Registration Dossier

Administrative data

Description of key information

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Skin Sens. 1.

No skin sensitisation was observed in the in vivo test with guinea-pigs.

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Referenceopen allclose all

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: Guideline study with GLP.
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Principles of method if other than guideline:
As the test substance was insufficiently soluble in inorganic or organic solvents, the "Buehler test" according to E.V. Buehler was used instead of the Maximisation Test (Magnusson and Kligman).
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
The LLNA-method was not available at the time of initiation the study.
Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen, Germany
- Age at study initiation: 6 weeks.
- Weight at study initiation: 315 - 396 g.
- Housing: single caged.
- Diet: Altromin Diet No 3022, ad libitum
- Water: acidified to pH 3 with HCl, ad libitum
- Acclimation period: ca. 2 weeks.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): mean of ca. 22
- Humidity (%): mean of 53.6
- Air changes (per hr): 12
- Photoperiod (hrs dark / hrs light): 12/12
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
Concentration of test material and vehicle used at induction: 50 % in white petrolatum for epicutaneous application. Concentration of test material and vehicle used for each challenge: 50 % in white petrolatum.
The concentration was derived from a preliminary experiment.

The test substance was incorporated in white petrolatum immediately before application. After clipping the hair, filter papers. about 2 cm x 2 cm, were covered with the test substance formulation and with white petrolatum, respectively, and were applied to the appropriate sites of administration. They were fixed with a strip of non-irritating tape ("Blenderm®"). The area of administration was then covered occlusively with aluminium foil which finally was fixed with "Fixomull® stretch" (self adhesive non woven fabric).
Applied amounts of the test substance formulation (per animal) were:
• approx. 0.5 g for the first induction exposure
• approx. 0.6 g for the second induction exposure
• approx. 0.6 g for the third induction exposure
• approx. 0.5 g for the challenge exposure.
Applied amounts of white petrolatum (per animal) were:
• approx. 0.5 g for each induction exposure and for the challenge exposure.
Route:
epicutaneous, occlusive
Vehicle:
petrolatum
Concentration / amount:
Concentration of test material and vehicle used at induction: 50 % in white petrolatum for epicutaneous application. Concentration of test material and vehicle used for each challenge: 50 % in white petrolatum.
The concentration was derived from a preliminary experiment.

The test substance was incorporated in white petrolatum immediately before application. After clipping the hair, filter papers. about 2 cm x 2 cm, were covered with the test substance formulation and with white petrolatum, respectively, and were applied to the appropriate sites of administration. They were fixed with a strip of non-irritating tape ("Blenderm®"). The area of administration was then covered occlusively with aluminium foil which finally was fixed with "Fixomull® stretch" (self adhesive non woven fabric).
Applied amounts of the test substance formulation (per animal) were:
• approx. 0.5 g for the first induction exposure
• approx. 0.6 g for the second induction exposure
• approx. 0.6 g for the third induction exposure
• approx. 0.5 g for the challenge exposure.
Applied amounts of white petrolatum (per animal) were:
• approx. 0.5 g for each induction exposure and for the challenge exposure.
No. of animals per dose:
Number of animals in test group: 20
Number of animals in negative control group: 10
One spare animal per group was kept and administered under the same conditions.
Details on study design:
The Buehler-Test was chosen, because the test substance was insufficiently soluble in appropriate inorganic or organic solvents. As the solubility of the test substance was less than 1% in deionised water, in corn oil, in ethanol, in DMSO and in acetone, white petrolatum was used as a vehicle. 50% of test substance in white petrolatum was the highest technically feasible concentration.

