2-anilinoethanol

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study, available as unpublished report, but only 5 animals tested.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

Cited as Directive 84/449/EEC, B.6

Deviations:

yes

Remarks:

only 5 animals exposed

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Study was conduceted well before LLNA guideline was available.

Test material

Test material information

Constituent 1

Reference substance name:

2-anilinoethanol

EC Number:

204-588-1

EC Name:

2-anilinoethanol

Cas Number:

122-98-5

Molecular formula:

C8H11NO

IUPAC Name:

2-(phenylamino)ethan-1-ol

Details on test material:

- Name of the test substance used in the study report: Hydroxyethylanilin
- Test substance No.: 89/89
- Physical state: yellowish liquid
- Storage: room temperature

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Lippische Versuchstierzucht, Hagemann GmbH & Co. KG, Extertal 1, FRG
- Age at study initiation: young adults
- Weight at study initiation: 300 - 350 g
- Housing: 5 animals per cage, Makrolon type IV, enriched with Granulat Typ 3/4 (dust-free), SSNIFF
- Diet: ad libitum, Kliba Laboratoriumdiet 341 (rabbit and guineapigs diet) from Firma Klingentalmühle AG, Kaiseraugst, Switzerland
- Water: ad libitum, tap water with 2 g ascorbic acid per 10 L water
- 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)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

olive oil

Induction 2

Concentration / amount:

- Intradermal induction: 5%
- Epicutaneous induction: unchanged
- Challenge: unchanged

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

olive oil

Challenge 2

Concentration / amount:

- Intradermal induction: 5%
- Epicutaneous induction: unchanged
- Challenge: unchanged

No. of animals per dose:

- Test material: 10
- Control: 5

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 and Fixomull Stretch (adhesive fleece) from Beiersdorf AG). The test filter paper strip was soaked in the test substance resp. in the test substance formulation; thus, the animals were exposed to about 0.15 g of the test substance resp. the test substance formulation.
- Exposure period: 2 times for 24 hours within a period of 96 hours in order to detect non-specific 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 row 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): 24 hours
- Control group: 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 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:

Identical procedure. A row of six intradermal injections in groups of two per animal 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 vehicle, C) back row: 2 injections each of 0.1 ml Freund's adjuvant/ 0.9% aqueous NaCl-solution (1 : 1) with vehicle. Epicutaneous exposure not performed, since the test substance was applied unchanged and thus no solvent was used.

Positive control substance(s):

yes

Remarks:

1-chlor-2,4-dinitro-benzol

Results and discussion

Positive control results:

In a positive control test with 1-chlor-2,4-dinitro-benzol, performed a year and 8 months before this study, in 55% of the animals sensitisation was observed.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

unchanged (100%)

No. with + reactions:

0

Total no. in group:

5

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: 5.0.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

unchanged (100%)

No. with + reactions:

0

Total no. in group:

5

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: 5.0.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

unchanged (100%)

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

originally 10 animals, but 5 died during epicutaneous induction

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: 5.0. Clinical observations: originally 10 animals, but 5 died during epicutaneous induction.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

unchanged (100%)

No. with + reactions:

0

Total no. in group:

5

Clinical observations:

originally 10 animals, but 5 died during epicutaneous induction

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: 5.0. Clinical observations: originally 10 animals, but 5 died during epicutaneous induction.

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-solution (1:1) was applied.

 - Injection of the test substance in olive oil DAB 9 resp. in Freund's adjuvant/0.9% aqueous NaCl-solution (1:1) caused necrotic skin changes and slight oedema in the animals of the test group.

 - The control group animals which were treated with olive oil DAB 9 showed well-defined erythema.

 - The epicutaneous induction was only carried out in the test group because the test substance was applied unchanged and thus no solvent was used.

 - Necrotic skin changes, partially open (caused by the intradermal induction) and slight oedema were observed in 5 test group animals.

 - 5 animals of the test group died during the epicutaneous induction. The death of the 5 test group animals is likely to be test substance related.

 - 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.