β,β-carotene

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2006-06-14 to 2006-07-21

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:

2006

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

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): beta carotene, technical grade
- Physical state: dark red crystals
- Analytical purity: 97.77%
- Purity test date: 4 May 2005
- Lot/batch No.: WC00504271
- Expiration date of the lot/batch: July 2007
- Stability under test conditions: stable under storage conditions
- Storage condition of test material: at room temperature (20 °C ± 5 °C), protected from humidity and light. Storage under inert gas (nitrogen)

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Harlan Netherlands BV, Horst, The Netherlands
- Age at study initiation: male: 13-14 weeks, female 8-9 weeks
- Weight at study initiation: 1460 - 2418 g
- Housing: Individually in stainless steel cages, equipped with feed hoppers and drinking water bowls. Wood blocks and haysticks were provided for gnawing.
- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 8/06) provided by Provimi Kliba AG, Kaiseraugst.
- Water (e.g. ad libitum): Community tap water from Füllinsdorf, ad libitum.
- Acclimation period: yes, under laboratory conditions after health examinations. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17-23 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 10-15 changes/hour
- Photoperiod (hrs dark / hrs light): 12 hours light, 12 hours dark (automatically controlled light cycle, with music played during the daytime)

Test system

Type of coverage:

semiocclusive

Preparation of test site:

other: clipped

Vehicle:

water

Controls:

no

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.5 g per animal moistened with purified water

VEHICLE
- Amount(s) applied (volume or weight with unit): 0.5 mL of water was used to moisten the substance

Duration of treatment / exposure:

4 hours

Observation period:

VIABILITY/MORTALITY
Daily from acclimatisation of the animals to the termination of the test

CLINICAL SIGNS
Daily from acclimatisation of the animals to the termination of the test

BODYWEIGHTS
At start of acclimatisation, on the day of application and at termination of observation

SKIN OBSREVATION
The skin reaction was assessed at approximately 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after exposure (removal of the dressing, gauze patch and test item)

Number of animals:

3 animals (1 male, 2 female)

Details on study design:

TEST SITE
- Area of exposure: left flank
- % coverage: not reported, 4 cm x 4 cm
- Type of wrap if used: surgical gauze

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the 4 hour treatment, the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site
- Time after start of exposure: 4 hours

SCORING SYSTEM:
The skin reaction was assessed according to the numerical scoring system listed in the Commission Directive 2004/73/EC, April 29, 2004. If evident, corrosive or straining properties of the test item were described and recorded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 2

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 3

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0.33

Reversibility:

fully reversible

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Reversibility:

other: not applicable

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score of the three animals was 0.00, 0.00 and 0.33 respectively and the mean oedema score was 0.00 for all the animals. Very slight erythema forming was observed in all animals, one hour after removal of the dressing and persisted in one female animal until the 24 hour reading.
Neither alterations of the treated skin were observed, nor were corrosive effects evident on the skin.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred. Slight red staining of the treated skin area produced by the test item was present in all animals from the 1 hour to the 7 day reading and persisted in one female animal until the 10-day reading. No abnormal findings were observed on the treated skin of animals 14 days after treatment, the end of the observation time. The body weights of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

Table 1: Bodyweights

Animal No.	Sex	First day of acclimatisation	Day of treatment	Last day of observation
31	Male	2130	2418	2841
32	Female	1304	1407	1936
33	Female	1249	1460	1864

Table 2: Skin irritation scores (individual values)

Animal number	Sex	Evaluation interval*	Erythema	Oedema
31 32 33	M F F	1 hour	1 1 1	0 0 0
31 32 33	M F F	24 hours	0 0 1	0 0 0
31 32 33	M F F	48 hours	0 0 0	0 0 0
31 32 33	M F F	72 hours	0 0 0	0 0 0
31 32 33	M F F	7 days	0 0 0	0 0 0
31 32 33	M F F	10 days	0 0 0	0 0 0
31 32 33	M F F	14 days	0 0 0	0 0 0

* examinations were performed at the specified times after removal of the dressing.

Note: observations continued for 14 days due to staining present on the skin

Table 3: Skin irritation scores (Individual mean values after 24, 48 and 72 hours)

Animal number	Sex	Erythema	N	Oedema	N
31 32 33	M F F	0.00 0.00 0.33	3 3 3	0.00 0.00 0.00	3 3 3

N = number of available data points

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on the referred classification criteria (Commission Directive 2001/59/EC of August 2001), the test item is considered to be "not irritating" to rabbit skin.

Executive summary:

The primary skin irritation potential of the test item was investigated under GLP according to OECD TG 404. The test item was applied by topical semi-occlusive application of 0.5 g substance moistened with 0.5 mL of water to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment under occlusion was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7, 10 and 14 days after removal of the dressing.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades separately. The mean erythema/eschar score of the three animals was 0.00, 0.00 and 0.33 respectively and the mean oedema score was 0.00 for all the animals.

The application of the test item to the skin resulted in very slight erythema in all animals 1 hour after removal of the dressing, persisting in one female animal until the 24 -hour reading. Red staining of the treated skin area produced by the test item was noted in all animals from the 1-hour reading to the 7-day reading and persisted in one female animal until the 10-day reading. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no clinical signs were observed. Thus, the test item did not induce significant or irreversible damage to the skin.

Based on the referred classification criteria (Commission Directive 2001/59/EC of August 2001), the test item is considered to be "not irritating" to rabbit skin.