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Description of key information

In the GLP compliant key Guinea Pig Maximisation Test according to OECD guideline 406, the test item induced no skin sensitisation under the conditions of the test.

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:
1994-07-29
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1981
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The in vivo study was initiated before the entry into force of the amendments to Annexes VII and VIII.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Females nulliparous and non-pregnant: Not specified
- Microbiological status of animals: SPF
- Weight at study initiation: 266 to 319 g
- Housing: In PPL (Type IV) cages, two or three animals per cage. The cages were cleaned and the bedding changed 3 times a week.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21 ± 3
- Humidity (%): 55 ± 15
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12/12

- IN-LIFE DATES: From day 0 to day 29
Route:
intradermal and epicutaneous
Vehicle:
arachis oil
Concentration / amount:
5% w/w
Day(s)/duration:
3 weeks: Intradermal injections on day 0, topical sodium lauryl sulphate treatment on day 6, topical treatment with 100% test article on day 7 until dressing was removed on day 9, another 12 days until challenge
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
other: 100% of the test item was used.
Remarks:
As control article, ethanol/diethylphthalate was used.
Concentration / amount:
100 % (w/w)
Day(s)/duration:
Day 27 (duration = 24 hours)
Adequacy of challenge:
highest non-irritant concentration
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: 100% of the test item was used.
Remarks:
As control article, ethanol/diethylphthalate was used.
Concentration / amount:
100% (w/w)
Day(s)/duration:
Day 20 (duration = 24 hours)
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
One dose group including 20 female guinea pigs
Details on study design:
RANGE FINDING TESTS:
Doses were selected on the basis of a range-finding test. Selected dose levels were 5% (w/w) test article in arachis oil for intradermal induction and 100% (w/w) test article for dermal induction, challenge and rechallenge application.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal injections and a closed patch topical application)
- Exposure period: 3 weeks (including the closed patch topical application of the test item one week after intradermal injections) until challenge procedure
- Test groups: One test group of 20 animals, receiving Freund's Complete Adjuvant (FCA) emulsified 1:1 in sterile water (1st pair), 5% (w/w) of the test substance in arachis oil (2nd pair) and 5% (w/w) of the test substance in arachis oil in a 1:1 mixture with FCA (3rd pair).
- Control group: A control group of 10 animals was treated with FCA emulsified 1:1 in sterile water (1st pair), arachis oil (2nd pair) and a 1:1 mixture of FCA and arachis oil (3rd pair)
- Site: An area of dorsal skin 4 x 6 cm in the scapular region (intradermal induction and topical induction)
- Frequency of applications: Single simultaneous application of three pairs of intradermal injections, following a single topical induction after 6 days
- Duration: 6 days until 0.5 g sodium lauryl sulphate (10% in petrolatum) was massaged into the skin to provoke a mild inflammatory reaction. 24 hours later, 0.4 mL of arachidis oil (control group) or 100% test article was spread over a 3x5 cm patch and placed on the skin and covered with impermeable bandage. The dressing was left in place for 48 hours.
- Concentrations: 5% (intradermal induction), 100% (dermal induction, challenge and rechallenge application)

