Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
The study was performed between 10 June 1996 and 17 July 1996.
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 not effect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Date of Inspection: 22/01/1996 Date of signature: 27/02/1996
Test type:
fixed dose procedure
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Valifast Black 3810
- Substance type: Black Powder
- Physical state: Solid
- Lot/batch No.: X-10543
- Stability under test conditions: Stable
- Storage condition of test material: Room temperature

Test animals

Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:
Charles River (UK) Ltd., Margate, Kent, UK.

- Age at study initiation:
five to eight weeks of age.

- Weight at study initiation:
Males weighed 150 to 178 g, and the females 146 to 168g.

- Fasting period before study:
overnight fast immediately before dosing

- Housing:
The animals were housed in groups of up to five by sex in suspended solid floor polypropylene cages furnished with woodflakes.

- Diet ad libitum):
(Rat and Mouse Expanded Diet No.1, special Diets Services Limited, Witham, Essex, UK) was allowed throughout the study.

- Water ad libitum

- Acclimation period: at least five days


ENVIRONMENTAL CONDITIONS
- Temperature (°C):
19 to 24°C

- Humidity (%):
48 to 57%

- Air changes (per hr):
The rate of air exchange was at least 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:
arachis oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 200mg/ml

- Amount of vehicle (if gavage):
10 ml

- Justification for choice of vehicle:
Not stated

- Lot/batch no. (if required):
Not stated

- Purity:
Not stated


MAXIMUM DOSE VOLUME APPLIED:
10 ml/kg


DOSAGE PREPARATION (if unusual):
Not applicable

CLASS METHOD (if applicable)

- Rationale for the selection of the starting dose:
Based on the results from the range finding study, a dose level of 2000 mg/kg bodyweight was selected for the main study.
Doses:
Following a sighting test at dose levels of 2000 mg/kg, a further group of five fasted females and five fasted males was given a single oral dose of test material at a dose level of 2000 mg/kg bodyweight.
No. of animals per sex per dose:
6 females at 2000 mg/kg
6 males at 2000 mg/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 then 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:
At the end of the study the animals were killed by cervical dislocation and subjected to gross pathological examination. This consisted of an external examination and opening of the abdominal and thoracic cavities for examination of major organs. The appearance of any macroscopic abnormalities was recorded. No tissues were retained.




Statistics:
Not applicable

Results and discussion

Preliminary study:
There were no deaths or clinical signs of toxicity. Based on this information, a dose level of 2000 mg/kg bodyweight was selected for the main study.
Effect levels
Sex:
female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Mortality:
There were no deaths.
Clinical signs:
Black-coloured staining of fur was noted in all animals up to six days after dosing. Black-coloured staining of the faeces was noted in all animals four hours and one day after dosing. The eyes, tail and feet of all animals appeared grey in colour two to nine days after dosing. The eyes, tail and feet or tail only, of all animals, appeared grey in colour ten days after dosing. The tail only, of all animals, appeared grey in colour eleven to fourteen days after dosing.

No signs of systemic toxicity were noted during the study.
Body weight:
All animals showed expected gains in bodyweight.
Gross pathology:

No abnormalities were noted at necropsy.
Other findings:

- Organ weights:
Not recorded

- Histopathology:
Not recorded

- Potential target organs:
Not recorded

- Other observations:
None

Any other information on results incl. tables

Results are shown in Tables 1-4 which are attached.

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 Sprague Dawley CD strain rat was estimated to be greater than 2000 mg/kg bodyweight.
Executive summary:

A study was performed to assess the acute oral toxicity of the test material in the Sprague Dawley CD strain rat. The method followed that in the OECD Guidelines for Testing of Chemicals No.401 "Acute Oral Toxicty" (adopted 24 February 1987) and Method B1 of Commission Directive 92/69/EEC (which constitutes Annex Y of council Directive 67/548/EEC).

Following a range finding study, a group of ten fasted animals ( five males and five females) was given a single oral dose of test material as a suspension in arachis oil BP at a dose level of 2000 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed and subjected to gross pathological examination.

There were no deaths. Black coloured staining of the faeces was noted. The eyes, tails and feet of all animals appeared grey in colour. No signs of systemic toxicity were noted.

All animals showed an expected gain in bodyweight during the study.

No abnormalities were noted at necropsy.