Reaction mass of 1,5-dihydroxy-4-nitro-8-(phenylamino)anthraquinone and 1,8-dihydroxy-4-nitro-5-(phenylamino)anthraquinone

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1987

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

adopted May 12, 1986

Deviations:

yes

Remarks:

Positive control not reported

GLP compliance:

no

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Data from a reliable in vivo test conducted before the enforcement of Commission Regulation (EU) 640/2012 of 06 July 2012 amending, for the purpose of its adaptation to technical progress, Regulation (EC) No 440/2008 laying down test methods pursuant to Regulation (EC) No 1907/2006 of the European Parliament and of the Council on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) are available.

Test material

Test material information

Constituent 1

Reference substance name:

1,8-dihydroxy-4-nitro-5-(phenylamino)anthraquinone

EC Number:

243-632-4

EC Name:

1,8-dihydroxy-4-nitro-5-(phenylamino)anthraquinone

Cas Number:

20241-76-3

Molecular formula:

C20H12N2O6

IUPAC Name:

1-anilino-4,5-dihydroxy-8-nitro-9,10-anthraquinone

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: EN 37761.62
- Expiration date of the lot/batch: October 1991

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Solubility of the test substance in the solvent/vehicle: 1g/l water

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

Remarks:

Pirbright White Strain (Tif: DHP)

Sex:

male/female

Details on test animals and environmental conditions:

Test System
The albino guinea pig is the recommended species for skin sensitization studies.
- Animal strain: Pirbright White Strain (Tif: DHP)
- Source: CIBA-GEIGY LTD. Tierfarm, 4334 Sisseln, Switzerland
- Date of acclimatization: January 5, 1987

Husbandry
- The test was performed on 10 male and 10 female guinea pigs per group weighing between 334 to 417 g (approx. 10 weeks old).
- The animals were housed individually in Macrolon cages (Type 3), assigned to the different groups by means of random numbers generated by the random number generator, identified with individual ear tags, kept at a constant room temperature of 22±3°C, at a relative humidity of 30 to 70 % and a 12 h light cycle day.
- The animals received ad libitum standard guinea pig pellets - NAFAG No.847, Gossau SG and fresh water.
- All batches of the diet are assayed for nutritive ingredients and contamination level by the manufacturer. Analytical results are available at the animal supply office.
- The drinking water is examined periodically by the IWB (Industrielle Werke Basel), results are available to CIBA-GEIGY Ltd.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

physiological saline

Concentration / amount:

0.1 ml per injection
- adjuvant and saline (1:1)
- 1 % test compound FAT 40278/A in physiological saline
- 1 % test compound FAT 40278/A in the adjuvant saline mixture

Adequacy of induction:

highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

0.4 g paste of 10 % FAT 40278/A in vaseline

Day(s)/duration:

one week after intradermal induction

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: vaseline

Concentration / amount:

approx. 0.2 g paste of 3 % FAT 40278/A in vaseline

Day(s)/duration:

Two weeks after the epidermal induction

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10 males and 10 females per group

Details on study design:

RANGE FINDING TESTS:
The concentration of the test compound for the induction and challenge periods were determined on separate animals.

MAIN STUDY
A. INDUCTION EXPOSURE (Intradermal induction)
- No. of exposures: 1
- Test groups: 1
- Control group: 1
- Site: shaved neck
- Frequency of applications: once
- Concentrations: 1 %

B. INDUCTION EXPOSURE (Epidermal induction)
- No. of exposures: 1
- Test groups: 1
- Control group: 1
- Site: neck of the animals
- Frequency of applications: once
- Duration of exposure: 48 h
- Concentrations: 10 % (with prior application of SDS 10%)

C. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: two weeks after epidermal induction
- Exposure period: 24 hours
- Test groups: 1
- Control group: 1
- Site: flank
- Concentrations: 3 %
- Evaluation (hr after challenge): 24 h after removing the dressing

Positive control substance(s):

not specified

Results and discussion

Positive control results:

None

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Intradermal induction: 1 %, Epidermal induction: 10 %, challenge: 3 %

No. with + reactions:

19

Total no. in group:

20

Clinical observations:

clinical observations not specified

Remarks on result:

positive indication of skin sensitisation

Results 2

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Intradermal induction: 1 %, Epidermal induction: 10 %, challenge: 3 %

No. with + reactions:

19

Total no. in group:

20

Clinical observations:

clinical observations not specified

Remarks on result:

positive indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

no indication of skin sensitisation

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

no indication of skin sensitisation

Results 5

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Remarks on result:

not measured/tested

Results 6

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Remarks on result:

not measured/tested

Any other information on results incl. tables

Under the experimental conditions employed, 95 % of the animals of the test group showed skin reactions 24 and 48 h after removing the dressings.

Applicant's summary and conclusion

Interpretation of results:

Category 1A (indication of significant skin sensitising potential) based on GHS criteria

Conclusions:

FAT 40278/A was considered to be a skin sensitiser.

Executive summary:

The skin sensitisation potential of FAT 40278/A (Disperse Blue 077 content: 45 %) has been investigated in a study conducted using Magnusson and Kligman maximation method as described in OECD Guideline 406. In this study, a group of guinea pigs containing 10 males and 10 females, received FAT 40278/A at 1 % concentration in the intradermal induction phase and at 10 % concentration during the epidermal induction phase. The day before epidermal induction, the application sites were pretreated with 10 % sodium lauryl sulfate as open application. Two weeks after the epidermal induction application the animals were challenged with the occluded epidermal application for 24 h on the flank with FAT 40278/A (concentration 3%) in vaseline and the vehicle alone. 95 % of the animals of the test group showed skin reactions 24 and 48 h after removing the dressings. Hence, FAT 40278/A was considered to be a skin sensitiser.