Registration Dossier

Administrative data

Description of key information

Testing for sensitising properties of the test item was performed in female guinea pigs according to the adjuvant sensitisation test by Magnusson and Kligman (Guinea pig Maximisation Test) according to OECD TG 406 and under GLP conditions. Intradermal induction was performed using 5% test item in PEG 400 or in a mixture of Freund's Adjuvant and physiological saline (1:1 (v/v)) or PEG 400 alone. Epidermal induction was carried out with 25 % test item in PEG 400.

Using 5% test item in PEG 400 for the challenge treatment resulted in a 0 % incidence of animals with skin reactions at the 24 and 48 hour reading in both the test group as well as the control group.

The positive control from the test lab showed distinctive positive results under the conditions applied in the lab (up to 100% of animals with positive skin reactions weras the control animals showed no skin reactions).

Under the conditions of the test, the test item was not a skin sensitiser.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 13 SEP 1999 to 14 OCT 1999
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
29 APR 1993
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
30 JUL 1996
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
According to Annex VII, Column 2 from Regulation (EC) No 1907/2006 (including amendments up to Commission regulation (EU) 2016/1688 of 20 September 2016) in vivo skin sensitisation studies that were carried out or initiated before 11 October 2016, and that meet the requirements set out in Article 13(3), first subparagraph, and Article 13(4) shall be considered appropriate to address standard information requirement for the endpoint skin sensitization. This applies for the guinea pig maximization test presented here.
Species:
guinea pig
Strain:
other: Ibm: GOHI; SPF quality (synonym: Himalayan spotted)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Biotechnology & Animal Breeding Division, CH-4414-Füllinsdorf / Switzerland
- Females: nulliparous and non-pregnant
- Microbiological status of animals, when known: SPF animals
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 322-335 g
- Housing: Individually in Makrolon type-4 cages with standard softwood bedding
- Diet: Pelleted standard Nafag Ecosan 845 25W4, batcg nos 64/99 and 79/99, guinea pig breeding / maintainance diet, ad libitum
- Water: Community tap water
- Acclimation period: 7 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +- 3
- Humidity (%): 40-70
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal
Vehicle:
other: PEG 400
Concentration / amount:
5%
Day(s)/duration:
Single application
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: PEG 400
Concentration / amount:
25 %
Day(s)/duration:
48 h
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: PEG 400
Concentration / amount:
5 %
Day(s)/duration:
24 h
Adequacy of challenge:
highest non-irritant concentration
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
other: PEG 400
Concentration / amount:
5%
Day(s)/duration:
24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
10 animals in test group, 5 animals in control group
Details on study design:
RANGE FINDING TESTS: yes
1) Intradermal pretest: Four intradermal injections (0.1 ml/site in 1:1 Freud´s complete adjuvant/physiological saline) in the neck of one guinea pig. One week later intradermal injections were made (0.1 ml/site in PEG 400) into the flank of the same animal at concentrations of 5, 3 and 1 %. Dermal reactions were assessed 24 h later.
2) Epidermal pretest: Four intradermal injections (0.1 ml/site in 1:1 Freud´s complete adjuvant/physiological saline) in the neck of two guinea pig. One week later, test substance was applied to the flanks of the animal (25, 15, 10 or 5 % in PEG 400). After 24 h substance was removed, 24 and 48 h after removal, reaction sited were assessed for erythema and oedema.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: Three pairs of intradermal injections and an epidermal application on test day 8
- Exposure period: 3 weeks (1 week after the inreadermal injections epidermal applicationwas done. Two weeks after that, challenge was performed)

- Exposure group:
Three pairs of intradermal injections (0.1 ml/site) were made at the border of a 4 x 6 cm area in the clipped region as follows:
1) 1:1 (v/v) mixture of Freud´s Complete Adjuvant and physiological saline
2) Test item at 5% in PEG 400
3) Test item at 5 % in a 1.1 (v/v) mixture of freud´s Adjunat and physiological saline

-Control group:
Three pairs of intradermal injections (0.1 mL/site) were made at the border of a 4 x 6 cm area in the clipped region as follows:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) PEG 400
3) 1:1 (w/w) mixture of PEG 400 in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.

- Site: scapular region
- Duration: single intradermal injection, occlusive dressing from epidermal application was left for 48 h.
- Control animals were not treated with the test item

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: day 22, two weeks after epidermal induction
- Exposure period: 24 h
- Site: flank (right flank treated with PEG 400, left flank treated with 5% test substance in PEG 400)
- Concentrations: 5 %
- Evaluation (hr after challenge): 24 and 48 h after removal, reaction sited were assessed for erythema and oedema.
- Control animals were treated with the test item as well.

Reading and Scoring:
The scoring system was performed by visual scoring of erythema, oedema and other clinical changes of skin conditions. They were assessed using the following Magnusson and Kligman gradng scale:
0 = no visible change
1 = discrete or patchy erytheman
2 = moderate and confluent erythema
3 = intense erythema and swelling
Positive control substance(s):
yes
Remarks:
2-mercaptobenzothiazole (analysed in a separate experiment)
Positive control results:
90% (24 h reading) and 100% (48 h reading) of treated animals were observed with very slight to moderate/severe erythematous reactions.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of systemic toxicity were observed in the animals.
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of systemic toxicity were observed in the animals.
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of systemic toxicity were observed
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of systemic toxicity were observed
Remarks on result:
no indication of skin sensitisation
Interpretation of results:
GHS criteria not met
Conclusions:
Tested in this study according to OECD test guideline no. 406 and GLP conditions the test item was not a skin sensitiser.
Executive summary:

Testing for sensitising properties of the test item was performed in female guinea pigs according to the adjuvant sensitisation test by Magnusson and Kligman (Guinea pig Maximisation Test) according to OECD TG 406 and under GLP conditions. Intradermal induction was performed using 5% test item in PEG 400 or in a mixture of Freund's Adjuvant and physiological saline (1:1 (v/v)) or PEG 400 alone. Epidermal induction was carried out with 25 % test item in PEG 400.

Using 5% test item in PEG 400 for the challenge treatment resulted in a 0 % incidence of animals with skin reactions at the 24 and 48 hour reading in both the test group as well as the control group.

The positive control from the test lab showed distinctive positive results under the conditions applied in the lab (up to 100% of animals with positive skin reactions weras the control animals showed no skin reactions).

Under the conditions of the test, the test item was not a skin sensitiser.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

No classification

In an in vitro study according to Magnusson & Kligman the test item did not induce skin sensitisation.