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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
12. October to 27 November 1992
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted to EU test guidance in compliance with GLP.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1993
Report date:
1993

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
84/449/EWG, B.6 (Magnusson-Kligman-Test)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (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

Constituent 1
Chemical structure
Reference substance name:
4,4'-[(6-chloro-1,3,5-triazine-2,4-diyl)diimino]bis[5-hydroxy-6-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonic] acid, sodium salt
EC Number:
303-152-9
EC Name:
4,4'-[(6-chloro-1,3,5-triazine-2,4-diyl)diimino]bis[5-hydroxy-6-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonic] acid, sodium salt
Cas Number:
94158-79-9
Molecular formula:
C39H32ClN9O26S8.xNa C39H26ClN9Na6O26S8 as hexasodium salt
IUPAC Name:
4,4'-[(6-chloro-1,3,5-triazine-2,4-diyl)diimino]bis[5-hydroxy-6-[[4-[[2-(sulphooxy)ethyl]sulphonyl]phenyl]azo]naphthalene-2,7-disulphonic] acid, sodium salt
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
Reaktiv-Rot F-67787

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
- Weight at study initiation: 240 to 288 g; mean: 257 g
- Housing: 5/cage
- Diet: Altromin 3112 guinea pig maintenance diet ad libitum
- Water: tap water ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25
- Humidity (%): 30 to 70
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 12. October to 27 November 1992

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
physiological saline
Concentration / amount:
Concentration of test material and vehicle used at induction:
a) Intradermal: 5 % in 0.9 % NaCl-solution
b) Dermal: 25 % in 0.9 % NaCl-solution
Concentration of test material and vehicle used for each challenge: a) 25 % in 0.9 % NaCl-solution
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
Concentration of test material and vehicle used at induction:
a) Intradermal: 5 % in 0.9 % NaCl-solution
b) Dermal: 25 % in 0.9 % NaCl-solution
Concentration of test material and vehicle used for each challenge: a) 25 % in 0.9 % NaCl-solution
No. of animals per dose:
Determination of primary non-irritating concentration: 6
Determination of the intradermal tolerance: 3
Number of animals in attending group: 5
Number of animals in test group: 10 Number of animals in negative control group: 5
Details on study design:
RANGE FINDING TESTS:
- Determination of primary non-irritating concentration: dermal-occlusive exposure for 24 hours - 3 concentrations (25%, 5%, 1%)
- Determination of the intradermal tolerance: intradermal injection: 2 x 3 concentrations (5 %, 1 %, 0.2 %)

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 8
- Site: shoulder

B. DERMAL INDUCTION EXPOSURE
- No. of exposures: one
- Exposure period: 48 hours
- Test groups: 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: TS + water
- Control group: TS + 0.9% NaCl
- Site: right flank: TS; left flank: 0.9% NaCl
- Concentrations: 25%
- Evaluation (hr after challenge): 24 and 48 hours
Challenge controls:
In addition to the control group, 5 further guinea pigs (attending group) were used to confirm that challenge exposure with 25% TS would not lead to dermal irritation in animals pre-treated with 50 % FCA.
Positive control substance(s):
yes
Remarks:
bi-annual validation of assay

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
skin slightly reddish stained by TS
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: 10.0. Clinical observations: skin slightly reddish stained by TS.
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
skin slightly reddish stained by TS
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: 10.0. Clinical observations: skin slightly reddish stained by TS.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
skin slightly reddish stained by TS
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: 5.0. Clinical observations: skin slightly reddish stained by TS.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
skin slightly reddish stained by TS
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: 5.0. Clinical observations: skin slightly reddish stained by TS.

Any other information on results incl. tables

Body weight gains and clinical signs

The treated animals showed no clinical signs of intoxication throughout the study. The intradermal injections with Freund's Adjuvant (with and without test substance) caused moderate oedema. Well defined up to moderate erythema was observed at the application sites without test substance, whereas evaluation of erythema formation was not possible at the sites treated with the test compound

in Freund's Adjuvant due to red discolouration of the skin. Additionally, the application sites treated with the test substance in Freund's Adjuvant were indurated and encrusted. The application sites treated with the test substance in the vehicle exhibited very slight oedema as well as encrusted skin. Intradermal applications of the vehicle caused no signs of irritation. The application sites treated with the test substance were discoloured red.

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 oedema, indurated and encrusted skin as well as necrosis were observed at the sites previously treated with Freund's Adjuvant. The injection sites treated with the test substance in the vehicle and with the vehicle alone showed no signs of irritation. Additionally, red discoloured skin was noted in the animals of the treatment group.

The body weight gains of the treated animals were not impaired.

Challenge treatment

No signs of irritation were observed in the control and treatment group 24 and 48 hours after removal of the occlusive bandage. The treated area was discoloured slightly red.

Assessment

Under the conditions of the present study, none of ten animals of the treatment group showed a positive skin response after the challenge procedure.

Based on the results of this study Reaktiv-Rot F-67 787 FW showed no evidence for sensitizing properties

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information not classified Criteria used for interpretation of results: EU
Conclusions:
There was no evidence of a positive reaction in animals after challenge treatment with Reaktiv-Rot F-67787 FW in the present study.
According to the classification criteria of Directive 83/467/EEC, Reaktiv-Rot F-67787 FW is not sensitizing in the guinea pig maximization test and therefore not subject to labelling requirements.
Executive summary:

The skin sensitizing potential of Reaktiv-Rot F-67787 FW was examined in female guinea pigs with the maximization test.

Intradermal induction was performed using 5% Reaktiv-Rot F-67787 FW in 0.9 % NaCl.solution.

Dermal induction and challenge treatment were carried out with 25 % Reaktiv-Rot F-67787 FW in 0.9 % NaCl.solution.

Based on the results of the present study, Reaktiv-Rot F-67787 FW showed no evidence for sensitizing properties and is not subject to labelling requirement.