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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:
1998-03-10 to 1998-04-10
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
This study was conducted when the LLNA has not already been the standard method for in vivo testing.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
see below

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Remarks:
Pirbright-White
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: HARLAN WINKELMANN Gartenstr. 27 D-33178 Borchen SPF breeding colony
- Strain: HsdPoc:DH
- Weight at study initiation: mean 356g (n=15)
- Housing: fully air-conditioned rooms in macrolon cages (type 4) on soft wood granulate in groups of 5 animals
- Diet (e.g. ad libitum): ssniff Ms-H (V2233), ad libitum
- Water (e.g. ad libitum): tap water in plastic bottles, ad libitum
- Acclimation period: at least 7 d

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 3 degrees C
- Humidity (%): 50± 20%
- Photoperiod (hrs dark / hrs light): 12 hrs lighting time daily

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
water
Concentration / amount:
5% / 2 * 0.1 mL in water
5% / 2 * 0.1 mL in 50% Freund Adjuvant
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:
25% / 0.5 mL
Day(s)/duration:
Day 8 / 48 h
Adequacy of induction:
highest technically applicable concentration used
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
25%
An amount of 0.5 ml of the test substance preparation was administered to a 2 x 2 cm cellulose patch. The administration area was then kept for 24 hours under an occlusive bandage with an impermeable film and an elastic bandage.
Day(s)/duration:
Day 22 / 24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Test Groups:
- Number of animals in test group: 10
- Number of animals in negative control group: 5
- Number of animals to determine the primary non-irritant concentration: 3
- Number of animals to determine the tolerance of the intradermal injections: 2

10 animals in the treatment group and 5 animals in the control group were used. In case of a questionable result additional animals will be tested to give a total of 20 test and 10 control animals.
Details on study design:
RANGE FINDING TESTS:
- Determination of the primary non-irritant concentration:
Prior to the determination of the primary non-irritation concentration in a dermal occlusive test the animals received 4 intradermal injections of a 50% Freund's
Complete Adjuvant emulsion (4 x 0.1 ml) into the dorsal area. Two guinea pigs then received either 25%, 5%, or 1% test substance in dionized water. Flank hair was mechanically removed and 0.5 ml of the test substance preparation was administered to a 2 x 2 cm cellulose patch covered occlusive for 24 hours. 24 hours after removal of the patches, the treated skin areas were examined for erythema and edema.
- Determination of the tolerance of the intradermal injections:
To determine the tolerance of intradermal injections, 5%, 1%, or 0.2% of the test substance was administered twice by intradermal injection to 2 guinea pigs. 24, 48, 72 and 96 hours after administration the injection sites were examined for local tolerance.


MAIN STUDY
A1. INTRADERMAL INDUCTION EXPOSURE
- No. of exposures: 2 intradermal injections per animal
- Test groups: 5% test substance in dionized water or in 50% Freund's Adjuvant
- Control group: 50% Freund's Adjuvant
- Site: within a dorsal area of 2 x 4 cm
- Concentrations: 5% test substance
A2. DERMAL INDUCTION EXPOSURE
- Exposure period: 48 h
- Test groups: 25.0 % test substance in deionized water
- Control group: deionized water
- Site: The cellulose patch including the test substance covered the area where the intradermal injection had been placed
- Concentrations: 25.0 % test substance

B. CHALLENGE EXPOSURE
- Exposure period: 24h
- Test groups: 25.0 % test substance
- Site: One area of approx. 5 x 5 cm on the left flank, shaved mechanically
- Concentrations: 25.0 % test substance
- Evaluation (hr after challenge): 24h and 48h; visual skin reaction, fold thickness and histophathology after 48 h
Positive control substance(s):
yes
Remarks:
benzocain

Results and discussion

Positive control results:
Benzocain caused sensitization by skin contact.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
body weight not impaired, no clinical signs of intoxication throughout study
Remarks on result:
no indication of skin sensitisation
Remarks:
based on skin fold measurement; see below
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
body weight not impaired, no clinical signs of intoxication throughout study
Remarks on result:
no indication of skin sensitisation
Remarks:
based on skin fold measurement; see below
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
5
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25 %
No. with + reactions:
2
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
no indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
24
Group:
positive control
Dose level:
25 %
No. with + reactions:
5
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Reading:
rechallenge
Hours after challenge:
48
Group:
positive control
Dose level:
25 %
No. with + reactions:
4
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation

Any other information on results incl. tables

Due to the red color of the test substance the treated skin of the animals could not be assessed for erythema 24 and 48 hours after removal of the occlusive bandage.

No significant difference in the fold thickness of the treated skin was detected between the animals of the control and of the treatment group.

In addition, no significant difference was found between the animals of the control and of the treatment group with respect to the histopathological evaluation of the treated skin area. The findings in both the control and the treatment group reveal a slight

reaction of the skin tissue indicating an irritant response to the occlusion of the skin.

Thus, neither measurement of the fold thickness nor histopathological examination of the treated skin showed any evidence for sensitizing properties of the test substance. No skin reactions were observed in the control group 24 and 48 hours after removal of the occlusive bandage.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Reactive Red FC73270 showed no evidence for sensitizing properties.
Executive summary:

Testing for sensitizing properties of Reaktiv Rot FC-73270 was performed in female guinea pigs according to the method of MAGNUSSON & KLIGMAN. Intradermal induction was performed using 5 % Reaktiv Rot FC-73270 in deionized water. Dermal induction and challenge treatment were carried out with 25 % Reaktiv Rot FC-73270 in deionized water.

The validity of the test system is confirmed by the periodically conducted positive control test using benzocain for the maximization test.

Due to the red color of the test substance the treated skin of the animals could not be assessed for erythema. Based on both measurement of fold thickness and histopathological examination of the treated skin Reaktiv Rot FC-73270 showed no evidence for sensitizing properties.