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Toxicological information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1987
Report Date:
1987

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
Tested according to GPMT as described by B. Magnussen and A.M. Kligman (allergic contact dermatitits in the guinea pig. Publ. Ch.C. Tomas: Springfiled, Illinois, USA, 1970)
Deviations:
not applicable
GLP compliance:
no
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The LLNA method was not adopted at the time of testing (1987)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
Constituent
Test material form:
liquid: viscous

In vivo test system

Test animals

Species:
guinea pig
Strain:
other:
Remarks:
albino (DHPW)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: F. Winkelmann, Instutute ofr Breeding of LAboratory Animals GmbH and Co. KG, Borchen, Germany
- Females nulliparous and non-pregnant: not specified
- Microbiological status of animals, when known: not specified
- Age at study initiation: not specified
- Weight at study initiation: 330-390 gram (male) and 280-375 gram (female)
- Housing: The animal were kept individually under conventional conditions un suspended, stainless steel cages, fitted with wire mesh floor and front
- Diet: Pelleted, natural ingredient diet, ad libitum
- Water: Unfluorinated tap water, ad libitum
- Acclimation period: 8 days
- Indication of any skin lesions: No visible skin abnormalities observed

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18±2 ºC
- Humidity (%): 40-70
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 / 12
- IN-LIFE DATES: From: Marc 18, 1987 To: April 11, 1987

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
propylene glycol
Concentration / amount:
10% dilution (v/v)
Day(s)/duration:
7
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
intradermal
Vehicle:
other: FCA and PG (1:1)
Concentration / amount:
10% dilution (v/v)
Day(s)/duration:
7
Adequacy of induction:
not specified
Route:
epicutaneous, occlusive
Vehicle:
other: vaseline
Concentration / amount:
25% mixture (w/w)
Day(s)/duration:
2
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:
10% mixture (w/w)
Day(s)/duration:
1 and 2
Adequacy of challenge:
not specified
No. of animals per dose:
20 (10 males and 10 females)
Details on study design:
RANGE FINDING TESTS:
The irritation response to intradermal injection of various concentrations was examined in 3 guinea pigs. A sufficiently large area of the scapular region was clipped free from hair with electric clippers. Subsequeantly 0.1 mL portions of 5, 10 and 25% (v/v) dilutions of the test substance in propylene glycol, were applied to each animal by intradermal injection.

The irritation response to topical treatment was tested in 2 guinea pigs were clipped free from hair with electric clippers. Subsequantly several 2x2 pacthes of filter paper (Whatman N0. 3 MM) were loaded with different concentrations of the test substance in vasilne, viz. 10 or 25% (w/w)/ The patched were placed on the clipped skin and covered with a piece of PVC foil and hypo-allergenic paper bandage (Leukopor) which was then secured by elastic adhesive bandage (Tensoplast), 7.5 cm in width, wound aroun the torso of the animal. two concentrations were tested per animal. The dressing was left in place for 24 h. Then the animal were examined for signs of irritation.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: two, one intradermal and one topical
- Exposure period: the topical application was performed 7 days after the intradermal injections, the dressing was removed after 2 days, the resting period after the topical applicatin was 14 days
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: scapular region
- Frequency of applications: one intradermal and one topical
- Duration: the topical application was performed 7 days after the intradermal injections, the dressing was removed after 2 days, the resting period after the topical applicatin was 14 days
- Concentrations: 10% (v/v) for intradermal injections and 25% (w/w) for topical application

B. CHALLENGE EXPOSURE
- No. of exposures: one
- Day(s) of challenge: one
- Exposure period: 24 h
- Test groups: 10 males and 10 females
- Control group: 5 males and 5 females
- Site: scapular region
- Concentrations: 10% (w/w)
- Evaluation (hr after challenge): 24 and 48
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10% (w/w)
No. with + reactions:
6
Total no. in group:
20
Clinical observations:
All animal remained in good health during the experimental period
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10 % (w/w)
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
All animal remained in good health during the experimental period
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10% (w/w)
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
All animal remained in good health during the experimental period
Remarks on result:
other:
Remarks:
Since, only very slight skin reactions were observed in controles these observations are considered to be due to iritation rather than to sensitization.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 % (w/w)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
All animal remained in good health during the experimental period
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
NA
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
NA
Remarks on result:
other: no positive control was added in the study design.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
NA
No. with + reactions:
0
Total no. in group:
0
Clinical observations:
NA
Remarks on result:
other: no positive control was added in the study design.

Any other information on results incl. tables

Dermal reactions elicited by the challenge application of 10% test substance in vaseline

 

 

Animal number

Scores at 24 and 48 h after removal of dressing

Males

 

Animal number

Females

24 h

48 h

24 h

48 h

ER

ED

ER

ED

ER

ED

ER

ED

Control group

7400

0

0

0

0

7401

0

0

0

0

7402

0

0

0

0

7403

0

0

0

0

7404

1

0

0

0

7405

1

0

0

0

7406

0

0

0

0

7407

0

0

0

0

7408

0

0

0

0

7409

0

0

0

0

Test group

7380

2

2

2

2

7381

1

0

1

1

7382

2

1

2

1

7383

1

1

1

1

7384

1

1

0

0

7385

1

1

0

0

7386

2

1

1

1

7387

1

1

0

0

7388

1

1

1

1

7389

0

0

0

0

7390

0

0

0

0

7391

2

2

0

0

7392

0

0

0

0

7393

0

0

0

0

7394

1

1

0

0

7395

2

2

1

1

7396

1

1

1

1

7397

2

2

1

1

7398

1

1

0

0

7399

1

0

1

0

ER = erythema ED = oedema

Individual skin reactions observed during the induction phase of the study

Animal number

Weight (g)

