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Description of key information

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:
2001 - 2002
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
not specified
Qualifier:
according to
Guideline:
other: Directive 96/54/EEC, B.6.
Deviations:
not specified
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
When this test was ordered, the LLNA was not yet a standard method.
Species:
guinea pig
Strain:
other: Ibm: GOHI; SPF-quality (Himalayan spotted)
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: RCC Ltd, Biotechnology & Animal Breeding Division, CH-4414 Fullinsdorf / Switzerland
- Age at study initiation: 5 - 6 weeks
- Weight at study initiation: 338 - 401 g
- Housing: Individually in Makrolon type-4 cages with standard softwood bedding
- Diet: Pelleted standard Provimi Kliba 3418, batch no. 92/01, guinea pig breeding / maintenance diet, containing Vitamin C (Provimi Kliba AG, CH-4303 Kaiseraugst), ad libitum.
- Water: Community tap water from Fullinsdorf, ad libitum.
- Acclimation period: One week for the control and test group under test conditions after health examination. No acclimatization for the animals of the pretest

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22±3°C
- Humidity (%): 30-70%
- Air changes (per hr): 10-15 air changes per hour
- Photoperiod (hrs dark / hrs light): controlled light cycle of 12 hours light and 12 hours dark. Music was played during the light period.

Route:
intradermal
Vehicle:
polyethylene glycol
Remarks:
300
Concentration / amount:
10% test material / / 0.1 ml
Day(s)/duration:
Day 1
Adequacy of induction:
not specified
Route:
intradermal
Vehicle:
other: PEG 300 in a 1:1 mixture of FCA/physiological saline
Concentration / amount:
10% test material / / 0.1 ml
Day(s)/duration:
Day 1
Adequacy of induction:
not specified
Route:
intradermal
Vehicle:
other: FCA/physiological saline
Concentration / amount:
0% test material (control) / / 0.1 ml
Day(s)/duration:
Day 1
Adequacy of induction:
not specified
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Remarks:
PEG 300
Concentration / amount:
50% test material
Day(s)/duration:
Day 22 / / 24-hour duration
Adequacy of challenge:
other: non-irritant substance, pre-treated with 10% SLS (Day 8)
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Remarks:
PEG 300
Concentration / amount:
0% test material
Day(s)/duration:
Day 22 / / 24-hour duration
Adequacy of challenge:
other: non-irritant substance, pre-treated with 10% SLS (Day 8)
No. of animals per dose:
Number of animals in test group: 10
Number of animals in negative control group: 5
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
INTRADERMAL INJECTIONS / PERFORMED ON TEST DAY 1
An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair. 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

EPIDERMAL APPLICATIONS / PERFORMED ON TEST DAY 8
On test day 7 and approximately 23 hours prior to the epidermal application the scapular area (approximately 6 x 8 cm) of the animals of the control and test group was clipped, shaved free of hair and the test area was pretreated with 0.5 ml of 10% Sodium-Lauryl- Sulfate (SLS) in paraffinum perliquidum as no primary irritation had been observed in the pretest. The SLS was massaged into the skin with a glass rod without bandaging. This 10 % concentration of SLS enhances sensitization by provoking a mild inflammatory reaction. On test day 8, a 2 x 4 cm patch of filter paper was saturated with the test item (50 % in PEG 300) and placed over the injection sites of the test animals. The amount of test item preparation applied was approximately 0.3 g. The patch was covered with aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The occlusive dressings were left in place for 48 hours. The epidermal application procedure described ensured intensive contact of the test item. The guinea pigs of the control group were treated as described above with PEG 300 only, applied at a volume of approximately 0.3 ml. 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


B. CHALLENGE EXPOSURE
CHALLENGE / PERFORMED ON TEST DAY 22
The test and control guinea pigs were challenged two weeks after the epidermal induction application and were treated in the same way. Hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea pig just prior to the application. Two patches (3x3 cm) of filter paper were saturated with the test item at the highest tested non-irritating concentration of 50 % (applied to the left flank) and the vehicle only (PEG 300 applied to the right flank) using the same method as for the epidermal application. The amount of test item preparation applied was approximately 0.2 g and a volume of approximately 0.2 ml was used for the vehicle. The dressings were left in place for 24 hours.

