Registration Dossier

Administrative data

Description of key information

Reaktivblau 1483 does not have a sensitizing effect on the skin of the guinea pig

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1990
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Method was not available
Species:
guinea pig
Strain:
Pirbright-Hartley
Remarks:
Pirbright White, Dunkin Hartley HOE DHPK [SPF-LAC] BÖ
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Lippische Versuchstierzucht, Hagemann GmbH & Co. KG, Extertal 1, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: SPF
- Age at study initiation: young adults
- Weight at study initiation: 273 - 337 g
- Housing: 5/cage
- Diet (e.g. ad libitum): Kliba Labordiät 341 (Kaninchen-Meerschweinchen-Haltungsdiät) ad libitum
- Water (e.g. ad libitum): Water ad libitum (tap water; about 2 g of ascorbic acid per 10 L water was added to the drinking water twice a week)
- Acclimation period: at least 7 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 30-70
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light): 12/12
- In-life dates: From March 27, 1990 To April 27, 1990.
Route:
intradermal
Vehicle:
water
Concentration / amount:
5%: four injections a 0.1 mL
Day(s)/duration:
Day 1
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
50%
Day(s)/duration:
on Day 8 - 48 hours
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
25%
Day(s)/duration:
on Day 15 - for 24 hours
Adequacy of challenge:
highest non-irritant concentration
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
25%
Day(s)/duration:
on Day 22 - for 24 hours
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Main test
Number of animals per control group: 10
Number of animals of the test group: 20
Details on study design:
RANGE FINDING TESTS:
Determination of the primary non-irritant concentration
Determination of the tolerance of the intradermal injections

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 1 intradermal + 1 epicutaneous
- Exposure period: intradermal: 24 hours; epicutaneous: 48 hours
- Test groups: 1
- Control group: 2
- Site: shoulder
- Frequency of applications: Day 1; Day 8
- Duration: 24 and 48 hours
- Concentrations: 5 and 50%

B. CHALLENGE EXPOSURE
- No. of exposures: 2
- Day(s) of challenge: Day 22 and Day 29
- Exposure period: 24 hours
- Test groups: 1
- Control group: 1 + 1
- Site: flank
- Concentrations: 25%
- Evaluation (hr after challenge): 24, 48 and 72 h after the beginning of application

1st challenge:
- treatment of the test group and of control group 1 with the test substance formulation (control group 2 remained untreated)
2nd challenge:
- treatment of the test group and of control groups 1 and 2 with the test substance formulation


OTHER:
Weight check of the individual animals: before intradermal induction and before the end of the study.
Clipping of the test animals: if required, about 3 to 5 hours before each reading and before each test substance application at the appropriate application sites.
Clinical examinations: no detailed examinations; check for sick animals and for those showing a deteriorated general state each workday.
Challenge controls:
yes
Positive control substance(s):
yes
Remarks:
a separate study is performed twice a year
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25%
No. with + reactions:
1
Total no. in group:
18
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
1
Total no. in group:
18
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
19
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
19
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
25%
No. with + reactions:
1
Total no. in group:
18
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
18
Remarks on result:
no indication of skin sensitisation

Body weights: The expected body weight gain has generally been observed in the course of the study.

Mortality: One animal of control group 1 and 2 animals of the test group died 9, 10 or 12 days after the beginning of the study. Macroscopic examination revealed that the animals suffered from pneumonia.

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance does not have a sensitizing effect on the skin of the guinea pig in the Maximization Test under the test conditions chosen
Executive summary:

The substance Reaktivblau 1463 was tested for its sensitizing effect on the skin of the guinea pig in the maximization test based on the method of Magnusson and Kligman.

After intradermal induction distinct erythema and oedema were observed at the injection sites of the control and the test animals at which only Freund's adjuvant/0.9% aqueous NaCl-solution (1 : 1) was applied. Injection of the test substance preparation in 0.9% aqueous NaCl-solution or in Freund's adjuvant/0.9% aqueous NaCl-solution (1 : 1) caused distinct oedema in the test animals. The irritation index for erythema could not be determined because of staining due to the colour of the test substance. The control animals injected with 0.9% aqueous NaCl-solution (vehicle) did not show any skin reactions.

The percutaneous induction was only carried out in the test group, because with aqua dest. a vehicle was used that was not expected to influence the result of the study. Incrustation, partially open (caused by the intra-dermal induction) could be observed in addition to distinct oedema. The index for erythema could not be determined because of staining due to the colour of the test substance.

The number of animals with skin findings after the 1st challenge (20 days after intradermal induction) and after the 2nd challenge (27 days after intradermal induction) is summarized in the following table:

 

 

 

1stchallenge

2ndchallenge

25% in aqua dest.

25% in aqua dest.

Control group

1

0/10

0/10

Control group

2*

no application of test substance

0/9

Test group**

 

1/18

1/18

x/y:  number of positive reactions/number of animals tested; readings 48 h after the beginning of application

*      one animal died 9 days after intradermal induction cause of death: pneumonia

**    two animals died 10 and 12 days after intradermal induction; cause of death: pneumonia

 

After the first challenge with the 25% test substance preparation in aqua dest. one test animal exhibited distinct erythema. Control group animals did not show any skin reactions.

The second challenge with the 25% test substance preparation in aqua dest. caused slight erythema in one animal of the test group. Animals of control groups one and two did not show any skin reactions.

Under these test conditions and following the results described above Reaktivblau 1483 does not have a sensitizing effect on the skin of the guinea pig.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:

The substance Reaktivblau 1463 was tested for its sensitizing effect on the skin of the guinea pig in the maximization test based on the method of Magnusson and Kligman.

After intradermal induction distinct erythema and oedema were observed at the injection sites of the control and the test animals at which only Freund's adjuvant/0.9% aqueous NaCl-solution (1 : 1) was applied. Injection of the test substance preparation in 0.9% aqueous NaCl-solution or in Freund's adjuvant/0.9% aqueous NaCl-solution (1 : 1) caused distinct oedema in the test animals. The irritation index for erythema could not be determined because of staining due to the colour of the test substance. The control animals injected with 0.9% aqueous NaCl-solution (vehicle) did not show any skin reactions.

The percutaneous induction was only carried out in the test group, because with aqua dest. a vehicle was used that was not expected to influence the result of the study. Incrustation, partially open (caused by the intra-dermal induction) could be observed in addition to distinct oedema. The index for erythema could not be determined because of staining due to the colour of the test substance.

The number of animals with skin findings after the 1st challenge (20 days after intradermal induction) and after the 2nd challenge (27 days after intradermal induction) is summarized in the following table:

 

 

 

1stchallenge

2ndchallenge

25% in aqua dest.

25% in aqua dest.

Control group

1

0/10

0/10

Control group

2*

no application of test substance

0/9

Test group**

 

1/18

1/18

x/y:  number of positive reactions/number of animals tested; readings 48 h after the beginning of application

*      one animal died 9 days after intradermal induction cause of death: pneumonia

**    two animals died 10 and 12 days after intradermal induction; cause of death: pneumonia

 

After the first challenge with the 25% test substance preparation in aqua dest. one test animal exhibited distinct erythema. Control group animals did not show any skin reactions.

The second challenge with the 25% test substance preparation in aqua dest. caused slight erythema in one animal of the test group. Animals of control groups one and two did not show any skin reactions.

Under these test conditions and following the results described above Reaktivblau 1483 does not have a sensitizing effect on the skin of the guinea pig.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Based on a local lymph node assay in mice, the test substance does not require classification for sensitization according to CLP (EC 1272/2008) criteria.