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Description of key information

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
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: In according to OECD guideline and GLP without deviations. With complete substance informations
Justification for type of information:
The choice of a non LLNA study was done in consideration of literature data on similar substances reporting some uncertain results from LLNA studies
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Data in the art are available taht substances with the same functional groups pf the target one can act as a confounder
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
Animal Supply and Acclimatisation
- Females: nulliparous and non-pregnant
- Age: 4 to 5 weeks
- Weight: 250 to 300 grams
- Supplier: Charles River Italia, S.p.A, Calco (Lecco), Italy
- Breeder: Charles River France Laboratories, Domaine des Oncins B.P. 0109, F 69592 L'Arbresle Cedex, France
- Date of arrival: 23 january 2014
- Weight range at arrival: 210 to 311 grams
- Acclimatisation: at least 5 days
- Veterinary health check: during the acclimatisation period
- Identification: Permanent by tatoo on the ear, following randomisation at arrival

Animal husbandry:
- Animals per cage: up to 5
- Housing: noryl cages measuring 74,3x54,3x25cm
- Cage control: daily inspected and changed as necessary (at least 3 times/week)
- Water: drinking water supplied to each cage via a water bottle
- Water supply: Ad libitum
-Diet: 8GP17 (Mucedula S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI) Italy)
- Diet supply: Ad libitum throughout the study
- Room lighting: artificial (fluorescent tubes); daily light/dark cycle of 12/12 hours
- Air change: approximately 15 to 20 air change per hour
- Temperature range: 22°C +- 2°C
- Relatively humidity: 55% +- 15%
Route:
intradermal and epicutaneous
Vehicle:
corn oil
Concentration / amount:
Dose-range screening: undiluated, 50, 20, 10, 5 and 1% of test item in corn oil
Dose for main study: 20% of test item in corn oil
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Concentration / amount:
Dose-range screening: undiluated, 50, 20, 10, 5 and 1% of test item in corn oil
Dose for main study: 20% of test item in corn oil
No. of animals per dose:
test group: 10 animals
control group: 5 animals
Details on study design:
The study was divided in 2 phases, phase 1 for screen the dose and phase 2 the main study
1.DOSE SCREENING:
1.1. Intradermal injection
- Allocation: 1 group of 2 animals to the intradermal injection tolerance test
- Preparation on the site (day 1): hair over the scapulae was removed using an electric clipper with suitable blade
- Dosage: 6 sites were selected on each animals over the shaved scapulae. Each site was injected with the single concentration of the item test. Undiluated test item and concentration of 50, 20, 10, 5, 1 % in corn oil were selected.
- Dosing method: A single intradermal injection of 0,1ml of each concentration was administrated with a suitable graduated syringe.
- Observations: observation of the treated sites was performed at day 2 (approximatively 24hours after dosing) and day 7 for any signs reactions.
1.2. Topical application:
- Allocation: 1 group of 5 animals was allocated to the topical application tolerance test
- Preparation of the site and F.C.A. injection (day 1): hair over scapulae was removed. Each animal was then injected intradermally at the prepared site with 2 injections, each of 0,1 ml, of emulsified Freund's complete adjuvant.
- Dosage: 7 days after adjuvant injection, the flank of each animal was clipped free of hair. Each animal was dosed with 2 concentrations of test items, one of each site. The undiluated test item and concentrations of 50, 20, 10 and 5 % in corn oil were selected.
- Dosing method: on the day of dosing, a gauze patch measuring at lest 20x20 mm was soaked with 0,2ml of each selected doses and placed onto the selected treatment site and covered by a trip of synthetic film and the trunk wrapped with an elastic adhesive bandage to maintain the test item in contact with the skin. After 24hours the treated sites is gently cleaned by washing with lukewarm water.
- Observations: 24 and 48 hours after removal of the dressings, the treated site were examined for signs of reactions ot treatment.

