Registration Dossier

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991-02-25 to 1991-04-05
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
cyclooctene of Hüls AG, Sample from B 803, ID No. 3630/81 288, produced December 1990.
Purity 96.2 % (GC-FID area).

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS: 
- Strain: Dunkin-Hartley, Pirbright White, BOR:DHPW (SPF)
- Sex: female
- Source: Fa. Winkelmann, Borchen (Germany)
- Age: healthy young adults
- Weight at study initiation: 464 g (mean test); 460 g, 457 g (means  control groups 1 and 2)
- Controls: 10 each for 1st and 2nd challenge; treatment: vehicle

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: corn oil (MEH 56)
Concentration / amount:
1st challenge: 100 % (0.20 ml) with patch on left side (control group:  1)  
2nd challenge: 10 % (0.12 ml) with patch on right side (control group:  2)
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: corn oil (MEH 56)
Concentration / amount:
1st challenge: 100 % (0.20 ml) with patch on left side (control group:  1)  
2nd challenge: 10 % (0.12 ml) with patch on right side (control group:  2)
No. of animals per dose:
test: 20
control: 10
Details on study design:
1st application: Induction 2.5 % intracutaneous
2nd application: Induction undiluted occlusive epicutaneous
3rd application: Challenge undiluted occlusive epicutaneous
ADMINISTRATION/EXPOSURE 
- Preparation of test substance for induction: clear solution in vehicle
- Induction schedule:   
day 0 injection,   
day 6 induction of slight to moderate skin irritation (10 % sodium  dodecyl sulfate in petrolatum)   
days 7-9 patch treatment (0.25 ml, 48 hours), readings 1 and 24 hours  after patch removal
- Injection details: 0.1 ml each at 6 positions on shoulders:   
2 x Freund's Complete Adjuvant / deionized water (50:50)   
2 x test substance 2.5 % in corn oil   
2 x test substance 2.5 % in Freund's Complete Adjuvant / corn oil  (50:50)   simultaneous and symmetrical application of each solution   
controls: corn oil instead of test substance
- Challenge schedule: 1st challenge on day 21, 2nd challenge on day 28   occlusive patch treatment for 24 hours, 
readings 24, 48, 72 hours after  administration
- Concentrations used for challenge:   
1st challenge: 100 % (0.20 ml) with patch on left side (control group:  1)   
2nd challenge: 10 % (0.12 ml) with patch on right side (control group:  2)
- Positive control: 1-chloro-2,4-dinitrobenzene
EXAMINATIONS
- Grading system: as usual for skin irritation, 0-8 scores possible
- Pilot study: dose finding (intracutaneous toxicity and skin irritation)   
intracutaneous doses: 0.1 ml x 0; 0.25; 0.50; 1.0; 2.5; 5.0; 10.0 %  w/w,    
2 animals each   dermal doses: 0.1 ml x 2.5; 25; 50; 100 % w/w  
4 animals each with 4 different concentrations at different sites   
24 hour occlusive patch test, readings 24 / 48 / 72 hours after  administration
Positive control substance(s):
yes
Remarks:
1-chloro-2,4-dinitrobenzene

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
100 %
No. with + reactions:
20
Total no. in group:
20
Clinical observations:
see below
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 100 %. No with. + reactions: 20.0. Total no. in groups: 20.0. Clinical observations: see below.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
100 %
No. with + reactions:
8
Total no. in group:
10
Clinical observations:
see below
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 100 %. No with. + reactions: 8.0. Total no. in groups: 10.0. Clinical observations: see below.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
100 %
No. with + reactions:
3
Total no. in group:
20
Clinical observations:
see below
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 100 %. No with. + reactions: 3.0. Total no. in groups: 20.0. Clinical observations: see below.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
100 %
No. with + reactions:
3
Total no. in group:
10
Clinical observations:
see below
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 100 %. No with. + reactions: 3.0. Total no. in groups: 10.0. Clinical observations: see below.
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no.
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 48.0. Group: negative control. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no.
Reading:
rechallenge
Hours after challenge:
72
Group:
test group
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
no
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 72.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: no.
Reading:
rechallenge
Hours after challenge:
72
Group:
negative control
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
no
Remarks on result:
other: Reading: rechallenge. . Hours after challenge: 72.0. Group: negative control. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: no.

Any other information on results incl. tables

RESULTS OF PILOT STUDY:
  2.5 % was chosen for intracutaneous induction
  100 % test substance caused no dermal irritation
RESULTS OF TEST
- Sensitization reaction: 
  1st challenge: inconclusive results due to irritation in both test and  control groups.
  2nd challenge: 0/20 animals positive at 48 hours and at 72 hours after  application = no sensitization, no animals positive in 

control group
- Clinical signs: No signs of systemic toxicity were observed.
  1st induction, FCA + water application sites: severe erythema, severe  edema, and deep wounds within 24 hours.
  1st induction, test substance + vehicle application sites: distinct  erythema, very slight edema within 1 hour; very slight erythema 

within 24  hours, edema were absent in 9/20 animals.
  1st induction, vehicle application sites: distinct erythema, very  slight edema within 1 hour; very slight erythema left in only 5/20  

animals and no more edema within 24 hours.
  1st induction, FCA + test substance application sites: distinct to  severe erythema, severe edema and (in 8/20 animals) deep 

wounds within 24  hours.
  1st induction, FCA + vehicle application sites: severe erythema and  edema and necrosis within 24 hours.
  2nd induction, FCA + water application sites: severe erythema and edema  wih deep wounds within 1 hour after patch removal; 

eschar formation  within 24 hours
  2nd induction, test substance + vehicle application sites: eschar  formation 1 and 24 hours after patch removal
  2nd induction, vehicle application sites: eschar formation 1 and 24  hours after patch removal
  2nd induction, FCA + test substance application sites: eschar formation  1 hour after patch removal, 1 animal with deep wounds 

and severe edema;  24 hours after patch removal eschar in 14/20 animals, severe edema and  crusts on heavy erythema with 

deep wounds in 6/20 animals.
  2nd induction, FCA + vehicle application sites: 1 hour after patch  removal eschar in 14/20 animals, severe erythema and edema with deep  wounds in 6/20 animals. 24 hours after patch removal eschar formation at  injection sites of 9/20 animals, 

severe edema and incrustation on severe  erythema / deep wounds in other 11 animals.
  1st challenge, test group: 24 hours after patch removal very slight to  distinct erythema and edema in all animals; 24 hours later 

the edema had  disappeared in 3/20 animals.
  1st challenge, control group 1: 24 hours after patch removal very  slight to distinct erythema and (in 8/10 animals) edema; 24 hours 

later  the edema were absent in 3/10 animals.
  2nd challenge: no signs of irritation in test group and control group 2.
- Other: Body weight development was similar in test and control groups.

Applicant's summary and conclusion

Conclusions:
No skin sensitization was induced by the test material. In this study, cyclooctene is not a dermal sensitizer.
Executive summary:

In a dermal sensitization study with cyclooctene in corn oil, was tested in a group of young Dunkin-Hartley guinea pigs using the method of Magnusson and Kligman. 1-Chloro-2,4-dinitrobenzene served as positive control material. No signs of systemic toxicity were noted.

No skin sensitization was induced by the test material. In this study, cyclooctene is not a dermal sensitizer.