Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From 09 june 1995 to 02 nov 1995
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: The study was performed by following standardised method and GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
All data come from the draft report. No final report was established by the laboratory. Only one dose tested (1%).
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Species: guinea pig
- Strain: Hartley
- Sex: male/female

Details on test animals and environmental conditions
- Source: Charles River France (Saint-Aubin-Lès-Elbeuf, 76410 Cléon, France)
- Age at study initiation: young adult (about 6 weeks old)
- Weight at study initiation: 250-550 g
- Housing: in air-conditionned building. Animals housed in groups according to EEC/86/609 in stainless steel mesh cages (internal dimensions: 500*600*200 mm)
- Diet: pelleted complete guinea-pig diet ad libitum, (Diet ref. R14 C-10. Usine d'alimentation rationnelle, Villemoisson, 91360 Epinay-sur-Orge, France), sterilised by irradiation and analysed for the absence of chemical and bacterial contaminants.
- Water: Softened and filtered mains drinking water, ad libitum analysed at least once a year for chemical contaminants, and at least twice a year for bacterial contaminants (Laboratoire de Chimie de l'environnement du Dépt d'Ecologie Urbaine de la ville de Lyon).
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19+/-3°C
- Humidity (%): >=45% R.H. (target values)
- Air changes (per hr): >= 22
- Photoperiod (hrs dark / hrs light): 12 h light (artificial) / 12 h dark

IN-LIFE DATES: main study: 27 nov 1995 treatment begining for treated animals.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: absolute ethanol
Concentration / amount:
1% (v/v)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: absolute ethanol
Concentration / amount:
1% (v/v)
No. of animals per dose:
10 (5M/5F) for control group, 20 (10M/10F) for treated group.
Details on study design:
1st application: Induction 1 % intracutaneous
2nd application: Induction 10 % occlusive epicutaneous
3rd application: Challenge 1 % occlusive epicutaneous

RANGE FINDING TESTS:
1- Induction
*intradermal route: 2M and 2F received 0.1 ml intradermal injections of 100%, 50%, 10% of the test substance in absolute ethanol. Evaluation of animals reactions were made 24h and 48h after.

*topical and occlusive application for 48h: 2M and 2F (differents than those for intradermal route) received 0.5 ml per area of 8 cm2 of the article non-diluted or at 50% in absolute ethanol. Evaluation of cutaneous lesions (erythema) were made 1h after removal of the bandage.

*Induction preliminary study: 2M and 2F received 3 injections of 0.1 ml of 1% of p-trifluoromethylaniline in anterior area and 0.5 ml of sodium lauryl sulfate at 10% in Codex paraffin. The day after the animals received 10% of substance by topical application. Erythema, oedema and other anomalies were noted after the injections and 1h after topical application.

2-Challenge
*2M and 2F received 0.5 ml of 100%, 50% of the test substance in absolute ethanol by topical application for 24h under an occlusive bandage to an area of 4cm².
Cutaneous macroscopic examinations were evaluated for each concentration 24h and 48h after removal of the bandage (erythema and oedema).
*2M and 2F received 0.5 ml of 10% of the test substance in absolute ethanol by topical application for 24h under an occlusive bandage to an area of 4cm².
Cutaneous macroscopic examinations (erythema and oedema) were evaluated at 24h and 48h after removal of the bandage.
*2M and 2F received 0.5 ml of 1% and 0.1% of the test substance in absolute ethanol by topical application for 24h under an occlusive bandage to an area of 4cm².
Cutaneous macroscopic examinations (erythema and oedema) were evaluated at 24h and 48h after removal of the bandage .

22 animals were used during preliminary study and permit to estimate a minimal irritant concentration for induction phases and a minimum non-irritant concentration for challenge phase.


MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 pairs of intradermal injections to the retroscapular region left and right of the spire on day 1. A topical application on area used for intradermal injections on day 9.
- type of epicutaneous induction: occlusive
- SLS application: yes, 0.5 ml at concentration of 10% (w/w)
- Exposure period: on day 8
- Test groups: test substance in FCA
- Control group: absolute ethanol
- Site: back, flanks
- Duration: 9 days
- Concentrations: 3 series of 2* 0.1 ml injections
* Freund's complete adjuvant at 50% (v/v) in an isotonic injectable solution;
* test article in a 1% (v/v) solution in absolute ethanol
* mixture 50/50 (v/v): test article in a 1% (v/v) solution in absolute ethanol + Freund's complete adjuvant at 50% (v/v) in an isotonic injectable solution, i.e. a final 0.5% concentration of test article.
- The topical application was done by topical occlusive route for 48 h: 0.5 ml in 10% in absolute ethanol

B. CHALLENGE EXPOSURE
- No. of exposures: 1: test article applied under occlusive dressing (2*2 cm) filter paper kept in contact with the skin using an occlusive patch.
- Day(s) of challenge: 22
- Exposure period: 24 h
- Test groups: Test substance
- Control group: Test substance: 1% in absolute ethanol, 0.5 ml applied
- Site: right flanks
- Concentrations: 1% in absolute ethanol
- Evaluation: 24 and 48 hours, cutaneous macroscopic reactions (erythema, oedema, other abnormalities).
Challenge controls:
Test substance: 1% in absolute ethanol
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Results
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
1%
No. with + reactions:
2
Total no. in group:
20
Clinical observations:
Burnt appearance of the skin on a surface less than one quarter of the application area was noted for 2 males. 1 male and 2 females presented scratches to the application area.
Remarks on result:
other: see Remark
Remarks:
Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: Burnt appearance of the skin on a surface less than one quarter of the application area was noted for 2 males. 1 male and 2 females presented scratches to the application area. .

Any other information on results incl. tables

-Preliminary study: 

Mortality was noted during a first series of  preliminary studies, at the following dose levels (considering guinea  pigs weighing 

about 350 g):
 *intradermal injections of the test article as supplied or at 50% or  10%, i.e. about 100 mg + 50 mg + 10 mg 

of p-trifluoromethylaniline, i.e.  460 mg/kg.
 *topical application for 48h of the test article as supplied or 50%,  i.e. 500 mg + 250 mg of p-trifluoromethylaniline, i.e. 2143 

mg/kg.

Therefore the choice of the concentration for the main study was based on the systemic toxicity of the test article rather than 

on the irritant potential. A third preliminary study for induction was performed at the following dose levels :
3 x 0.1 ml injections of the test article at 1 % = 3 mg active
+ 0.5ml of the test article at 10 % = 50 mg active
+ sodium lauryl sulphate
i.e. about 151 mg/kg. These dose levels were well tolerated.

For challenge, the dose levels of the preliminary study were :
topical application for 24 hours of the test article as supplied or at 50  %, i.e. 500 mg + 250 mg of para- trifluoromethylaniline, 

i.e. 2143 mg/kg.  All animals died. With 10 % test article only (i.e. 125 mg/kg) no deaths  were noted but irritation was severe.

At 1% and 0.1%, no abnormalities were observed in the third challenge preliminary study.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Classification: not sensitizing
Executive summary:

In a dermal sensitization study (Chrysalis, 1995) with p-trifluoromethylaniline in absolute ethanol, Hartley guinea pigs (5/sex) were tested using the method of maximization.

2/20 animals presented positive reaction. It is not sufficient to consider p-TFMA for sensitization. 30% of positive reaction is necessary to classify a test item as skin sensitizer.

Burnt appearance of the skin on a surface less than one quarter of the application area was noted for 2 males. 1 male and 2 females presented scratches to the application area.

At dose of 1%, p-trifluoromethylaniline was not considered as dermal sensitizer, according to the EC classification criteria (67/548/EC directive) and CLP criteria.