Cyclopentane, methoxy-

Endpoint summary

Description of key information

Acute toxicity, oral, (rat): 200 mg/kg bodyweight < LD50 < 2000 mg/kg bodyweight

Acute toxicity, inhalation (rat): 4h LC50 >21.5 mg/L

Acute toxicity, dermal (rat): LD50 > 2000 mg/kg bodyweight

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

18 January 2002 - 30 September 2002

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.1 tris (Acute Oral Toxicity - Acute Toxic Class Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

acute toxic class method

Limit test:

no

Specific details on test material used for the study:

- Analytical purity: 99.8%
- Lot/batch No.: 010912

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 5 - 7 weeks
- Weight at study initiation: 103 - 130g
- Fasting period before study: Yes (overnight before dosing and for 4 hours after dosing)
- Housing: metal cages with wire mesh floors
- Diet (e.g. ad libitum): Adlibitum
- Water (e.g. ad libitum): Ad libitum
- Acclimation period: Minimum of five days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 40 - 70%
- Photoperiod (hrs dark / hrs light): 12 hours light : 12 hours dark

IN-LIFE DATES: From: 11 February 2002 To:08 March 2002

Route of administration:

oral: gavage

Vehicle:

other: 1% Aqueous methylcellulose

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 20 mg/mL or 200 mg/mL

MAXIMUM DOSE VOLUME APPLIED: 10 mL/kg

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 2000 mg/kg chosen as the starting dose for compliance with the guidelines.

Doses:

Group 1: 2000 mg/kg (3 females)
Group 2: 200 mg/kg (3 females)
Group 3: 200 mg/kg (3 males)

No. of animals per sex per dose:

3 (see above)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: recorded on day 1 prior to dosing, day 8, and day 15. Body weights were also recorded on death if applicable.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight, macropathology.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 200 - < 2 000 mg/kg bw

Mortality:

Female: 2000 mg/kg bw; Number of animals: 3; Number of deaths: 2
Female: 200 mg/kg bw; Number of animals: 3; Number of deaths: 0
Male: 200 mg/kg bw; Number of animals: 3; Number of deaths: 0

Clinical signs:

Signs of toxicity related to dose levels: Two females dosed at 2000 mg/kg died within six and a half- hours of dosing.

Clinical signs of reaction to treatment comprised of piloerection, seen in all rats at both dosages. These signs were accompanied in all females at 2000 mg/kg by salivation, abnormal gait, lethargy, reduced body temperature, prostration, shallow respiration and lacrimation with prominent eyes and hunched posture in one female at 2000 mg/kg. Among rats at 200 mg/kg signs include hunched posture in all males and one female and abnormal gait in all females and one male. Recovery of surviving rats, as judged by external appearance and behaviour, was complete by either Day 2 (all rats at 200 mg/kg) or Day 3 (female at 2000 mg/kg).

Body weight:

All surviving animals were considered to have achieved satisfactory bodyweight gains throughout the study.

Gross pathology:

Macroscopic examination of both decedents revealed congestion (characterised by blood vessels injected) in the brain and thickened tissues and atrophy of the heart. Congestion and fluid contents were noted in the stomach and along the alimentary tract.

Effects on organs: No abnormalities were revealed for the remaining surviving animals at the macroscopic examination at study termination on Day 15.

Interpretation of results:

Category 4 based on GHS criteria

Conclusions:

The acute median lethal oral dose (LD50) to rats of the substance was demonstrated to be between 200 and 2000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

200 mg/kg bw

Quality of whole database:

Guideline study, under GLP.

Acute toxicity: via inhalation route

Link to relevant study records

Reference

Endpoint:

acute toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2002-12-16 to 2003-02-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 403 (Acute Inhalation Toxicity)

Version / remarks:

1981

Deviations:

no

GLP compliance:

yes

Test type:

traditional method

Limit test:

no

Specific details on test material used for the study:

Lot No.: 020618
Purity: 99.99%

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories
- Females nulliparous and non-pregnant: yes
- Age at study initiation: Approximately 8 weeks
- Weight at study initiation: MaIe: 277 - 319 g; Female: 210 - 233 g
- Housing: group housed (2-6/cage) during acclimation, and then individually in suspended stainless steel wire mesh cages during the study.
- Diet: Certified Rodent Diet, No. 5002; (Meal), available without restriction
- Water: available without restriction via an automated watering system
- Acclimation period: approximately 1 week.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 to 23°C
- Humidity (%): 13 to 69%
- Photoperiod (hrs dark / hrs light): twelve-hour light/dark cyde controlled via an automatic timer

Route of administration:

inhalation: vapour

Type of inhalation exposure:

nose only

Vehicle:

other: nitrogen

Mass median aerodynamic diameter (MMAD):

