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Description of key information

Skin sensitisation (equivalent to OECD 406): not sensitising

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:
7 May - 1 Jun 1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
Also in accordance with GLP principles.
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
adopted 19. Sept. 1984
Deviations:
no
Qualifier:
equivalent or similar to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Test was done before LLNA as first-choice method for in-vivo testing was set into force.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: D. Hall, Newchurch, Staffordshire, England
- Age at study initiation: 6 to 7 weeks of age
- Weight at study initiation: 327 - 389 g
- Housing: in groups of ten in suspended cages with wire mesh floors
- Diet: vitamin C-enriched guinea pig diet F.D.1., ad libitum; hay was given weekly
- Water: ad libitum
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): 30-70
- Air changes (per hr): approximately 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 7 May To 01 June 1991
Route:
intradermal and epicutaneous
Vehicle:
other: Alembicol D
Concentration / amount:
Induction: intradermal - 2.5% w/w; epicutaneous - 70% w/w
Challenge: epicutaneous - 25 and 50% w/w
Route:
epicutaneous, semiocclusive
Vehicle:
other: Alembicol D
Concentration / amount:
Induction: intradermal - 2.5% w/w; epicutaneous - 70% w/w
Challenge: epicutaneous - 25 and 50% w/w
No. of animals per dose:
10 (controls), 20 (in test group)
Details on study design:
RANGE FINDING TESTS: The intradermal and topical irritancy of a range of dilutions of T-4113 was investigated to identify where possible (a) irritant test concentrations suitable for induction phase of the main study and (b) non-irritant concentrations by the topical route of administration for the challenge phase. Concentration ranges tested in two animals each were: intradermal - 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, 7.5, 10.0% (w/w); topical - 10, 30, 50, 70% (w/w)

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single intradermal injection and 48 hours topical application
- Test groups:
Intradermal - 3 pairs of injections: (1) a 1:1 mixture (v/v) Freund's Complete adjuvant (FCA)/water; (2) test substance in Alembicol D; (3) test substance in a 1:1 mixture of Freund's complete adjuvant and Alembicol D
Epicutaneous - test substance in Alembicol D
- Control group:
Intradermal - 3 pairs of injections: (1) a 1:1 mixture (v/v) FCA/water; (2) Alembicol D; (3) 1:1 mixture Freund's Complete adjuvant and Alembicol D
Epicutaneous - Alembicol D
- Site: scapular region, intradermal and epicutaneous
- Frequency of applications: every 7 days
- Duration: Days 0-9
- Concentrations: intradermal 2.5% w/w, epicutaneous 70% w/w

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 21
- Exposure period: 24 hours
- Test groups: test substance at 25 and 50% w/w in Alembicol D
- Control group: test substance at 25 and 50% w/w in Alembicol D
- Site: anterior and posterior site on the left flank
- Concentrations: 25 and 50% w/w
- Evaluation (hr after challenge): 24, 48 and 72 hours

