Cuprate(4-), [2-[[[[2-hydroxy-3-sulfo-5-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]phenylmethyl]azo]-4-sulfobenzoato(6-)]-, sodium

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

since October 12,1992 to November 13,1992

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP compliance with international guideline

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

Test guideline 2

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:

Test was not valid at time of test conduct

Test material

Test material information

Constituent 1

Reference substance name:

Cuprate(4-), [2-[[[[2-hydroxy-3-sulfo-5-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]phenylmethyl]azo]-4-sulfobenzoato(6-)]-, sodium

EC Number:

291-103-1

EC Name:

Cuprate(4-), [2-[[[[2-hydroxy-3-sulfo-5-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]phenylmethyl]azo]-4-sulfobenzoato(6-)]-, sodium

Cas Number:

90341-71-2

Molecular formula:

C22H14CuN4Na4O15S4

IUPAC Name:

Cuprate(4-), [2-[[[[2-hydroxy-3-sulfo-5-[[2-(sulfooxy)ethyl]sulfonyl]phenyl]azo]phenylmethyl]azo]-4-sulfobenzoato(6-)]-, sodium

Details on test material:

- Name of test material:Remazol Brillantblau BB neu

In vivo test system

Test animals

Species:

guinea pig

Strain:

Pirbright-Hartley

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: HOECHST AG, Kastengrund, SPF breeding colony
- Weight at study initiation: 318 ± 30 g
- Housing: in fully air-conditioned rooms in Makrolon cages (Type 4) on soft wood granulate, in groups of 5 animals
- Diet : Altromin 3112 for guinea pigs and rabbits, ad libitum
- Water : tap water in plastic bottles, ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 20 %
- Photoperiod: 12 hours daily

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

other: isotonic saline solution and Freud's Complete adjuvant

Induction 2

Concentration / amount:

5% of test substance (Intradermal induction treatment)
25% of the test substance (Dermal induction treatment)
25% of the test substance (Dermal challenge treatment)

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

other: isotonic saline solution and Freud's Complete adjuvant

Challenge 2

Concentration / amount:

5% of test substance (Intradermal induction treatment)
25% of the test substance (Dermal induction treatment)
25% of the test substance (Dermal challenge treatment)

No. of animals per dose:

Determination of primary not irritating concentration: 6
Determination of intradermal tolerability: 3
Sentinel group: 5
Control group: 5
Treatment group: 10

Details on study design:

RANGE FINDING TESTS:
Determination of the primary non-irritant concentration:
In a dermal-occlusive test for primary skin irritation, each of the following test concentrations was applied to the left flank of two guinea pigs:
25.0 % TS in isotonic saline
5.0 % TS in isotonic saline
1.0 % TS in isotonic saline
The hair on the left flanks of the animals was removed mechanically. 0.5 mL of the test substance preparation was applied to a 2 x 2 cm cellulose patch, which was then fixed to the left flank and covered occlusively for 24 hours with a bandage and film. 24 hours after removal of the patches, the treated skin areas were examined for erythema and oedema

Determination of the tolerance of intradermal injections:
To determine the tolerance of intradermal injections, each of the following preparations (5.0%, 1.0%, 0.2% in isotonic saline) was administered twice by intradermal injection to 3 guinea pigs. The injection sites (sites 1, 2 and 3) were all within a dorsal area measuring 2 x 4 cm in the vicinity of the shoulder.


MAIN STUDY
A. INTRADERMAL INDUCTION
- No of Injections: 2 x 3 preparations: 50% FCA, 5% TS in 0.9% NaCl, 5% TS in 50% FCA - treatment group
50% FCA, 0.9% NaCl, 50% FCA - control and attending group
- Exposure period: Injection on Day 1, observation Day 1 to Day 7
- Site: shoulder

B. DERMAL INDUCTION EXPOSURE
- No. of exposures: one
- Exposure period: 48 hours
- Test groups: 25% TS in 0.9% NaCl
- Control group: 0.9% NaCl
- Site: shoulder
- Frequency of applications: single
- Duration: Day 8 to Day 22
- Concentrations: 25%

C. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22 (15 for attending group)
- Exposure period: 24 hours
- Test groups: 25% TS + 0.9% NaCl
- Control group: 25% TS + 0.9% NaCl
- Site: right flank: TS; left flank: 0.9% NaCl
- Concentrations: 25%
- Evaluation (hr after challenge): 24 and 48 hours

Positive control substance(s):

no

Study design: in vivo (LLNA)

Positive control substance(s):

not specified

Results and discussion

In vivo (non-LLNA)

Results

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

25%

No. with + reactions:

6

Total no. in group:

10

Clinical observations:

dry, rough and encrusted skin

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 25%. No with. + reactions: 6.0. Total no. in groups: 10.0. Clinical observations: dry, rough and encrusted skin.

Any other information on results incl. tables

The substance is considered to be sensitising if 30 % of the animals in the treated group definitely show a positive reaction.

Challenge treatment:

Very slight to moderate erythema and very slight edema as well as dry, rough and encrusted skin were observed in the treatment group 24 and 48 hours after removal of the occlusive bandage. No signs of irritation occurred in the control group. The skin surface of all animals was discolored light blue.

Clinical signs:

The intradermal injections with Freund's Adjuvant caused severe erythema and edema, indurations and encrustations (evaluation of erythema formation was not possible at the sites treated with the test substance due to intensive blue discolourations). The application sites treated with the test substance showed edema, indurations and encrustations. Evaluation of erythema formation was not possible. Injections of the vehicle alone did not cause any sign of irritation. Due to these strong irritation reactions of the skin, 10% sodium dodecylsulfate was not applied at day 7.

After the removal of the patch at day 10, erythema and edema, scabbed and encrusted skin as well as necrosis and open wounds were observed at the application sites. Additionally the application sites of the treatment group were discoloured light blue.

Assessment

Under the conditions of the present study, six of ten animals of the treatment group showed a positive skin response after the challenge procedure.

Based on the results of this study Remazol-Brillantblau BB neu may cause sensitisation by skin contact.

Applicant's summary and conclusion

Interpretation of results:

sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under the conditions of the present study, six of ten animals of the treatment group showed a positive skin response after the challenge procedure.
Based on the results of this study Remazol-Brillantblau BB neu may cause sensitisation by skin contact.

Executive summary:

Testing for sensitising properties of Remazol-Brillantblau BB neu was performed in female Guinea pigs according to the method of MAGNUSSON & KLIGMAN. Intradermal induction was performed using 5 % Remazol-Brillantblau BB neu in isotonic saline. Dermal induction and challenge treatment were carried out with 25 % Remazol-Brillantblau BB neu in isotonic saline.

Very slight to moderate erythema and very slight edema as well as dry, rough and encrusted skin were observed in the treatment group 24 and 48 hours after removal of the occlusive bandage. No signs of irritation occurred in the control group. The skin surface of all animals was discolored light blue.

Under the conditions of the present study, six of ten animals of the treatment group showed a positive skin response after the challenge procedure.

Based on the results of this study Remazol-Brillantblau BB neu may cause sensitisation by skin contact.