Registration Dossier

Diss Factsheets

Administrative data

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
Study period:
28 June - 05 Aug 2016
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
17 July 1992
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.6 (Skin Sensitisation)
Version / remarks:
30 May 2008
Deviations:
no
Qualifier:
according to guideline
Guideline:
EPA OPPTS 870.2600 (Skin Sensitisation)
Version / remarks:
March 2003
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
OGYÉI, Országos Gyógyszerészeti és Élelmezés-egészségügyi Intézet, Budapest, Hungary
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
none
Specific details on test material used for the study:
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature (15-25 ºC, below a room humidity level of 70%), protected from light and humidity
Species:
guinea pig
Strain:
other: CRL:HA Guinea pig
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH, Sulzfeld, Germany
- Females nulliparous and non-pregnant: yes
- Age at study initiation: about 7 weeks old
- Weight at study initiation: 324 – 361 g
- Housing: 5 animals per cage in macrolon cages (size IV) on certified wood chips bedding (Lignocel 3/4-S Hygienic Animal Bedding, J. Rettenmaier & Söhne GmbH + Co.KG, Rosenberg, Germany)
- Diet: Cunigra Diet for Rabbits with high vitamin D level (# NN16201718, Bonafarm-Bábolna Takarmány Ltd., Hungary), ad libitum
- Water: tap water (containing 50 mg/100 mL ascorbic acid), ad libitum
- Acclimation period: 19 days
- Indication of any skin lesions: no

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.7 - 25.0
- Humidity (%): 31 - 84%
- Air changes (per hr): 15 - 20
- Photoperiod (hrs dark / hrs light): 12/ 12
- IN-LIFE DATES: From: 28 June To: 05 Aug 2016
Route:
intradermal
Vehicle:
other: sesame oil
Concentration / amount:
5%
0.1 mL/ injection site
Day(s)/duration:
single injection
Adequacy of induction:
other: The test substance did not exhibit skin irritating properties in the screening test. Thus, undiluted test substance was used for intradermal injection.
Route:
epicutaneous, occlusive
Vehicle:
other: sesame oil
Concentration / amount:
100%
2.5x2.5 cm patch saturated with 0.5 g of test item
Day(s)/duration:
48 h
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
unchanged (no vehicle)
Concentration / amount:
100% and 50% (in sesame oil, safeguard dose)
2.5x2.5 cm patch saturated with 0.5 g of test item
Day(s)/duration:
24 h
Adequacy of challenge:
other: ?
No. of animals per dose:
Test group: 10 females (intradermal and epicutaneous main test)
Control group: 5 females (intradermal and epicutaneous main test)
6 females (intradermal and epicutaneous pretest)
Details on study design:
RANGE FINDING TESTS:
- intradermal injections:
site 1: 1% test substance (in sesame oil)
site 2: 2.5% test substance (in sesame oil)
site 3: 5% test substance (in sesame oil)
site 4: 5% test substance (in 1:1 mixture of TiterMax (TM) and physiological saline solution)
0.1 mL of each concentration were injected into shaved skin of two guinea pigs.

- epidermal applications: 25, 50, 75 % (in sesame oil) and 100% (undiluted). 0.5 mL of the test article (diluted) or 0.5 g (undiluted) were applied to hair-free scapular area (approximately 5x5 cm) of each of four guinea pigs via a sterile gauze patch (4 layers of porous gauze pads). A surrounding adhesive hypoallergenic plaster held the patches in place. The treated areas were covered for 48 hours with a fully occlusive foil (Closed Patch Test).
After the patch removal any remaining test substance was removed with 70% (w/v) ethanol using a gauze swab.
Local effects were examined and scored 1, 24, 48 and 72 hours after the treatment or after patch removal.

The highest concentration (5%) formulated in the vehicle caused no more than mild-to moderate erythema (score 1 or 2) during the observation period, therefore this concentration was chosen in the main study.
It was found that all the dermal treatments at the tested concentrations produced no reaction on the skin of guinea pigs.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous)
- Test groups:
Intradermal (three pairs of injections):
Injection 1: 1:1 mixture (v/v) of FCA and physiological saline solution
Injection 2: 5% test substance (in sesame oil)
Injection 3: 5 % test substance, emulsified in a 1:1 mixture (v/v) of FCA and physiological saline solution

Epicutaneous:
Since the undiluted test item was not skin irritant in the dermal dose range-finding study, the test area was painted with 0.5 mL of 10% sodium dodecyl sulphate in Vaseline
24 hours prior to the topical induction application. A 2.5x2.5 cm sterile gauze patch (4 layers of porous gauze pads) was saturated with approximately 0.5 g of the undiluted
test item and placed over the injection sites. A surrounding adhesive hypoallergenic plaster held the patches in place. The treated areas were covered for 48 hours with a
fully occlusive foil (Closed Patch Test).
After the patch removal any remaining test substance was removed with 70% (w/v) ethanol using a gauze swab.
Local effects were examined and scored 1, 24, 48 and 72 hours after the treatment or after patch removal.

