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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:
21 Oct 2014 to 05 Dec 2014
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study has been performed according to OECD and/or EC guidelines and according to GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2015
Report Date:
2015

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
1992
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
May 2008, including most recent amendments
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
March 2003
Deviations:
no
Qualifier:
according to
Guideline:
other: Japanese Ministry of Agriculture, Forestry and Fisheries (JMAFF), 12 Nousan, Notification No 8147, November 2000; including the most recent partial revisions
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Sulfosuccinates may cause false positive/negative results in the LLNA, therefore the GPMT is a golden standard test for these compounds.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
- Name of test material (as cited in study report): Disodium isodecyl sulfosuccinate
- Substance type: organic
- Physical state: white powder
- Storage condition of test material: At room temperature in a well-sealed container

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River France, L’Arbresle Cedex, France
- Age at study initiation: approx. 4-8 weeks old
- Weight at study initiation: 361-477 g
- Housing: Group housing of maximally 5 animals per cage
- Diet: Complete maintenance diet for guinea pigs (SSNIFF® Spezialdiäten GmbH, Soest, Germany), ad libitum
- Water: Free access to tap water
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 18-24
- Humidity (%): 40-70
- Air changes (per hr): at least 10
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From 21 Oct 2014 To 05 Dec 2014

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
water
Concentration / amount:
Preliminary test:
Intradermal injection: 40%, 20%, 10%, 5%, 2%, 1%, 0.5%, 0.2%.
Epidermal exposure: 40%, 20%, 10%, 5%.

Main study:
Intradermal injection: 0.5%
Epidermal application: 10%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Preliminary test:
Intradermal injection: 40%, 20%, 10%, 5%, 2%, 1%, 0.5%, 0.2%.
Epidermal exposure: 40%, 20%, 10%, 5%.

Main study:
Intradermal injection: 0.5%
Epidermal application: 10%
No. of animals per dose:
10 (test group)
5 (control group)
Details on study design:
RANGE FINDING TESTS:
For the intradermal injections a series of four test substance concentrations was tested; the highest concentration was the maximum concentration that could technically be injected. Two animals received two different concentrations in duplicate (0.1 mL/ site) in the clipped scapular region. The resulting dermal reactions were assessed 24 and 48 hours after treatment. Based on the results in the initially treated animals, two additional animals were treated in a similar manner with four lower concentrations at a later stage.
For the epidermal application, a series of four test substance concentrations was tested, the highest concentration being the maximum concentration that could technically be applied. Two different concentrations were applied (0.5 mL each) per animal semi-occlusively to the clipped flank (site of 2x3cm). The animals receiving intradermal injections were treated with the lowest concentrations and two other animals with the highest concentrations. After 24 hours, the dressing was removed and the skin cleaned of residual test substance using water. The treated skin areas were assessed for irritation 24 and 48 hours after exposure.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: one intradermal injection on day 1; epidermal application on day 8
- Exposure period (epidermal application): 24 hours
- Test group: 10 animals
- Control group:5 animals
- Site: scapular area
- Concentrations: 0.5% (intradermal injection), 10% (epidermal application)

B. CHALLENGE EXPOSURE
- No. of exposures: single
- Day of challenge:day 22
- Exposure period: 24 hours
- Test groups: 10% test substance
- Control group: vehicle only
- Site: flank
- Evaluation (hr after challenge): at 24 and 48 hours after removal of the dressing
Challenge controls:
Historical control data are provided.
Positive control substance(s):
yes
Remarks:
Alpha- hexylcinnamaldehyde

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
None
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
None
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
None
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
None
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
20% Alpha- hexylcinnamaldehyde
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
The skin reactions observed in nine experimental animals in response to the 20% test substance concentration in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals.
Remarks on result:
positive indication of skin sensitisation
Remarks:
90% sensitisation rate in reliability check performed in July/August 2014
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
20% Alpha- hexylcinnamaldehyde
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
The skin reactions observed in nine experimental animals in response to the 20% test substance concentration in the challenge phase were considered indicative of sensitisation, based on the absence of any response in the control animals.
Remarks on result:
positive indication of skin sensitisation
Remarks:
90% sensitisation rate in reliability check performed in July/August 2014

Any other information on results incl. tables

No mortality occurred and no symptoms of systemic toxicity were observed in the animals of the main study. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period.

On day 3 after intradermal injection of 1:1 Mixture of FCA and water (and a 1:1 Mixture of FCA and vehicle for injection), all animals had erythema grade 3. One control animal had slight erythema (grade 1) related to injection of vehicle. This reaction was also found in 6 guinea pigs of the test group after injection with 0.5% test substance. Additionally, one animal in the test group had well-defined erythema (grade 2). One animal from the test group had signs of necrosis (1 mm in diameter). Signs of necrosis were seen in all animals of the test group at the injection site of 1:1 Mixture of FCA and a 20% test substance concentration (size ranging form 3 to 5 mm, on average 3.8 mm); No skin reaction was seen after epidermal exposure to the vehicle or to 10% test substance in any of the animals.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Criteria used for interpretation of results: EU
Conclusions:
Based on the results of a guinea pig maximisation test, performed according to OECD/EC guidelines and GLP principles, disodium isodecyl sulfosuccinate was found to be not sensitising.
Executive summary:

A guinea pig maximisation test was performed with disodium isodecyl sulfosuccinate according to OECD/EC guidelines and GLP principles. Reliable positive and negative controls were included. Based on a preliminary irritation study, the test substance concentrations selected for the main study were a 0.5% concentration for the intradermal induction and a 10% concentration for the epidermal induction exposure. A 10% test substance concentration was selected for the challenge phase. During the main study, no mortality occurred and no symptoms of systemic toxicity were observed. Body weights and body weight gain of experimental animals remained in the same range as controls over the study period. Epidermal exposure to the vehicle or to 10% test substance did not result in a skin reaction in any of the animals. Since no responses were observed in the experimental animals in response to a 10% test substance concentration in the challenge phase, it is concluded that disodium isodecyl sulfosuccinate has no sensitising properties according to CLP Regulation (EC) No. 1272/2008.