Decahydro-1,1,7-trimethyl-3a,7-methano-3aH-cyclopentacyclooct-3-yl formate

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11.01.2018 to 06.02.2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes

Test type:

acute toxic class method

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Decahydro-1,1,7-trimethyl-3a,7-methano-3aH-cyclopentacyclooct-3-yl formate

EC Number:

261-118-8

EC Name:

Decahydro-1,1,7-trimethyl-3a,7-methano-3aH-cyclopentacyclooct-3-yl formate

Cas Number:

58096-47-2

Molecular formula:

C16H26O2

IUPAC Name:

decahydro-1,1,7-trimethyl-3a,7-methano-3aH-cyclopentacyclooct-3-yl formate

Test material form:

liquid

Specific details on test material used for the study:

CAS No: 58096-47-2
EC No: 261-118-8
Chemical Name: Decahydro-1,1,7-trimethyl-3a,7-methano-3aH-cyclopentacyclooct-3-yl formate
Trade Name: Clovanyl-3-Formate (CARYOLAN)
Molecular formula: C16H26O2
Molecular weight: 250.379

Test animals

Species:

rat

Strain:

Wistar

Sex:

female

Details on test animals or test system and environmental conditions:

Details on Test Animals
Species : Rat (Rattus norvegicus)

Strain : Wistar rats

Source : Geniron Biolabs Pvt. Ltd.
No.93, Solur, Anekal-Thally Road, Anekal
Bengaluru – 562106, India

No. of groups : One treatment group – 4 steps
(G1-FTS & STS and G2-FTS & G2-STS)

No. of animals : 3 animals (females) / treatment step

Age at treatment : 9 to 12 Weeks

Body weight range at treatment : 197.5 to 228.4 g
Note: At the time of selection of animals for each treatment step, the weight variation did not exceed ± 20 per cent of the mean body weight of any previously dosed animals.

Identification : By rat accession number. Identification of individual rats was by cage card and turmeric colour body markings. The rat accession number was allotted during the course of the study. The temporary body marking during acclimatization period was done with crystal violet.

Acclimatization : After physical examination for good health and suitability for experiment, the animals were acclimatized six days for G1-FTS, eight days for G1-STS, 10 days for G2-FTS and 12 days for G2-STS before treatment. Animals were observed once daily during acclimatization period. Females were nulliparous and non-pregnant.

Details on Environmental Conditions
Rats were housed under standard laboratory conditions, air conditioned with adequate fresh air supply (13.4 air changes/hour). Environment: with temperature 21 to 24°C, relative humidity 65 to 68%, with 12 hours light and 12 hours dark cycle.

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

The undiluted test item as supplied by the sponsor was administered a dose of 300 mg/kg bodyweight (0.29 mL//kg) for the treatment steps (G1-FTS & G1STS) and a dose of 2000 mg/kg (1.95 mL//kg) for the treatment steps (G2-FTS & G2-STS) as a single oral gavage to rats fasted for 16 to 18 hours (access to water will not be interrupted) based on the specific gravity of the test item which is 1.024 (as per TIDS provided by the sponsor).
Each animal was administered the test item orally by gavage using disposable plastic syringe attached with metal feeding canula. Animals were fed approximately 3 to 4 hours after dosing.

Doses:

300 mg/kg bodyweight and 2000 mg/kg bodyweight

No. of animals per sex per dose:

3 females per dose

Control animals:

yes

Details on study design:

To determine the acute toxicity, a stepwise procedure was employed with the use of three animals of a single sex (female), at each step. Sufficient information was obtained on the acute toxicity of the test item for its classification. The test item was administered orally to a group of experimental animals at one of the defined doses (i.e. 300 mg/kg body weight) as a first step (G1-FTS). As all the rats survived at this step, the test was continued at the same dose of 300 mg/kg bodyweight (G1-STS), all the rats survived at this step, the test was continued at the next higher dose of 2000 mg/kg bodyweight (G2-FTS), all the rats survived at this step, hence the test was confirmed with three additional animals with the same dose of
2000 mg/kg body weight (G2-STS). No test item-related mortality was observed, and hence testing was stopped and the LD50 cut-off value was arrived.

Results and discussion

Effect levels

Key result

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Based on:

test mat.

Mortality:

None observed

Clinical signs:

other: Reported in table below

Gross pathology:

Reported in table below

Any other information on results incl. tables

Table 1: Body weight, bidy weight changes and pre-terminal deaths

Group and Dose (mg/kg body weight)	Rat No.	Sex	Body weight (g)						Day of Death  (Time of Death)	No. dead/  No. tested	Pre-terminal deaths (%)
			Initial (Day 1)	8^thday	Weight change (day 8 – Initial)	15^thday	Weight change           (day 15 – Initial)	At Death
G1 (FTS) 300	Rm7691	F	204.8	209.8	5.0	219.6	14.8	-	NA	0/3	0
	Rm76982	F	228.4	234.9	6.5	242.1	13.7	-	NA
	Rm7693	F	201.4	210.6	9.2	219.3	17.9	-	NA
G1 (STS) 300	Rm7694	F	227.8	232.4	4.6	238.4	10.6	-	NA	0/3	0
	Rm7695	F	204.7	211.6	6.9	218.1	13.4	-	NA
	Rm7696	F	203.4	210,5	7.1	215.6	12.2	-	NA

Applicant's summary and conclusion

Interpretation of results:

Category 5 based on GHS criteria

Conclusions:

Based on the results of the present study, the test item Decahydro-1,1,7-trimethyl-3a,7-methano-3aH-cyclopentacyclooct-3-yl formate (Clovanyl-3-formate)LD50 was determined to be 5000 mg/kg body weight or Unclassified as per LD50 cut-off value.

Executive summary:

The acute oral toxicity study with Clovanyl-3-Formate (Caryolan)in Wistar rats was conducted to assess the toxicological profile of the test item. 

The undiluted test item was administered as a single oral gavage to overnight fasted (16 to 18 hours) three female rats (G1-FTS) at the dose of 300 mg/kg (0.29 mL/kg) body weight. There were no clinical signs of toxicity and pre-terminal deaths. Hence, three additional female rats were tested at the same dose of 300 mg/kg (0.29 mL/kg) body weight (G1-STS). There were no clinical signs of toxicity and pre-terminal deaths. Based on the scheme - Annex 2c of the guideline OECD 423, three additional female rats were tested at the next higher dose of 2000 mg/kg 1.99 mL/kg) body weight (G2-FTS). There were no clinical signs of toxicity and pre-terminal deaths. Hence, three additional female rats were tested at the same dose of 2000 mg/kg (1.99 mL/kg) body weight (G2-STS). There were no clinical signs of toxicity and pre-terminal deaths, dosing was stopped.

The rats were observed for mortality and clinical signs for 14 days post treatment. Body weights were recorded prior to dosing on day 1 and again on days 8 and 15. Necropsy was performed for all the rats at termination. All survived rats gained weight during experimental period. There were no gross pathological changes at necropsy.

Based on the results of the present study, The LD₅₀is 5000 mg/kg or unclassified as per the as per LD₅₀cut-off value.