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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:
February 19, 1997 through April 4, 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
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
RCC Holding Company Ltd.; 4414 Füllinsdorf, Switzerland
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study was performed before LLNA was formally validated and available as OECD guideline in 2002.

Test material

Constituent 1
Details on test material:
- Name of test material (as cited in study report): CFG-C-1607
- Physical state: yellow solid
- Analytical purity: >98%
- Lot/batch No.: 002
- Expiration date of the lot/batch: 31 Aug 1998
- Stability under test conditions: stable in PEG 400 for at least 48 hours
- Storage condition of test material: in the original container at room temperature away from direct sunlight

In vivo test system

Test animals

Species:
guinea pig
Strain:
Himalayan
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: BRL, Biological Research Laboratories Ltd., Wolferstrasse 4, CH-4414 Fullinsdorf / Switzerland
- Age at study initiation: 5 - 7 weeks
- Weight at study initiation: 328 - 406 g
- Housing: Individually in Makrolon type-3 cages with standard softwood bedding ("Lignocel", Schill AG, CH-4132 Muttenz)
- Diet (e.g. ad libitum): Pelleted standard Nafag Ecosan 845 25W4, batch nos. 01/97 and 10/97 guinea pig breeding 1 maintenance diet ("Nafag II , Nahr- und Futtermittel AG, CH-9202 Gossau), ad libitum.
- Water (e.g. ad libitum): Community tap water from Itingen, ad libitum. Once weekly additional supply of ascorbic acid (approx. 1 gil) via the drinking water was provided.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 40-70 %
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12-hour light, 12-hour dark cycle

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Concentration / amount:
3%
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Concentration / amount:
30%
Day(s)/duration:
48h
Adequacy of induction:
other: Non irritant test substance but animals were pretreated with 10% SLS
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
polyethylene glycol
Remarks:
PEG 400
Concentration / amount:
30%
Day(s)/duration:
24h
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
5 animals in control group, 10 animals in test group, 1 animal in intradermal pretest, 2 animals in epidermal pretest
Details on study design:
RANGE FINDING TESTS:
A pretest was performed during the acclimatization period. The concentrations tested were for the epicutaneous application to assure an optimum technical application procedure and for the intracutaneous injection the selected concentrations were tested up to 5 % (intradermal) and up to 30% (epicutaneous).

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epidermal)
- Test groups: Intradermal induction: 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline, test article diluted to 3% with PEG 400, the test article diluted to 3% by emulsion in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline; Epidermal induction: 30%
- Control group: Intradermal induction: 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline, PEG 400, 1:1 (w/w) mixture of PEG 400 in a 1:1 (v/v) mixture of Freund's Complete Adjuvant and physiological saline; Epidermal induction: PEG 400
- Site: dorsal skin in the scapular region
- Frequency of applications: intradermal injection once on day 1 and epidermal application once on day 8
- Duration:epidermal application for 48 hours
- Concentrations: 3% in intradermal injection and 30% in epicutaneous application

B. CHALLENGE EXPOSURE
- No. of exposures: one epidermal exposure
- Day(s) of challenge: Day 22
- Exposure period: challenge was two weeks after the epidermal induction application for 24h.
- Test groups: 30% and PEG 400
- Control group: 30% and PEG 400
- Site: left and right flank of each guinea pig
- Concentrations:30%
- Evaluation (hr after challenge): 24 hours and 48 hours after removal of the dressing
Challenge controls:
- Control group: 30% and PEG 400
Positive control substance(s):
yes
Remarks:
alpha-hexylcinnamaldehyde

Results and discussion

Positive control results:
In this study 70 % of the animals of the positive control group were observed with positive skin reactions after treatment of the positive control in 25 % polyethylene glycol (PEG 400). No skin reactions were observed in the control group.

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
30% in PEG 400
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
30% in PEG 400
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25% in PEG 400
No. with + reactions:
7
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
positive control
Dose level:
25% in PEG 400
No. with + reactions:
7
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
5
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
vehicle
No. with + reactions:
0
Total no. in group:
5

Any other information on results incl. tables

As there were no deaths during the course of the treatment period no necropsies were performed.

No symptoms of systemic toxicity were observed in the animals.

The body weight of the animals was within the range of physiological variability known for this strain and age.

Pretest:

Table 1. Reaction Readings from intradermal injection - Pretest.

Animal No.

Sex

Concentration (%)

Reaction Readings after 24 hours

Erythema

Oedema

Diameter (mm)

552

M

5

1

1

7 x 7

 

 

3

1

1

7 x 7

 

 

1

1

1

7 x 8

Table 2. Reactions from epidermal application - Pretest.

Animal No.

Sex

Concentration (%)

Reaction Readings after Removal of Bandage

After 24 hours

After 48 hours

E

Oe

E

Oe

553

M

30

0

0

0

0

25

0

0

0

0

15

0

0

0

0

10

0

0

0

0

554

M

10

0

0

0

0

30

0

0

0

0

25

0

0

0

0

15

0

0

0

0

E = Erythema

Oe = Oedema

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Conclusions:
In this study 0 % of the animals of the test group were observed with positive skin reactions after treatment with a non-irritant test article concentration of 30 % in PEG 400. No skin reactions were observed in the control group. Therefore, the test article CGF-C-1607 applied at a concentration of 30 % 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 CGF-C-1607, the Maximization-Test in accordance with OECD Guideline No. 406 and the Directive 96/54IEEC, B.6 was carried out in 15 (10 test and 5 control) male Himalayan guinea pigs. The intradermal induction of sensitization was carried out with a 3 % dilution of the test article in PEG 400 and in an emulsion with Freund's Complete Adjuvant (FCA) / physiological saline. The epicutaneous induction of sensitization was conducted under occlusion with the test article at 30 % in PEG 400. Two weeks after the epicutaneous induction application the challenge was completed by epicutaneous application of the test article at 30 % in PEG 400 under occlusive dressing. The animals of the control group were induced with PEG 400 and FCA/physiological saline and challenged similarly as 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 of the dressing. No toxic symptoms were evident in the guinea pigs of the control or test group. No deaths occurred. In this study 0 % of the animals of the test group were observed with positive skin reactions after treatment with a non-irritant test article concentration of 30 % in PEG 400. No skin reactions were observed in the control group.