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Days of challenge: On day 20 (three weeks after intradermal induction) and on day 27
- Exposure period: From day 20 to day 27 (first challenge), from day 27 to day 29 (rechallenge)
- Test groups: One test group of 20 animals, receiving a volume of 0.1 mL of test article (100%) or control article (ethanol/diethylphthalate)
- Control group: One test group of 20 animals, receiving a volume of 0.1 mL of test article (100%) or control article (ethanol/diethylphthalate)
- Site: Left anterior flank (test item), left posterior flank (vehicle)
- Concentrations: 100% test item
- Evaluation (hr after challenge): 24 hours and 48 hours after challenge and rechallenge
Challenge controls:
The left posterior flank was treated with ethanol/diethylphthalate in both test and control groups.
Positive control substance(s):
no
Positive control results:
No positive control was included.
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
test chemical
Dose level:
100% test item
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test chemical
Dose level:
100% test item
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Remarks:
Slight and discrete erythema were seen in one animal (grade 1). One animal showed moderate and confluent erythema (grade 2).
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
positive control
Dose level:
Not tested
Clinical observations:
Not tested
Remarks on result:
positive indication of skin sensitisation
Remarks:
No positive control was inlcuded in the study
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
positive control
Dose level:
Not tested
Clinical observations:
Not tested
Remarks on result:
positive indication of skin sensitisation
Remarks:
No positive control was inlcuded in the study
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
100% test item
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
100% test item
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Remarks:
Slight and discrete erythema were seen in one animal (grade 1)
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
100% test item
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Remarks:
Slight and discrete erythema were seen in 4 animals (grade 1). One animal showed moderate and confluent erythema (grade 2).
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
100% test item
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Remarks:
Slight and discrete erythema were seen in 8 animals (grade 1). One animal showed moderate and confluent erythema (grade 2).
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
Not tested
Clinical observations:
Not tested
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
Not tested
Clinical observations:
Not tested
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100% test item
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No clinical signs were recorded.
Remarks on result:
no indication of skin sensitisation
Remarks:
Slight and discrete erythema were seen in 2 animals (grade 1).
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100% test item
No. with + reactions:
1
Total no. in group:
10
Clinical observations:
No clincial signs were recorded.
Remarks on result:
no indication of skin sensitisation
Remarks:
Slight and discrete erythema were seen in 5 animals (grade 1). One animal showed moderate and confluent erythema (grade 2).
Interpretation of results:
GHS criteria not met
Conclusions:
In a GLP compliant Guinea Pig Maximisation Test according to OECD guideline 406, the test item induced no skin sensitisation under the conditions of the test.
Executive summary:

The allergenic potential of the test item was investigated according to one of the methods recommended in the OECD Guidelines No. 406, "Skin Sensitization", 1992 under GLP conditions. The delayed contact hypersensitivity test used was the Guinea Pig Maximization Test described by B. Magnusson and A. M. Kligman. Thirty animals divided into a test group of 20 animals and a control group of 10 animals were included in the study. The study included an induction and a challenge phase. The animals in the test group were induced with the test article, whereas the animals in the control group were induced with oleum arachidis. The induction procedure included intradermal injections and a closed patch topical application one week apart. All animals were challenged by a closed patch topical application of the test article for 24 hours. The challenge procedure included a closed patch topical treatment of the test article an the flank 3 weeks after the intradermal injection. The skin reactions were evaluated 24 and 48 hours after the challenge application.  A 5% (w/w) test article concentration in oleum arachidis was used for the intradermal induction. Undiluted test article was used for the topical induction and for the challenge and rechallenge application. Under these experimental conditions no evidence of delayed contact hypersensitivity was seen after treatment with the test item.

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

Skin sensitisation in vivo, RL1


The allergenic potential of the test item was investigated according to one of the methods recommended in the OECD Guidelines No. 406, "Skin Sensitization", 1992 under GLP conditions. The delayed contact hypersensitivity test used was the Guinea Pig Maximization Test described by B. Magnusson and A. M. Kligman. Thirty animals divided into a test group of 20 animals and a control group of 10 animals were included in the study. The study included an induction and a challenge phase. The animals in the test group were induced with the test article, whereas the animals in the control group were induced with oleum arachidis. The induction procedure included intradermal injections and a closed patch topical application one week apart. All animals were challenged by a closed patch topical application of the test article for 24 hours. The challenge procedure included a closed patch topical treatment of the test article an the flank 3 weeks after the intradermal injection. The skin reactions were evaluated 24 and 48 hours after the challenge application.  A 5% (w/w) test article concentration in oleum arachidis was used for the intradermal induction. Undiluted test article was used for the topical induction and for the challenge and rechallenge application. Under these experimental conditions no evidence of delayed contact hypersensitivity was seen after treatment with the test item.

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 the available data, the test substance is not considered to be a skin sensitiser and is thus not classified according to Regulation (EC) No 1272/2008 (CLP), as amended for sixteenth time in Regulation (EU) No 2021/743.