Registration Dossier

Administrative data

Description of key information

A Guinea Pig Maximisation Test (GPMT) was performed to examine the sensitizing potential of the substance. In this study it was determined that the test material is not a skin-sensitizer (GLP, OECD

406; RCC Ltd, 2002)

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:
2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Currently no LLNA study is available for assessment. The Guinea Pig Maximization Test (GPMT) has been carried out as an animal test to predict human sensitization for over a decade and is recommended by international test guidelines such as OECD.
Species:
guinea pig
Strain:
other: Ibm: GOHI; SPF-quality guinea pigs (synonym: Himalayan spotted)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: RCC Ltd, Biotechnology & Animal Breeding Division, Fuellinsdorf / Switzerland- Age at study initiation: 3 - 6 weeks- Weight at study initiation: 379 - 426 g- Housing: Individually in Makrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, Muttenz)- Diet (e.g. ad libitum): Pelleted standard Provimi Kliba 3418, batch no. 93/01, guinea pig breeding / maintenance diet, containing Vitamin C (Provimi Kliba AG, Kaiseraugst), ad libitum.- Water (e.g. ad libitum): Community tap water from Fiillinsdorf, ad libitum- Acclimation period: One weekENVIRONMENTAL CONDITIONS- Temperature (°C): 20 ± 3- Humidity (%): 30-70- Air changes (per hr): 10-15- Photoperiod (hrs dark / hrs light): 12 hours light and 12 hours dark.
Route:
intradermal and epicutaneous
Vehicle:
polyethylene glycol
Remarks:
PEG 300
Concentration / amount:
intradermal induction: 10% test itemepidermal induction: 25% test item
Route:
epicutaneous, occlusive
Vehicle:
propylene glycol
Remarks:
PEG 300
Concentration / amount:
0.2 mL of 1% test item
Day(s)/duration:
24 hours, two weeks after epidermal induction
No. of animals per dose:
15 males for main study5 males for pretests
Details on study design:
RANGE FINDING TESTS: To determine the different concentrations one intradermal and two epidermal pretests were performed. Intradermal injections (0.1 ml/site) were made into the clipped flank of the same guinea pig at concentrations of 10 %, 5 % and 3% of the test item in PEG 300. The three concentrations were determined during non-GLP formulation trials performed before the study initiation date. The concentration of 10 % was considered to be the highest technically applicable concentration which could be injected into the intra-cellular space in spite of the high viscosity of the application dilution and the obstacle caused by the tissues. For intradermal injections4 patches of filter paper (3x3 cm) were saturated with the test item at 25 % (technically the highest possible concentration to be applied sufficiently), 15 %, 10 % and 5 % in PEG 300 and applied to the clipped and shaved flanks. The amount of test item preparation applied was approximately 0.2 g for the test item at 25 % and a volume of approximately 0.2 ml was applied for the remaining test item concentrations. The patches were covered by a strip of aluminum foil and firmly secured by elastic plaster wrapped around the trunk and covered with impervious adhesive tape. This procedure ensured the intensive contact of the test item. The dressings were removed after an exposure period of 24 hours.Twenty-one hours after removal of the dressing the application site was depilated with an approved depilatory cream (VEET Cream, Reckitt & Colman AG, Allschwil) in order to visualize any resulting erythema. The depilatory cream was placed on the patch sites and surrounding areas, and left on for 3-5 minutes. It was then thoroughly washed off with a stream of warm, running water. Thereafter, the animals were dried with a disposable towel, and returned to their cages. The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman.MAIN STUDYA. INDUCTION EXPOSUREIntradermal inductionTest Group: 1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.2) The test item, at 10 % in PEG 300.3) The test item at 10 % in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.Control Group: 1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.2) PEG 3003) 1:1 (w/w) mixture of PEG 300 in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.Epidermal induction on test day 8:- Time point of exposure: One week after the intradermal induction- Exposure period: 48 hours- Site: scapular area- Exposure area: 2 x 4 cm patch of filter paper was saturated with the test item and placed over the injection sites of the test animals- Amount applied: approximately 0.3 g- Concentrations: 25 % test item in PEG 300B. CHALLENGE EXPOSURE- Time point of exposure: test and control guinea pigs were challenged two weeks after the epidermal induction- Exposure period: 24 hours- Site: on the left and right flank of each guinea pig just prior to the application; test item was applied to the left flank, vehicle only (PEG 300) was applied to the right flank- Exposure area: 3x3 cm; two patches of filter paper were saturated with the test item at the highest tested non-irritating concentration- Concentrations: 1% test item in vehicle and the vehicle only (PEG 300) using the same method as for the epidermal application.- Amount applied: approximately 0.2 ml- Evaluation (hr after challenge): 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman.
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle alone
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No findings noted.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle alone
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No findings noted.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
1% test item in PEG
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No findings noted. But animal no. 52 died.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
1% test item in PEG
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No findings observed. But animal no. 62 died.
Interpretation of results:
GHS criteria not met
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

In a GLP conform study the cutaneous allergenic potential of the test substance (purity: > 98 weight- %), was assessed using the Maximization-Test (GPMT) in accordance with OECD Guideline No. 406 (RCC Ltd, 2002). The test was performed in 15 (10 test and 5 control) male albino guinea pigs. The intradermal induction of sensitisation in the test group was performed in the nuchal region with a 10% dilution of the test item in PEG 300 and in an emulsion of Freund's Complete Adjuvant (FCA) / physiological saline. The epidermal induction of sensitization was conducted for 48 hours under occlusion with the test item at 25 % in PEG 300 one week after the intradermal induction. The animals of the control group were intradermally induced with PEG 300 and FCA/physiological saline and epidermally induced with PEG 300 under occlusion. Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test item at 1 % in PEG 300 and PEG 300 alone under occlusive dressing. Cutaneous reactions were evaluated at 24 and 48 hours after removal of the dressing. The sensitising potential of the test substance was classified according to the grading of Magnusson and Kligman. No toxic symptoms were evident in the guinea pigs of the control or test group. None of the control and test animals showed skin reactions after the challenge treatment with the test item at 1 % (w/w) in PEG 300.


Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) 1272/2008. Based on this data the substance does not need to be classified for skin sensitization under Regulation (EC) No. 1272/2008.