Test principle
Induction exposures: Three epicutaneous administrations to the left flanks of the animals, test substance for the test substance group, reference substance for the negative control group.
Challenge exposure: All animals of both groups are treated in the same way: Administration of the test substance to the posterior right flanks, administration of the reference substance to the anterior right flanks.
Skin reactions after the challenge exposure are compared between the test substance group and the negative control group

Procedure and time table:
Day 0: recording of body weight, clipping of hair, first 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: ca. 21 hours after removing the patch cleaning of the area of administration, ca. 3 hours later scoring.
Day 30: scoring, recording of body weight, sacrifice of animals, end of test.
Positive control substance(s):
yes
Positive control results:
To check the sensitivity of the strain of animals used to detect a possible sensitising potential of a test substance as well as the reliability of the experimental technique the relevant OECD- and EC-guideline requires periodic checks with known sensitisers. The last check was performed with hexyl cinnamic aldehyde. 8/10 animals, i.e. 80 % of the animals of the positive control group, were regarded as sensitised.
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50 %
No. with + reactions:
0
Total no. in group:
10
Reading:
other: 1st
Hours after challenge:
24
Group:
positive control
Dose level:
20 %
No. with + reactions:
8
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
20 %
No. with + reactions:
8
Total no. in group:
10

Skin reactions after the induction exposures:

Group K (negative control): No adverse skin reactions in any animal at any time.

Group A (test substance): No adverse skin reactions in any animal at any time.

 

Skin reactions after the challenge exposure:

Negative control group:

Vehicle site: no positive skin reaction in any animal at any reading time.

Test substance site: no positive skin reaction in any animal at any reading time.

Test substance group:

Vehicle site: no positive skin reaction in any animal at any reading time.

Test substance site: no positive skin reaction in any animal at any reading time.

 

Other observations:

One animal of the test substance group died spontaneously on day 10 and was replaced by a spare animal, kept and administered under the same conditions as the other animals of this group. The cause of death was not test substance related.

The body weight development was inconspicuous.

Except for reactions due to the dressing, no other animal observations were made.

 

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The test substance is not regarded as a sensitizer in this test.
Executive summary:

Investigations to detect a sensitising potential of the test substance were performed according to OECD-Guideline 406 and the Directive 96/54/EC, 8.6. with the Buehler Test. Twenty female guinea pigs were used as a test substance group and another 10 females were used as a negative control group. There were three epicutaneous induction exposures and one epicutaneous challenge exposure. The concentration of the test substance in white petrolatum was 50% (w/w) for all three induction exposures and for the challenge exposure. The areas of administration were covered occlusively for 6 hours.

No adverse skin reactions were noted after the challenge exposure in any animal of any group, neither 24 nor 48 hours after the end of the exposure. All test substance treated skin areas were normal and no animal was regarded as irritated or sensitised.

 

Endpoint:
skin sensitisation, other
Remarks:
Experience with the chemical class of the substance and from observations of workers.
Type of information:
other: Experience with the chemical class of the substance and from observations of workers.
Adequacy of study:
other information
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Experience with the chemical class of the substance and from observations of workers.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Experience with the chemical class and from observations of workers.
GLP compliance:
no
Type of study:
other: Experience with the chemical class of the substance and from observations of workers.
Species:
other: Humans
Interpretation of results:
Category 1 (skin sensitising) based on GHS criteria
Conclusions:
Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Skin Sens. 1.
Executive summary:

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Skin Sens. 1.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)

Respiratory sensitisation

Link to relevant study records
Reference
Endpoint:
respiratory sensitisation, other
Remarks:
Experience with the chemical class of the substance and from observations of workers.
Type of information:
other: Experience with the chemical class of the substance and from observations of workers.
Adequacy of study:
other information
Study period:
2018
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Experience with the chemical class of the substance and from observations of workers.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Experience with the chemical class of the substance and from observations of workers.
GLP compliance:
no
Species:
other: Humans
Interpretation of results:
Category 1 (respiratory sensitising) based on GHS criteria
Conclusions:
Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Resp.Sens. 1.
Executive summary:

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Resp.Sens. 1.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

Experience with the chemical class of the substance and from observations of workers advises to classify the substance as Resp.Sens. 1.

Justification for classification or non-classification

Experience advises to classify the substance as Skin Sens. 1 and Resp.Sens. 1.