Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2000
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
Directive 96/54/EC, B.6 Maximization test
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study have been performed before the REACh legislation enters into force.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
None

In vivo test system

Test animals

Species:
guinea pig
Strain:
Himalayan
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source:RCC Ltd, Biotechnology & Animal Breeding Division, Wölferstrasse 4, CH-4414 Füllinsdorf / Switzerland
- Age at study initiation:4 - 6 weeks
- Weight at study initiation:Pretest groups: 328 - 347 g;
- Housing:Individually in Makrolon type-4 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz).
- Diet (e.g. ad libitum):Pelleted standard Nafag Ecosan 845 25W4, batch nos. 33/00 and 45/00, guinea pig breeding / maintenance diet, containing Vitamin C ("Nafag", Nähr- und Futtermittel AG, CH- 9202 Gossau), ad libitum. Results of analyses for contaminants are archived at RCC Ltd, Hingen.
- Water (e.g. ad libitum):Community tap water from Füllinsdorf, ad libitum. Results of bacteriological, chemical and contaminant analyses are archived at RCC Ltd, Hingen.
- Acclimation period:One week for the control and test group under test conditions after health examination. No acclimatization for the animals of the pretest. Only animals without any visible signs of illness were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C):20-22.5
- Humidity (%):between 45-73
- Air changes (per hr):10-15 air changes per hour
- Photoperiod (hrs dark / hrs light):12 hours light and 12 hours dark.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Polyethylene glycol 300 (PEG 300)
Concentration / amount:
Concentration of test material and vehicle used at induction:
intradermal: 0.1 ml 1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and
physiological saline 2) test substance diluted to 10% (w/w) in PEG300
3) test substance diluted to 10% (w/w) by emulsion in a
1:1 (v/v) mixture of Freund's Complete Adjuvant and
physiological saline epidermal:
test substance 40% in PEG300 Concentration of test material and vehicle used for each challenge: epidermal: test substance 40% in PEG300
Challengeopen allclose all
Route:
epicutaneous, open
Vehicle:
other: Polyethylene glycol 300 (PEG 300)
Concentration / amount:
Concentration of test material and vehicle used at induction:
intradermal: 0.1 ml 1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and
physiological saline 2) test substance diluted to 10% (w/w) in PEG300
3) test substance diluted to 10% (w/w) by emulsion in a
1:1 (v/v) mixture of Freund's Complete Adjuvant and
physiological saline epidermal:
test substance 40% in PEG300 Concentration of test material and vehicle used for each challenge: epidermal: test substance 40% in PEG300
No. of animals per dose:
Number of animals in test group: 10
Number of animals in negative control group: 5
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
INTRADERMAL INJECTIONS / PERFORMED ON TEST DAY 1
Three pairs of intradermal injections (0.1 ml/site) were made at the border of a 4 x 6 cm area in the clipped region as follows:
- Test groups:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) The test item, at 10 % in PEG 300.
3) The test item at 10 % in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
- Control group:
1) 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
2) PEG 300
3) 1:1 (w/w) mixture of PEG 300 in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline.
- Site:An area of dorsal skin from the scapular region (approximately 6 x 8 cm) was clipped free of hair.

EPIDERMAL APPLICATIONS / PERFORMED ON TEST DAY 8
On test day 7 and 23 hours prior to the epidermal application the scapular area (approximately 6 x 8 cm) of the animals of the control and test group was clipped, shaved free of hair and the test area was pretreated with 0.5 ml of 10 % Sodium-Lauryl-Sulfate (SLS) in paraffinum perliquidum as no primary irritation had been observed in the pretest. The SLS was massaged into the skin with a glass rod without bandaging. This 10 % concentration of SLS enhances sensitization by provoking a mild inflammatory reaction (Magnusson and Kligman 1970).
On test day 8, a 2 x 4 cm patch of filter paper was saturated with the test item (40 % in PEG 300) and placed over the injection sites of the test animals. The amount of test item preparation applied was approximately 0.3 g. The patch was covered with aluminum foil and firmly secured by an elastic plaster wrapped around the trunk of the animal and secured with impervious adhesive tape. The occlusive dressings were left in place for 48 hours. The epidermal application procedure described ensured intensive contact of the test item. The guinea pigs of the control group were treated as described above with PEG 300 only, applied at a volume of approximately 0.3 ml. The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman

