Registration Dossier

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Sep 1996 - Oct 1996
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Version / remarks:
Jul 1992
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
No justification given.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source: TH Goldschmidt AG, Essen, Germany
- Batch No.of test material: 571003
- Active component: 35%
- Physical state: clear, viscous liquid
- Density: approx. 1.00 g/ml

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: ambient
- Stability under test conditions: March 1997

OTHER SPECIFICS:
- Solvent/vehicle: intradermal injection and dermal application: aqua ad iniect.

In vivo test system

Test animals

Species:
guinea pig
Strain:
Pirbright-Hartley
Remarks:
white. Substrain: Hsd/Poc: DH
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan-Winkelmann GmbH, 33178 Borchen, Germany
- Date of receipt: August 7, 1996 (range finding); September 18, 1996 (main test)
- Weight at study initiation: male 308 - 349 g, female 295 - 339 g (main test)
- Housing: collective housing up to a maximum of 10 animals per cage
- Diet: ad libitum, Ssniff-G (Ssniff Spezialdiäten GmBh, Soest, Germany)
- Water: ad libitum
- Acclimation period: 35 days (range finding), 8 days (main test)

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3
- Humidity (%): 30 - 70
- Photoperiod (hrs dark / hrs light): 12/12 (illumination from 7 am to 7 pm)
- Measurement: twice daily

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
water
Concentration / amount:
2.5%
Day(s)/duration:
7 days
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
50%
Day(s)/duration:
48 h
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
5%
Day(s)/duration:
24 h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
20 animals used for the test substance and 10 control animals
Details on study design:
RANGE FINDING TESTS:
A. INTRADERMAL INJECTION
- Concentration: 5%, dilution with aqua ad iniect. and Freund's complete adjuvant (FCA). Since this concentration produced immoderate irritation such as grey-black discoloration, lower concenrations were tested.
- 4 animals were employed and skin reactions were recorded 48 h post application

B. DERMAL APPLICATION
- the test article was used undiluted. Since the maximum concentration was irritating, lower concentrations were tested.
- occlusive bandage was used
- 10 animals were employed and skin reactions were recorded 48 h post application

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 1 intradermal, 1 epicutaneous
- Exposure period: - (id), 48 hours (epic)
- Test groups: Test subsstance, Test substance + FCA, FCA (id); Test substance (epic.)
- Control group: FCA, water+FCA, water (id); water (epic.)
- Site: shoulder
- Frequency of applications: 0 (id), 7d (epic.)
- Duration: 0-7d
- Concentrations: 2.5% (id), 50% (epic.)

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 14d
- Exposure period: 24 h
- Test groups: test substance
- Control group: test substance
- Site: left flank test substance, right flank water
- Concentrations: 5%
- Evaluation (hr after challenge): 24, 48 h
Challenge controls:
Control animals were treated with the test substance concurrently with the test groups in the challenge phase.
Positive control substance(s):
no
Remarks:
The reaction to the positive control substance (benzocaine) is tested periodically. The last test was performed in June 1996.

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
5%
No. with + reactions:
12
Total no. in group:
20
Clinical observations:
Skin reactions were observed
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
13
Total no. in group:
20
Clinical observations:
Skin reactions were observed
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
5%
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
Skin reactions were observed
Remarks on result:
no indication of skin sensitisation
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
5%
No. with + reactions:
2
Total no. in group:
10
Clinical observations:
Skin reactions were observed
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

Table 1: Challenge exposure in the test group - results

 

24h

 

Animal

 

Initial

Test

 

Test

 

Final

no.

