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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP guideline study with acceptable restrictions. Split Adjuvant Test due to limited solubility, but no positive control included.
Justification for type of information:
Study was undertaken prior to the acceptance and development of the LLNA test.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report Date:
1986

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
no positive control substance included
GLP compliance:
yes
Type of study:
split adjuvant test
Justification for non-LLNA method:
Study was undertaken prior to the acceptance and development of the LLNA test.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: solid, not further specified

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Olac, Great Britain
- Age at study initiation: 2 months
- Weight at study initiation: 320-412 g
- Housing: 4 per cage in metal cages with wire-mesh floors (RUCO, Valkenswaard). Distal parts of both rear feet were wrapped with adhesive tape (Sleek, Smith and Nephew, England) to prevent skin injuries.
- Diet: Standard guinea pig diet including ascorbic acid (1600 mg/kg), obtained from Hope Farms, Woerden (LC 23-B, pellet diameter 4 mm), availability not specified. In addition, once a week hay was provided.
- Water: Not specified.
- Acclimation period: 11 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19.5-21.0
- Humidity (%): 50-80
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
corn oil
Remarks:
SAT: test substance in corn oil only applied epicutaneously, not injected
Concentration / amount:
Induction: 0.2 mL of 25% (w/v) on day 0 and 2; 5% (w/v) on day 4 and 7
Challenge: 50 µL of 25, 5, 1 and 0% (w/v) on day 21
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Remarks:
SAT: test substance in corn oil only applied epicutaneously, not injected
Concentration / amount:
Induction: 0.2 mL of 25% (w/v) on day 0 and 2; 5% (w/v) on day 4 and 7
Challenge: 50 µL of 25, 5, 1 and 0% (w/v) on day 21
No. of animals per dose:
20 females in test group, 10 in control group
Details on study design:
RANGE FINDING TESTS:
Before challenge phase a primary irritation test was carried out with 4 guinea pigs in order to find a concentration for challenge not inducing substantial irritation. Four guinea pigs were shaved and their left flank was exposed for 24 hours to 25, 10, 2.5 and 1% (w/v) of the test substance in corn oil under occlusive conditions; application was done in plastic Square chambers (v.d. Bend, Brielle, The Netherlands) provided with aluminium foil to avoid contact with the compound. Two animals died from bandage-induced stress, but the remaining animals showed no skin response to any of the tested concentrations.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 4
- Test groups: test substance in corn oil epicutaneously under occlusive conditions on day 0, 2, 4 and 7 (on day 0 treatment of skin with dry ice for 5 seconds prior to application); additionally 2 i.d. injections of 0.1 mL FCA on day 4 on either side of sensitisation area.
- Control group: 2 i.d. injections of 0.1 mL FCA on day 4 on either side of sensitisation area.
- Site: Shaved area of 2x2 cm on the back just behind the right shoulder girdle, injections were done on either side of the sensitisation area. The animals were wrapped in a "window dressing" consisting of Lenkelast elastic bandage (Lohmann, Neuwied, FRG) into which a 2x2 cm opening had previously been cut. This opening was placed over the sensitisation site. After applications the site was covered with a Metalline-patch (Lohmann), which was held in place by covering with water-impermeable tape (Sleek, Smith and Nephew, England).
- Frequency of applications: 2 days
- Duration: 8 days
- Concentrations: Test group: 0.2 mL of 25% (w/v) on day 0 and 2, and 5 % (w/v) on day 4 and 7

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 21
- Exposure period: 24 hours
- Test groups: test substance in corn oil under occlusive conditions
- Control group: test substance in corn oil under occlusive conditions
- Site: Application of test substance to the shaved left flank by patches consisting of 1 cm² filter paper (Schleicher & Schull), attached to a piece of aluminium foil with petrolatum, the aluminium foil being mounted on non-irritating Micropore tape (3M Co. St. Paul, USA). The animals were then wrapped in Coban elastic bandage (Smith and Nephew, England). The dressing was fixed in place for 24 hours.
- Concentrations: 50 µL of 25, 5, 1 and 0% (w/v)
- Evaluation (hr after challenge): 48 and 72 hours (24 and 48 hours after removal of the bandage and a close shave of the skin)
Challenge controls:
Control group also serves as challenge control, as injection of these animals with FCA alone on day 4 of the induction should not elicit any sensitivities.
Positive control substance(s):
no

Results and discussion

Positive control results:
No positive control was included.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
5%
No. with + reactions:
1
Total no. in group:
20
Clinical observations:
Moderate and diffuse reaction; two other animals demonstrated red spots (scattered reaction) which were not considered positive.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 5%. No with. + reactions: 1.0. Total no. in groups: 20.0. Clinical observations: Moderate and diffuse reaction; two other animals demonstrated red spots (scattered reaction) which were not considered positive..
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
1%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
5%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
1%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
0%
No. with + reactions:
0
Total no. in group:
20
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 20.0.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
Two animals demonstrated red spots (scattered reaction), which were not considered positive.
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: Two animals demonstrated red spots (scattered reaction), which were not considered positive..
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
One animal demonstrated red spots (scattered reaction) which were not considered positive. A second one could not be evaluated due to reddening from shaving
Remarks on result:
other: see Remark
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 9.0. Clinical observations: One animal demonstrated red spots (scattered reaction) which were not considered positive. A second one could not be evaluated due to reddening from shaving.
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
One animal could not be evaluated due to reddening from shaving.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 9.0. Clinical observations: One animal could not be evaluated due to reddening from shaving..
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 5%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
1%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 1%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 0%. No with. + reactions: 0.0. Total no. in groups: 10.0.

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Remarks:
Migrated information