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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:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1994
Report Date:
1994

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A valid GPMT was performed according to guideline and GLP - no further data is necessary

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Physical state: tan powder
- Stability under test conditions: stable
- Storage condition of test material: room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Sprague Dawley Inc., Houston, Texas
- Age at study initiation: no data
- Weight at study initiation: Males 432-476 g; Females 380-495 g (Test Group); Males 420-495 g; Females 405-491 g (Control Group)
- Housing: single housing (cage: suspended, wire bottom, stainless steel)
- Diet: Purina Guinea Pig Chow; ad libitum
- Water: municipal water, ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22+/-2
- Humidity (%): 30-80
- Air changes (per hr): 10-12
- Photoperiod: 12 hrs dark / 12 hrs light

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
cotton seed oil
Concentration / amount:
5%
Route:
epicutaneous, occlusive
Vehicle:
cotton seed oil
Concentration / amount:
patch saturated with moistened test substance
Adequacy of induction:
non-irritant substance, but skin pre-treated with 10% SDS
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
cotton seed oil
Concentration / amount:
100%
No. of animals per dose:
Control: 5 animals per sex
Dose groups: 10 animals per sex
Details on study design:
RANGE FINDING TESTS:
A screening test was conducted with 3 males and 3 females prior to the definitive study. Five percent (w/v) test material in cottonseed oil was selected for intradermal injection. For the topical insult, a dose of 500 mg of test material moistened with petrolatum was selected because it was not irritating in the screening test.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and epicutaneous, respectively)
- Exposure period: single injection (intradermal) and 48 h (epicutaneous, occlusive)
- Test group:
Intradermal (3 pairs of injections): 1:1 mixture (v/v) FCA/water, test substance (5%) in cottonseed oil, test substance (5%) in a 1:1 mixture FCA/water and cottonseed oil
Epicutaneous: 2 x 4 cm patch of filter paper was saturated with the moistened test material
- Control group:
Intradermal (3 pairs of injections): 1:1 mixture (v/v) FCA/water, Cottonseed oil, 1:1 mixture (v/v) FCA/water and cottonseed oil
Epicutaneous: dry filter paper
- Site: upper back

B. CHALLENGE EXPOSURE
- No. of exposures: 1(epicutaneous, occlusive)
- Day of challenge: day 21
- Exposure period: 24 h
- Test group:
Epicutaneous: 500 mg on a filter paper moistened with petrolatum
- Control group:
Epicutaneous: dry filter paper
- Site: on the flank
- Concentrations: 0 and 500 mg (0 and 100%)
- Evaluation (hrs after challenge): 24 and 48 h

OTHER:
The application site was pretreated with 0.5 mL of a 10% sodium-laurylsulfate (open application) 24 hours prior to the epidermal induction application.

Results and discussion

Positive control results:
No positive controls were included in this study. The reference values with DNCB were reported (experimental date May-June 1994).
DNCB (Intradermal induction: Concentration of compound: 0.1%; Vehicle: cottonseed oil; Epidermal induction: Concentration of compound: 1%; Vehicle: vaseline; Epidermal challenge: Concentration of compound: 0.1%; Vehicle: vaseline) induced reactions in 18/18 animals, thus meeting the reliability criteria (>= 15% positive responders).

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
500 mg (100%)
No. with + reactions:
0
Total no. in group:
10
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
500 mg (100%)
No. with + reactions:
0
Total no. in group:
10
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
0
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
0
No. with + reactions:
0
Total no. in group:
20
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
500 mg (100%)
No. with + reactions:
0
Total no. in group:
20
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
500 mg (100%)
No. with + reactions:
0
Total no. in group:
20

Any other information on results incl. tables

Erythema and edema reactions (score 1-2) have been observed after intradermal induction (day 6) at the sites treated with Freund´s Complete Adjuvant. No reactions have been observed at the sites treated with test substance alone, neither during induction nor challenge period.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Since none of the test animals (Group I) or control animals (Group II) exhibited skin reactions after the challenge treatment, the test material was rated non-sensitizing according to the methods of Magnusson and Kligman.
Executive summary:

A maximization test for topically applied test materials was conducted on 30 short-haired male and female albino guinea pigs to determine if the test mnaterial produced a sensitizing reaction. This study was designed and performed in accordance with OECD Guideline 406, EPA FIFRA Guideline 81-6, EPA TSCA (40 CFR 798.4100), and EPA Good Laboratory Practice Standards (40 CFR 792). Methods and grading of sensitization were based on Magnusson and Kligman (J. Invest. Dermat. 52: 268-276 (1969)). Twenty guinea pigs (Group I) each received three pairs of intradermal injections (adjuvant, a 5 % w/v solution of the test material, and a mixture of adjuvant and 5% w/v solution of the test material) followed one week later by a single topical application of 500 mg of test material moistened with petrolatum. The 5% dose was determined from pretest screening and is the maximum level suggested (Magnusson and Kligman, 1969), and the 500 mg of test material moistened with petrolatum was chosen for the topical applications because the screening did not indicate excessive irritation. Ten additional animals served as a control group. Control animals (Group II) were treated at the same time periods and locations as the test group but with the vehicle used in place of the test material for the intradermal injections. The control animals received a topical application of dry filter paper only. Two weeks after the topical application, all animals were challenged with a topical application of 500 mg of test material moistened with petrolatum at a virgin test site. The percentage of test animals exhibiting erythema and/or edema after the challenge treatment was used to assign the test material a sensitization potency rating. Since none of the test animals (Group I) or control animals (Group II) exhibited skin reactions after the challenge treatment, the test material was rated non-sensitizing according to the methods of Magnusson and Kligman.