1,4,5,6,7,7-hexachloro-8,9,10-trinorborn-5-ene-2,3-dicarboxylic anhydride

Repeated dose toxicity: dermal

Administrative data

Endpoint:

short-term repeated dose toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 13 January 1978 to 3 February 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:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 410 (Repeated Dose Dermal Toxicity: 21/28-Day Study)

Deviations:

yes

Remarks:

information on temperature and humidity were not recorded. Food consumption was not monitored.

GLP compliance:

no

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

1,4,5,6,7,7-hexachloro-8,9,10-trinorborn-5-ene-2,3-dicarboxylic anhydride

EC Number:

204-077-3

EC Name:

1,4,5,6,7,7-hexachloro-8,9,10-trinorborn-5-ene-2,3-dicarboxylic anhydride

Cas Number:

115-27-5

Molecular formula:

C9H2Cl6O3

IUPAC Name:

1,4,5,6,7,7-hexachloro-8,9,10-trinorborn-5-ene-2,3-dicarboxylic anhydride

Details on test material:

- Name of test material (as cited in study report): Chlorendic anhydride- Physical state: White somewhat chunky powder- Analytical purity: 93.81%- Lot/batch No.: 3-12-206

Test animals

Species:

rabbit

Strain:

New Zealand White

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Sweetwater Farms, Hillsboro, Ohio
- Weight at study initiation: Male 2124 to 2781 g. Female 2187 to 2602 g
- Diet: Purina Rabbit Chow ad libitum
- Water: ad libitum
- Acclimation period: Approximately 2 weeks.

Administration / exposure

Type of coverage:

occlusive

Vehicle:

physiological saline

Details on exposure:

TEST SITE
- Area of exposure: Dorsal
- % coverage: Approximately 10% of body surface
- Type of wrap if used: None

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 100, 500 and 2500 mg/kg

VEHICLE
- Justification for use and choice of vehicle (if other than water): physiological saline used as control
- Amount(s) applied (volume or weight with unit): 1.2 ml/kg/day

USE OF RESTRAINERS FOR PREVENTING INGESTION: yes

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

5 days per week for 3 weeks for a total of 15 applications

Frequency of treatment:

Daily for 5 weekdays, break at week-ends for 3 weeks.

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

100 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

500 mg/kg bw/day (nominal)

Doses / concentrations 3

Dose / conc.:

2 500 mg/kg bw/day (nominal)

No. of animals per sex per dose:

4

Control animals:

yes, concurrent vehicle

Positive control:

None

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: daily

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

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: All

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At termination of study
- Animals fasted: No data
- How many animals: All

URINALYSIS: Yes
- Time schedule for collection of urine:
- Metabolism cages used for collection of urine: No data

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: YesHISTOPATHOLOGY: Yes

Statistics:

All statistical analyses compared the treatment groups with the control group by sex. At termination of the study, body weights, hematologic, biochemical and urinalysis parameters and absolute and relative organ weights 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:

no effects observed

Description (incidence and severity):

Incidental findings (primarily at the 2500 mg/kg/day dosage level) included: diarrhea, nasal or ocular discharge, hypoactivity, anorexia and dehydration.

Dermal irritation:

effects observed, treatment-related

Description (incidence and severity):

One or more signs of dermal irritation were present for all treated rabbits. The level of intensity and time of onset were primarily dose related. At the 100 mg/kg/day dosage level, a barely perceptible erythema was noted for all rabbits beginning in weeks 2 or 3 of treatment. At the 500 mg/kg/day dosage level, the onset of erythema (barely perceptible to slight) was usually evident by day 4 and persisted throughout most, or all or the treatment period. Other signs of dermal irritation (barely perceptible to slight) included: edema, atonia, desquamation, coriaceousness and fissuring. These signs were evident for most treated rabbits during either the second or third week of treatment. At the 2500 mg/kg/day dosage level, erythema (barely perceptible to moderate) was evident as early as day 2 and persisted throughout the study for most rabbits. Other signs of dermal irritation as noted previously (barely perceptible to moderate) were evident by, or on, day 7 and for most rabbits persisted throughout the treatment period.

Mortality:

no mortality observed

Description (incidence):

None of the rabbits died.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Female rabbits in the 2500 mg / kg / day group had decreased body weight gains when compared to controls. Males had slight but statistically significant weight losses.