>= 1.204 - <= 3.775 µm

Geometric standard deviation (GSD):

> 1.649 - <= 2.342

Remark on MMAD/GSD:

Group 1 - 10.5 mg/L: Mean MMAD: 2.334 µm, mean GSD: 1.891
Group 2 - 21.5 mg/L: Mean MMAD: 2.149 µm, mean GSD: 1.896

Details on inhalation exposure:

- Pre-Study Trials: Trials were performed to evaluate the optimal set of conditions and equipment to generate stable atmospheres at the targeted exposure level.
- Chamber Operation:
The flow-past nose-only exposure chamber was operated at a flow rate of 10 Liters per minute. The final airflow was set to provide at least one air change in 5.0 minutes (12 air changes/hour) and a T99 equilibrium time of approximately 3 minutes. The T 99 equilibrium time was added to the 4 hours; i.e., the atmosphere was generated for 4 hours and T 99 minutes. This chamber size and airflow rate is considered adequate to maintain the oxygen level at least 19%. The chamber was exhausted via a 1" tubing through the in-house filtering system, which consisted of a coarse filter, a HEPA filter and an activated charcoal bed. The nose-only exposure chamber was contained within a 10 m3 chamber.
At the end of the exposure, all animals remained in chamber for a minimum of 30 minutes. During this time the chamber was operated at the same flow rate as used during the exposure using clean air only.
Recordings of total chamber flow and static pressure were made every half-hour during the exposure.
- Environmental conditions: Temperature: 20 to 23°C, Relative Humidity 17 to 23%

Analytical verification of test atmosphere concentrations:

yes

Duration of exposure:

4 h

Concentrations:

Group 1: 10 mg/L (Trarget)
Group 2: 20 mg/L (Trarget)

No. of animals per sex per dose:

5 animals per sex per dose

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Viability Checks: observed for mortality and general condition twice daily (once in the morning and once in the afternoon).
- Physical Examinations: Day 0 (Day of Exposure): Animals were observed individually prior to exposure, as a group, at fifteen minutes intervals during the first hour of exposure, and hourly for the remainder of the exposure. They were observed individually upon removal from the chamber and hourly for two hours post -exposure. Days 1 – 14: once daily.
- Body Weights: Weighed on Day 0 (prior to exposure), Days 7 and 14 (prior to termination).
- Necropsy: All survivors were euthanized by exsanguination following carbon dioxide inhalation at termination on Test Day 14.
- Macroscopic Examinations: Complete macroscopic postmortem examinations were performed on all animals.

Statistics:

No statistical analysis was performed.

Key result

Sex:

male/female

Dose descriptor:

LC50

Effect level:

> 21.5 mg/L air (analytical)

Based on:

test mat.

Exp. duration:

4 h

Mortality:

All animals survived

Clinical signs:

other: There were no signs of toxicity noted during the exposure periods. Clinical signs of toxicity noted immediately following the 10.5 mg/L exposure included lacrimation and red nasal discharge. Clinical signs of toxicity noted immediately following the 21.5

Body weight:

AlI animals gained weight during both weeks of observations after both exposures.

Gross pathology:

There was no effect of treatment evident from the macroscopic postmortem observations of the test animals from both exposures. All animals were within normallimits.

Interpretation of results:

GHS criteria not met

Conclusions:

The test item was considered to be relatively non-toxic to rats by nose-only inhalation exposure with a 4-hour LC50 greater than 21.5 mg/L (analytical) for males and females.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LC50

Value:

21 500 mg/m³

Quality of whole database:

Guideline study, under GLP.

Acute toxicity: via dermal route

Link to relevant study records

Reference

Endpoint:

acute toxicity: dermal

Type of information:

experimental study

Adequacy of study:

key study

Study period:

11 November 2002 - 01 April 2003

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was conducted according to EC and OECD test guidelines, and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

EU Method B.3 (Acute Toxicity (Dermal))

Deviations:

no

Qualifier:

according to guideline

Guideline:

OECD Guideline 402 (Acute Dermal Toxicity)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Limit test:

yes

Specific details on test material used for the study:

- Analytical purity: 99.99%
- Lot/batch No.: 020618

Species:

rat

Strain:

Sprague-Dawley

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 8 to 12 weeks.
- Weight at study initiation: 213 to 267g.
- Housing: Housed individually in metal cages until day 9, when the rats were returned to group housing.
- Diet (e.g. ad libitum): Ad libitum
- Water (e.g. ad libitum): Ad libitum access to drinking water
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 40 - 70%
- Photoperiod (hrs dark / hrs light): 12 hours artificial light / 12 hours darkness.

