2-anilinoethanol

Endpoint summary

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

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP compliant guideline study, available as unpublished report, but only 5 animals tested.

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.

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

Route:

intradermal and epicutaneous

Vehicle:

olive oil

Concentration / amount:

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

Route:

epicutaneous, occlusive

Vehicle:

olive oil

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

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.

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.

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.

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.

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.

- 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.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

The test substance was tested for its sensitizing effect on the skin of 5 guinea pig in the Maximization Test according to EU Method B.6 and in compliance with GLP (BASF 1994). The intradermal and percutaneous induction, 5% test substance preparation and unchanged test substance respectively, caused necrotic skin changes in addition to slight edema in the test group animals. The challenge with the 100% test substance preparation in olive oil did not cause any skin reactions.

Justification for selection of skin sensitisation endpoint:
One study on skin sensitization is available. The study which was performed most similar to OECD 406 whereas this study has been selected as key study.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the available information on the skin sensitizing potential of the test substance, classification for skin sensitization is not warranted in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.