- Control group:
Intradermal (three pairs of injections):
Injection 1: 1:1 mixture (v/v) of Freund's Complete Adjuvant abd physiological saline solution
Injection 2: sesame oil
Injection 3: sesame oil, emulsified in a 1:1 mixture (v/v) of Freund's Complete Adjuvant abd physiological saline solution

Epicutaneous:
The test area was painted with 0.5 mL of 10% sodium dodecyl sulphate in Vaseline 24 hours prior to the topical induction application. A 2.5x2.5 cm sterile gauze patch (4
layers of porous gauze pads) was saturated with 0.5 mL of sesame oil only and placed over the injection sites. A surrounding adhesive hypoallergenic plaster held the
patches in place. The treated areas were covered for 48 hours with a fully occlusive foil (Closed Patch Test).
Local effects were examined and scored 1, 24, 48 and 72 hours after the treatment or after patch removal.

- Site: scapular region
- Frequency of applications: every 7 days
- Duration: days 0-14
- Concentrations: 5% (intradermal) and 100% (epicutaneous)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: 24 h
- Test groups: test substance
- Control group: 100% test substance
- Site: shaved left and right side of respective animal
- Concentrations: 100% (left side) 50% (in sesame oil) (right side; safeguard dose)
- Evaluation (hr after challenge): 24 and 48 h
Challenge controls:
the control group is actually a challenge control
Positive control substance(s):
yes
Remarks:
2-Mercaptobenzothiazole (reliability study)
Positive control results:
The selection of dose levels for the reference item 2-Mercaptobenzothiazole was set on the basis of the previous reliability study.
Intradermal induction exposure: 1% (w/v) (in vehicle (1% methyl cellulose (in distilled water))
Dermal induction exposure: 75% (w/v) (in vehicle)
Challenge treatment: 50% (w/v) (in vehicle)

Challenge with reference item 2-Mercaptobenzothiazole resulted in a positive response in test animals previously sensitised. The net response values at the 24 and 48 hours observations represented an incidence rate of 80% and 70% and net score values of 0.80 and 0.70 respectively. In the control animals no visible changes were found after 24 or 48 hours of examinations following challenge with the reference item. The dermal scores represented discrete erythema (score 1) developed on the skin of sensitised guinea pigs.
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
intradermal induction: 0%; challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of systemic or local toxicity were observed in any animal. No mortality was observed during the study. No histopathological evaluation was performed after the sacrifice of the animals.
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
intradermal induction: 5% challenge: 100%
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
No signs of systemic or local toxicity were observed in any animal. No mortality was observed during the study. No histopathological evaluation was performed after the sacrifice of the animals.
Remarks on result:
other: Since the undiluted test item was not skin irritant in the dermal dose range-finding study, the test area was pre-treated with 0.5 mL of 10% SDS in Vaseline 24 hours prior to the topical induction application, in order to create a local irritation
Remarks:
Since the undiluted test item was not skin irritant in the dermal dose range-finding study, the test area was pre-treated with 0.5 mL of 10% SDS in Vaseline 24 hours prior to the topical induction application, in order to create a local irritation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
intradermal induction: 0% challenge: 100%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of systemic or local toxicity were observed in any animal. No mortality was observed during the study. No histopathological evaluation was performed after the sacrifice of the animals.
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
intradermal induction: 5% challenge: 100%
No. with + reactions:
10
Total no. in group:
10
Clinical observations:
No signs of systemic or local toxicity were observed in any animal. No mortality was observed during the study. No histopathological evaluation was performed after the sacrifice of the animals.
Remarks on result:
other: Since the undiluted test item was not skin irritant in the dermal dose range-finding study, the test area was pre-treated with 0.5 mL of 10% SDS in Vaseline 24 hours prior to the topical induction application, in order to create a local irritation
Remarks:
Since the undiluted test item was not skin irritant in the dermal dose range-finding study, the test area was pre-treated with 0.5 mL of 10% SDS in Vaseline 24 hours prior to the topical induction application, in order to create a local irritation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
intradermal induction: 1% challenge: 50%
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
no information
Remarks on result:
positive indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
intradermal induction: 1 challenge: 50%
No. with + reactions:
7
Total no. in group:
10
Clinical observations:
no information
Remarks on result:
positive indication of skin sensitisation

Pre-test

Table 1: Results after intradermal injection

Animal No.

Concentration (w/v) [%]

Reaction scores after [h]

1

24

48

72

E

O

E

O

E

O

E

O

1

5 (in FCA:saline)

2

0

2

0

2

0

2

0

2

2

0

2

0

2

0

1

0

1

5 (in vehicle = sesame oil)

1

0

1

0

1

0

1

0

2

1

0

2

0

2

0

1

0

1

2.5 (in vehicle)

1

0

1

0

0

0

0

0

2

1

0

1

0

0

0

0

0

1

1 (in vehicle)

0

0

0

0

0

0

0

0

2

0

0

1

0

0

0

0

0

E = Erythema

O = Oedema

Epicutaneous application

No local reactions (oedema or erythema) were visible after topical application of 25, 50, 75 or 100% test substance, neither after 1 nor after 24 or 48 or 72 h after the removal of the dressing.

