Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
From January 31, 2013 to June 27, 2013
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed in GLP following OECD official guidelines with no deviations

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2013
Report Date:
2013

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
Buehler test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
Females were nulliparous and non-pregnant

Animal supply and acclimatisation

Age and weight range (at order):4 to 5 weeks old, 250 to 300 grams
Supplier : Charles River Italia S.p.A., Calco (Lecco), Italy.
Breeder : Charles River Germany, Sandhofer Weg n° 7, D- 97633 Sulzfeld – Germany
Date of arrival : 07 February 2013
Weight range at arrival :251 to 310 grams (Order No. 111),248 to 287 grams (Order No. 112)
Acclimatisation period :At least 5 days
Veterinary health check :After arrival
Identification : Permanent by tattoo on the ear, following randomisation at arrival

Caging

No. of animals/cage :5 animals/cage
Housing : Noryl cages measuring 74.3x54.3x25 cm
Cage tray control :Daily inspected and changed as necessary (at least 3 times/week)

Water and diet

Water : drinking water supplied to each cage via a water bottle
Water supply : ad libitum
Diet : 8GP17 (Mucedola S.r.l., Via G. Galilei, 4, 20019, Settimo Milanese (MI) Italy)
Diet supply : ad libitum
Records of analyses of water and diet are kept on file at RTC. Components present in the drinking water or diet are not at a level likely to interfere with the purpose or conduct of the study.

Housing conditions (parameters set)

Room lighting : Artificial (fluorescent tubes), daily light/dark cycle of 12/12 hours
Air changes : Approximately 15 to 20 air changes per hour
Temperature range :22°C ± 2°C
Relative humidity range :55% ± 15%
Actual conditions were monitored and recorded, and records retained. No relevant deviations occurred.


Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Remarks:
Acetone for the challenge
Concentration / amount:
Induction: 50% in corn oil
Challenge: 20% v/v in acetone
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Remarks:
Acetone for the challenge
Concentration / amount:
Induction: 50% in corn oil
Challenge: 20% v/v in acetone
No. of animals per dose:
a test group of 20 animals and a control group of 10 animals
Details on study design:
The study was divided into 2 distinct phases. The first of these consisted of a preliminary screen which was used to determine suitable test item concentrations to be used in the second phase. This second phase constituted the main study: the determination of the sensitisation potential of the test item.

Preliminary screen

For each vehicle investigated (corn oil and acetone), 5 animals were selected from those available and the flanks clipped free of hair. Each animal was dosed with 2 concentrations of the test item, 1 on each flank. A gauze patch measuring at least 20x20 mm was soaked with 0.4 mL of the selected concentration of the test item. This was then placed onto the selected treatment site. A strip of synthetic film was placed over the treated sites and the whole assembly was secured in position by encircling the trunk of the animal with a length of adhesive strappin.
All animals were treated in this manner such that a total of 5 concentrations (100%, 50%, 20%, 10% and 5% v/v in corn oil or acetone) of the test item were dosed each in duplicate. The adhesive strapping and patches were removed after approximately 6 hour contact with the skin. The treated sites were then washed with lukewarm water to remove any remaining test item.
Approximately 24 and 48 hours after removal of the dressings and patches, the treated sites were examined for signs of reaction to treatment. Each site was assessed and scored

Main study - Induction

On the day of dosing (Day 0), animals were allocated to treatment to give a test group of 20 animals and a control group of 10 animals. Each animal was weighed and the hair was clipped from the anterior region of the left flank using an electric clipper and a razor.
Animals of the test group were treated with the test item at a concentration of 50% v/v in corn oil. A gauze patch measuring 20x20 mm was soaked with 0.4 mL of the test item and placed onto the selected skin site. A strip of synthetic film was placed over the treated sites and the whole assembly was secured in position by encircling the trunk of the animal with a length of adhesive strapping. Animals of the control group were similarly treated with the vehicle alone (corn oil).
After an exposure period of 6 hours, the dressings and patches were removed. The treated sites were cleaned of remaining test item or vehicle by washing with lukewarm water.
Approximately 24 and 48 hours after removal of the dressings and patches, the treated sites were examined for signs of reaction to treatment. Each site was assessed and scored


Challenge controls:
Main study - Challenge

On Day 28, the hair was removed with electric clippers and a razor from both the anterior and posterior regions of the right flank of all animals of both test and control groups.

A 0.4 mL aliquot of the test item at a concentration of 20% v/v in acetone was spread evenly over an absorbent patch measuring approximately 20x20 mm. This was placed onto the skin of the posterior region of the prepared site on the right flank. A similar patch, containing 0.4 mL of the vehicle (acetone), was placed onto the anterior region of the prepared site. A strip of synthetic film was placed over the treated sites and the whole assembly was secured in position by encircling the trunk of the animal with a length of adhesive strapping. All animals of the test and control groups were treated with both the test item and vehicle in this manner. After an exposure period of approximately 6 hours the dressings were removed and the treated sites cleaned of the remaining test item by washing with lukewarm water.
On Day 29, approximately 21 hours after removal of the dressing and patches, the treated sites were closely clipped (with electric clippers only) to remove any hair that may have grown. Approximately 3 hours later, 24 hours after removal of the dressing and patches, the treated sites were examined for any signs of reaction to treatment.

The degree of skin reaction was scored

Skin reaction on the treated sites was again assessed approximately 24 hours after the first examination (approximately 48 hours after removal of the patches)

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
20% conc.
No. with + reactions:
4
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 20% conc.. No with. + reactions: 4.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
20% conc.
No. with + reactions:
6
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 20% conc.. No with. + reactions: 6.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
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: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
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: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:
sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
These results indicate that the test item,Paraffin waxes and hydrocarbon waxes, chloro,sulfochlorinated saponified (C18-C30) SSP-SAMPLE 3, may elicit a sensitisation response in the guinea pig
Executive summary:

The potential of the test item, Paraffin waxes and hydrocarbon waxes, chloro, sulfochlorinated saponified (C18-C30) SSP-SAMPLE 3, to induce and elicit delayed dermal sensitisation was assessed by a guinea pig model using the methods of Buehler.

 

At challenge, response was observed in a total of 6/20 animals of the test group (30%) following treatment with the test item at 20% concentration in acetone. No reaction was observed in the control group animals at sites treated with the test item at the same concentration or in any animal at sites treated with the vehicle alone (acetone).

 

The validity of the test system was verified with periodic testing of the positive control.

 

These results indicate that the test item, Paraffin waxes and hydrocarbon waxes, chloro, sulfochlorinated saponified (C18-C30) SSP-SAMPLE 3, may elicit a sensitisation response in the guinea pig, since there was evidence of response at challenge following a period of induction exposure to the test item.