tetrasodium 4-{4-[(9,10-dioxo-4-{[4-(2-sulfophenoxy)phenyl]amino}-9,10-dihydroanthracen-1-yl)amino]phenoxy}benzene-1-sulfonate 4-{4-[(9,10-dioxo-4-{[4-(4-sulfophenoxy)phenyl]amino}-9,10-dihydroanthracen-1-yl)amino]phenoxy}benzene-1-sulfonate

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

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)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

GLP compliance:

yes (incl. QA statement)

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:

Reaction mass of Disodium 2-{4-[(9,10-dioxo-4-{[4-(4-sulfonatophenoxy)phenyl]amino}-9,10-dihydroanthracen-1-yl)amino]phenoxy}benzenesulfonate and Disodium 4,4'-[(9,10-dioxo-9,10-dihydroanthracene-1,4-diyl)bis(iminobenzene-4,1-diyloxy)]dibenzenesulfonate

EC Number:

941-784-2

Molecular formula:

C38H26N2O10S2.2Na

IUPAC Name:

Reaction mass of Disodium 2-{4-[(9,10-dioxo-4-{[4-(4-sulfonatophenoxy)phenyl]amino}-9,10-dihydroanthracen-1-yl)amino]phenoxy}benzenesulfonate and Disodium 4,4'-[(9,10-dioxo-9,10-dihydroanthracene-1,4-diyl)bis(iminobenzene-4,1-diyloxy)]dibenzenesulfonate

Test material form:

not specified

Details on test material:

- Test article: FAT 21030/D
- Batch N°: 16.36
- Additional specification: Irganol Gruen BLS roh Trocken
- Purity: 67%
- Physical properties: solid
- Storage conditions: Room temperature
- Validity: December, 1998
- Test article received: October 21, 1993

Specific details on test material used for the study:

Test article : FAT 21030/D
Batch No : 16.36
Additional specification: Irganol Gruen BLS roh trocken
Contents/Purity: 67%
Physical properties: solid
Storage conditions: room temperature
Validity: December, 1998
Test article received: October 21, 1993

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

male/female

Details on test animals and environmental conditions:

TEST ANIMALS
- Animal strain: Pirbright White Strain (Tif: DHP)
- Breeder: CIBA-GEIGY Limited, Animal Production, 4332 Stein / Switzerland
- Weight at study initiation: Between 326 to 400 g
- Housing: 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 by individual ear tags.
- Diet: standard guinea pig pellets - NAFAG No. 845, Gossau SG, Ad-libitum
- Water: Ad libitum
- Acclimation period: 23 February, 1994 (5 days before the starting date of the experiment)

ENVIRONMENTAL CONDITIONS
- Temperature: 22±3 °C
- Humidity: 30 to 70 %
- Photoperiod: 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:

physiological saline

Concentration / amount:

50 %

Day(s)/duration:

8

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:

10 %

Day(s)/duration:

21

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

According to the test guidelines cited under "Introduction" the test was started with the minimum number of animals (5 per sex for the test group and 5 of one sex for controls). After the challenge procedure, it was not possible to conclude if the test substance is a sensitiser or not and therefore testing in additional animals was performed to give a total of 20 test and 10 control animals.

Details on study design:

Test procedure and concentrations used:
*Pretests (Intradermal Induction)
The concentration for the intradermal injections was selected on account of the solubility of the test article in standard vehicles and its local and systemic tolerability in a pretest. The following concentration of test article has been used for intradermal injection:
5 % in physiological saline (w/v).
Since 5 % FAT 21030/D in physiological saline could be injected and was well tolerated, this concentration was used for the intradermal induction.

*Pretest (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 21030/D have been examined on separate animals for the determination of the maximum subirritant concentration:
- 10, 20, 30, and 50 % in physiological saline.
- 50 % was the highest possible concentration of the test article in physiological saline.
Reactions were observed with 20, 30, and 50 % FAT 21030/D in physiological saline.

*Test procedure (Induction, intraderma injection):
- DAY 0: 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 21030/D in physiological saline (w/v)
- 5% FAT 21030/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
In the test group FAT 21030/D was incorporated in physiological saline and applied on a filterpaper patch to the neck of the animals; (occluded
administration for 48 hours). The control group was treated with the vehicle only.
Test group:
- 50% FAT 21030/D in physiological saline
Control group:
- physiological saline only

DAY 21: Challenge
The test and control group animals were tested on one flank with FAT 21030/D in physiological saline and on the other flank with the vehicle alone (occluded administration for 24 hours).

Test and control group:
- 10% FAT 21030/D in physiological saline
- physiological saline only

Adjuvant: Bacto adjuvant, Complete, Freund (Difco Lab. Detroit, Michigan USA)
Physiological saline (0.9%), sterile solution (Hausmann, St Gallen, Switzerland)

Positive control substance(s):

yes

Remarks:

2-Mercaptobenzothiazole puriss. (Intradermal induction: 5 %, epidermal induction: 50 % and Epidermal challenge: 30 %)

Results and discussion

Positive control results:

100 % animals showed positive reaction.

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No signs of toxicity

Remarks on result:

no indication of skin sensitisation

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

No signs of toxicity

Remarks on result:

no indication of skin sensitisation

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

10 % Challenge (Intradermal - 5 %, Epidermal - 50 %)

No. with + reactions:

1

Total no. in group:

20

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

10 % Challenge (Intradermal - 5 %, Epidermal - 50 %)

No. with + reactions:

1

Total no. in group:

20

Results 5

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

30 % Challenge. (Induction - 5 % and Epidermal - 50 %)

No. with + reactions:

20

Total no. in group:

20

Remarks on result:

positive indication of skin sensitisation

Results 6

Reading:

2nd reading

Hours after challenge:

48

Group:

positive control

Dose level:

30 % Challenge. (Induction - 5 % and Epidermal - 50 %)

No. with + reactions:

20

Total no. in group:

20

Remarks on result:

positive indication of skin sensitisation

Any other information on results incl. tables

Observations:

Body weight were not affected by treatment.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

FAT 21030/D is not a skin sensitiser.

Executive summary:

FAT 21030/D was tested for skin sensitisation potential in albino guinea pigs using the maximisation test. This test was performed according to the OECD Guideline No 406 (GLP). Based on the findings of the pre-study, the 5, 50 and 10 % test concentrations were used for intradermal induction (day 0), epidermal induction (day 8) and epidermal challenge (day 21), respectively. 1/20 animals showed a positive reaction in the treated group, while no positive reactions were found in the control group. Hence, under experimental conditions employed, 5 % of the animals of the test group showed skin reactions 24 and 48 hours after removing the dressing. Hence, FAT 21030/D can be considered to be not a skin sensitiser.