IN-LIFE DATES: From: 11 November 2002 To: 27 November 2002

Type of coverage:

occlusive

Vehicle:

unchanged (no vehicle)

Details on dermal exposure:

TEST SITE
- Area of exposure: Approximately 50mm x 50mm
- Type of wrap if used: Porous gauze held in place with a non-irritant dressing.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Yes - washed with warm (30 - 40°C) water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 2.35 mL/kg bodyweight (specific gravity = 0.850 g/mL)
- Concentration (if solution): N/A - administered as supplied

Duration of exposure:

24 h

Doses:

2000 mg/kg bodyweight

No. of animals per sex per dose:

5 (5 males and 5 females)

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: All animals were observed twice daily for clinical signs of toxicity, and daily for signs of a dermal response. Bodyweights were recorded on days 1 (prior to dosing), 8, and 15.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, dermal responses, body weight, macroscopic pathology.

Sex:

male/female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Mortality:

Male: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0
Female: 2000 mg/kg bw; Number of animals: 5; Number of deaths: 0

Clinical signs:

Signs of toxicity related to dose levels: There was no systemic response to treatment in any animals throughout the study.

Body weight:

Low bodyweight gains were recorded for four females on day 8 and two females on day 15. All other animals were considered to have achieved satisfactory bodyweight gains throughout the study.

Gross pathology:

Effects on organs: No abnormalities were noted in any animal.

Other findings:

Signs of toxicity (local): Very slight to well-defined dermal irritation with or without very slight oedema was observed in nine animals following removal of the dressings, resolving completely by Day 5. In addition, dryness/exfoliation was observed in nine animals and spots and/or scabbing (confined to small area of dose site) was observed in one animal, resoving completely by Day 9.

Interpretation of results:

not classified

Conclusions:

The acute lethal dermal dose to rats of the substance was demonstrated to ber greater than 2000 mg/kg bodyweight.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

LD50

Value:

2 000 mg/kg bw

Quality of whole database:

Guideline study, under GLP.

Additional information

An acute oral toxicity study was conducted (Huntingdon Life Sciences, 2002, Study ZCE093) to assess the toxicity of the substance following a single oral administration. The study was conducted in accordance with OECD and EC test guidelines, and in compliance with GLP.

In accordance with the acute toxic class method, three fasted female rats were administered a single dose of 2000 mg/kg the substance by oral gavage. As two of the rats at this dose level died, a further three female rats were dosed at 200 mg/kg. No rats in the first group died at 200 mg/kg, and so three males were dosed at 200 mg/kg. No rats died at 200 mg/kg.

It was concluded that the acute median lethal dose in rats following oral administration of the substance was between 200 mg/kg and 2000 mg/kg.

An acute dermal toxicity study was conducted (Huntingdon Life Sciences, 2003, Study ZCE105) to assess the toxicity of the substance following a single dermal administration. The study was conducted in accordance with OECD and EC test guidelines, and in compliance with GLP.

Five male and five female rats were administered a single dermal dose of 2000 mg/kg; the application site was washed 24 hours after initial administration, then the rats observed for 14 days.

No rats died during the observation period, no systematic response to treatment was observed, and no abnormalities were seen in a macroscopic examination performed following study termination at day 15. The acute median lethal dose in rats following dermal administration of the substance was greater than 2000 mg/kg.

An acute inhalation toxicity study was conducted (Huntingdon Life Sciences, 2003) to assess the toxicity of the substance following a 4h inhalation administration. The study was conducted in accordance with OECD and EC test guidelines, and in compliance with GLP.

The test item was considered to be relatively non-toxic to rats by nose-only inhalation exposure with a 4-hour LC50 greater than 21.5 mg/L (analytical) for males and females.

Justification for classification or non-classification

The acute oral LD50 was determined to be between 200 - 2000 mg/kg, and on this basis the corresponding classification under the CLP Regulation (Commission Regulation 1272/2008) is H302 "Harmful if swallowed", and is calssified as acute toxicity category 4. It is acknowleged that under the CLP Regulation, Category 4 for acute oral toxicity applies when the Acute Toxicity Estimate (ATE) / LD50 falls between 300 - 2000 mg/kg (the substance LD50 between 200 - 2000 mg/kg), however as no deaths were seen at 200 mg/kg, it is assumed for the purpose of classification that the LD50 is above 300 mg/kg, and that classification in Category 4 is appropriate.

The acute dermal LD50 of the substance was found to be greater than 2000 mg/kg; on this basis the substance is not classified for acute dermal toxicity under the CLP Regulation (criteria LD50>2000 mg/kg).

The acute inhalation LC50 of the substance was found to be greater than 21.5 mg/L air, on this basis the substance is not classified for acute inhalation toxicity under the CLP Regulation (criteria LC50 >20 mg/L).