Main study

Table 2: Skin response after intradermal induction with vehicle (sesame oil, control group) or test substance (5 % in sesame oil, test group)

Animal No.

Group

Reaction scores after 24 h

E

O

1

control

0

0

2

0

0

3

0

0

4

0

0

5

0

0

6

test

1

0

7

2

0

8

1

0

9

2

0

10

2

0

11

1

0

12

2

0

13

1

0

14

1

0

15

1

0

E = Erythema

O = Oedema

Skin response after epidermal application of the vehicle (sesame oil) or test substance (5 %) during induction period

Animals were treated with a 10 % sodium-lauryl-sulphate 24 h prior to the epidermal induction application.

No local reactions (oedema or erythema) were visible after topical application of the vehicle or 100% test substance, neither after 1 nor after 24 or 48 or 72 h after the removal of the dressing.

Challenge

Table 3: Skin response (Erythema) after challenge (treated with 100% or 50% test substance)

 

Group

Reaction scores after removal of the dressing

Reading time point

24 h

48 h

Concentration [%]

100

50

100

50

Animal No.

 

1

control

0

0

0

0

2

0

0

0

0

3

0

0

0

0

4

0

0

0

0

5

0

0

0

0

6

test

1

1

1

1

7

1

0

1

1

8

2

1

2

1

9

1

1

1

1

10

1

0

1

1

11

2

1

2

1

12

2

1

2

1

13

1

1

2

0

14

1

1

1

0

15

1

1

2

1

Mortality /viability/systemic effects

No mortalities or signs indicative for systemic or local toxicity were observed.

No histopathological evaluation was performed after the sacrifice of the animals

Clinical observations showed no abnormalities.

Body weights

Table 4: Body weights [g]

 

Animal No.

Day(s) relative to start date

-1

7

14

25

Control group

1

359

394

429

451

2

338

371

388

424

3

349

430

459

497

4

324

376

410

430

5

346

444

462

511

Mean

 

343.2

403.0

429.6

462.6

SD

13.1

32.6

31.7

39.4

Test group

6

358

407

422

442

7

345

405

416

459

8

353

376

401

419

9

329

355

374

416

10

349

377

427

463

11

356

378

403

424

12

340

400

416

445

13

361

374

398

433

14

346

390

402

430

15

325

382

404

440

Mean

 

346.2

384.4

406.3

437.1

SD

12.0

16.2

15.0

15.8

The body weight gain of the animals was not affected adversely during the study.

Interpretation of results:
Category 1A (indication of significant skin sensitising potential) based on GHS criteria
Conclusions:
The skin sensitization potential of Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs was evaluated in accordance to OECD 406 using guinea pigs.

In relation to challenge, after intradermal injection with 5% formulated in sesame oil and challenge with 100%, after 24 and 48 hours 10/10 animals has positive reactions (Erythema - Draize score 1 -2). At 50%, after 24 and 48 hours 8/10 animals has positive reactions (Erythema - Draize score 1).

Based on the observed effects of Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs, the substance is a skin sensitizer. Applying the CLP/ EU GHS criteria according to Regulation (EC) No 1272/2008, Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs is categorized as a strong sensitizer (sub-category 1A).
Executive summary:

The skin sensitization potential of Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs was evaluated in accordance to OECD 406 using guinea pigs.

No signs of systemic or local toxicity were observed in any animal. No mortality was observed during the study. No histopathological evaluation was performed after the sacrifice of the animals.

In relation to skin response after epidermal application of the vehicle (sesame oil) or test substance (5 %) during the induction period, no local reactions (oedema or erythema) were visible after topical application of the vehicle or 100% test substance, neither after 1 nor after 24 or 48 or 72 h after the removal of the dressing. It is noted that animals were treated with a 10 % sodium-lauryl-sulphate 24 h prior to the epidermal induction application.

In relation to challenge, after intradermal injection with 5% formulated in sesame oil and challenge with 100%, after 24 and 48 hours 10/10 animals has positive reactions (Erythema - Draize score 1 -2). At 50%, after 24 and 48 hours 8/10 animals has positive reactions (Erythema - Draize score 1).

Based on the observed effects of Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs, the substance is a skin sensitizer. Applying the CLP/ EU GHS criteria according to Regulation (EC) No 1272/2008, Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs is categorized as a strong sensitizer (sub-category 1A).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)

Respiratory sensitisation

Endpoint conclusion
Endpoint conclusion:
no study available (further information necessary)

Justification for classification or non-classification

The skin sensitization potential of Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs was evaluated in accordance to OECD 406 using guinea pigs.

In relation to challenge, after intradermal injection with 5% formulated in sesame oil and challenge with 100%, after 24 and 48 hours 10/10 animals has positive reactions (Erythema - Draize score 1 -2). At 50%, after 24 and 48 hours 8/10 animals has positive reactions (Erythema - Draize score 1).

Based on the observed effects of Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs, the substance is a skin sensitizer. Applying the CLP/ EU GHS criteria according to Regulation (EC) No 1272/2008, Reaction product of 2-Propenoic acid and Oxirane, mono[(C12-16-alkyloxy)methyl] derivs is categorized as a strong sensitizer (sub-category 1A).