Trisodium 1-amino-4-[[3-[[4-chloro-6-[(sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-2,4,6-trimethyl-5-sulphonatophenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

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:

Trisodium 1-amino-4-[[3-[[4-chloro-6-[(sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-2,4,6-trimethyl-5-sulphonatophenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

EC Number:

276-481-8

EC Name:

Trisodium 1-amino-4-[[3-[[4-chloro-6-[(sulphonatophenyl)amino]-1,3,5-triazin-2-yl]amino]-2,4,6-trimethyl-5-sulphonatophenyl]amino]-9,10-dihydro-9,10-dioxoanthracene-2-sulphonate

Cas Number:

72214-18-7

Molecular formula:

C32H26ClN7O11S3.3Na

IUPAC Name:

hexasodium 1-amino-4-{[3-({4-chloro-6-[(3-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)-2,4,6-trimethyl-5-sulfonatophenyl]amino}-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate 1-amino-4-{[3-({4-chloro-6-[(4-sulfonatophenyl)amino]-1,3,5-triazin-2-yl}amino)-2,4,6-trimethyl-5-sulfonatophenyl]amino}-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate

Test material form:

other: liquid

Details on test material:

None

Specific details on test material used for the study:

Test article: FAT 41001/F
Batch No.: Op.Nr.2 2
Additional specification: Cibacron Blau P-3R
Contents/Purity: ca. 80 %
Physical properties: liquid
Storage conditions: room temperature
Validity: June, 1997
Test article received: June 25, 1992

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

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: between 312 to 412 g
- Housing: The animal s 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 by individual ear tags
- Diet (e.g. ad libitum): ad libitum standard guinea pig pellets - NAFAG No. 845, Gossau SG
- Water (e.g. ad libitum): ad libitum, fresh water.
- Acclimation period: November 19, 1992.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3.
- Humidity (%): 30 to 70.
- Photoperiod (hrs dark / hrs light): 12 hours light cycle day.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal

Vehicle:

other: physiological saline and adjuvant/saline mixture

Concentration / amount:

5 %

Day(s)/duration:

0

Adequacy of induction:

highest technically applicable concentration used

Induction 2

Route:

epicutaneous, occlusive

Vehicle:

other: Vaseline

Concentration / amount:

50%

Day(s)/duration:

7 days

Adequacy of induction:

other: non-irritant substance, but skin pre-treated with 10% SLS.

Challenge

No.:

#1

Route:

epicutaneous, occlusive

Vehicle:

other: Vaseline

Concentration / amount:

50%

Day(s)/duration:

5 weeks

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

Test groups: 10 male and 10 females
Control groups: 5 males and 5 females

Details on study design:

Test procedure

Induction procedure (weeks 1 and 2):
The induction was a two-stage operation. First, intradermal injections (into the neck region); second, closed patch exposure over the injection sites one week later.

First induction week, intradermal injection
Three pairs of intradermal injections (0.1 ml per injection) were made simultaneously into the shaved neck of the guinea pigs as follows:
- adjuvant/saline mixture 1:1 (v/v)
- test article FAT 41001/F in physiological saline (w/v)
- test article FAT 41001/F in the adjuvant/saline mixture (w/v)

Second induction week, epidermal application
In the second week of induction FAT 41001/F was incorporated in Vaseline (w/w) and applied on a filter paper patch to the neck of the animals (patch 2x4 cm; approx. 0.4 g paste per patch; occluded administration for 48 hours).

Rest period
During weeks 3 and 4 no treatments were performed.

Challenge (week 5)
The animals were teste d on the flank with FAT 41001/F in Vaseline (w/w) and the vehicle alone (patch 2x2 cm; approx. 0.2 g paste per patch; occluded administration for 24 hours).

Control group
A control l group of 10 animals (5 m/5 f) was treated d with adjuvant and the vehicle during the induction period. During the challenge period the group was treated with the vehicle as well as with the test t article e to check the maximum sub irritant concentration of the test article e in adjuvant treated animals.

Challenge controls:

No data

Positive control substance(s):

yes

Remarks:

Potassium dichromate

Results and discussion

Positive control results:

Not available

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Intradermal induction - 5%, Epidermal induction - 50% and Epidermal challenge - 50%

No. with + reactions:

20

Total no. in group:

20

Remarks on result:

positive indication of skin sensitisation

Results 2

Reading:

1st reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Intradermal induction - 5%, Epidermal induction - 50% and Epidermal challenge - 50%

No. with + reactions:

18

Total no. in group:

20

Remarks on result:

positive indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0%

No. with + reactions:

0

Total no. in group:

20

Results 4

Reading:

1st reading

Hours after challenge:

48

Group:

negative control

Dose level:

0%

No. with + reactions:

0

Total no. in group:

20

Results 5

Reading:

1st reading

Group:

positive control

Remarks on result:

not measured/tested

Applicant's summary and conclusion

Interpretation of results:

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

Conclusions:

FAT 41001/F is classified as sensitiser in albino guinea pigs according to the grading of Magnusson and Kligman.

Executive summary:

A sensitization test in albino guinea pigs was performed to determine the contact allergenic potency of FAT 41001/F in albino guinea pigs. This test was based on the OECD Guideline No. 406 and GLP. In a pre-test conducted for assessment of irritation potential, no skin irritation was observed. Therefore, the application site was pretreated with 10% sodium-lauryl sulfate (open application) 24 hours prior to the epidermal induction application. The concentration of 5% was selected for induction (day 0) and 50% was selected for epidermal induction (7 days) and epidermal challenge (5 weeks). The incidence of positive animals per group, the individual challenge reactions and the evaluation of the primary skin irritation potential was carried according to grading of Magnusson and Kligman.

Under the experimental conditions employed, 100 and 90% of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressings, respectively. No abnormality in increase of body weight was observed in test animals. According to the maximization grading FAT 41001/F showed an extreme grade of skin-sensitizing (contact allergenic) potential in albino guinea pigs.