B. CHALLENGE EXPOSURE
The test and control guinea pigs were challenged two weeks after the epidermal induction application and were treated in the same way.
Hair was clipped and shaved from a 5 x 5 cm area on the left and right flank of each guinea pig just prior to the application. Two patches (3x3 cm) of filter paper were saturated with the test item at the highest tested non-irritating concentration of 40 % (applied to the left flank) and the vehicle only (PEG 300 applied to the right flank) using the same method as for the epidermal application. The amount of test item preparation applied was approximately 0.2 g and a volume of approximately 0.2 ml was used for the vehicle. The dressings were left in place for 24 hours.
Twenty-one hours after removal of the dressing the test sites treated with the test item were depilated as described in the epidermal pretest.
The reaction sites were assessed 24 and 48 hours after removal of the bandage for erythema and oedema according to the method of Magnusson and Kligman
Challenge controls:
No data
Positive control substance(s):
not specified

Results and discussion

Positive control results:
None

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
40 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 40 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
40 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 40 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
40 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 40 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0 %. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
40 %
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 40 %. No with. + reactions: 0.0. Total no. in groups: 5.0.

Any other information on results incl. tables

Maximum concentration not causing irritating effects in preliminary test: 40 %

Signs of irritation during induction:
Control group:

Discrete/patchy erythema were observed in 4/5 animals at the 24- and 48-hour reading.

Test group:

Discrete/patchy erythema were observed in all animals at the 24- and 48-hour reading. No oedema, but yellow discoloration of the treated skin areas were observed.

Evidence of sensitisation of each challenge concentration: Concentration 40%: 0/9 (0%) animals showed sensitization.

Other observations:
MORTALITY/CLINICAL SIGNS:
One animal was found dead on test day 18. At necropsy, congestion of the lungs was noted. The cause of death could not be established. No clinical signs were noted in this study.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Remarks:
Migrated information
Conclusions:
Based on the above mentioned findings in an adjuvant sensitization test (M&K-test) in guinea pigs and in accordance to Commission Directive 96/54/EEC, FAT 60'253/A has not to be classified and labelled as a skin sensitizer.
Executive summary:

In order to assess the cutaneous allergenic potential of FAT 60'253/A, the Maximization-Test was performed in 15 (10 test and 5 control) male albino guinea pigs, in accordance with OECD Guideline No. 406 and the Directive 96/54/EEC, B.6.

The intradermal induction of sensitization in the test group was performed in the nuchal region with a 10 % dilution of the test item in PEG 300 and in an emulsion of Freund's Complete Adjuvant (FCA) / physiological saline. The epidermal induction of sensitization was conducted for 48 hours under occlusion with the test item at 40 % in PEG 300 one week after the intradermal induction and following pretreatment of the test areas with 10 % Sodium-Lauryl-Sulfate (SLS) approximately 23 hours prior to application of the test item. The animals of the control group were intradermally induced with PEG 300 and FCA/physiological saline and epidermally induced with PEG 300 under occlusion following pretreatment with 10% SLS.

Two weeks after epidermal induction the control and test animals were challenged by epidermal application of the test item at 40 % in PEG 300 and PEG 300 alone under occlusive dressing.

Cutaneous reactions were evaluated at 24 and 48 hours after removal of the dressing.

0% of skin reactions was observed after the challenge procedure.

No signs of toxicity were evident in the guinea pigs of the control or test group.

None of the 9 surviving test animals showed skin reactions after the challenge treatment with FAT 60'253/A at 40 % (w/w) in PEG 300. No skin effect was observed in the control group.

Based on the above mentioned findings in an adjuvant sensitization test (M&K-test) in guinea pigs and in accordance to Commission Directive 96/54/EEC, FAT 60'253/A has not to be classified and labelled as a skin sensitizer.