[No public or meaningful name is available]

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1994

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

None

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

adopted in May 12, 1981

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

92/69/EEC,

Deviations:

no

Principles of method if other than guideline:

None

GLP compliance:

yes

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:

[4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-3-hydroxynaphthalene-1-sulphonato(3-)]chromium

EC Number:

271-352-2

EC Name:

[4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-3-hydroxynaphthalene-1-sulphonato(3-)]chromium

Cas Number:

68541-72-0

Molecular formula:

C20H13CrN4O5S

IUPAC Name:

[4-[(4,5-dihydro-3-methyl-5-oxo-1-phenyl-1H-pyrazol-4-yl)azo]-3-hydroxynaphthalene-1-sulphonato(3-)]chromium

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

- Test article: FAT 20042/D
- Additional specification: Neolan Rosa BE ZP feucht
- Batch No.: 276
- Purity/Contents: ca. 50 %
- Physical properties: solid
- Storage conditions: room temperature
- Validity: October, 1998
- Test material received: November 19, 1993

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: Pirbright White Strain (Tif: DHP)

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: CIBA-GEIGY Limited, Animal Production, 4332 Stein / Switzerland
- Weight at study initiation: 324 to 417 g
- Housing: housed individually in Macrolon cages (Type 3)
- Diet: Standard guinea pig pellets- NAFAG No. 845, Gossau SG, ad libitum
- Water: Fresh water, ad libitum
- Acclimation period: 1 week


SENSITIVITY OF STRAIN
- The sensitivity of the strain is checked once or twice a year with a known mild to moderate sensitiser , such as mercaptobenzothiazole, hexyl cinnamic aldehyde or potassium dichromate

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3 °C
- Humidity: 30-70 %
- Photoperiod: 12 h light/12 h dark

IN-LIFE DATES: From 14 February, 1994 to April 28, 1994

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

physiological saline

Concentration / amount:

Test group:
- adjuvant/saline mixture 1:1 (v/v)
- 5 % FAT 20042/D in physiological saline (w/v)
- 5 % FAT 20042/D in the adjuvant/saline mixture (w/v)

Day(s)/duration:

Test Day 0

Adequacy of induction:

highest technically applicable concentration used

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

Test group:
- 50 % FAT 20042/D in physiological saline

Day(s)/duration:

Test Day 08

Adequacy of induction:

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

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

Test and control group:
- 30 % FAT 20042/D in physiological saline
- physiological saline only

Day(s)/duration:

Test day 21

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Main test: 10 per sex for the test group and 5 of one sex for controls

Details on study design:

RANGE FINDING TEST:
Intradermal Induction
Since 5 % FAT 20042/D in physiological saline could be injected and was well tolerated, this concentration was used for the intradermal induction.

Epidermal Applications (induction and challenge)
The concentrations for the epidermal applications were selected on account of the primary irritation potential of the test article. The following concentrations of FAT 20042/D have been examined on separate animals for the determination of the maximum subirritant concentration.

- 30 and 50 % in physiological saline.

50 % was the highest possible concentration of the test article in physiological saline. Reactions were observed with 50% FAT 20042/D in physiological saline.

MAIN STUDY
DAY 0: INDUCTION, intradermal injections
Three pairs of intradermal injections (0.1 ml per injection) were made simultaneously into the left and right side of the shaved neck of the test and control group animals.
Test group:
- adjuvant/saline mixture 1:1 (v/v)
- 5 % FAT 20042/D in physiological saline (w/v)
- 5 % FAT 20042/D in the adjuvant/saline mixture (w/v)
Control group:
- adjuvant/saline mixture 1:1 (v/v)
- adjuvant/saline mixture 1:1 (v/v)
- physiological saline

DAY 8: INDUCTION, epidermal application
The application site of all animals was pretreated with 10 % sodium-laurylsulfate (open application) 24 hours prior to the epidermal induction application. In the test group FAT 20042/C was incorporated in physiological saline and applied on a filterpaper patch to the neck of the animals (patch 2x4 cm; approx. 0.4 g per patch; occluded administration for 48 hours). The control group was treated with the vehicle only.
Test group:
- 50 % FAT 20042/C in physiological saline
Control group:
- physiological saline only

DAY 21: Challenge
The test and control group animals were tested on one flank with FAT 20042/D in physiological saline and on the other flank with the vehicle alone (patch 2x2 cm; approx. 0.2 g per patch; occluded administration for 24 hours).