Twenty-one hours after removal of the dressing the test sites treated with the test item were depilated as described in the epidermal pretest. The reaction sites were assessed approximately 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:
test group
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
0.5%
No. with + reactions:
10
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: positive control. Dose level: 0.5%. No with. + reactions: 10.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
0.5%
No. with + reactions:
10
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: positive control. Dose level: 0.5%. No with. + reactions: 10.0. Total no. in groups: 10.0.

SKIN EFFECTS AFTER INTRADERMAL INDUCTION -PERFORMED ON TEST DAY 1

The expected and common findings were observed in the control and test group after the different applications using FCA intradermally. These findings consisted of erythema, oedema, necrotizing dermatitis, encrustation and exfoliation of encrustation.

 

SKIN EFFECTS AFTER EPIDERMAL INDUCTION -PERFORMED ON TEST DAY 8

CONTROL GROUP

Discrete/patchy erythema were observed in all animals treated with PEG 300 only.

TEST GROUP

Discrete/patchy erythema were observed in all animals at the 24- and 48-hour reading after treatment with the test item at 50 % in PEG 300. The reactions observed in both groups occurred following pretreatment with 10 % SLS in paraffinum perliquidum.

 

SKIN EFFECTS AFTER THE CHALLENGE - PERFORMED ON TEST DAY 22

CONTROL GROUP

No skin reactions were observed in the animals when treated with either PEG 300 only or when treated with the test item at 50 % in PEG 300.

TEST GROUP

No skin reactions were observed in the animals when treated with either PEG 300 only or when treated with the test item at 50 % in PEG 300.

 

VIABILITY / MORTALITY / MACROSCOPIC FINDINGS

There were no deaths during the course of the study, hence no necropsies were performed.

 

CLINICAL SIGNS, SYSTEMIC

No signs of systemic toxicity were observed in the animals.

 

BODY WEIGHTS

One animal of the test group showed a loss of body weight (1.6 %) during the acclimatization period. It recovered between treatment start and the end of the observation period. The body weight of the other animals was within the range commonly recorded for animals of this strain and age.

Based on the above mentioned findings in an adjuvant sensitization test (M&K-test) in guinea pigs the test substance does not have to be classified and labeled as a skin sensitizer.

 

Interpretation of results:
GHS criteria not met
Conclusions:
The test material failed to elicit a skin sensitizing response in the guinea pig under the conditions of the test. No treatment related irritation was observed over the duration of the exposure and observation periods.
Executive summary:

In this guideline (OECD 406) study conducted with GLP certification, the test material (EC 438-340-0) did not induce skin sensitisation in the test animals (Guinea pig). The test was conducted using the Guinea pig maximisation method, with an induction of 10% test material via intradermal injection. 10% Sodium-Lauryl-Sulphate (SLS) in paraffinum perliquidum was applied topically to test sites on day 8. A challenge exposure of 50% test material was applied under occlusive conditions, 24 hour exposure, two weeks after topical application of SLS. Test animals showed no signs of sensitisation to the test material during the challenge and observation periods. Based upon the results of the test, the test material does not meet the criteria for classification under the EU Classification, Labelling, and Packaging (CLP) regulation (1272/2008).

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

The skin sensitising potential was tested in a guinea pig maximisation test according to OECD 406 and GLP (RCC, 2002). 15 female guinea pigs were used (10 animals in test group, and 5 animals in control group). During the induction procedure the test group was exposed to 10% via intradermal injection and 50 % test substance via topical application. The control group was exposed to vehicle only during this procedure. All animals were challenged with 50% test substance and vehicle. After the challenge with 50% test substance and vehicle, 0/10 (0%) positive responses were noted in the test group and 0/5 (0%) positive response were noted in the control group (24 and 48 h past treatment). Under the experimental conditions employed, none of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressings. In this study the test substance is not a dermal sensitizer.


Migrated from Short description of key information:
Sensitization
- skin: not sensitizing (OECD 406)

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

Migrated from Short description of key information:
No data available

Justification for classification or non-classification

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

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for skin sensitization under Regulation (EC) No. 1272/2008, fifth time in Directive EC 944/2013.