2. MAIN STUDY:
2.1. Induction-Intradermal injection
- Allocation: 10 animals for test item and 5 animals for control group
- Preparation of site: hair over the scapulae was removed using an electric clipper with suitable blade.
- Dosage: 3 pairs of intradermal injection were made at the prepared skin site of each animal. All injections were made at the edge of the prepared site and the anterior and median injections were positioned close together and distant from the posterior injections. A volume of 0,1ml was injected at each site.
- Dosing method: the animals were administrated by intradermal injection with graduated syringe on day 1
- observation: skin reactions at the injection site was assessed 24 hours after injection
2.2. Induction- Topical application
- Preparation of the site (day8): hair was removed on the area surrounding the injection sites
-Treatment: animals of test group were treated with 0,4 ml of the undiluted test item at 100% concentrations and those of the control group with the vehicle alone (corn oil). A gauze patch was soaked wigth the selected concentrations of the test item or the vehicle and then placed over the injection site. The treatment site was covered with a strip of synthetic film and the trunk wrapped with an elastic adhesive bandage to maintain the test item in contact with the skin. After 48 hours, the treated sites was gently cleaned by washing with lukewarm water.
- Observation: reaction to treatment was assessed 24 hours after removal of the dressings.
2.3. Challenge:
- preparation of the site (day 22) : 3 weeks after preparing the animals in the first induction phase of the main study, all animals were prepared for challenge by clipping the flanks free of hair to expose area of approximatively 50 x 50 mm on each flank.
- dosages: all animals (test and control groups) were treated with 0,2 ml aliquots of test item at 20% concentrations in corn oil on the right flank. The left flank of each animal was treated with 0,2 ml of the vehicle alone.
- Dosing method: A gauze patch measuring 20×20 mm was soaked with the test item (or vehicle) and then placed over the centre of the right (or left) flank. When both sites of the animal had been treated, they were covered with a strip of synthetic film and the trunk wrapped with an elastic adhesive bandage to maintain the test item and vehicle in contact with the skin (occlusive barrier). After a contact period of 24 hours, the dressings and patches were removed and the treated sites were washed with lukewarm water.
- Observations: The treated sites were closely clipped to remove any hair that may have grown approximately 21 hours after the removal of the dressings. Observation of the treated sites was carried out approximately 24 hours and 48 hours after removal of the dressings.
Positive control substance(s):
yes
Remarks:
alpha-Hexylcinnamaldehyde
Positive control results:
A positive control check using alpha-Hexylcinnamaldehyde was performed at approximately 6 monthly intervals. Sensitisation reactions were observed in 60% of the animals in the reliability
check indicated in report. These results are considered to be acceptable for a weak sensitiser like the alpha-Hexylcinnamaldehyde.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
20% test item on corn oil
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no visible change
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 20% test item on corn oil. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no visible change.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle (corn oil)
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
no visible change
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle (corn oil). No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: no visible change.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
20% of alpha-Hexylcinnamaldehyde in corn oil (injection) and 50% for topical application
Remarks on result:
positive indication of skin sensitisation
Remarks:
in 60% of the tested animals

MAIN STUDY-INJECTION INDUCTION- INDIVIDUAL RESULTS:

Group function

Animal number

Dermal response

Anterior site:

Median site:

Posterior site:

(FCA emulsion)

 (Vehicle)

(Vehicle/FCA)

Left

Right

Left

Right

Left

Right

5
CONTROL

29

3

3

1

1

3

3

31

3

3

1

1

3

3

33

3

3

1

1

3

3

35

3

3

1

1

3

3

37

2

2

1

1

3

3

6
TEST

39

3

3

2

2

3

3

41

3

3

1

1

3

3

43

3

3

1

1

3

3

45

3

3

1

1

3

3

47

3

3

2

2

3

3

49

3

3

2

2

3

3

51

3

3

2

2

3

3

53

3

3

1

1

3

3

55

3

3

1

1

3

3

57

3

3

2

2

3

3

0= No erythma

1= Very slight erythema

2= Well defined erythma

3= Moderate to severe erythema

4= Severe erythma (beef redness) to eschar formation preventing grading of erythema

MAIN STUDY-TOPICAL INDUCTION-INDIVIDUAL RESULTS:

Group function

Animal number

Dermal response

5
CONTROL

29

0

31

0

33

0

35

0

37

0

6
TEST

39

2

41

2

43

1

45

1

47

2

49

1

51

2

53

2

55

2

57

2

KEY:
0= No visible change

1 = Discrete or patchy erythema

2= Moderate and confluent erythema
3= Intense erythema and swelling

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
Trimethylolpropane Tripelargonate is not sensitising to the skin.
Executive summary:

In accordance to the OECD guideline 406 and the European Directive B6 this study showed that Trimethylolpropane Tripelargonate does not induce and elicit delayed dermal sensitisation in the guinea pig.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (not sensitising)
Additional information:
Migrated from Short description of key information:
There is one delayed dermal sensitisation study available (OECD406), which indicates that Trimethylolpropane Tripelargonate does not induce and elicit delayed dermal sensitisation in guinea pigs.

Justification for selection of skin sensitisation endpoint:
This study is a well documented, recent GLP study according to international guidelines.

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available

Justification for classification or non-classification

Skin sensitisation

As no sensitisation response was observed during the study (OECD406), the substance is not to be classified for skin sensitisation effects according to the criteria described in Regulation No. 1272/2008 on the Classification, Labelling and Packaging of Substances and Mixtures (CLP) or Directive 67/548/EEC (Dangerous Substances Directive).