β,β-carotene

Acute Toxicity: oral

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 guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 401 (Acute Oral Toxicity)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.1 (Acute Toxicity (Oral))

GLP compliance:

yes (incl. QA statement)

Test type:

standard acute method

Limit test:

yes

Test material

Test material information

Constituent 1

Reference substance name:

β,β-carotene

EC Number:

230-636-6

EC Name:

β,β-carotene

Cas Number:

7235-40-7

Molecular formula:

C40H56

IUPAC Name:

β,β-carotene

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 or test system 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:

other: 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.

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.