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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
1989-01-18 till 1989-02-17
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (OECD) with GLP

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1989
Report Date:
1989

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
(adopted 1981)
GLP compliance:
yes
Remarks:
(BASF AG, Department of Toxicology )
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The Guinea Pig Maximazitation Test met the previous requirements before the entry into force of REACH. The GPMT is suitable and reliable to cover this endpoint. For this reason and for animal welfare reasons, no further in vivo study (LLNA test) needs to be performed.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Trioxan-Schuppen
- Physical state: solid, crystalline, white
- Analytical purity: 99.9%
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Lippische Versuchstierzucht, Hagermann GmbH & Co.
- Weight at study initiation: 259-319 g
- Housing: 5 per cage
- Diet (e.g. ad libitum): Kliba 341.4 mm (Klingentalmühle AG, Kaiseraugst)
- Water (e.g. ad libitum): tap water, addition of about 2g ascorbic acid/10 L water twice every week
- Acclimation period: at least 8 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Photoperiod (hrs dark / hrs light): 12/12


Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
PRETEST
- The animals were exposed to about 0.15 g of the test substance formulation applied to the skin under occlusive conditions

MAIN TEST
- Intradermal induction: 5% test substance formulation in either aqua dest. or Freund's adjuvant/ aqua dest. (1:1)
- Epicutaneous induction: 50% test substance formulation in either aqua dest.
- First Challenge: 50% test substance formulation in either aqua dest.
- Second Challenge: 50% test substance formulation in either aqua dest.

Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
PRETEST
- The animals were exposed to about 0.15 g of the test substance formulation applied to the skin under occlusive conditions

MAIN TEST
- Intradermal induction: 5% test substance formulation in either aqua dest. or Freund's adjuvant/ aqua dest. (1:1)
- Epicutaneous induction: 50% test substance formulation in either aqua dest.
- First Challenge: 50% test substance formulation in either aqua dest.
- Second Challenge: 50% test substance formulation in either aqua dest.

No. of animals per dose:
20 animals in the test group
10 animals in each of two control groups
Details on study design:
PRETEST
- No. of exposures: 2 times for 24 hours within a period of 96 hours (about 0.15 g of the test substance formulation under occlusive condition)
- Exposure period: 24h
- Test groups: 4 animals per test concentration
- Site: Flank, second application on the same spot
- Frequency of applications: 24h after initial application, second application was administered
- Reading: about 24h and 48h after the begining of application (assessment of skin findings)
- Duration: 96h

MAIN STUDY

INDUCTION
Intradermal Application
- No. of exposures: 6 intradermal injections in groups of two per animal
- Exposure period: 24h
- Test groups: 20 animals
- Control group: 10 animals
- Site: Shoulder
- Frequency of applications: one time
- Concentrations: 2 injections each of 0.1 ml Freund's adjuvant without test substance emulsified with water (1:1) followed by 2 injections each of 0.1 ml of the test substance formulation, followed by 2 injections each of 0.1 ml Freund's adjuvant/water (1:1) with test substance in the test group. Control group were given the same injections but without test substance, only with the formulating agent.

Percutaneous Application
- Time of application: one week after intradermal application
- No. of exposures: single dermal application
- Exposure period: 48h
- Test groups: 20
- Control group: 10
- Site: Shoulder, same area as preceding intradermal application
- Frequency of applications: one time
- Concentrations: The test animals were exposed to about 50% test substance formulation in aqua dest (0.3 g of the test substance formulation) under an occlusive dressing. The control animals were treated since with distilled water in similar manner.

CHALLENGE
Two challenges were conducted:
- First challenge: treatment of the test group and of control group 1 with the test substance formulation (control group 2 remained untreated);
- Second challenge: treatment of the test group and of control groups 1 and 2 with the test substance formulation
- Day(s) of challenge: (first challenge: 14d after percutaneous exposure, second challenge: 7d after first challenge)
- Exposure period: 24h
- Test groups: 20
- Control group: 10
- Site: Intact clipped flank
- Concentrations: The test animals were exposed to about 50% test substance formulation in aqua dest under an occlusive dressing.
- Evaluation (hr after challenge): 24, 48 and 72h after begin of application
Challenge controls:
10 guinea pigs
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
50% test substance in aqua dest.
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50% test substance in aqua dest.. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
50% test substance in aqua dest.
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% test substance in aqua dest.. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
50% test substance in aqua dest.
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 50% test substance in aqua dest.. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
50% test substance in aqua dest.
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 50% test substance in aqua dest.. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
No test substance application
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: No test substance application. No with. + reactions: 0.0. Total no. in groups: 10.0.

Any other information on results incl. tables

Skin irritation findings during induction:

After intradermal induction, distinct erythema and edema were observed at the injection sites of both control and test animals, at which only Freund's adjuvant/aqua dest. (1:1) was applied. Injection of test substance preparation in aqua dest caused distinct erythema in the test animals. The application areas which were injected with test substance preparation in Freund's adjuvant/aqua dest. (1:1) exhibited distinct erythema and edema. The control animals injected with aqua dest. (vehicle) did not show any skin reactions.

 

Result of the maximization test:   

No skin reactions could be observed in the control and treated groups after both, the first and the second challenge.

Applicant's summary and conclusion