Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
4 April to 13 May 1995
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline and GLP-compliant study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1995
Report Date:
1995

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): BEPD
- Physical state: colourless solid
- Analytical purity: 99.5%
- Storage conditions: at room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
Male albino guinea-pigs obtained from D. Hall, Newchurch. Staffordshire, England. They were approximately four to five weeks old and in a weight range of 301 to 339 g on arrival. The animal room temperature was maintained at 21°C with a relative humidity at 30-70%. Air exchange was maintained at 15 air changes/hour and lighting was controlled by a time switch to give 12 hours of artificial light (7:00 - 19:00) in each 24 hour period.

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Alembicol D
Concentration / amount:
2.5% v/v in Alembicol D (intradermal induction application)
100% (topical induction application)
100 % and 50% v/v in Alembicol D (topical challenge)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Alembicol D
Concentration / amount:
2.5% v/v in Alembicol D (intradermal induction application)
100% (topical induction application)
100 % and 50% v/v in Alembicol D (topical challenge)
No. of animals per dose:
10 test animals
5 control animals
Details on study design:
MAIN STUDY
A. INDUCTION EXPOSURE
3 pairs of intradermal injections: 1). Freund's complete adjuvant diluted with equal volume of water for irrigation (Ph. Eur.)
2). 2.5% BEPD v/v in Alembicol D
3). 2.5% BEPD v/v in a 50:50 mixture of Freund's complete adjuvant and Alembicol D.
6 days after the injections, the same site was shorn and pre-treated with 0.5 mL 10% w/w sodium lauryl sulphate in petroleum. 24 hours later, a patch saturated with 0.4 mL BEPD was applied and held in place for a further 48 hours.
- Test group: 10 animals
- Control group: 5 animals

B. CHALLENGE EXPOSURE
0.2 mL BEPD was applied to the anterior site. 50% BEPD v/v in Alembicol D was applied to the posterior site. The patches were held in place for 24 hours.
Challenge controls:
None
Positive control substance(s):
yes
Remarks:
hexyl cinnamic aldehyde

Results and discussion

Positive control results:
The results of regular studies with the positive control HCA (reliability checks) confirm the sensitivity of the test method (80-100% response).

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
100%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
100%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
50%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: No signs of ill health or toxicity were recorded.
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
50%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
No signs of ill health or toxicity were recorded
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 50%. No with. + reactions: 0.0. Total no. in groups: 5.0. Clinical observations: No signs of ill health or toxicity were recorded.

Any other information on results incl. tables

No reactions were observed in any group after challenge application with BEPD.

Dermal reactions observed after each induction:

Site

Intradermal injections

Topical applications

 

Test animals

Control animals

Test animals

Control animals

1

Necrosis

Necrosis

 

Slight erythema

 

Slight erythema

2

Slight irritation

Slight irritation

3

Necrosis

Necrosis

1: 0.1 mL Freund’s complete adjuvant 50:50 with sterile water for irrigation (Ph. Eur.)

2: 0.1 mL 2.5% BEPD v/v in Alembicol D

3: 0.1 mL 2.5% v/v in a 50:50 mixture of Alembicol D and Freund’s complete adjuvant

Bodyweights

Group

Guinea-pig number

Day 1

18 April 1995

Last observation day

13 May 1995

 

 

Control

1205

488

759

1206

440

585

1207

450

638

1208

453

597

1209

492

728

 

 

 

 

 

Test

1210

477

723

1211

462

638

1212

464

665

1213

477

686

1214

443

639

1215

441

648

1216

475

644

1217

433

626

1218

436

603

1219

454

650

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
No evidence of sensitisation was seen under the conditions of this study.  BEPD does not require classification for skin sensitisation.
Executive summary:

The potential of BEPD to induce delayed contact hypersensitivity (skin sensitisation) was investigated in a Maximisation test using male Dunkin-Hartley guinea-pigs. Concentrations used for induction and challenge applications were based on the results of a preliminary study. Intradermal injection was performed on test animals (10) using paired injections of the test material (2.5% BEPD v/v in Alembicol D, 2.5% BEPD v/v in a 50:50 mixture of Freund's Complete Adjuvant and Alembicol D) and FCA diluted with equal volume of water. Topical application was performed using 48 -hour occlusive application of the test material as supplied; local dermal irritation at the application site had previously been induced by topical application of 10% w/w sodium lauryl sulphate in petroleum. Control animals (5) were similarly treated during the induction phase, with vehicle in place of the test material. All test and control animals were challenged by 24 -hour occlusive application of the test material as supplied and as dilution (50% BEPD v/v in Alembicol D). No dermal reactions were observed at 24, 48 and 72 hours following the challenge application in test or control animals. No evidence of sensitisation was seen under the conditions of this study.  BEPD does not require classification for skin sensitisation.