Registration Dossier

Diss Factsheets

Toxicological information

Skin sensitisation

Currently viewing:

Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11.06.80- 12.11.81
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Meets generally accepted scientific standards and acceptable for assessment.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1981
Report date:
1981

Materials and methods

Test guideline
Qualifier:
equivalent or similar to guideline
Guideline:
other: Magnusson, B. and Kligman, A.M. 1970. Allergic Contact Dermatitis in the Guinea Pig: Identification of Contact Allergens. C.C. Thomas, Springfield, Illinois, U.S.A
Principles of method if other than guideline:
Sensitisation was induced in guinea pigs by intradermal injections of both test substance and Complete Freunds Adjuvant and the induction process supplemented 7 days later by test substance applied to the shoulder injection sites under occluded patch: further challenges were made at weekly intervals as required.
GLP compliance:
not specified
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
LLNA not available at time of testing

Test material

Constituent 1
Chemical structure
Reference substance name:
Pentyl salicylate
EC Number:
218-080-2
EC Name:
Pentyl salicylate
Cas Number:
2050-08-0
Molecular formula:
C12H16O3
IUPAC Name:
pentyl salicylate
Constituent 2
Chemical structure
Reference substance name:
2-methylbutyl salicylate
EC Number:
256-972-3
EC Name:
2-methylbutyl salicylate
Cas Number:
51115-63-0
Molecular formula:
C12H16O3
IUPAC Name:
2-methylbutyl 2-hydroxybenzoate
Test material form:
liquid
Specific details on test material used for the study:
- Name of test material (as cited in study report): amyl salicylate
- Structural formula attached as image file (if other than submission substance): see Fig.

In vivo test system

Test animals

Species:
guinea pig
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Environmental Safety Division
- Weight at study initiation: approximately 344 g- 364 g
- Water (e.g. ad libitum): ad libitum

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
other: Injection induction: 0.01 % dobs/ saline. Application induction and application challenge: acetone.
Concentration / amount:
Induction (intradermal injection): 1%
Induction (covered patch application): 40%
challenge (covered patch application): 10%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
other: Injection induction: 0.01 % dobs/ saline. Application induction and application challenge: acetone.
Concentration / amount:
Induction (intradermal injection): 1%
Induction (covered patch application): 40%
challenge (covered patch application): 10%
No. of animals per dose:
6 males and 4 females
Details on study design:
RANGE FINDING TESTS: Animals were treated by intradermal injections in the shoulder region to induce sensitisation and 7 days later the sensitisation was boosted by an occluded patch placed over the injection site. Fourteen days later the animals were challenged on 1 flank by occluded patch. Seven days after this a further confirmatory challenge was given on the opposite flank using the same method.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2, intradermal injection followed 1 week later by topical application
- Exposure period: topical application 48 hrs
- Test groups: 1
- Control group: 2 types of control groups are used. Treated controls: 4 animals of the same sex are selected and treated controls for the first challenge. They are given a mock induction treatment at the same time and in the same way as for the test animals except that test substance is omitted from the injection and application preparations. Untreated control: 4 previously untreated animals of the same sex and weighing approximately the same as the test animals.
- Site: 2x4 cm are in the dorsal shoulder region.
- Frequency of applications: Intradermal injection: 3 pairs of intradermal injections are made within the site as follows (i) two 0.1 mL injections of 50 % Freund's complete adjuvant (FCA) in the solvent chosen for the test substance. (ii) two 0.1 mL injections of test substance at the concentration selected for the induction from the preliminary irritation test. (iii) two 0.1 mL injections of the test substance in solvent mixed 50/50 with FCA such that the final concentration of test substance injected is the same as that in (ii). Topical application: 1 week after the injections the same 2x4 cm area was clipped and shaved. A 2x4 cm filter paper patch attached by double-sided adhesive tape to a 4x6 cm piece of thin polythene was saturated with test substance and placed over the shaved site. The patch was held in place by adhexive plaster wrapped around the trunk behind the forelimbs.
- Duration: 13 to 14 days
- Concentrations: intradermal injection 1 %, covered patch application 40 %

