2'-anilino-6'-[ethyl(3-methylbutyl)amino]-3'-methylspiro[isobenzofuran-1(3H),9'-[9H]xanthene]-3-one

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

Study period:

6 April to 1 May 1982

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study undertaken to internationally accepted guidelines but before the implementation of GLP regulations.

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

a local irritation was not created in the induction phase 24 hr before topical application

GLP compliance:

not specified

Type of study:

guinea pig maximisation test

Species:

guinea pig

Strain:

Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: reputable breeder
- Age at study initiation: 4 to 6 weeks old
- Weight at study initiation: 400 ± 50 g
- Housing: in suspended cages with wire mesh floors
- Diet (e.g. ad libitum): free access to Vitamin C-enriched Guinea-Pig Diet
- Water (e.g. ad libitum): free access to tap water
- Acclimation period: yes but no data on time

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21°C
- Humidity (%): 30-70%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: 6 April To: 1 May 1982

Route:

intradermal and epicutaneous

Vehicle:

other: Alembicol D

Route:

epicutaneous, occlusive

Vehicle:

other: Alembicol D

No. of animals per dose:

10 guinea pigs treated with S-2055
5 guinea pigs with vehicle only

Details on study design:

RANGE FINDING TESTS:
The intradermal and topical irritancy of a range of suspensions of S-205 in Alembicol D was investigated to identify
(a) irritant test substance concentrations suitable for the induction phase of the main study and
(b) non-irritant concentrations by the topical route of administration for the challenge phase.

The following concentrations of S-205 were selected:
Induction
- Intradermal injection: 5% w/w in Alembicol D.
- Topical application: 60% w/w in Alembicol D.
Challenge
- 15% and 10% w/w in Alembicol D.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 1
- Site: a 4 x 6 cm area of dorsal skin on the scapular region of the guinea-pig.
- Applications to test groups:
Intradermal injections: three pairs of intradermal injections were made simultaneously into the site
1. 0.1mL Freund's complete adjuvant 50:50 with water for injection.
2. 0.1mL S-205, 5% w/w in Alembicol D.
3. 0.1mL S-205, 5% w/w in a 50 : 50 mixture of Freund's complete adjuvant and Alembicol D.
Topical application: one week after the injection, a 2 x 4 cm patch of Whatman No. 3 paper saturated with S-205, 60% w/w in Alembicol D was placed on the same area of the skin and covered by an occlussive dressing
- Control group: treated similarly to the test animals with the exception that S-205 was omitted from the intradermal injections and topical application.
- Exposure period: 48 h

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 1
- Exposure period: 24 h
- Test and control groups: 2 x 2 cm Whatman No.3 papers saturated with approximately 0.2 ml of S-205, 15% w/w in Alembicol D or
S-205, 10% w/w in Alembicol D were applied to an anterior or a posterior sites, respectively, on left flank of the guinea pigs and covered by an
occlussive dressing
- Evaluation (hr after challenge): 24, 48 and 72 h after removal of the patches

SCORING SYSTEM:
Erythema and eschar formation:
0 No erythema
1 Very slight erythema (barely perceptible)
2 Well-defined erythema
3 Moderate to severe erytryema
4 Severe erythema (beet redness) to slight eschar formation (injuries in depth)

Oedema formation:
0 No oedema
1 Very slight oedema (barely perceptible) 1
2 Slight oedema (edges of area well-defined by definite raising)
3 Moderate oedema (raised approximately 1 mm)
4 Severe oedema (raised more than 1 millimeter and extending beyond the area of exposure)

Positive control substance(s):

not specified

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

15% w/w in Alembicol D

No. with + reactions:

3

Total no. in group:

10

Clinical observations:

Erythemas scoring 2 (localised for 2/3 guinea pigs)

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 15% w/w in Alembicol D. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: Erythemas scoring 2 (localised for 2/3 guinea pigs).

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

10% w/w in Alembicol D

No. with + reactions:

2

Total no. in group:

10

Clinical observations:

Erythemas scoring 1 and 2 (localised reaction)

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10% w/w in Alembicol D. No with. + reactions: 2.0. Total no. in groups: 10.0. Clinical observations: Erythemas scoring 1 and 2 (localised reaction).

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

15% w/w in Alembicol D

No. with + reactions:

3

Total no. in group:

10

Clinical observations:

Erythemas scoring 1 and 2 (localised for 2/3 guinea pigs). Dryness and sloughing of the epidermis observed in animal 285 and 288

Remarks on result:

other: see Remark

Remarks:

Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 15% w/w in Alembicol D. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: Erythemas scoring 1 and 2 (localised for 2/3 guinea pigs). Dryness and sloughing of the epidermis observed in animal 285 and 288.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10% w/w in Alembicol D

No. with + reactions:

1

Total no. in group:

10

Clinical observations:

Localised erythema scoring 1

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10% w/w in Alembicol D. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: Localised erythema scoring 1.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

15% w/w in Alembicol D

No. with + reactions:

3

Total no. in group:

10

Clinical observations:

Localised erythemas scoring 1 with some dryness and sloughing of the epidermis in animal 288

Remarks on result:

other: see Remark

Remarks:

Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 15% w/w in Alembicol D. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: Localised erythemas scoring 1 with some dryness and sloughing of the epidermis in animal 288.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

10% w/w in Alembicol D

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 10% w/w in Alembicol D. No with. + reactions: 0.0. Total no. in groups: 10.0.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

In this screening test, performed in ten albino guinea-pigs, S-205 produced an inconclusive result in one guinea-pig.
No evidence of delayed contact hypersensitivity was seen in the remaining nine animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Justification for selection of skin sensitisation endpoint:
No other study available.

Justification for classification or non-classification

S-205 was shown to not be a sensitiser and will therefore not be classified as such.