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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation, other
Remarks:
in vivo (non-LLNA).
Type of information:
experimental study
Adequacy of study:
key study
Study period:
August-December 2003
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Well-documented and corresponded to the requirements of the recommended Annex V Test Guidelines.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
There was one deviation from the study protocol which was concerned with the relative humidity section 6.2 Husbandry). On some days of the study the relative humidity was higher than 70%.
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Appropriate guinea pig maximisation test available which would not justify conducting an additional LLNA due to animal welfare.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Specific details on test material used for the study:
- Name of test material (as cited in study report): PEBETAL dibasic lead phthalate
- Physical state: fine, white powder
- Analytical purity: 74.15% Pb= 97.6% Pebetal
- Impurities (identity and concentrations):
- Composition of test material, percentage of components: ash: 79.3; water: 0.2
- Lot/batch No.: D1/4296
- Expiration date of the lot/batch: July 31, 2004
- Radiochemical purity (if radiolabelling):
- Specific activity (if radiolabelling):
- Locations of the label (if radiolabelling):
- Expiration date of radiochemical substance (if radiolabelling): July 31, 2004
- Storage condition of test material: At roome temperature. Shelf-life: at least one year after dispatch.

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: pirbright white
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Deutschland GmbH
- Age at study initiation:
- Weight at study initiation: males: 436-504 g; females: 395-481 g
- Housing: The guinea pigs were kept in collective housing up to a maximum of 5 animals per cage in a battery of cages, equipped with a paper disposal system.
- Diet (e.g. ad libitum): The animals received 2040 Teklad Global Guinea Pig Diet offered ad libitum.
- Water (e.g. ad libitum):Tap water as for human consumption was continuously available ad libitum via drinking nipples.
- Acclimation period: 19 days (range finding); 22 days ( main test)


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/-3 degrees centigrade
- Humidity (%): 37-76%
- Air changes (per hr): 16 times per hour and filtered adequately
- Photoperiod (hrs dark / hrs light): 12 hours dark/ 12 hours light with light on at 7:00 AM


IN-LIFE DATES: From: To:

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
other: Intradermal application: The solid test article was diluted with aqua ad iniectabilia and Freund's complete adjuvant to a final concentration of 5%. Dermal Application: the solid test article was used 50% in petrolatum
Concentration / amount:
For the intradermal injection, the test article was diluted with aqua ad iniectabilia and Freund's complete adjuvant to a final concentration of 5%. For the dermal application, the solid test article was used 50% in petrolatum.
Challenge
Route:
epicutaneous, occlusive
Vehicle:
other: Intradermal application: The solid test article was diluted with aqua ad iniectabilia and Freund's complete adjuvant to a final concentration of 5%. Dermal Application-the solid test article was used 50% in petrolatum
Concentration / amount:
For the intradermal injection, the test article was diluted with aqua ad iniectabilia and Freund's complete adjuvant to a final concentration of 5%. For the dermal application, the solid test article was used 50% in petrolatum.
No. of animals per dose:
10 test and 5 control animals
Details on study design:
RANGE FINDING TESTS: The range finding test was performed to determine the concentrations of the test article to be used in the main test. For the intradermal injection, the test article was diluted with aqua ad iniectabilia and Freund's complete adjuvant to a final concentration of 5%. Two animals were employed, skin reactions being recorded 48 and 72 hours after treatment. For the dermal application, the solid test article was used 50% in petrolatum. A closed patch exposure was effected by means of an occlusive bandage using swabs, Leukosilk, and non-irritating tape Elastoplast which enveloped the whole animal's trunk. Two animals were employed and skin reactions were recorded 48 and 72 hours post application.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: Seven days after the injection, the injection sites were covered occlusively for 48 hours.
- Test groups: 10 guinea pigs
- Control group: 5 guinea pigs
- Site: Injections were made in the clipped intracapsular region on either side of the spine.
- Frequency of applications: Three pairs of intradermal injections (0.1 ml)
- Duration: Seven days after the injection, the injection sites were covered occlusively for 48 hours with a patch carrying the test article (50%) or, in control animals, the control article petrolatum.
- Concentrations: Test group: 1) FCA 50% (v/v) diluted in aqua ad iniectabilia, 2) test article 5% in peanut oil, 3) test article 5% in aqua iniectabilia/FCA. Control group: 1) FCA 50% (v/v) diluted in aqua ad iniectabilia, 2) peanut oil, 3) peanut oil 50% (v/v) diluted in FCA.




