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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1993
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report Date:
1993

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
Deviation from Standard Protocol: Due to technical reasons the acclimatisation period lasted 4 instead of 5 days.The challenge was performed during experimental week 4 (day 21) instead of week 5.
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Data from a reliable in vivo test conducted before the enforcement of Commission Regulation (EU) 640/2012 of 06 July 2012 amending, for the purpose of its adaptation to technical progress, Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) are available.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
other:
Details on test material:
None

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright White Strain (Tif: DHP)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: CIBA-GEIGY Limited, Animal Production, 4332 Stein / Switzerland
- Weight at study initiation: 307 to 387 g
- Housing: Macrolon cages (Type 3)
- Diet: NAFAG No. 845, Gossau SG ad libitum
- Water: fresh water ad libitum
- Acclimation period: 4 days

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3 °C
- Humidity: 30 to 70 %

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
physiological saline
Concentration / amount:
5 %
Day(s)/duration:
Day 0
Adequacy of induction:
highest technically applicable concentration used
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline
Concentration / amount:
50 %
Day(s)/duration:
Day 8
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline
Concentration / amount:
30 %
Day(s)/duration:
Day 21
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
The test was performed on 10 male and 10 female guinea pigs in the test group and 5 males and 5 females in the control group.
Details on study design:
PRETESTS
- Intradermal Induction
The concentration for the intradermal injections was selected on account of the solubility of the test article in standard vehicles and its local and systemic tolerability in a pretest. The following concentration of test article has been used for intradermal injection:
5 % in physiological saline (w/v).

- Epidermal Applications (induction and challenge)
The concentrations for the epidermal applications were selected on account of the primary irritation potential of the test article. The following concentrations of FAT 36038/F have been examined on separate animals for the determination of the maximum subirritant concentration:
30 % and 50 % in vaseline.
50 % in vaseline was the highest applicable concentration of the test article.

TEST PROCEDURE
DAY 0: INDUCTION, intradermal injections
Three pairs of intradermal injections (0.1 ml per injection) were made simultaneously into the left and right side of the shaved neck of the test and control group animals.
*Test group:
- adjuvant/saline mixture 1:1 (v/v)
- 5 % FAT 36038/F in physiological saline (w/v)
- 5 % FAT 36038/F in the adjuvant/saline mixture (w/v)

*Control group:
- adjuvant/saline mixture 1:1 (v/v)
- adjuvant/saline mixture 1:1 (v/v)
- physiological saline


DAY 8: INDUCTION, epidermal application
The application site of all animals was pretreated with 10 % sodium-laurylsulfate (open application) 24 hours prior to the epidermal induction applica tion. In the test group FAT 36038/F was incorporated in vaseline (w/w) and applied on a filterpaper patch to the neck of the animals (patch 2x4 cm; approx. 0.4 g per patch; occluded administration for 48 hours). The control group was treated with the vehicle only.
*Test group: 50 % FAT 36038/F in vaseline
*Control group: vaseline only

DAY 21: Challenge
The test and control group animals were tested on one flank with FAT 36038/F in vaseline (w/w) and on the other flank wit the vehicle alone (patch 2x2 cm; approx. 0.2 g per patch; occluded administration for 24 hours).
*Test and control group:
- 30 % FAT 36038/F in vaseline
- vaseline only
Positive control substance(s):
yes
Remarks:
The sensitivity of the strain is checked once or twice a year with a known mild to moderate sensitiser, such as mercaptobenzothiazole, hexyl cinnamic aldehyde or potassiumdichromate.

