Registration Dossier

Diss Factsheets

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Not reported.
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to GLP in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2001
Report date:
2001

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
: The environmental conditions deviated from the guideline as the ranges used were larger.
GLP compliance:
yes
Type of study:
Buehler test

Test material

Constituent 1
Reference substance name:
Hydrocarbon waxes (petroleum), oxidized
EC Number:
265-205-1
EC Name:
Hydrocarbon waxes (petroleum), oxidized
Cas Number:
64743-00-6
Molecular formula:
too complex
IUPAC Name:
Hydrocarbon waxes (petroleum), oxidized
Test material form:
other: greasy solid
Details on test material:
- Physical appearance: Brown solid.
- Storage: at room temperature.

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Hilltop Lab Animals, Inc., Scottdale, PA.
- Animal husbandry was based on the standards recommended by the Guide for the Care and Use of Laboratory Animals.
- Age at study initiation: Males ~ 6 weeks old, females ~ 8 weeks old.
- Weight at study initiation: Males ~ 366-439 g, females ~ 319-380 g.
- Housing: Individually in suspended stainless steel cages.
- Diet (e.g. ad libitum): PMI Certified Guinea Pig Chow #5026 (Purina Mills, Inc.), provided ad libitum.
- Water (e.g. ad libitum): Municipal tap water treated by reverse osmosis, provided ad libitum.
- Acclimation period: At least 5 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 60-77 ºF (16-25ºC).
- Humidity (%): 23-49%.
- Air changes (per hr): 10-15 air charges per hour.
- Photoperiod (hrs dark / hrs light): 12 hours light/dark cycle.

IN-LIFE DATES: From: 05/12/2000 To: 11/01/2001

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Mineral oil
Concentration / amount:
Induction exposure: 50% or 40 % w/v in the vehicle.
1st Challenge and challenge control: 25% w/v in the vehicle.
2nd Challenge: 20 and 10% w/v in the vehicle.
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Mineral oil
Concentration / amount:
Induction exposure: 50% or 40 % w/v in the vehicle.
1st Challenge and challenge control: 25% w/v in the vehicle.
2nd Challenge: 20 and 10% w/v in the vehicle.
No. of animals per dose:
- Twenty animals were used in the test group, 10 males and 10 females.
- Ten animals were used for the positive control, 5 males and 5 females.
- Ten animals were used for the challenge control group, 5 males and 5 females.
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Three, on days 1, 7 and 14.
- Exposure period: 6 hours, after which the site was wiped down with gauze moistened in deionised water to remove any residue.
- Site: Left dorsal side of the animal, called site 1.
- Test groups: 50% in the first induction then 40% for the second and third induction exposures, which were all applied at site 1.
- Control group: Animals were exposed three times to 0.1% concentration of the test material in the same manner as the test group.
- Frequency of applications: Every seven days until all three consecutive applications have been administered.

B. CHALLENGE EXPOSURE
- Rest time before challenge: Approximately 2 weeks.
- No. of exposures: Two, the Challenge and the Rechallenge.
- Exposure period: 6 hours, after which the site was wiped down with gauze moistened in deionised water to remove any residue.
- Day(s) of challenge: Application on Day 28 and Day 35 for the challenge and rechallenge respectively.
- Site: Three sites were used, for exposure, on the right dorsal side of the animal, called site 2, 4 and 6.
- Test groups: 25% w/v of the test material was applied to site 2 in the challenge, then 20% and 10% w/v was applied at sites 4 and 6 during the rechallenge.
- Challenge control group: The challenge control was performed concurrently with the test and applied in the same manner as the test material.
- Positive Control group: The DNCB control and DNCB challenge were performed on Day 28 only, at site 2 and 4. 0.1% was applied at site 2 and 0.05% was applied at site 4.
- Wrapping: Following application of the test material, challenge control or positive control the trunk of the animal was wrapped in an occlusive manner.

C. OBSERVATIONS
-Dermal reactions were recorded at 24 and 48 hours following application of the test material during induction exposure and both the challenge and rechallenge.
-Clinical observations and mortality were recorded twice daily.
-Body weights were recorded on the day before dosing in the induction, challenge and rechallenge.
- All animals were euthanized after the final scoring.
Positive control substance(s):
yes
Remarks:
DNCB (1-chloro-2,4-dinitrobenzene) in acetone/ethanol: 0.1% w/v for the induction exposure and then 0.1% and 0.05% w/v in the challenge exposure.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
other: Challenge
Hours after challenge:
24
Group:
test chemical
Dose level:
25% w/v
No. with + reactions:
6
Total no. in group:
20
Remarks on result:
other: Reading: other: Challenge. . Hours after challenge: 24.0. Group: test group. Dose level: 25% w/v. No with. + reactions: 6.0. Total no. in groups: 20.0.
Reading:
other: Challenge
Hours after challenge:
48
Group:
test chemical
Dose level:
25% w/v
No. with + reactions:
9
Total no. in group:
20
Remarks on result:
other: Reading: other: Challenge. . Hours after challenge: 48.0. Group: test group. Dose level: 25% w/v. No with. + reactions: 9.0. Total no. in groups: 20.0.
Reading:
other: Challenge
Hours after challenge:
24
Group:
other: Challenge control
Dose level:
25% w/v
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
other: Reading: other: Challenge. . Hours after challenge: 24.0. Group: other: Challenge control. Dose level: 25% w/v. No with. + reactions: 2.0. Total no. in groups: 10.0.
Reading:
other: Challenge
Hours after challenge:
48
Group:
other: Challenge Control
Dose level:
25% w/v
No. with + reactions:
1
Total no. in group:
10
Remarks on result:
other: Reading: other: Challenge. . Hours after challenge: 48.0. Group: other: Challenge Control. Dose level: 25% w/v. No with. + reactions: 1.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
20% w/v
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 20% w/v. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
20% w/v
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 20% w/v. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
other: rechallenge control
Dose level:
20% w/v
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: other: rechallenge control. Dose level: 20% w/v. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
other: rechallenge control
Dose level:
20% w/v
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: other: rechallenge control. Dose level: 20% w/v. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
10% w/v
No. with + reactions:
2
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: test group. Dose level: 10% w/v. No with. + reactions: 2.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
10% w/v
No. with + reactions:
1
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 10% w/v. No with. + reactions: 1.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
24
Group:
other: rechallenge control
Dose level:
10% w/v
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 24.0. Group: other: rechallenge control. Dose level: 10% w/v. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
other: rechallenge control
Dose level:
10% w/v
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: other: rechallenge control. Dose level: 10% w/v. No with. + reactions: 0.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Under the conditions of the test there is no significant difference in the dermal response after the challenge or rechallenge applications of the test material. The test material was determined to be not sensitising and therefore does not require classification according to Regulation (EC) 1272/2008.
Executive summary:

In a GLP compliant Buehler test, the skin sensitisation potential was assessed according to OECD guideline 406. Twenty Guinea pigs were exposed to the test material first using three induction exposures at 50 and 40 % w/v, then in a challenge and rechallenge exposure at 25, 20 and 10 % w/v. None of the test animals’ displayed a dermal reaction greater than the challenge control during the challenge or rechallenge. Under the conditions of the test, the test material was determined to be not sensitising and therefore does not require classification according to Regulation (EC) 1272/2008.