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Diss Factsheets

Administrative data

Description of key information

Guinea pig maximization test (OECD 406); GLP; intradermal induction 1%, epicutaneous (occlusive) induction 25%, challenge 3 and 25%; not skin sensitizing

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:
June 1987
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted 1992
Deviations:
yes
Remarks:
animals housed in groups of 5 instead alone
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
adopted 1981
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was initiated prior to the implementation of the LLNA method.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Laboratory animal breeder Winkelmann, Borchen, Germany
- Age at study initiation: 5 - 8 weeks
- Weight at study initiation: 309 - 403 g
- Housing: in groups of 5 in Makrolon type IV cages, equipped with low-dust wood granules as bedding material (Bogner GmbH, Solingen, Germany)
- Diet: Altromin 3022 Diet for Guinea Pigs (Altromin GmbH, Lage, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 7 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 - 24
- Humidity (%): approx. 50
- Air changes (per hr): approx. 10
- Photoperiod (hrs dark / hrs light): 12 / 12
- IN-LIFE DATES: the study was carried out in June 1987
Route:
intradermal
Vehicle:
other: Cremophor EL (2% v/v) in sterile physiological saline solution
Concentration / amount:
1%
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
other: Cremophor EL (2% v/v) in sterile physiological saline solution
Concentration / amount:
25%
Day(s)/duration:
2 x 24h
Adequacy of induction:
highest technically applicable concentration used
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
other: Cremophor EL (2% v/v) in sterile physiological saline solution
Concentration / amount:
3% / 25%
Day(s)/duration:
3 weeks after induction for 24h
Adequacy of challenge:
other: highest concentration that could be suspended (25%) and a concentration of a factor of 10 lower (3%)
No. of animals per dose:
10 (control), 20 (test substance)
Details on study design:
RANGE FINDING TESTS:
The choice of dose used in the treatment for induction and challenge was based on results obtained in preliminary studies, as reported in the annex. In these studies local and systemic tolerance following intradermal and epicutaneous treatment was tested, and the threshold concentration for the primary irritant effect of the test article determined.
The following concentrations were determined:
Intradermal induction: 1%
Topical induction: 25%
Challenge: 3% / 25%
The highest concentration that could be suspended (25%), as well as a concentration a factor of 10 lower (3%) was used. As the challenge was performed using two concentrations simultaneously, the second challenge was not necessary and the second control group was not used.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: single injection (intradermal), 2 x 24h (epicutaneous)
- Test groups:
Intradermal (3 injection sites)
1) Freund's complete adjuvant 1:1 diluted with sterile physiological saline
2) 1% test substance formulated with Cremophor EL (2% v/v) in sterile physiological saline
3) 1% test substance formulated as above and Freund's complete adjuvant in equal parts
Epicutaneous: 25% test substance

- Control group:
1) Freund's complete adjuvant 1:1 diluted with sterile physiological saline
2) Cremophor EL (2% v/v) in sterile physiological saline
3) Cremophor EL (2% v/v) in sterile physiological saline and Freund's complete adjuvant in equal parts
Epicutaneous: sterile physiological saline

- Site: backs and flanks
- Frequency of applications: once each
- Duration: 2 x 24h (epicutaneous)


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: 24h
- Test groups: 3 and 25 % test item on the left flank, vehicle only on the right flank
- Control group: 3 and 25 % test item on the left flank, vehicle only on the right flank
- Concentrations: 3 and 25%
- Evaluation (hr after challenge): 24 and 48 h

Additional information on cutaneous exposure
The animals were teated with 0.5 mL sodium lauryl sulfonate (10%, in vaseline) one day prior to epicutaneous induction, in order to induce local skin irritation.
Positive control substance(s):
yes
Remarks:
laboratory control data for formaldehyde are provided in the study report
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
induction: 5% challenge: 2%
No. with + reactions:
9
Total no. in group:
18
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
24
Group:
positive control
Dose level:
induction: 5% challenge: 0.2%
No. with + reactions:
3
Total no. in group:
18
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
induction: 0% challenge: 25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
induction: 25% challenge: 3%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
induction: 0% challenge: 3%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
induction: 25% challenge: 3%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
induction: 25% challenge: 25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
induction: 0% challenge: 25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
induction: 25% challenge: 25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
no indication of skin sensitisation

Body weight

The body weights of the treatment group animals corresponded to those of the control animals. A slight reduction in body weight of the animals was found in both groups at the end of the study, this is considered due to the treatment technique used (occlusive dressing).