OTHER:
Justification for choice of vehicle: Alembicol D# is a product of coconut oil. By fractionating the fatty acids of coconut oil and re-esterifying with glycerine, a Medium Chain Triglyceride (MCT) Oil is produced. Alembicol D is such an oil. It is advantageous in this type of test in that it is stable, has a low viscosity and surface tension and is easily absorbed into the skin. These properties make it ideal as a vehicle for intradermal injections and topical applications.
Positive control substance(s):
yes
Remarks:
Formalin (checked periodically)
Positive control results:
Formalin is used as positive control and checked regularly to test the stability of the assay;
concentrations used: intradermal - 0.05 -0.1%; epicutaneous - 5 -10%; challenge - 1 -5%; usually 50 - 100% of the tested animals showed sensitizing reactions in all studies conducted
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
Challenge 25 and 50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Skin reactions following intradermal injections Necrosis at sites receiving FCA (test and control animals). Slight irritation in test animals for 2.5% w/w test substance in Alembicol D and control animals for Alembicol D. No dermal reaction at challenge.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
Challenge 25 and 50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Skin reactions following intradermal injections Necrosis at sites receiving FCA (test and control animals). Slight irritation in test animals for 2.5% w/w test substance in Alembicol D and control animals for Alembicol D. No dermal reaction at challenge.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
negative control
Dose level:
Challenge 25 and 50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Skin reactions following intradermal injections Necrosis at sites receiving FCA (test and control animals). Slight irritation in test animals for 2.5% w/w test substance in Alembicol D and control animals for Alembicol D. No dermal reaction at challenge.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
Challenge 25 and 50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Skin reactions following intradermal injections Necrosis at sites receiving FCA (test and control animals). Slight irritation in test animals for 2.5% w/w test substance in Alembicol D and control animals for Alembicol D. No dermal reaction at challenge.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
Challenge 25 and 50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Skin reactions following intradermal injections Necrosis at sites receiving FCA (test and control animals). Slight irritation in test animals for 2.5% w/w test substance in Alembicol D and control animals for Alembicol D. No dermal reaction at challenge.
Reading:
other: 3rd reading
Hours after challenge:
72
Group:
test chemical
Dose level:
Challenge 25 and 50%
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
Skin reactions following intradermal injections Necrosis at sites receiving FCA (test and control animals). Slight irritation in test animals for 2.5% w/w test substance in Alembicol D and control animals for Alembicol D. No dermal reaction at challenge.
Reading:
1st reading
Group:
positive control
Dose level:
Challenge 5% and 1%
No. with + reactions:
10
Total no. in group:
10
Interpretation of results:
GHS criteria not met
Conclusions:
In this adjuvant type guinea pig test method for skin sensitisation according to OECD 406 guideline and GLP principles, the substance did not produce evidence of skin sensitization in any of the twenty test animals.
Executive summary:

Assessment of the skin sensitization potential of the test substance using the guinea-pig according to OECD 406 guideline and GLP principles. No signs of ill health or toxicity were recorded. 


Induction:

Intradermal injections: Necrosis was recorded at sites receiving FCA in test and control animals. Slight irritation was seen in test animals at sites receiving the 2.5% test substance in Alembicol D and similar signs of irritation were observed in control animals receiving Alembicol D.

Topical application: Slight erythema was observed in test animals following topical application with the 70% test substance in Alembicol D and similar signs of irritation were seen in the controls.


Challenge:

There were no dermal reaction seen in any of the test or control animals.


Based on the results, the substance did not produce evidence of skin sensitization in any of the twenty test animals.

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

Skin sensitisation

The skin sensitising potential of 4,4'-(9H-fluoren-9-ylidene)bis(2-chloroaniline) (CAS 107934-68-9) was evaluated in a guinea pig maximisation test performed according to OECD 406 (Parcell and Healing, 1991). 20 female guinea pigs in the test group and 10 females in the control group were induced by intradermal and epicutaneous exposure to the test substance. Intradermal induction was performed with 2.5% test solution, and epicutaneous induction was performed with 70% test solution. Skin reactions (necrosis) were observed after intradermal induction at application sites receiving FCA. In addition, slight irritation was observed at sites receiving 2.5% (intradermal induction) test substance in Alembicol D. Slight erythema were observed following topical induction. The challenge was performed by open epicutaneous exposure to a 25% and 50% formulation of test substance in Alembicol D. The animals in the treatment group and the control group did not show any signs of a sensitising effect at the reading time points (24, 48, and 72 h after challenge). In conclusion 0/10 and 0/20 animals of the control and test group showed any reaction after the challenge period, thus indicating that the test substance is not sensitising under the conditions of the test.


Justification for selection of skin sensitisation endpoint:
There is only one study available.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available
Additional information:

No data available.


Justification for selection of respiratory sensitisation endpoint:
The study is not required according to Annex VII - X of Regulation (EC) No 1907/2006.

Justification for classification or non-classification

The available data on skin sensitisation of 4,4´-(9H-fluoren-9 -ylidene)bis(2 -chloroaniline) do not meet the criteria for classification according to Regulation (EC) 1272/2008, and are therefore conclusive but not sufficient for classification.