Induction period

Initial score at the sites of intradermal injections

Topical application

al

ar

bl

br

cl

cr

D

C

H

D

C

H

D

C

H

D

C

H

D

C

H

D

C

H

ER

ED

Control group - males

7400

355

7

P

2

7

P

1

4

PA

0

5

PA

0

7

PA

2

8

PA

2

0

0

7402

355

7

P

2

8

P

2

4

PA

0

5

PA

0

8

PA

2

7

PA

1

0

0

7404

356

6

P

2

6

P

1

5

PA

0

5

PA

0

7

PA

2

6

PA

1

0

0

7406

360

7

P

2

5

P

1

5

PA

0

6

PA

0

4

PA

1

4

PA

1

0

0

7408

385

8

P

2

7

P

2

5

PA

0

7

PA

0

4

PA

1

-

0

0

Control group - females

7401

330

8

P

2

7

P

2

8

PA

0

5

PA

0

9

PA

2

6

PA

1

0

0

7403

355

8

P

2

8

P

2

5

PA

0

5

PA

0

8

PA

2

8

PA

2

0

0

7405

345

-

7

P

2

5

PA

0

4

PA

0

8

PA

2

5

PA

2

0

0

7407

295

7

P

2

7

P

2

6

PA

0

-

5

PA

2

7

PA

2

0

0

7409

350

8

P

2

9

P

2

5

PA

0

-

9

PA

2

8

PA

2

0

0

Test group – males

7380

363

8

P

2

8

P

2

7

RA

0

6

RA

0

9

RA

1

9

RA

1

0

0

7382

330

8

P

2

8

P

2

5

PA

0

7

RA

0

9

RA

1

10

RA

1

0

0

7384

375

7

P

2

8

P

2

8

RA

0

7

RA

0

9

RA

1

10

RA

1

0

0

7386

345

7

P

2

-

7

PA

0

8

RA

0

9

RA

1

9

RA

1

0

0

7388

380

9

P

2

7

P

2

6

PA

0

7

RA

0

8

RA

1

6

RA

1

0

0

7390

380

8

P

2

7

P

2

7

RA

0

7

RA

0

9

RA

1

9

RA

1

0

0

7392

390

8

P

2

8

P

2

7

RA

0

6

RA

0

9

RA

1

9

RA

1

0

0

7394

380

8

P

2

8

P

2

7

RA

0

7

RA

0

10

RA

1

0

RA

1

0

0

7396

355

8

P

2

7

P

2

6

RA

0

7

RA

0

9

RA

1

10

RA

1

0

0

7398

385

7

P

2

8

P

2

6

RA

0

7

RA

0

7

RA

1

8

RA

1

0

0

Test group – females

7381

310

5

P

2

7

P

2

7

RA

0

8

RA

0

-

11

RA

1

0

0

7383

305

8

P

2

8

P

2

7

RA

0

9

RA

0

9

RA

0

8

RA

0

0

0

7385

280

8

P

2

-

6

RA

0

8

RA

0

9

RA

1

5

RA

0

0

0

7387

320

7

P

2

8

P

2

7

RA

0

7

RA

0

6

RA

1

6

RA

1

0

0

7389

365

8

P

2

8

P

2

7

RA

0

7

RA

0

8

RA

1

9

RA

1

0

0

7391

320

7

P

2

7

P

2

8

RA

0

7

RA

0

10

RA

1

9

RA

1

0

0

7393

350

8

P

2

8

P

2

7

RA

0

7

RA

0

9

RA

1

9

RA

1

0

0

7395

375

7

P

2

6

P

2

7

RA

0

7

RA

0

9

RA

1

10

RA

1

0

0

7397

310

8

P

2

8

P

2

7

RA

0

7

RA

0

9

RA

1

9

RA

1

0

0

7399

365

7

P

2

6

P

2

8

RA

0

7

RA

0

7

RA

1

10

RA

1

0

0

 

al and ar: FCA

bl and br: PG

cl and cr: PC and FCA (1:1)

-              = not scored

D = diameter of skin reaction

H = height of skin reaction

C = color of skin reaction (PP = pale pink; P = pink; R = red; A = abscess; N = necrosi

Applicant's summary and conclusion

Interpretation of results:
Category 1B (indication of skin sensitising potential) based on GHS criteria
Conclusions:
From the results of the challenge treatment with the 10% dilution it appeared that the test substance induced skin reactions in 16 out of 20 test animals, while also in 2 out of 10 controls showed dermal reactions. Since, only very slight skin reactions were observed in controls the very slight skin reactions seen in the test animals are considered due to irritation rather than the senisitzation. However, the more severe reactions observed in 6 out of 20 animals are possible due to sensitization. Therefore, the test substance is considered to be a moderate sensitizer.
Executive summary:

The test substance was examined for possible sensitizing properties by a maximization test in guinea pigs using 20 test animals and 10 controls.

The test comprised of:

a. induction treatment by intradermal injection of a 10% dilution of the test substance in propylene glycol, followed one week later by a topical treatment with 25% test substance in vaseline;

b. challenge treatment, 14 days after the last induction, by topical application of 10% test substance in vaseline.

The challengde treatment with the 10% dilution showed evidence of delayed contact hypersensitivity. It was concluded that the test substance is a moderate sensitizer.