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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
September to December 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1997
Report Date:
1997

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study already available

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): Javanol
- Physical state: Pale yellow viscous liquid
- Analytical purity: 88.4% (GC, sum of two peaks)
- Lot/batch No.: 008825
Specific details on test material used for the study:
Name of test material (as cited in study report): Javanol
- Physical state: Pale yellow viscous liquid
- Analytical purity: 88.4% (GC, sum of two peaks)
- Lot/batch No.: 008825

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
Age at delivery: 4 - 6 weeks
Age at beginning of pre-test/acclimatization period: 5-7 weeks
Body weight at pretest start: 361-369g (pretest group)
Body weight at beggining of acclimatization period: 310-431g (control and test group)
Acclimatization: One week for the control and test group under test conditions after health examination. No acclimatization for the animals of the pretest. Only animals without any visible sigs of illness were used.
Standard laboratory conditions: 10-15 air changes per hour, temeprature target at 22 +/- 3°C and relative humidity between 40-70% (values above 70% during cleaning process possible). 12 hour dark/light cycle. Recorded music was played for approximately 8 hours during the light period.
Accomodation: indivudally in Makrolon type-3 cages with standard softwood bedding.
Diet: Pelleted standard Nafag Ecosan 845 25W4, batch nos 58/97, 69/97 and 80/97 guinea pig breeding / maintenance diet, ad libitum.
Water: Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid (approx. 1 g/l) via the drinking water was provided.

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
polyethylene glycol
Remarks:
400
Concentration / amount:
intradermal: 5%
epicutaneous: 100%
Day(s)/duration:
intradermal at day 1, epicutaneous at day 8
Challengeopen allclose all
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Concentration / amount:
25%
Day(s)/duration:
At day 22
Adequacy of challenge:
highest non-irritant concentration
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Concentration / amount:
5%
Day(s)/duration:
At day 29
No.:
#2
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Concentration / amount:
10%
Day(s)/duration:
At day 29
No. of animals per dose:
15 females for main study and 3 females for pretest
Details on study design:
The intradermal induction of sensitization was carried out with a 5% dilution of the test article in polyethylene glycol (PEG400) and in an emulsion with Freund's Complete Adjuvant (FCA) / physiological saline. The epidermal induction of sensitization was conducted under occlusion with the undiluted test article (100%). Two weeks after the epidermal induction application the first challenge was completed by epidermal application of the test article at 25% in PEG 400 under occlusive dressing. The animals of the control group were induced with PEG400 and FCA/physiological saline and challenged similarly to those of the test group. Cutaneous reactions, i.e. erythema and eschar, as well as oedema formation were evaluated at 24 and 48 hours after removal the dressing.
As positive skin reactions were observed in the control group after the first challenge, a second challenge was performed in the control and test group one week after the first challenge using two lower test article concentrations of 10 and 5%.
Positive control substance(s):
yes

Results and discussion

Positive control results:
A positive skin reactions were observed in the control group after the first challenge. After the second challenge performed one week after the first one, no toxic symptoms were evident in the guinea pigs of the control or test group.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25%
No. with + reactions:
7
Total no. in group:
10
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
7
Total no. in group:
10
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25%
No. with + reactions:
4
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25%
No. with + reactions:
5
Total no. in group:
5
Reading:
rechallenge
Hours after challenge:
24
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Reading:
rechallenge
Hours after challenge:
48
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
5
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
5
Key result
Reading:
rechallenge
Hours after challenge:
24
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
10
Key result
Reading:
rechallenge
Hours after challenge:
48
Group:
test group
Dose level:
10%
No. with + reactions:
0
Total no. in group:
10

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Positive skin reactions were observed in the control and test group after the first challenge performed with the test article at 25% in PEG 400, considered to be of primary toxic origin (irritation).
At the second challenge no skin reactions were observed in the same control and test group treated with two lower concentrations of 10% and 5% in PEG 400.
Therefore, the test article Javanol applied at 10% and 5% in PEG 400 is considered not to be a sensitizer when used under the described test conditions.
Executive summary:

In order to assess the cutaneous allergenic potential of Javanol, the Maximization-Test in accordance with OECD Guideline No. 406 and the directive 96/54 /EEC, B.6 was carried out in 15 (10 test and 5 control) female Albino guinea pigs.

The intradermal induction of sensitization was carried out with a 5% dilution of the test article in polyethylene glycol (PEG400) and in an emulsion with Freund's Complete Adjuvant (FCA) / physiological saline. The epidermal induction of sensitization was conducted under occlusion with the undiluted test article (100%). Two weeks after the epidermal induction application the first challenge was completed by epidermal application of the test article at 25% in PEG 400 under occlusive dressing. The animals of the control group were induced with PEG400 and FCA/physiological saline and challenged similarly to those of the test group. Cutaneous reactions, i.e. erythema and eschar, as well as oedema formation were evaluated at 24 and 48 hours after removal the dressing.

Erythematous reactions after the first challenge procedure:

 positive / total after 24 hours  positive / total after 48 hours
 Control group - Javanol 25%  4/5  5/5
 Control group - PEG 400 only  0/5  0/5
 Test group - Javanol 25%  7/10  7/10
 Test group - PEG 400 only  0/10

 0/10

As positive skin reactions were observed in the control group after the first challenge, a second challenge was performed in the control and test group one week after the first challenge using two lower test article concentrations of 10 and 5%.

Erythematous reactions after the second challenge procedure:

 positive / total after 24 hours  positive / total after 48 hours
 Control group - Javanol 10%  0/5  0/5
 Control group - Javanol 5%  0/5  0/5
 Test group - Javanol 10%  0/10  0/10
 Test group - Javanol 5%  0/10

 0/10

No toxic symptoms were evident in the guinea pigs of the control or test group.

No deaths occured.

Therefore, the test article Javanol applied at 10% and 5% in PEG 400 is considered not to be a sensitizer when used under the described test conditions.