Tetrasodium 4-amino-6-[[2,5-dimethoxy-4-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]-5-hydroxy-3-[[4-[[2-(sulphonatooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonate

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15 April to 22 May 1998

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1998

Report date:

1993

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

Buehler test

Test material

Test material information

Constituent 1

Reference substance name:

Reactive Blue 203

IUPAC Name:

Reactive Blue 203

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Test item: Reactive Blue 203

In vivo test system

Test animals

Species:

guinea pig

Strain:

other: pirbright-white

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: HOECHST AG, Kastengrund, SPF breeding colony
- Age at study initiation: approx. 10 weeks
- Weight at study initiation: 283 g to 397 g
- Housing: 5 animals/group
- Diet (ad libitum): Altromin 3112 for guinea pigs and rabbits
- Water (ad libitum): tap water
- Acclimation period: at least 5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 55 ± 20 %
- Air changes (per hr): -
- Photoperiod (hrs dark / hrs light): 12/12


IN-LIFE DATES: From: 26 of July To: 01 of September 1993

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

epicutaneous, occlusive

Vehicle:

water

Induction 2

Concentration / amount:

25%

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

water

Challenge 2

Concentration / amount:

25%

No. of animals per dose:

Determination of primary non-irritant concentration: 6
Control group: 10
Treatment group 20

Details on study design:

RANGE FINDING TESTS:
25 %, 2.5 %, 0.25 %

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3
- Exposure period: 6 hours occlusive
- Test groups: 25% in deionized water
- Control group: 0.5 mL deionized water
- Frequency of applications: once/week
- Duration: Day 1 to 15
- Site: left flank
- Concentrations: 25%


B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: Day 29
- Exposure period: 6 hours
- Test groups: 25% in deionized water
- Control group: 25% in deionized water
- Site: right flank
- Concentrations: 25 %
- Evaluation (hr after challenge): 24 and 48

Challenge controls:

NA

Positive control substance(s):

yes

Remarks:

alpha-hexyl cinnamic aldehyde in PEG 400

Results and discussion

Positive control results:

30% positive = valid

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

25%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

skin discoloured blue

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: skin discoloured blue.

Results 2

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

25%

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

skin discoloured blue

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: skin discoloured blue.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

25%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

skin discoloured blue

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: skin discoloured blue.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

25%

No. with + reactions:

0

Total no. in group:

20

Clinical observations:

skin discoloured blue

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: skin discoloured blue.

Any other information on results incl. tables

Determination of the primary non-irritant concentration:

No signs of irritation occurred after application of the different test concentrations.

Based on these results, 25 %Remazol-Marineblau GG was selected for the sensitization treatments.

Body weight gains and clinical signs:

There were no recognizable differences in body weight gains between the control and treated animals.

No clinical signs of intoxication were observed at any time during the study.

No signs of irritation occurred on the treated skin areas during the sensitization phase from days 1 to 19. The skin on the treated areas was slightly discoloured blue from the test substance. Despite this blue discolouration, it would have been possible to recognize any signs of skin irritation. The animals of the control group showed no signs of irritation.

Challenge treatment

At both of the designated examination times it was found that challenge treatment did not cause any effects on the treated skin in either the control or the treatment group. The skin on the treated areas was slightly discoloured blue from the test substance.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Reactive Blue 203 showed no evidence for sensitizing properties.

Executive summary:

Testing for sensitizing properties of Reactive Blue 203 was performed in female guinea pigs according to the method of BUEHLER. Dermal induction was performed using 25 % Reactive Blue 203 in deionised water. The control group was exposed to deionised water only.

Determination of the primary non-irritant concentration:

No signs of irritation occurred after application of the different test concentrations.

Based on these results, challenge treatment was carried out with 25 % Reactive Blue 203 in deionised water.

Challenge treatment At both of the designated examination times it was found that challenge treatment did not cause any effects on the treated skin in either the control or the treatment group. The skin on the treated areas was slightly discoloured blue from the test substance. Based on the results of this study Reactive Blue 203 showed no evidence for sensitizing properties.

Body weight gains and clinical signs:

There were no recognizable differences in body weight gains between the control and treated animals.

No clinical signs of intoxication were observed at any time during the study.

No signs of irritation occurred on the treated skin areas during the sensitization phase from days 1 to 19. The skin on the treated areas was slightly discoloured blue from the test substance. Despite this blue discolouration, it would have been possible to recognize any signs of skin irritation. The animals of the control group showed no signs of irritation.

Under the conditions of the present study, none of twenty animals of the treatment group showed a positive skin response after the challenge procedure. Based on the results of this study Rective Blue 203 showed no evidence for sensitizing properties.