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 28 MAR 2011 to 4 MAY 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study (EU method B.6; OECD TG 406) (Performance of the otherwise preferred LLNA test was technically not feasible: the test material was insoluble in any of the solvents recommended to be used for the LLNA)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
Buehler test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):C-Wax Type S
- Substance type: yellowisch to white powder
- Physical state: solid
- Analytical purity: >=99.5 to <=100% (w/w)

In vivo test system

Test animals

Species:
guinea pig
Strain:
other: Dunkin Hartley, HsdPoc:DH
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Germany
- Age at study initiation: 5 - 7 weeks at the first application
- Weight at study initiation: 304 g to 362 g at the first application
- Housing: Group caging in plastic containers (48 cm x 115 cm bottom area, 36 cm height), partly shaded, 10 animals per container
- Diet: Ssniff Ms-H (Guinea Pig Maintenance Diet V2233), includes ascorbic acid (2400 mg/kg), ad libitum,
- Water: Tap water from Makrolon-bottles with stainless steel canules, ad libitum.
- Acclimation period: 6 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): Mean of 20.8
- Humidity (%): Mean of 42.4 %
- Air changes (per hr): approx. 12
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: white petrolatum
Concentration / amount:
50 % (w/w) (i.e. highest technically feasible concentration)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: white petrolatum
Concentration / amount:
50 % (w/w) (i.e. highest technically feasible concentration)
No. of animals per dose:
control: 10
treatment group: 20
Details on study design:
RANGE FINDING TESTS:
To obtain the appropriate concentrations of the test substance for the definitive study, a preliminary test was carried out with 3 female guinea pigs. 4 different concentrations of the test substance were administered epicutaneously.
The test substance concentrations were: 50 %, 25 %, 10 % and 5 % (w/w) in white petrolatum. 50 % of test item in white petrolatum was the highest technically feasible concentration.
The modes of application were the same as in the definitive study. The duration of the epicutaneous exposure was 6 hours.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 (day 0, day 7 and day 14)
- Exposure period: 6 hours
- Test groups: test substance in white petrolatum
- Control group: white petrolatum
- Site: left flank
- Frequency of applications: once per week
- Duration: 14 days
- Concentrations: 50 % (w/w)

B. CHALLENGE EXPOSURE
- No. of exposures:1
- Day(s) of challenge: day 28
- Exposure period:6 hours
- Test groups and control group (exposure and site): All animals from both groups are treated in the same way: Administration of 50 % test substance (w/w) in white petrolatum to the posterior right flanks, administration of white petrolatum to the anterior right flanks.
- Evaluation (hr after challenge): Skin was examined 24 hours after the start of each induction exposure and 24 and 48 hours after the end of the challenge exposure (blind reading of test and control animals). Findings on the test substance and on the control areas were scored.

OTHER:
Time schedule of the main study:
Day 0 was 4 April 2011.
Day 0: recording of body weight, clipping of hair, 1st induction exposure, removal of dressings 6 hours after application
Day 1: scoring
Day 7: clipping of hair, second induction exposure, removal of dressings 6 hours after application
Day 8: scoring
Day 14: clipping of hair, third induction exposure, removal of dressings 6 hours after application
Day 15: scoring
Day 28: clipping of hair, challenge exposure, removal of dressings 6 hours after application
Day 29: approx. 21 hours after removing the patch the hair is clipped on the application area (if necessary), approx. three hours later scoring
Day 30: scoring, recording of body weight, sacrifice of animals, end of test

Scoringen scheme of skin reactions:
Skin reactions Score Graded as
no reactions 0 normal
very slight erythema 1 mild
well defined erythema 2 moderate
severe erythema and/or oedema 3 severe
Positive control substance(s):
yes
Remarks:
ALPHA-HEXYLCINNAMIC ALDEHYDE

Results and discussion

Positive control results:
6/10 animals (60 %) had a positive response in this test, which is markedly more than the minimum of 15 %, the threshold for classification requested by the guideline. Thus the results confirm both the sensitivity and the reliability of the experimental techniques.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no effects
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. Clinical observations: no effects.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50 % (w/w)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no effects
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50 % (w/w). No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no effects.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no effects
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. Clinical observations: no effects.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50 % (w/w)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no effects
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50 % (w/w). No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no effects.

Any other information on results incl. tables

- all animals survived until the end if the study

- no statistically differences in the mean body weights between the test group and the control group on days 0 and 30

- Immediately after the beginning of all exposures (inductions, challenge) the motor activity of all animals was increased. This was considered to be due to the dressings which restrict the freedom of movement. Soon afterwards the behaviour was normal again.

- skin reactions after induction treatments as well as after challenge treatment:

Neither in the negative control group nor in the test substance group skin reactions were noted in any animal at any time.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance was not sensitising in concentrations up to 50% (w/w) in white petrolatum in this guinea pig test according to Buehler. This test was considered sufficient to fulfil the information requirements.
Executive summary:

Testing for sensitizing properties of the test item was performed in female Dunkin Hartley guinea pigs according to the method of Buehler (OECD 406). Three epicutaneous induction treatments and one challenge treatment were performed using 50% (w/w; i.e. highest technically feasible concentration) test item in white petrolatum. Based on the results of this study the test item showed no evidence of sensitizing properties. Therefore, the test item has not to be classified as skin sensitiser according to Regulation (EC) No 1272/2008.