Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP compliant guideline study, available as unpublished report, no restrictions, fully adequate for assessment

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994
Reference Type:
other: Amendment
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of the test substance used in the study report: Asta-C6-Acetal
- Physical state: liquid, achromatic
- Storage: refrigerator, exclusion of light

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River GmbH - Wiga, Sulzfeld, FRG
- Age at study initiation: young adults
- Weight at study initiation: 278 - 331 g
- Housing: 5 animals per cage, Makrolon type IV cages, Granulat typ 3/4 (staubfrei) SSNIFF bedding
- Diet: ad libitum, Kliba Labordiät 341 (Kaninchen-Meerschweinchen-Haltungsdiät), from firma Klingentalmühle AG, Kaiseraugst, Switzerland
- Water: ad libitum, tap water (about 2 g of ascorbic acid per 10 L water was added twice a week)
- Acclimation period: 7 days

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

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
olive oil
Concentration / amount:
- Intradermal induction: test substance 5% in olive oil DAB 10 or in Freund’s adjuvant/0.9% aqueous NaCl-solution (1:1) or olive oil DAB 10
- Epicutaneous induction: unchanged
- Challenge: unchanged
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
olive oil
Concentration / amount:
- Intradermal induction: test substance 5% in olive oil DAB 10 or in Freund’s adjuvant/0.9% aqueous NaCl-solution (1:1) or olive oil DAB 10
- Epicutaneous induction: unchanged
- Challenge: unchanged
No. of animals per dose:
- Test material: 20
- Control: 10
Details on study design:
RANGE FINDING TESTS:
- Amount applied: 2 x 2 cm filter paper strips were applied to the skin of the flanks under an occlusive dressing (the bandage consists of rubberized linenpatches 4 x 4 cm from Russka, test patch 5 x 5 cm of Idealbinde from Pfälzische Verbandstoff-Fabrik, and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG). The test filter paper strip was soaked in the test substance or test substance formulation; thus, the animals were exposed to about 0.15 g of the test substance or test substance formulation.
- Exposure period: The test substance was applied 2 times for 24 hours within a period of 96 hours in order to detect nonspecific phenomena that are not caused by a sensitization reaction but could possibly be attributed to a shift in the irritation threshold.
- Site of application: flank, respective on the same area
- Number of test animals: 4 per test concentration
- Readings: 24 and 48 h after the beginning of application

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (day 0 intradermal injections + day 7 epicutaneous exposure)
- A total of six intradermal injections in groups of two per animal was made. These injections were: A) front row: 2 injections each of 0.1 mL Freund's adjuvant without test substance emulsified with 0.9% aqueous NaCl-solution in a ratio of 1:1, B) middle row: 2 injections each of 0.1 mL of the test substance formulation, C) back row: 2 injections each of 0.1 mL Freund's adjuvant/ 0.9% aqueous NaCl-solution (1 : 1) with test substance.
- Epicutaneous exposure: 2 x 4 cm filter paper strips were applied to the skin of the shoulder under an occlusive dressing (the bandage consists of rubberized linenpatches 6 x 4 cm from Russka and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG). The filter paper strip was soaked in the test substance; thus, the animals were exposed to about 0.3 g of the test substance.
- Exposure period (epicutaneous): 48 hours
- Control group 1 and 2: The animals were given the same injections (A, B, C) but without test substance, only with the formulating agent. For epicutaneous exposure the control groups were not treated.
- Site: shoulder
- Evaluation: 24 h after intradermal injections, and 48 h after epicutaneous exposure.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Exposure: 2 x 2 cm filter paper strips were applied to the skin of the flank under an occlusive dressing (the bandage consists of rubberized linenpatches 4 x 4 cm from Russka, test patch 5 x 5 cm of Idealbinde from Pfälzische Verbandstoff-Fabrik, and Fixomull® Stretch (adhesive fleece) from Beiersdorf AG). The test filter paper strip was soaked in the test substance; thus the animals were exposed to about 0.15 g of the test substance.
- Day(s) of challenge: 21
- Exposure period: 24 h
- Site: intact flank
- Evaluation (hr after challenge): 24 and 48 after the removal of the patch
Challenge controls:
Two control groups were used:
- Both control groups received the same injections (A, B, C) but without test substance, only with the formulating agent.
- For epicutaneous exposure both control groups were not treated.
- Challenge: Treatment of control group 1 with the test substance and control group 2 remained untreated.
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene

Results and discussion

Positive control results:
1-Chloro-2,4-dinitrobenzene has a sensitizing effect an the skin of the guinea pig.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
unchanged (100%)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: unchanged (100%). No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
unchanged (100%)
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: unchanged (100%). No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
unchanged (100%)
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: unchanged (100%). No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
unchanged (100%)
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: unchanged (100%). No with. + reactions: 0.0. Total no. in groups: 20.0.

Any other information on results incl. tables

- After the intradermal induction well-defined erythema and slight oedema were observed at the injection sites of the control group animals and test group animals at which only Freund's adjuvant/0.9% aqueous NaC1-solutian (1:1) was applied. Injection of the test substance in olive oil DAB 10 or in Freund's adjuvant/0.9% aqueous NaCl-solution (1:1) caused well-defined erythema and slight oedema in the test group animals. The control group animals which were treated with olive oil DAB 10 exhibited well-defined erythema.

- The percutaneous induction was only carried out in the test group because the test substance was applied unchanged and thus no solvent was used. Incrustation, partially open (caused by the intradermal induction) in addition to well-defined erythema and slight oedema could be observed in the animals of the test group.

- The challenge with the unchanged test substance did not cause any skin reactions neither in the test group animals nor in the animals of control group 1.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information
Conclusions:
Under the conditions of the test the substance is considered to be not sensitising to the skin.
Executive summary:

The substance was tested for its sensitizing effect in an OECD 406 guideline study (in compliance with GLP), on the skin of the guinea pig in the Maximization Test. The intradermal induction with the 5% test substance preparations caused slight to well-defined sings of irritation in the test group animals. After the percutaneous induction with the unchanged test substance the animals of the test group exhibited incrustation, partially open (cause by the intradermal induction) in addition to well-defined erythema and slight edema. No skin effects were observed, in both the test as the control animals, after the challenge (21 days after the intradermal induction). Based on this it was concluded that the test substance does not have a sensitizing effect on the skin of the guinea pig in the Maximization Test.