 

General examination/clinical signs

The treatment was tolerated by all animals without any signs occurring.

 

Mortality

There were no mortalities.

 

Positive control data (M-027579-01-1)

Formaldehyde (reported on 01 Feb 1985)

1st challenge: 2%
Number of animals with skin reactions (corrected values): 7/20 (test article group); 0/10 (control group)

2nd challenge: 0.5%
Number of animals with skin reactions (corrected values): 7/20 (test article group); 0/10 (control group)

The results were evaluated by subtracting the number of reacting animals on the control side from the number of reacting animals on the test article side. This was performed for the animals in the test article group, as well as for those in the control group. The resulting values represent the corrected values as given above.

 

Positive control data (M-121846-01-1)

Formaldehyde (reported on 20 Jul 1987 )

1st challenge: 2%
Number of animals with skin reactions (corrected values): 8/18 (test article group); 0/10 (control group)

2nd challenge: 0.2%
Number of animals with skin reactions (corrected values): 3/18 (test article group); 0/10 (control group)

The results were evaluated by subtracting the number of reacting animals on the control side from the number of reacting animals on the test article side. This was performed for the animals in the test article group, as well as for those in the control group. The resulting values represent the corrected values as given above.

Interpretation of results:
other: CLP/EU GHS criteria not met, no classification required according to Regulation (EC) No 1272/2008
Conclusions:
The study is in accordance to OECD TG 406, was performed under GLP conditions and is considered valid and reliable. Under the conditions tested, the test item did not induce skin sensitization in guinea pigs. According to criteria of the CLP Regulation (EU) No. 1272/2008, no classification of the test item for skin sensitisation is required.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

Sensitisation

To evaluate skin sensitising properties of the test substance, data on a GLP-conform skin sensitisation study according to OECD Guideline 406 is available.

 

In the study, skin sensitising properties of the test substance (powder with a purity of 94.2%) were evaluated in guinea pigs according to the maximization method of Magnusson and Kligman (OECD 406) in compliance with GLP (M-027579-01-1). Concentrations of 1 and 25% (w/w) were chosen for intradermal and epicutaneous induction, respectively, in regard to the results of a range finding study. The test group comprised 20 and the control group 10 male guinea pigs (strain DHPW). On Day 1, 3 pairs of intradermal injections were performed in the interscapular region of all animals:

1) Freund's complete adjuvant (FCA) diluted at 50% (v/v) with 0.9% NaCl (both groups),

2) test substance at a concentration of 1% in physiological saline (treated group) or vehicle alone (control group),

3) test substance (1%, (w/w)) in a mixture FCA/0.9% NaCl 50/50 (v/v) (treated group) or physiological saline (50%, w/v) in a mixture FCA/0.9% NaCl 50/50 (v/v) (control group).

On Day 7, the skin of animals was treated with 0.5 mL sodium lauryl sulfonate (10%, in vaseline) one day prior to epicutaneous induction, in order to induce local skin irritation. On Day 8, the test substance (treated group) or the vehicle (control group) was applied topically and covered by an occlusive dressing for 2 x 24 hours. The challenge with 3 and 25% test substance (w/w) occurred on the left flank of all animals on Day 22. The right flank received the vehicle only and served as control. Test substance and vehicle were held in place by an occlusive dressing for 24 hours. No mortality or signs of clinical symptoms were recorded until the end of the observation period and body weight gain was comparable among the groups. No cutaneous reactions were observed in control and test substance treated animals.

 

Thus, the test substance did not induce skin sensitisation in guinea pigs under the experimental conditions chosen.

 

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

no study available

Justification for classification or non-classification

Under the conditions tested, the test item did not induce skin sensitization in guinea pigs. No classification in regard to skin sensitisation is required according to CLP Regulation (EC) No. 1272/2008.