Test and control group:
- 30 % FAT 20042/D in physiological saline
- physiological saline only

Observations and records
Induction reactions
After the intradermal and the epidermal induction application skin irritation is normally induced by the adjuvant, the high test article concentration, or the sodium lauryl sulfate pretreatment. Because most of the reactions are treatment related and not test article related, the reactions were generally not described in the results section.

Challenge reactions
Twenty four and forty eight hours after removing the dressings, the challenge reactions were graded according to the Draize scoring scale.

General
The body weight was recorded at start and end of the test.

Interpretation of results
The sensitising potential of FAT 20042/D was classified according to the grading of Magnusson and Kligman. According to the guide to the labelling of dangerous substances and the criteria for the choice of sentences indicating particular hazards (R sentences) attributed to dangerous substances (Commission Directive 93/21/EEC, April 27, 1993), a test article was classified as a sensitiser in the case where a positive response was noted in at least 30 % of the animals.

Challenge controls:

Not available

Positive control substance(s):

yes

Remarks:

2-Mercaptobenzothiazole puriss

Results and discussion

Positive control results:

Twenty out of 20 test animals, were found to have positive reactions after challenge when tested with positive control 2-Mercaptobenzothiazole puriss.

In vivo (non-LLNA)

Results

Results 1

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

Intradermal Induction: 5 %, epidermal induction: 50 %, challenge (epidermal): 30 %

No. with + reactions:

20

Total no. in group:

20

Remarks on result:

other: Reference values 03.01.1994

Results 2

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

Intradermal Induction: 5 %, epidermal induction: 50 %, challenge (epidermal): 30 %

No. with + reactions:

20

Total no. in group:

20

Remarks on result:

other: Reference values 03.01.1994

Results 3

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

Intradermal Induction: 5 %, epidermal induction: 50 %, challenge (epidermal): 30 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Results 4

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

Intradermal Induction: 5 %, epidermal induction: 50 %, challenge (epidermal): 30 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Results 5

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Intradermal Induction: 5 %, epidermal induction: 50 %, challenge (epidermal): 30 %

No. with + reactions:

3

Total no. in group:

20

Results 6

Key result

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Intradermal Induction: 5 %, epidermal induction: 50 %, challenge (epidermal): 30 %

No. with + reactions:

5

Total no. in group:

20

Any other information on results incl. tables

Body weight and body weight gains of the test as well as control animals were found to have not been affected.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 20042/D is a non-sensitiser.

Executive summary:

A guinea pig maximization study was conducted to evaluate the skin sensitisation potential of FAT 20042/D (at ca. 50 % purity) according to OECD Guideline 406 and EU Method B.6. Since 5% FAT 20042/D in physiological saline could be injected and was well tolerated, this concentration was used for the intradermal induction. Based on the results of a preliminary study, 50 % concentration was selected for epidermal induction, while 30 % concentration was used for epidermal challenge. The main test consisted of 3 phases i.e. intradermal induction on day 0 at 5 % conc., epidermal induction on Day 8 at 50 % conc. and epidermal challenge on Day 21 at 30 % conc. Skin reactions were graded using Draize scoring at 24 and 48 h after removal of dressing at each phase. Under the experimental conditions employed, 15 and 25 % of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressings when challenged. FAT 20042/D is , therefore, classified as a mild sensitiser in albino guinea pigs according to the grading of Magnusson and Kligman. According to the EEC classification criteria (Commission Directive 93/21/EEC , April 27, 1993) FAT 20042/D did not show a skin sensitising (contact allergenic) potential in albino guinea pigs.