B. CHALLENGE EXPOSURE
- No. of exposures: 3
- Day(s) of challenge:
- Exposure period: 24 hrs
- Test groups: 1
- Control group: Treated controls: At first challenge they are treated in exactly the same way as the test animals. Untreated controls: At every challenge in the test are treated in exactly the same way as the test animals.
- Site: flank. For each animal, an 8 mm diameter filter paper patch in am 11 mm aluminium patch test cup was saturated with the test substance at the selected challenge concentration and applied to the site. The patch was held in place by adhesive plaster wound around the trunk.
- Concentrations: 10 %
- Evaluation (hr after challenge): 24 and 48 hrs
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
4 animals were noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: 4 animals were noted to have barely perceptible erythema..
Reading:
1st reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
1 animal was noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: 1 animal was noted to have barely perceptible erythema..
Reading:
2nd reading
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
1 animal was noted to have spots of erythema.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: 1 animal was noted to have spots of erythema..
Reading:
2nd reading
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: Challenge 3
Hours after challenge:
24
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: Challenge 3. . Hours after challenge: 24.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
other: Challenge 3
Hours after challenge:
48
Group:
test chemical
Dose level:
10 %
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: other: Challenge 3. . Hours after challenge: 48.0. Group: test group. Dose level: 10 %. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
other: Treated controls
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
3 animals were noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: Treated controls. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: 3 animals were noted to have barely perceptible erythema..
Reading:
1st reading
Hours after challenge:
48
Group:
other: Treated control
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
2 animals were noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: other: Treated control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: 2 animals were noted to have barely perceptible erythema..
Reading:
1st reading
Hours after challenge:
24
Group:
other: Untreated control
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
1 animal was noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: other: Untreated control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: 1 animal was noted to have barely perceptible erythema..
Reading:
1st reading
Hours after challenge:
48
Group:
other: Untreated control
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
1 animal was noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 48.0. Group: other: Untreated control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: 1 animal was noted to have barely perceptible erythema..
Reading:
2nd reading
Hours after challenge:
24
Group:
other: Untreated control
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Clinical observations:
1 animal was noted to have barely perceptible erythema.
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 24.0. Group: other: Untreated control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0. Clinical observations: 1 animal was noted to have barely perceptible erythema..
Reading:
2nd reading
Hours after challenge:
48
Group:
other: Untreated control
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: other: Untreated control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0.
Reading:
other: Challenge 3
Hours after challenge:
24
Group:
other: Untreated controls
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Remarks on result:
other: Reading: other: Challenge 3. . Hours after challenge: 24.0. Group: other: Untreated controls. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0.
Reading:
other: Challenge 3
Hours after challenge:
48
Group:
other: Untreated control
Dose level:
0
No. with + reactions:
0
Total no. in group:
4
Remarks on result:
other: Reading: other: Challenge 3. . Hours after challenge: 48.0. Group: other: Untreated control. Dose level: 0. No with. + reactions: 0.0. Total no. in groups: 4.0.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The test substance was assessed for skin sensitisation according to the method described in Magnusson and Kligman (1970). Sensitisation was induced in guinea pigs by intradermal injections and supplemented 7 days later by an occluded patch. Three challenges were then given by occluded patch. None of the animals sensitised after 3 challenges and so the test substance is not considered to be a sensitiser by this method.
Executive summary:

Male and female Dunkin-Hartley guinea pigs were purchased from Environmental Safety Division. A preliminary, dose range-finding study was conducted on 4 male guinea pigs weighing 360 g- 428 g with the test substance being administered via intradermal injection. A preliminary, dose range-finding study was conducted on 4 male guinea pigs weighing 454 g- 546 g with the test substance being administered via an occluded patch.

The main study was conducted using 1 group of 10 guinea pigs, composed of 6 males and 4 females. Sensitisation was induced by intradermal injections and supplemented 7 days later by an occluded patch at dose levels 1 % and 40 % respectively. Three challenges were then given by occluded patch at the dose level 10 %. None of the animals sensitised after 3 challenges and so the test substance is not considered to be a sensitiser by this method.

All animals were observed at 24 and 48 hours after each challenge; reactions were noted.

There were no positive results for sensitization after 3 challenges. Four animals were noted to have barely perceptible erythema at 24 hours after the first challenge. One animal was noted to have barely perceptible erythema at 48 hours after the first challenge. One animal was noted to have barely perceptible erythema at 24 hours after the second challenge.

None of the animals sensitised after 3 challenges and so the test substance is not considered to be a sensitiser by this method.