Sex

BW(g)

article

E Oe

Vehicle

E Oe

article

E Oe

Vehicle

E Oe

BW(g)

  

1

m

321

0

0

0

0

0

0

0

0

489

2

m

343

0

0

0

0

0

0

0

0

580

3

m

349

1

0

0

0

1

0

0

0

543

4

m

316

1

0

0

0

1

0

0

0

490

5

m

317

2

0

0

0

1

0

0

0

517

6

m

320

0

0

0

0

1

0

0

0

532

7

m

326

0

0

0

0

1

0

0

0

476

8

m

329

1

0

0

0

0

0

0

0

516

9

m

308

1

0

0

0

2

0

0

0

484

10

m

322

0

0

0

0

0

0

0

0

489

11

f

303

1

0

0

0

1

0

0

0

441

12

f

337

1

0

0

0

1

0

0

0

518

13

f

295

2

0

0

0

2

0

0

0

411

14

f

323

0

0

0

0

0

0

0

0

459

15

f

314

2

0

0

0

1

0

0

0

468

16

f

326

0

0

0

0

0

0

0

0

485

17

f

322

0

0

0

0

0

0

0

0

480

18

f

313

1

0

0

0

1

0

0

0

446

19

f

318

1

0

0

0

1

0

0

0

436

20

f

305

2

1

0

0

2

1

0

0

430

 

Reaction rate at 24 h: 60%

Sensitization rate at 24 h: 20%

 

Reaction rate at 48 h: 65%

Sensitization rate at 48 h: 15%

Severity index:

test article (leftflank):

mean value (E) 24 h: 0.80

mean value (Oe) 24 h: 0.05

mean value (E) 48 h: 0.80

mean value (Oe) 48 h: 0.05

 

Severity index: vehicle (rightflank):

mean value(E)24 h: 0.00

mean value (Oe) 24 h: 0.00

mean value(E)48 h: 0.00

mean value (Oe) 48 h: 0.00

 

m=male; f=female; BW=body weight; E=erythema; Oe=oedema

Table 2: Challenge exposure in the control group - results

 

Animal no.

 

Initial SexBW(g)

24hTest

article

E Oe

 

Vehicle

E Oe

48hTest

article

E Oe

 

Vehicle

E Oe

 

Final BW(g)

1

m

324

0

0

0

0

0

0

0

0

522

2

m

320

0

0

0

0

0

0

0

0

518

3

m

317

0

0

0

0

0

0

0

0

471

4

m

314

0

0

0

0

0

0

0

0

484

5

m

310

0

0

0

0

0

0

0

0

468

6

f

303

0

0

0

0

0

0

0

0

509

7

f

339

1

0

0

0

1

0

0

0

511

8

f

317

0

0

0

0

0

0

0

0

508

9

f

327

1

0

0

0

1

0

0

0

500

l0

f

315

0

0

0

0

0

0

0

0

525

Table 2.2.: Results belonging to results presented in table 2

Reaction rate at 24 h:

 

Severity index:

test article (leftflank):

20%

Reaction rate at 48 h:

 

Severity index: vehicle (rightflank):

20%

mean value (E) 24 h:

0.20

mean value(E)24 h:

0.00

mean value (Oe) 24 h:

0.00

mean value (Oe) 24 h:

0.00

mean value(E)48 h:

0.20

mean value(E)48 h:

0.00

mean value (Oe) 48 h:

0.00

mean value (Oe) 48 h:

0.00

 

m=male; f=female; BW=bodyweight; E=erythema; Oe=oedema

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
According to the maximization scheme of Magnusson and Kligman (1996), the test substance is classified as a mild sensitzier (The sensitization rate at 24 h was 20 % and at 48 h 15 %).
According to GHS, EU criteria the substance has not to be classfied as skin sensitizer.
Executive summary:

The potential skin sensitizing potential of the test substance was assessed in the guinea pig maximization test using 20 test and 10 control animals. Following induction exposure to the test substance or the vehicle, the animals were subjected to two weeks later to a challenge a challenge exposure with the test substance. The treated skin areas were evaluated 24 and 48 hours after the end of the exposure period. According to Magnusson and Kligman (J. Invest. Dermatol. 52, 268 -276, 1969), the test substance is classified as a "mild sensitizer".