compared by analysis of variance (one way classification) Bartlett's test for homogeneity of variances and the appropriate t-test (for equal or unequal variances) as described by Steel and Torrie using Dunnett's multiple comparison tables to judge significance of differences.

Results and discussion

Results of examinations

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
There were no signs of overt toxicity observed for the treated rats. Some incidental signs seen in a few control and or treated rats were malaligned upper incisors, soft stools, skin lesions, hair loss, lacrimation, corneal opacity, redness around eye.
Mortality:
mortality observed, treatment-related
Description (incidence):
Three high dose females died between the 5th and 13th week of study no other rats died during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
The mid and high dose males and all three groups of treated females had a decreased rate of weight gain that was compound related. The group mean body weights of the mid and high dose males and high dose females were significantly less than controls at week 13.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Mid and high dose males and high dose females had decreased food consumption values over the 90-day study when compared with controls however only the food consumption of the high dose females was significantly less than the controls.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
effects observed, treatment-related
Description (incidence and severity):
Eye problems such as pale coloration of eye, decrease in size of eyeball, dilated pupil unresponsive to light, clear or white internal eye and increased distance between pupil and cornea, occurred most frequently in rats that had had blood drawn via the orbital sinus technique.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
No compound-related effects were observed in the results of the hematologic tests. An incidental finding was the slightly elevated number of leucocytes seen for the high dose males at 1, 2 and 3 months of study. Other occasional statistically significant values were of no physiologic importance.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Both treated males and females had elevated SAP activities at 1, 2 and 3 months of study (only the 2 and 3 month values of the high dose females and the 2 month value of the mid dose females were significantly greater than controls). At 3 months of study, all groups had greater SAP activities than controls. Mid and low dose males and females had SGOT activities significantly lower than controls at 3 months of study. However, these activities as well as other occasional statistically significant values were of no physiologic importance. No other compound-related effects were seen in the results of the biochemical tests.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
No compound-related effects were seen in the results of the urinalyses. An incidental finding at 3 months of study was the elevated urinaly pH of two of five high dose females tested. The few statistically significant values (specific gravity) were of no physiologic importance.
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Statistically significant (p< 0.05) decreases in the mean absolute weight of spleens of female rats at the 2500 ppm dosage level, mean absolute weight of kidneys of male rats at the 2500 ppm dosage level, mean absolute weight of hearts of male rats at all dosage levels and the female rats at the 500 ppm dosage level and an increase in the mean weight of testes of male rats at the 500 ppm dosage level were noted. In respect of liver, statistically significant (p< 0.01) decreases in the mean absolute and relative weights at all dosage levels were observed. Of these variations decreases in the mean absolute weights of heart of male rats (p< 0.05) and in the mean absolute and relative weights of livers (p< 0.01) at all dosage levels, appeared to be treatment related. In the absence of any significant histomorphologic changes in these organs in the test groups, these decreases probably might be due to the overall reduction in the body weights resulting from a reduction of body fat and or extracellular body fluid.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
The three rats from the 2500 ppm group that died during the course of the study did not show any compound related lesions. None of the rats that were sacrificed at the termination of the study had any compound related lesions.
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
No compound-related microscopic lesions were observed in any of the tissues from rats that were examined from the 2500 ppm group. The microscopic lesions seen were considered spontaneous and incidental in nature.
Histopathological findings: neoplastic:
not examined

Effect levels

Key result
Dose descriptor:
NOAEL
Effect level:
> 2 500 ppm
Based on:
test mat.
Sex:
male/female
Basis for effect level:
clinical signs
mortality
body weight and weight gain
food consumption and compound intake
ophthalmological examination
haematology
clinical biochemistry
urinalysis
organ weights and organ / body weight ratios
gross pathology
histopathology: non-neoplastic

Target system / organ toxicity

open allclose all
Critical effects observed:
no
Lowest effective dose / conc.:
2 500 ppm
System:
immune system
Organ:
spleen
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Critical effects observed:
no
Lowest effective dose / conc.:
2 500 ppm
System:
hepatobiliary
Organ:
liver
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Critical effects observed:
no
Lowest effective dose / conc.:
2 500 ppm
System:
male reproductive system
Organ:
testes
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Critical effects observed:
no
Lowest effective dose / conc.:
2 500 ppm
System:
urinary
Organ:
kidney
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified
Critical effects observed:
no
Lowest effective dose / conc.:
2 500 ppm
System:
cardiovascular
Organ:
heart
Treatment related:
yes
Dose response relationship:
not specified
Relevant for humans:
not specified

Applicant's summary and conclusion

Conclusions:
No compound related gross lesions were seen in any of the rats from the treatment groups. No compound related microscopic lesions were seen in any of the tissues from rats that were examined from the 2500 ppm group.
Executive summary:

Chlorendic Anhydride was fed in the diet at levels of 100, 500, and 2S00 ppm to three groups of Charles River CD rats in a 90-day subacute toxicity study performed according to a method similar to OECD Testing Guideline 408 (non GLP) with deviations. There were 15 rats/sex/group. Control rats received the basal laboratory diet only. The rats were observed twice daily for signs of overt toxicity and mortality. Detailed observations, individual body weights and individual food consumption were recorded weekly. Ophthalmic examinations were conducted during the pretest period and at 3 months of study. Hematologic and biochemical tests and urinalyses were performed at 1, 2 and 3 months of study for five rats/sex/groups. Mid and high dose males and all three groups of treated females had decreased group mean body weights when compared with controls. Mid and high dose males and high dose females had decreased food consumption over the 90-day study when compared with controls. Both treated males and females had elevated SAP activities at 1, 2 and 3 months of study. Statistically significant decreases in the mean absolute weights of hearts of male rats and in the mean absolute and relative weights of livers of male and female rats at all dosage levels, appeared to be treatment related. No compound-related gross lesions were seen in any of the rats from the treatment groups. No compound-related microscopic lesions were seen in any of the tissues from rats that were examined from the 2500 ppm group.