disodium 7-{[4-chloro-6-(dodecylamino)-1,3,5-triazin-2-yl]amino}-4-hydroxy-3-[{4-[(4-sulfonatophenyl)diazen-1-yl]phenyl}diazen-1-yl]naphthalene-2-sulfonate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation, other

Remarks:

in vivo (guinea pig maximisation test)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from August 10 to September 16, 1993

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test conducted according to internationally accepted guidelines.

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:

EU Method B.6 (Skin Sensitisation)

Version / remarks:

(1992)

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Version / remarks:

(1981)

Principles of method if other than guideline:

The study refers also to the publication: 'Allergic contact dermatitis in the guinea pig: identification of contact allergenes'; Magnusson B. and Kligman A.M., 1970

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

Red GS 3848

IUPAC Name:

Red GS 3848

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

In vivo test system

Test animals

Species:

guinea pig

Strain:

Himalayan

Sex:

female

Details on test animals and environmental conditions:

Test animalsSource: BRL Ltd., Basel, SwitzerlandAge at study initiation: approx. 9 weeksWeight at study initiation: 343 - 484 gHousing: 2 animals per cageDiet: ad libitumWater: ad libitumAcclimation period: 5 days before start of the treatmentEnvironmental conditionsTemperature: 21 °CHumidity: 55 %Air changes: 15 per hourPhotoperiod: 12 hours artificial fluorescent light

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: water or physiological saline

Induction 2

Concentration / amount:

Preliminary studyintradermal: 5 % w/w in distilled waterEpidermal: 50 % w/w in distilled waterEpidermal: 5 %, 10 %, 25 % and 50 % w/w in distilled waterMain studyInductionI) intradermal: 1 % w/w in physiological salineII) epidermal: 50 % w/w in distilled waterChallenge and re-challengeEpidermal: 50 %, 25 % and 10 % w/w in distilled water

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: water or physiological saline

Challenge 2

Concentration / amount:

Preliminary studyintradermal: 5 % w/w in distilled waterEpidermal: 50 % w/w in distilled waterEpidermal: 5 %, 10 %, 25 % and 50 % w/w in distilled waterMain studyInductionI) intradermal: 1 % w/w in physiological salineII) epidermal: 50 % w/w in distilled waterChallenge and re-challengeEpidermal: 50 %, 25 % and 10 % w/w in distilled water

No. of animals per dose:

20.

Details on study design:

Preliminary studyIntradermal 5 % w/w in water (1 animal)Epidermal 50 % w/w in water (same animal as above)Epidermal 50 %, 25 %, 10 % and 5 % w/w in water (4 animals)Main studyA. Induction exposure (intradermal and epidermal)No. of exposures: 2Days of induction: day 1 and day 8Test groups: test substance in physiological saline (intradermal) or water (epidermal)Control group: physiological saline (intradermal) or water (epidermal)Site: clipped scapular region Duration (epidermal): 48 hoursConcentrations: 1 % (intradermal) and 50 % (epidermal)B. Challenge and re-challenge exposure (epidermal)No. of exposures: 2Days of challenge: day 22 and day 28Duration of exposure: 24 hoursTest groups: test substance in water Control group: test substance in waterSite: clipped and shaved flank (challenge); contralateral shaved flank (re-challenge)Concentrations: 50 %, 25 % and 10 %Evaluation: 24 and 48 hours after removal of the dressing

Positive control substance(s):

yes

Remarks:

formaldehyde

Results and discussion

Positive control results:

Skin reaction to the second challenge with formaldehydeconc. of formaldehyde 0.2 %0.1 %0.05 %0%experimental animals with skin reaction121160control animals with skin reaction1210Although skin reactions were observed in experimental and control animals in response to all three concentrations, it was clearly noted that a higher number of experimental animals showed a skin reaction than control animals and an higher degree of skin reaction in a few experimental animals in response to the three concentrations tested. A similar difference was noted on the first challenge skin sites and at histopathology examination. Therefore, it can be concluded that a clear sensitisation effect had occurred.

In vivo (non-LLNA)

Results

Results 1

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

50 %

No. with + reactions:

2

Total no. in group:

20

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 50 %. No with. + reactions: 2.0. Total no. in groups: 20.0.

Results 2

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

25 %

No. with + reactions:

2

Total no. in group:

20

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 25 %. No with. + reactions: 2.0. Total no. in groups: 20.0.

Results 3

Reading:

rechallenge

Hours after challenge:

48

Group:

test chemical

Dose level:

10 %

No. with + reactions:

2

Total no. in group:

20

Remarks on result:

other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 2.0. Total no. in groups: 20.0.

Any other information on results incl. tables

Preliminary study

No signs of systemic toxicity were observed during the preliminary study. However, body-weight loss was noted in one of the five animals.

Test substance concentrations used in the main study were based on the findings in the preliminary study and in accordance with Magnusson and Kligman (1969).

Main Study - Induction

The experimental animals showed no marked erythema or oedema after the 48 hours occluded epidermal induction exposure. The treated area was discoloured red/pink by test substance, which made skin reading difficult.

Main study - Challenges - Control group

In the first challenge, five, five and three animals showed redness in response to 50 %, 25 % and 10 % test substance concentrations, respectively. One control animal also showed swelling in response to the 25 % and 10 % concentrations. Test substance treated skin sites showed a red/pink discolouration, which made skin reading difficult.

In the second challenge, only one animal showed red spots in response to the 50 %, 25 % and 10 % concentrations. Majority of test substance treated skin sites showed a red/pink discolouration, which made skin reading difficult.

Other observations

One experimental animal was found dead on day 14 without previously showing adverse clinical signs. At necropsy, the animal was found with clotted blood in the abdominal cavity. No further mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study during the study period.

The average body weight gain over the study period of experimental and control animals was similar.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information according to the CLP Regulation (EC 1272/2008) Criteria used for interpretation of results: EU

Conclusions:

Taking into account the intensity of macroscopic responses and comparing these with skin reactions seen in control animals, none of experimental animals showed a positive reaction in response to any of test substance concentrations tested.Since no difference was noted microscopically in the type and incidence of the skin reactions between the experimental and control animals, it was considered that a 50 % concentration of substance induced no sensitisation reaction. Therefore, the substance is considered as non-sensitising.

Executive summary:

Method

Guinea pig maximisation test. Preliminary study (5 animals) and main study (20 animals in the experimental group + 10 animals in the control group), female guinea pigs. In the main study, first induction(day 1) via intradermal exposure at 1 % w/w; second induction (day 8) at 50 % w/w, challenge (day 22) and rechallenge (day 28) at 50 %, 25 % and 10 % via epidermal exposure.

Result

No signs of systemic toxicity in the preliminary study.

In most of the cases, the treated area was discoloured red/pink by test substance. No marked erythema or oedema after epidermal induction exposure was seen. One experimental animal was found dead on day 14 without previously showing adverse clinical signs. At necropsy, this animal

was found with clotted blood in the abdominal cavity. No further mortality occurred and no signs of systemic toxicity were observed in the study period.

Taking into account the intensity of the macroscopic responses and comparing these with the skin reactions seen in the control animals, none of the

experimental animals showed a positive reaction in response to any of test substance concentrations tested.

Moreover, no difference was noted microscopically in the type and incidence of the skin reactions between the experimental and control animals with a 50 % test substance concentration.