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:

no effects observed

Description (incidence and severity):

No changes considered to be related to compound were seen in the hematologic studies.

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Description (incidence and severity):

No urine was collected from one male and one female rabbit in the 2500 mg kg day treatment group. All other urinalyses were similar to the control.

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

No compound related organ weight variations were observed

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

Six rabbits from the 2500 mg/kg/day group and 2 rabbits from the 500 mg/kg/day group had stomach lesions which may have been compound related. These stomach changes were described as ulcerations, erosions and yellow gray or white foci or areas in the mucosa. They were not seen in rabbits from the control or 100 mg/kg/day groups.

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Stomach erosions noted grossly and confirmed microscopically in several rabbits from the 2500 mg/kg/day group were shallow and did not extend the full thickness of the mucosa. The stomach mucosa away from the grossly noted erosions was completely normal. Erosions or other stomach alterations could not be confirmed microscopically in all stomachs in which a gross description of a lesion was made. Stomach lesions were not seen microscopically in rabbits from the control or 100 mg/kg/day groups; their occurrence in rabbits from the 500 and 2500 mg/kg/day groups was probably compound related.
Evidence of very slight to slight compound-related dermal irritation was seen in most rabbits from the 2500, 500 and 100 mg/kg/day groups. These skin changes included epidermal acanthosis and hyperkeratosis and inflammatory cell infiltrate in the dermis. The severity of these skin changes appeared somewhat dose related and the overall skin response to this compound could best be characterized as mild. Other microscopic lesions were considered spontaneous and unrelated to treatment and were typical of the usual lesions seen in untreated rabbits.
Brain lesions characterized by perivascular lymphocytic cuffing, glial nodules and lymphocytic meningitis were considered due to infestation by Encephalitozoan cunniculli, a common protozoan parasite of laboratory rabbits.

Histopathological findings: neoplastic:

not examined

Effect levels

Key result

Dose descriptor:

NOEL

Effect level:

> 2 500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: observed effects were not considered as adverse and significant

Target system / organ toxicity

Critical effects observed:

no

Lowest effective dose / conc.:

500 mg/kg bw/day (nominal)

System:

gastrointestinal tract

Organ:

stomach

Treatment related:

yes

Dose response relationship:

not specified

Relevant for humans:

not specified

Applicant's summary and conclusion

Conclusions:

Evidence of mild skin irritation, characterized by hyperkeratosis, acanthosis and dermal inflammatory cell infiltrate was seen at the application site in most rabbits from the 100, 500 and 2500 mg / kg / day groups and was considered compound related. Overall skin response based on microscopic examination of the application site was characterized as mild.

Executive summary:

Chlorendic Anhydride was administered to the backs of New Zealand White rabbits at dosage levels of 100 500 and 2500 mg/kg/day 5 days a week during this 3 week dermal study according to a method similar to OECD Testing Guideline 410 (non GLP) with deviations. Four male and four female rabbits were used at each dosage level and in the control group. The rabbits were observed daily for signs of overt toxicity, general behavior, dermal irritation, moribundity or mortality. Body weights were recorded weekly Hematologic, biochemical and urinalysis studies were conducted during the control period and following the 2l day treatment period. One or more of the following signs of dermal irritation were noted for all treated rabbits: erythema, edema, atonia, desquamation coriaceousness and fissuring. The number of signs observed, severity of the conditions barely perceptible to moderate and duration were dose related. Incidental findings primarily at the 2500 mg/kg/day dosage level included diarrhea, nasal or ocular discharge, hypoactivity, anorexia and dehydration. Male and female rabbits at the high dosage level had decreases in weight when compared with the controls. All rabbits survived the treatment period. No changes considered related to compound were seen in the hematologic and biochemical studies. Urinalyses were considered normal. Stomach mucosal lesions described as erosions ulcerations or light foci and areas at necropsy in rabbits from the 2500 and 500 mg/kg/day were the only gross findings at terminal sacrifice which were considered compound related. No compound related organ weight variations were observed. Microscopically grossly described stomach changes were confirmed in several rabbits from the 500 and 2500 mg/kg/day groups. These changes were attributed to compound effect. Evidence of mild skin irritation characterized by hyperkeratosis, acanthosis and dermal inflammatory cell infiltrate was seen at the application site in most rabbits from the 100, 500 and 2500 mg/kg/day groups and was considered compound related. Overall skin response based on microscopic examination of the application site was characterized as mild.