Registration Dossier

Administrative data

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1994
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline-conform study performed under GLP without deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
GLP compliance:
yes (incl. certificate)
Test type:
standard acute method
Limit test:
yes

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): β-Carotene
- Substance type:
- Molecular weight: 536.8
- Physical state: dark-redcrystals
- Analytical purity: 98.2%
- Purity test date: 15 Dec 1993
- Lot/batch No.: 312131
- Expiration date of the lot/batch: December 1994
- Storage condition of test material: closed cans, at -20 °C. Once cans are opened, stored at 4 °C, dry and away from light, and covered with an inert gas

Test animals

Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL (Biological Research Laboratoies), Füllinsdorf, Switzerland
- Weight at study initiation: M: 190 g, F: 153 g
- Fasting period before study: overnight (also fasted overnight before necropsy)
- Housing: Macrolon cages, with dust-free wood-shaving and feed containers integrated in cage lid, air-conditioned animal room, under periodic bacteriological control
- Diet (e.g. ad libitum): pelleted complete rodent diet (KLIBA 343) (animals were fasted overnight before treatment and access to feed was ca 3 hours after treatment)
- Water (e.g. ad libitum): tap water ad libitum (water bottles)


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 °C ± 2 °C
- Humidity (%): 55 % ± 15 %
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle (fluorescent tubes)

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
other: Standard Suspending Vehicle (SSV): 5 g Sodium-carboxymethylcellulose (CMC), 4 mL TWEEN 80, 5 mL Benzylalcohol, ad 1000 mL physiological sodium chloride (0.9%)
Details on oral exposure:
VEHICLE
Standard Suspending Vehicle (SSV):
5 g Sodium-carboxymethylcellulose (CMC)
4 mL TWEEN 80, 5 mL Benzylalcohol
1000 mL physiological sodium chloride (0.9%)

The test material was suspended in SSV. The constant application volume was increased to 15 mL/kg body weight because of the low water solubility of the test item.


Doses:
(single dose): 2000 mg/kg bw

The low acute oral toxicity of the test item has long been known. A dose of 2000 mg/kg weight is accepted as a "limit dose" in accordance with OECD and EU guidelines.
No. of animals per sex per dose:
5 male and 5 female
Control animals:
no
Details on study design:
- Duration of observation period following administration: 14 days

- Frequency of observations and weighing:

Mortalities and clinical signs - The animals were observed for mortality and signs of toxicity at least three times at the day of treatment (Day 0) and then at least once daily for 5 days/week.

Body weights: Individual body weights were recorded on Day 0 (before treatment), one, two and three days after treatment and then at least twice per week.

Weight recordings and calculations of doses were made on an on-line data acquisition system (DATATOX) with electronic balance. This system defines the study commencement as time-point "0". The weight recorded at that time is considered as that of day 0 (actually the beginning of study day 1). The weight recorded the day after study commencement is considered to be that of day 1 (actually the end of study day 1 and beginning of study day 2). In case of technical problems, animal weights may occasionally not be recorded on-line.


- Necropsy of survivors performed: yes
At the end of the 14-day observation period the rats were sacrified with carbon dioxide, exsanguinated and grossly necropsied. In case animals were sacrificed moribund or found dead during the study, they were grossly necropsied, either. All organs and tissues with macroscopic findings at necropsy were examined microscopically and the relevant findings reported.

Results and discussion

Effect levels
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 2 000 mg/kg bw
Based on:
test mat.
Mortality:
No mortalities occurred after a single oral dose of 2000 mg/kg body weight of the test material.
Clinical signs:
The test substance was well tolerated and no clinical signs indicative of reduced health or behavioural changes were observed in the animals. About one day after administration of the test item, the faeces of all animals were discoloured to reddish-brown.
Body weight:
Body weight gains were within the expected range for male and female Han Wistar rats of this age. The reduced body weights on day 14 are due to overnight fasting before necropsy.
Gross pathology:
No findings were reported.

Any other information on results incl. tables

Table 1: Mean body weight of test animals during the observation period of 14 days

Male

 

Day 0

Day 1

Day 2

Day 3

Day

Day 10

Day 14

Mean

189.6

212.9

221.0

224.1

251.3

266.6

262.0

SD

10.6

10.3

11.1

10.0

8.4

7.2

7.5

 Female:

 

Day 0

Day 1

Day 2

Day 3

Day

Day 10

Day 14

Mean

152.5

167.7

169.4

167.7

174.0

181.4

173.5

SD

3.3

4.0

3.3

5.7

4.2

5.1

6.5

Table 2: Mortality of test animals during the observation period of 14 days.

 

Animals per dosage

Within 6 hours

7 - 24 hours

Day 2 - 7

Day 8 – end of observation

Male

5

0

0

0

0

Female

5

0

0

0

0

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Single oral dose administration of 2000 mg/kg body weight of B-carotene, was well tolerated. No mortalities occurred and no findings were noted during the 14-day observation period and at scheduled necropsy.
Executive summary:

The acute oral toxicity of the test item was investigated under GLP in Han Wistar rats of both sexes (10 animals) according to OECD TG 401. Single oral dose administration of 2000 mg-kg body weight of the test item was well tolerated. No mortalities occurred and no clinical signs indicative of reduced health or behavioural changes were observed in the animals. No macroscopic findings were noted at scheduled necropsy. According to OECD and EU guidelines, the test substance is considered to present no significant acute toxic risk if swallowed.