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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
Study period:
1980-09 to 1980-10
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Remarks:
No data on positive control.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1980
Report Date:
1980

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
Guinea pig maximization test as described by B. Magnusson and A.M. Kligman (1970), the later released OECD Guideline 406 (1981) is based to a great extent on this publication.
GLP compliance:
no
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A valid Guinea pig maximization test conducted comparable to guideline with acceptable restrictions is available, which is reliable with restrictions and adequate for classification and labelling purposes. Potency estimation is not mandatory when existing guideline and GLP conforming data are available, which were conducted before the new annex of the REACH Regulation entered into force. Moreover, no indication for skin sensitisation was observed in this study, thus, no dose response information is needed. For this reason and for reasons of animal welfare no additional LLNA was conducted.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid - liquid: aqueous solution

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Pirbright white
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 4 - 6 weeks
- Weight at study initiation: 400 +/- 50 g
- Housing: suspended cages
- Diet: ad libitum, Guinea-pig diet (Ssniff G)
- Water: ad libitum, tap water
- Acclimation period: no data


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 2
- Humidity (%): 55 +/- 5 % RH
- Air changes (per hr): 16
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Induction:
Intradermal injection:0.5 % v/v,
Topical application after 7 days: epicutaneous 60 % v/v
Day(s)/duration:
1 week after injections; topical application 48 h
Challenge
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Challenge: 10 % v/v
Day(s)/duration:
2 weeks after the induction; 24 h exposure
No. of animals per dose:
15 test animals,
5 control animals
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 times
- Exposure period: 0 and 1 week after injections topical application for 48 h
- Test groups: 15
- Control group: 5
- Site: a 4 x 6 cm area of dorsal skin on the scapular region was clipped free of hair.

First stage:: 3 pairs of intradermal injections were made simultaneously. A volume of 0.1 ml was injected into both infection sites (left and right site)
- Concentrations:
Test group:
1. 0.1 ml of Freund´s complete adjuvant 50 : 50 with water for injection
2. 0.1 ml of 0.5 % v/v test item in sterile isotonic saline
3. 0.1 ml of 0.5 % v/v test item in a 50 : 50 mixture of isotonic saline and Freund´s complete adjuvant
Control group:
1. 0.1 ml of Freund´s complete adjuvant 50 : 50 with water for injection
2. 0.1 ml of sterile isotonic saline
3. 0.1 ml of Freund´s complete adjuvant 50 : 50 with isotonic saline
Control animals: during the induction period control animals were treated similarly to the test animals but the test substance was omitted form intradermal injections and topical applications.

Second stage: 7 days after the first induction the same area was clipped and shaved free of hair. A 2 x 4 cm patch of filter paper was saturated with 60 % v/v test item in distilled water. Patch was placed on the skin and covered by a impermeable plastic adhesive tape ("Dermicel"). Tape
was secured by a elastic adhesive bandage ("Elastoplast"). Occulsive dressing was left in place for 48 h.

B. CHALLENGE EXPOSURE
- No. of exposures: 1 time
- Day(s) of challenge: 2 weeks after the induction
- Exposure period: 24 h
- Test groups:
2 x 2 cm filter paper was saturated with the test sample in a similar as used for second stage induction. Occlusive dressing for 24 h.
- Control group: The 5 animals of the control group were similarly challenged
- Site: hair was removed by clipping and then shaving a 5 x 5 cm area on the left flank.
- Concentrations: 10 % v/v test item in distilled water
- Evaluation (hr after challenge): 24, 48, 72 h after patch removal

Scoring:
Erythema and eschar formation:
No erythema 0
Slight erythema (barely perceptible): 1
Well-defined erythema 2
Moderate erythema 3
Severe erythema (beet. redness) to slight eschar formation (injuries in depth) 4

Oedema formation:
No oedema 0
Slight oedema (barely perceptible): 1
Well-defined oedema (edges of area well-defined by definite raising) 2
Moderate oedema (raised app. 1 mm 3
Severe oedema (raised more than 1 mm and extending beyond the area 4
of exposure



Challenge controls:
5
Positive control substance(s):
not specified

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10 %
No. with + reactions:
2
Total no. in group:
15
Clinical observations:
erythema score 1 (Slight erythema (barely perceptible))
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
15
Remarks on result:
no indication of skin sensitisation
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test group
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
15
Remarks on result:
no indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
5

Any other information on results incl. tables

Induction:

Intradermal injections containing Freund´s complete adjuvant: elicited well-defined dermal irritation

Intradermal injection of 0.5 % v/v test item in all test animals: temporary slight dermal irritation

Second stage (topical application) of 60 % v/v test item:slight dermal reactions

Challenge (10 % v/v Test item):

Test animals:

After 24 h: slight erythema (1): 2 animals

Control animals: no reactions

All animals (test and control animals) showed slight bandage reactions

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Migrated information
Conclusions:
In this guinea-pig maximization test, performed in 20 albino guinea-pigs, Coco AAPB (30 % a.i.) did not produce evidence of delayed contract hypersensitivity.
Executive summary:

In a dermal sensitization study with Coco AAPB (commercial product) 20 male Pirbright white guinea pig were tested using the MAXIMIZATION TEST described by b. Magnusson and A.M. Kligman (1970). The test method is similar to OECD Guideline 406 (Skin Sensitisation).

The analytical purity of the test item is not stated in study report, according to producer information test material has 30 % a.i., impurities relevant to sensitisation and referred to a.i. are 8.3 % Alkylamidopropylamine and 33 -50 ppm DMPA (3-dimethylaminopropylamine).

At the first scoring, 24 h after challenge, 2/15 showed a skin reaction score 1 (slight erythema (barely perceptible)). The animals were free of erythema and edema at the second (48 h) and the third (72 h) scoring. None of the five negative animals showed up to and including the third scoring a positive skin reaction to challenge treatment.

In this study, Coco AAPB (30 % a.i.) is not a dermal sensitizer.