Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
gas under pressure: refrigerated liquefied gas
Details on test material:
Identification: HEOX-CN
Chemical Name: 1-0xa-4-azaspiro[4.5]decane-4-ethanol
Cas-number : 2359-11-7
Description Clear liquid at 30°C
Batch: 0029504011005
Purity: ( purity: 98.9+/- 0.4 % m/m; cyclohexanone: 0.50 +/- 0.15% m/m; diethanolamine: 0.15 +/- 0.10 % m/m)
Test substance storage: In refrigerator, dry and in the dark
Stability under storage
conditions: Stable
Expiry date: December 01, 1995
Density: 1050 kg/m3 (40°C)
Preparation The test substance was dosed undiluted as delivered by the sponsor.
.

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
Rat, Wistar strain Crl:(WI) BR (outbred, SPFQuality).
Source : Charles River, Germany.
Age: Approx. 6 weeks.
WBody weight: ithin ± 20% of the sex mean.
5 males and 5 females
Identification: Earmark

Air-conditioned room with approximately 15 air changes per hour and the environment controlled with optimal conditions considered as being a temperature of 21°C and a relative humidity of 50%. Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity. Lighting was 12 hours artificial fluorescent light and 12 hours dark per day.

Accommodation
Group housing of 5 animals per sex per cage in labelled polycarbonate cages containing purified sawdust as bedding material (Woody SPF, supplied by B.M. I., Helmond, The Netherlands). Certificates of analysis were examined and then retained in the NDTDX a rchives.

Acclimatisation period was at least 5 days before start of treatment under laboratory conditions.

Diet
Free access to standard pelleted laboratory animal diet (from Carfil Quality BVBA, Dud-Turnhout, Belgium). Certificates of analysis are examined and then retained in the NOT OX archives.

Water
Free access to tap-water. Certificates of analysis (performed quarterly) are examined and then retained in the NOTOX archives

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
not specified
Details on oral exposure:
Food was withheld overnight prior to dosing until approximately 3-4 hours after administration of the test substance.
Oral gavage, Once, on day 1: 2000 mg/kg (1.905 ml/kg) body weight.
Dose volume calculated as follows: dose level (g/kg): density (g/mol).
Doses:
2000 mg/kg (1.905 ml/kg) body weight.
Control animals:
no
Details on study design:
Mortality/Viability: Twice daily. The time of death was recorded as precisely as possible.
Body weights: Days 1 (pre-administration), 8 and 15. and at death (if found dead after day 1).
Clinical signs: At periodic intervals on the day of dosing (day 1) and once daily thereafter, until day 15. The time of onset,
degree and duration were recorded.
Necropsy: All animals surviving to the end of the observation period (day 15) were sacrificed by oxygen/carbon dioxide asphyxiation.
All animals assigned to the study were sUbjected to necropsy and descriptions of all macroscopic abnormalities recorded.

Statistics:
No statistical analysis was performed.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
Mortality: One female was found dead on day 9.
Clinical signs:
Lethargy and uncoordinated movements were observed in all males on day 1.
Lethargy, uncoordinated movements, hunched posture, ventro-lateral recumbency and/or piloerection were observed in the females on day 1.
From day 6 onwards, signs of toxicity were again observed in three of the females and consisted of tremors, uncoordinated movements, hunched posture, pale skin, piloerection and/or emaciation. These signs of toxicity resulted in the death of one animal on day 9. The symptoms had completely disappeared by day 14 in the other two animals.
Body weight:
Body weight loss was observed in two females and slight body weight gain was noted in a third female over the first week. Body weight gain shown by the males and other females over the study period was considered to be normal.
Gross pathology:
Macroscopic Findings:
Partial cannibalism prohibited complete post mortem examination of the female that died during the study. No abnormalities were found in the organs present. Enlarged kidneys were found in one surviving female at macroscopic post mortem examination.

Applicant's summary and conclusion

Interpretation of results:
not classified
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
The oral LD50 value of HEOX-CN in rats was established as exceeding 2000 mg/kg body weight. The sensitivity of the females to HEOX-CN after oral administration appeared to be higher than in the males.
Executive summary:

The study was carried out in accordance with OECD Guideline No. 401, "Acute Oral Toxicity' and EEC Directive 92/69/EEC, Part B.1, 'Acute Toxicity-Oral'. This study should provide part of a rational basis for risk assessment in man. The oral route was selected as it is a possible route of human exposure during manufacture, handling or use of HEOX-CN.

HEOX-CN (CAS 2359 -11 -7) was administered by oral gavage to five rats of each sex per group, at 2000 mg/kg body weight. Animals were subjected to daily observations. Body weights were determined weekly and at death. Macroscopic examination was performed on the day of death or at the end of the experimental period (day 15). Clinical signs of toxicity were observed in all animals on the day of treatment. The severity and number of symptoms were increased in the females. From day 6 onwards, signs of toxicity were again observed in three females, resulting in the death of one animal on day 9. The symptoms had completely disappeared by day 14 in the other two animals. Enlarged kidneys were found in one of the latter females at macroscopic post mortem examination. Body weight loss or reduced body weight gain were observed in these three females over the first week. The sensitivity of the females to HEOX-CN after oral administration appeared to be higher than in the males.

The oral LD50 value of HEOX-CN in rats was established as exceeding 2000 mg/kg body weight. According to the Guidance to Regulation (EC) No. 1272/2008 on classification, labelling and packaging (CLP) of substances and mixtures, November 2012, the test substance is not classified.