Registration Dossier

Administrative data

Description of key information

In the key study a guinea pig maximization test was performed and it was concluded that the test substance was non sensitising.
An additional open epicutaneous test in guinea pigs resulted in the overall conclusion that the test substance was non sensitising (supporting study).

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:
1985-05-10 to 1985-06-08
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
No LLNA study was conducted as an in vivo study comparable to OCED 406 was conducted, before the first version of the LLNA (OECD 429) was issued in 2002.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: 5-8 weeks
- Weight at study initiation: 300-389 g
- Housing: up to 4/cage, solid-floor polypropylene cages (supplied by NKP Limited, Dartford, Kent)
- Diet: ad libitum, Guinea Pig FD1 Diet (supplied by Special Diet Services Limited, Witham, Essex)
- Water: ad libitum, tap water
- Acclimation period: min. 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 45-65
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12
Route:
intradermal and epicutaneous
Vehicle:
arachis oil
Remarks:
for injection
Concentration / amount:
Intradermal Induction: 5 % in arachis oil B.P.
Topical Induction: 100 %
Day(s)/duration:
48 h for topical induction
Adequacy of induction:
other: maximum non-irritant concentration of the test material
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
Topical Challenge: 100 %
Day(s)/duration:
24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20
Details on study design:
RANGE FINDING TESTS (called Preliminary Siting test):
- Injection with 0.1 mL of 1 and 5 % test substance with observation after 24, 48 and 72 h and 7 days.
- 2 cm* 2 cm patches with 50 and 100 % concentration were applied to clipped skin of guinea pigs that were injected with adjuvant 1-3 weeks before. Patches were removed after 24 h. After 1, 24 and 48 h the skin was evaluated. The maximum non-irritant concentration of the test material was selected for the main study.


MAIN STUDY
A. INDUCTION EXPOSURE (Injection)
- No. of exposures: 1 (3 injection sides)
- Exposure period: 1 injection and 6 days observation
- Test group: 20 animals received 3 pairs of injections: (1) 0.1mL of Freunds complete adjuvant (Difco Laboratories, Detroit, Michigan USA) (2) 0.1mL of a 5 % concentration of the test material (3) 0.1mL of a 50:50 mixture of a 5 % concentration of the test material emulsified in the adjuvant
- Control group: 10 animals received 3 pairs of injections: (1) 0.1mL of Freunds complete adjuvant (2) 0.1mL of vehicle alone (3) 0.1mL of a 50:50 mixture of vehicle emulsified in Freunds complete adjuvant
- Site: 4 cm x 6 cm in the shoulder region
- Frequency of applications: 1 application
- Duration: single injection
- Concentrations: 5 % and 50:50 mixture of a 5 % concentration of the test material emulsified in the adjuvant

A. INDUCTION EXPOSURE (topical)
- No. of exposures: 1
- Day after intradermal injection: day 7
- Exposure period: 48 h
- Test groups: 100 % test substance on Whatman No 4 Filter paper, fixed with Sleek waterproof adhesive strapping in a cross and 3.5 cm x 25 cm length of Elastoplast elastic adhesive bandage which was wound in a double layer around the torso of the animal
- Control group: similar to test group but only vehicle
- Site: 2 cm x 4 cm in the shoulder region
- Concentrations: 100 %

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 21
- Exposure period: 24 h
- Test group: on clipped right flank 100 %, on clipped left flank vehicle
- Control group: on clipped right flank 100 %, on clipped left flank vehicle
- Site: 5 cm x 5 cm an both flanks
- Concentrations: 100 %
- after 24 h the patches were removed and challenge sites were cleaned using cotton wool soaked in lukewarm water or ether
- Evaluation: was done 24 h after removal (48 h after application) and after 48 h (72 h)

OTHER:
on day 0 and day 24 body weights were recorded
Challenge controls:
yes, see above
Positive control substance(s):
no
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
none
Remarks on result:
no indication of skin sensitisation
Interpretation of results:
GHS criteria not met
Conclusions:
A guinea pig maximization test was conducted. The challenge with 100 % of the test substance resulted in no reactions and therefore it was conclueded that the test substance was non sensitising.
Executive summary:

A guinea pig maximization test was conducted. The test group consisted of 20 and the control group of 10 guinea pigs. Intradermal induction was done with 3 injection pairs into different sides. The injections contained 1) Freunds complete adjuvant, 2) 5 % concentration of the test material or vehicle only for the control group and 3) 50:50 mixture of a 5 % concentration of the test material emulsified in the adjuvant or 50:50 mixture of vehicle emulsified in Freunds complete adjuvant for the control animals. The topical induction was done with 100 % test substance or vehicle for the control group. The challenge with 100 % of the test substance resulted in no reactions in any of the 20 guinea pigs both at the 24 h and 48 h observations. The animals of the control group showed to reactions either. Therefore it was concluded that the test substance was non sensitising.

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

Skin sensitisation

In the key study a guinea pig maximization test was conducted. The test group consisted of 20 and the control group of 10 guinea pigs. Intradermal induction was done with 3 injection pairs into different sides. The injections contained 1) Freunds complete adjuvant, 2) 5 % concentration of the test material or vehicle only for the control group and 3) 50:50 mixture of a 5 % concentration of the test material emulsified in the adjuvant or 50:50 mixture of vehicle emulsified in Freunds complete adjuvant for the control animals. The topical induction was done with 100 % test substance or vehicle for the control group. The challenge with 100 % of the test substance resulted in no reactions in any of the 20 guinea pigs both at the 24 h and 48 h observations. The animals of the control group showed to reactions either. Therefore it was concluded that the test substance was non sensitising.

The supporting study showed the results of an open epicutaneous test in guinea pigs using 0.3, 1, 3, 10, 30 and 100 % of the test substance for induction and 1, 3, 10 and 30 % for the challenge. The challenge concentrations included the maximal non-irritant and minimal irritant concentrations and two challenges were done. A reaction in the skin of all animals including the vehicle control animals challenged with 30 % for the first time was detected. After the second challenge with 30 % test substance not all animals reacted. The first 10 % challenge caused a reaction in 3/36 previously treated animals and 1/6 control animals while the second challenge caused no skin reactions. No effects to lower concentrations of the test substance were observed. The effect in the pretreated animals did not exceed the effect in control animals. Due to these observations the overall conclusion of the experiments was that the test substance was non sensitising.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Classification, Labelling, and Packaging Regulation (EC) No 1272/2008

The available experimental test data is reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. Based on available data on skin sensitisation, the test item is not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for the eighth time in Regulation (EU) No 2016/918.