Results and discussion

Positive control results:
Reference value with 2-Mercaptobenzothiazole puriss. Test No 930012.
- Experimental starting date: July 5, 1993
- Experimental completion date: August 5, 1993

Concentrations of the reference compound and vehicles used:
*Intradermal induction
Concentration of compound: 5 %
Vehicle: Oleum arachis

*Epidermal induction
Concentration of compound: 50 %
Vehicle: vaseline

*Epidermal challenge
Concentration of compound: 30 %
Vehicle: vaseline

- Number of positive animals per group after occlusive epidermal application: 0/10 animal after 24 and 48 hours in the control group. In test group, 0/10 animal in vehicla control after 24 and 48 hours. In test article group 9/10 animals after 24 and 48 hours.
- Challenge reactions after epidermal application (control group): Score for erythema and edema is 0.0 (in males and females for the vehicle control and the test article control).
- Draize score 48 hours after removal of the dressing: Score for erythema and edema in vehicle control for male and female is 0.0. Score for erythema and edema for the test article control in male and female is 0.0.
- Challenge reactions after epidermal application (test group); Draize score 24 hours after removal of the dressing: Erythema and edema score is 0.0 in male and female for the vehicle contro group. For the test article group in males: 2 males have a score 1.0 and 2.0 and for edema 3 males scored with 1.0. In females 3 females have a score 1 and 2 females scored 2 for erythema parameter, for edema only 2 females are scored with 1.0.
- Challenge reactions after epidermal application (test group); Draize score 48 hours after removal of the dressing: Erythema and edema score is 0.0 in male and female for the vehicle contro group. For the test article group in males: 2 males have a score 1.0 and 2 other males with score 2.0 and for edema 1 male scored with 1.0. In females 3 females have a score 1 and 2 females scored 2 for erythema parameter, for edema only 1 female are scored with 1.0.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
other: Control group (vehicle control)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group:
Reading:
1st reading
Hours after challenge:
24
Group:
other: Control gorup ( test article)
Dose level:
30 % and 50 % FAT 36038/F in vaseline
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group:
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Control group (vehicle control)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group:
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Control group (test article)
Dose level:
30 % and 50 % FAT 36038/F in vaseline
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group:
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
- 30 % FAT 36038/F in vaseline (test group); - vaseline only (negative control)
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
- 30 % FAT 36038/F in vaseline (test group); - vaseline only (negative control)
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50 % FAT 36038/F in vaseline
No. with + reactions:
12
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50 % FAT 36038/F in vaseline
No. with + reactions:
7
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
vaseline (vehicle control)
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
Vaseline (vehicle control)
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
Intradermal: 5 %, Epidermal: 50 % and Challenge: 30 % (2-Mercaptobenzothiazole puriss)
No. with + reactions:
9
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
Intradermal: 5 %, Epidermal: 50 % and Challenge: 30 % (2-Mercaptobenzothiazole puriss)
No. with + reactions:
9
Total no. in group:
10

Any other information on results incl. tables

Results in the pretest:

Intradermal induction: Since 5 % FAT 36038/F in physiological saline could be injected and was well tolerated, this concentration was used for the intradermal induction.

Epidermal application: Slight erythema reactions were observed with 50 % FAT 36038/F in vaseline in 1/2 animals at the 24 hours evaluation only.

Evaluation of the primary skin irritation potential:

Procedure: On each animal 2 concentrations of FAT 36038/F were applied simultaneously on the left and right flank. A naive skin site served as control (not reported). 7 days before application of FAT 36038/F two pairs of intradermal injections of an adjuvant/sa line mixture 1:1 (v/v; 0.1 ml per injection) were made simultaneously into the shaved neck of the guinea pigs.

Applicant's summary and conclusion

Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria
Conclusions:
FAT 36038/F is classified as a skin sensitiser 1B.
Executive summary:

A key study to determine the contact allergenic potency of FAT 36038/F test in albino guinea pigs was performed. This test was based on the OECD Guideline No. 406, adopted May 12, 1981, adapted July 17, 1992, by the OECD council, and on Annex V, Part B of Council Directive 67/548/EEC (Commission Directive 92/69/EEC of July 31, 1992).

The test was performed on 10 male and 10 female guinea pigs in the test group and 5 males and 5 females in the control group, respectively. The doses used were 5 % FAT 36038/F in physiological saline for intradermal induction, 50 % FAT 36038/F in vaseline for epidermal induction and 30 % FAT 36038/F in vaseline for epidermal challenge. 60 % and 35 % of the animals of the test group showed skin reactions in the challenge after 24 and 48 hours, respectively. Hence, FAT 36038/F is classified as a skin sensitiser 1B.