B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 14 days after the second stage of the induction
- Exposure period: 24 hours
- Test groups: 10 test animals
- Control group: 5 control animals
- Site: The challenge test was performed on a 5 x 5 cm clipped skin area on each flank.
- Concentrations: The test article was applied at the concentration of 50% to the left flank and the control article petrolatum to the right in a volume
of 0.5 g using the patch technique. In each case, the duration of exposure was 24 hours under an occlusive dressing.
- Evaluation (hr after challenge): 24 and 48 hours after patch removal the appearance of the challenge and re-challenge skin sites was observed and skin responses were gradfed on the basis of the classification system according DRAIZE. The animals werer weighed before treatment and at the end of the study. Al findings were recorded with respect to duration and severity on special report forms.




OTHER:
Challenge controls:
The control article petrolatum was applied to the left flank of the guinea pig in a volume of 0.5 g using the patch technique for 24 hours under an occlusive dressing.
Positive control substance(s):
no
Remarks:
The sensitivity of the test system and the reliability of the experimental technique is assessed at least at every 6 months by use of "4-aminobenzoic acid ethyl ester (benzocaine)" which is known to induce skin sensitisation in guinea pigs.

Study design: in vivo (LLNA)

Vehicle:
other: petrolatum
Concentration:
50% and 25% in Vaseline (petrolatum)
No. of animals per dose:
10
Details on study design:
The sensitivity of the test system and the reliability of the experimental technique is assessed at least at every 6 months by use of "4-aminobenzoic acid ethyl ester (benzocaine)" which is known to induce skin sensitisation in guinea pigs. This test was conducted as a Magnusson & Kligman test according to the OECD Guidelines for Testing of Chemicals, OECD 406, 17 July 1992 and to the EEC Directive 2001/59/EEC, 6 August 2001. The last test that was performed with acceptable levels of responses to this substance was performed from April 8 to May 2, 2003.

Results and discussion

Positive control results:
Sensitisation rate (50%) at 24 h: 70% Sensitisation rate (50%) at 48h: 70%
Sensitisation rate (25%) at 24 h: 30% Sensitisation rate (25%) at 24h: 30%

In vivo (non-LLNA)

Results
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50% in petrolatum
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
None

Any other information on results incl. tables

There were no skin reactions after the dermal application in the induction phase. No allergic skin reactions were observed in the test animals after the challenge exposure 24 and 48 h after patch removal. No findings were observed in control animals. The sensitisation rate, i.e. the number of animals showing an allergic response expressed as a percentage of the total number of animals, was determined 24 and 48 h after patch removal and was 0% in test animals in each case. The corresponding reaction rate in control animals was 0%.

Applicant's summary and conclusion

Interpretation of results:
other: Not classified under CLP criteria
Conclusions:
According to the EEC Directive 2001/59/EEC, 6 August 2001 and the Gefahrstoffverordnung (GefStoffV) of 15 November 1999 (BGB1. I, p. 2233), the test article PEBETAL dibasic lead phthalate can be classified as a "non-sensitiser" since no allergic responses were observed in test animals after the adjuvant test according to Magnusson & Kligman.
Executive summary:

The potential skin sensitising properties of the test article PEBETAL dibasic lead phthalate were assessed in the guinea pig maximisation test method, using 10 test and 5 control animals in the main test. Following the induction exposure to the test article (50% in petrolatum) or the vehicle petrolatum (control article), the animals of both groups were subjected two weeks later to a challenge with the test article (50% in petrolatum) as well as the control article. Responses to the challenge procedure were evaluated 24 and 48 h after the end of the exposure period.

Results: - No allergic skin reactions occurred in test animals 24 and 48 h after the end of the challenge procedure. The sensitisation was 0%.

-No findings were observed in control animals (reaction rate: 0%).

According to the EEC Directive 2001/59/EEC, 6 August 2001 and the Gefahrstoffverordnung (GefStoffV) of 15 November 1999 (BGB1 I. p. 2233), the test article PEBETAL dibasic lead phthalate can be classified as a "non-sensitiser" since no allergic responses were observed in test animals under the experimental conditions described.