C-M5

Acute Toxicity: oral

Administrative data

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

12 April 2007 and 02 May 2007

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to GLP and in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do no effect the quality of the relevant results.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2007

Report date:

2007

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 420 (Acute Oral Toxicity - Fixed Dose Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test type:

fixed dose procedure

Limit test:

yes

Test material

Reference

Name:

Unnamed

Type:

Constituent

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River (UK) Ltd, Margate, Kent, UK.
- Age at study initiation: Eight to twelve weeks of age.
- Weight at study initiation: Weights are not recorded, however the bodyweight variation did not exceed ± 20% of the initial mean bodyweight of any previously dosed animal(s).
- Fasting period before study: Overnight fast immediately before dosing and for approximately three to four hours after dosing.
- Housing:The animals were housed in groups of up to four in suspended solid floor polypropylene cages furnished with woodflakes.
- Diet (e.g. ad libitum): Certified Rat and Mouse Diet.
- Water (e.g. ad libitum):Free access to mains drinking water was allowed throughout the study.
- Acclimation period:Five days.


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25°C
- Humidity (%): 30 to 70%
- Air changes (per hr): Fifteen changes per hour.
- Photoperiod (hrs dark / hrs light): Lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.


IN-LIFE DATES: From:Day 1 To:Day 14

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

VEHICLE
- Concentration in vehicle: 200 mg/ml
- Amount of vehicle (if gavage):Not available
- Justification for choice of vehicle: For the purpose of the study the test material was freshly prepared, as required, as a solution in distilled water.
- Lot/batch no. (if required): Not available
- Purity: Not available.


MAXIMUM DOSE VOLUME APPLIED: 10ml/kg


DOSAGE PREPARATION (if unusual): Not applicable


CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose:Following a sighting test in which there was no mortality at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a solution in distilled water, at a dose level of 2000 mg/kg bodyweight.

Doses:

dose level of 2000 mg/kg.

No. of animals per sex per dose:

5 females at 2000mg/kg

Control animals:

no

Details on study design:

- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Clinical observations were made ½, 1, 2, and 4 hours after dosing and subsequently once daily for fourteen days. Morbidity and mortality checks were made twice daily.
Individual bodyweights were recorded on Day 0 (the day of dosing) and on Days 7 and 14.
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight,organ weights, histopathology, other:External examination and opening of the abdominal and thoracic cavities. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.

Statistics:

None reported.

Results and discussion

Preliminary study:

In the absence of data suggesting the test material was toxic, 2000 mg/kg was chosen as the starting dose.
In the absence of toxicity at a dose level of 2000 mg/kg, an additional group of animals was treated.

Effect levels

Sex:

female

Dose descriptor:

LD50

Effect level:

> 2 000 mg/kg bw

Remarks on result:

other: 95% CL not determined

Mortality:

There were no deaths.

Clinical signs:

No signs of systemic toxicity were noted.
Purple stained faeces was noted in all animals one day after dosing and four animals two days after dosing. Purple stained urine was noted in all animals one day after dosing, two animals two days after dosing and one animal three and four days after dosing.

Body weight:

Individual bodyweights and bodyweight changes are given in Table 2.
All animals showed expected gains in bodyweight over the observation period.

Gross pathology:

Individual necropsy findings are given in Table 3.
Red stained kidneys were noted at necropsy of one animal that was killed at the end of the study. No abnormalities were noted at necropsy of the remaining four animals that were killed at the end of the study.

Other findings:

None.

Any other information on results incl. tables

RESULTS

Individual clinical observations and mortality data are given in Table 1.

Mortality

There were no deaths.

Clinical Observations

No signs of systemic toxicity were noted.

Purple stained faeces was noted in all animals one day after dosing and four animals two days after dosing.  Purple stained urine was noted in all animals one day after dosing, two animals two days after dosing and one animal three and four days after dosing.

Bodyweight

Individual bodyweights and bodyweight changes are given in Table 2.

All animals showed expected gains in bodyweight over the observation period.

Necropsy

Individual necropsy findings are given in Table 3.

Red stained kidneys were noted at necropsy of one animal that was killed at the end of the study.  No abnormalities were noted at necropsy of the remaining four animals that were killed at the end of the study.

CONCLUSION

The acute oral median lethal dose (LD₅₀) of the test material in the female Sprague-Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System - Unclassified).

Table 1                Individual Clinical Observations and Mortality Data

Dose Level mg/kg	Animal Number and Sex	Effects Noted After Dosing (Hours)				Effects Noted During Period After Dosing (Days)
		½	1	2	4	1	2	3	4	5	6	7	8	9	10	11	12	13	14
2000	1-0 Female	0	0	0	0	0FU*	0U*	0U*	0U*	0	0	0	0	0	0	0	0	0	0
	2-0 Female	0	0	0	0	0FU*	0F	0	0	0	0	0	0	0	0	0	0	0	0
	2-1 Female	0	0	0	0	0FU*	0F	0	0	0	0	0	0	0	0	0	0	0	0
	2-2 Female	0	0	0	0	0FU*	0F	0	0	0	0	0	0	0	0	0	0	0	0
	2-3 Female	0	0	0	0	0FU*	0FU	0	0	0	0	0	0	0	0	0	0	0	0

Table 2                Individual Bodyweights and Bodyweight Changes

Dose Level mg/kg	Animal Number and Sex	Bodyweight (g) at Day			Bodyweight Gain (g) During Week
		0	7	14	1	2
2000	1-0 Female	239	269	282	30	13
	2-0 Female	247	286	312	39	26
	2-1 Female	233	260	282	27	22
	2-2 Female	201	233	259	32	26
	2-3 Female	223	269	276	46	7

Table 3                Individual Necropsy Findings

Dose Level mg/kg	Animal Number and Sex	Time of Death	Macroscopic Observations
2000	1-0 Female	Killed Day 14	Kidneys: stained red
	2-0 Female	Killed Day 14	No abnormalities detected
	2-1 Female	Killed Day 14	No abnormalities detected
	2-2 Female	Killed Day 14	No abnormalities detected
	2-3 Female	Killed Day 14	No abnormalities detected

0 = No signs of systemic toxicity

F = Faeces stained purple

U = Urine stained purple

* = Bedding stained purple

Applicant's summary and conclusion

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The acute oral median lethal dose (LD50) of the test material in the female Sprague Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System- Unclassified).

Executive summary:

Introduction. The study was performed to assess the acute oral toxicity of the test material in the Sprague‑Dawley CD strain rat. The method was designed to meet the requirements of the following:

§        OECD Guidelines for Testing of Chemicals No 420 “Acute Oral Toxicity - Fixed Dose Method” (adopted)

§        Method B1 bis Acute Toxicity (Oral) of Commission Directive 2004/73/EC

Method. Following a sighting test in which there was no mortality at a dose level of 2000 mg/kg, an additional four fasted female animals were given a single oral dose of test material, as a solution in distilled water, at a dose level of 2000 mg/kg bodyweight. Clinical signs and bodyweight development were monitored during the study. All animals were subjected to gross necropsy.

Mortality. There were no deaths.

Clinical Observations. There were no signs of systemic toxicity.

Purple-coloured staining of the faeces and urine was noted in all animals during the observation period.

Bodyweight. All animals showed expected gains in bodyweight.

Necropsy. Red stained kidneys were noted at necropsy one animal that was killed at the end of the study. No abnormalities were noted at necropsy of the remaining four animals that were killed at the end of the study.

Conclusion. The acute oral median lethal dose (LD₅₀) of the test material in the female Sprague‑Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight (Globally Harmonised Classification System-Unclassified).