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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:
1992-04-08 to 1992-05-22
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was conducted before REACH came into force.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Trimethylorthoformiat, TMOF
- Substance type: pure active substance
- Physical state: liquid, colorless
- pH: 5.02
- Water solubility: 10 g/liter
- Analytical purity: 99.8 area-% (gas chromatography)
- Impurities (identity and concentrations): no data
- Stability under test conditions: Sensitive to hydrolysis
- Storage condition of test material: Closed container, in hood

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Strain: Pirbright White HSD/Win:DH (SPF)
- Age at study initiation: young adult
- Weight at study initiation: 310 - 381 g
- Housing: conventional, up to 5 animals/Macrolon cage type IV
- Diet: ad libitum, Ssniff G 4 Alleindiaet fuer Meerschweinchen
- Water: communal drinking water ad libitum
- Acclimation period: > 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 3 °C
- Humidity (%): 30 - 70 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12

IN-LIFE DATES: From: 1992-04-08 To: 1992-05-08

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
corn oil
Remarks:
Maiskeimoel MEH 56
Concentration / amount:
Intradermal pretest: 0.25, 0.5, 1.0, 2.5, 5, 10 % w/w in corn oil
Epicutaneous pretest: 2.5, 25, 50 % w/w in corn oil, 100% undiluted liquid test article
Epicutaneous induction: 100% undiluted liquid
Intradermal induction: 10% w/w in corn oil
Epicutaneous induction: 100% undiluted liquid
Challenge
Route:
epicutaneous, occlusive
Vehicle:
corn oil
Remarks:
Maiskeimoel MEH 56
Concentration / amount:
100% undiluted liquid
No. of animals per dose:
Intradermal pretest: 2 animals
Epicutaneous pretest: 4 animals
Main test: 20 animals
Controls to main test: 10 animals
Details on study design:
RANGE FINDING TESTS:
Formulation: Corn oil was found to be an appropriate vehicle, which formed homogeneous mixtures with the liquid test article
Tolerability intradermal: Injection of the test substance in corn oil (0.15 cm3 of the mixtures, 6 concentrations from 0.25 to 10 % w/w) to the shaved flanks of 2 animals. Recording of skin reactions at 24 hours after injection.
Tolerability epicutaneous: Application of the test substance in corn oil (0.1 cm3 each, 3 concentrations: 2.5, 25, and 50 % w/w, and 100% liquid test material without vehicle) to the shaved flanks of 4 animals (4 gauze patches per animal, area 2x2cm each), with occlusive dressing, for 24 hours. Recording of skin reactions after removal of the patch, and at 48 and 72 hours after the start of the application.

MAIN STUDY
A. INTRADERMAL INDUCTION EXPOSURE
- No. of exposures: 1
- Exposure period:
- Test groups: 1 group of 20 animals
- Control group:
- Site:
- Frequency of applications:
- Duration:
- Concentrations:

B. EPICUTANEOUS INDUCTION EXPOSURE
- No. of exposures: 1
- Exposure period:
- Test groups:
- Control group:
- Site:
- Frequency of applications:
- Duration:
- Concentrations:

B. CHALLENGE EXPOSURE
- No. of exposures:
- Day(s) of challenge:
- Exposure period:
- Test groups:
- Control group:
- Site:
- Concentrations:
- Evaluation (hr after challenge):
Challenge controls:
No naive control group. An FCA-treated control group (as described above) was challenged in the same was as the test group.
Positive control substance(s):
yes
Remarks:
Strain sensitivity to 1-chloro-2,4-dinitrobenzene tested in regular intervals

Results and discussion

Positive control results:
Strain sensitivity to 1-chloro-2,4-dinitrobenzene tested in regular intervals, no results given in present report

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
test group
Dose level:
0.15 cm3 of undiluted liquid test substance (100 %)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No erythema, no edema observable
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
test group
Dose level:
0.15 cm3 of undiluted liquid test substance (100 %)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No erythema, no edema observable
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
negative control
Dose level:
0.15 cm3 of undiluted liquid test substance (100 %)
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No erythema, no edema observable
Remarks on result:
no indication of skin sensitisation
Remarks:
Reading: 1st reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 0.15 cm3 of undiluted liquid test substance (100 %). No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: No erythema, no edema observable.
Key result
Reading:
2nd reading
Hours after challenge:
72
Group:
negative control
Dose level:
0.15 cm3 of undiluted liquid test substance (100 %)
No. with + reactions:
0
Total no. in group:
20
Clinical observations:
No erythema, no edema observable
Remarks on result:
no indication of skin sensitisation
Remarks:
Reading: 2nd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: 0.15 cm3 of undiluted liquid test substance (100 %). No with. + reactions: 0.0. Total no. in groups: 20.0. Clinical observations: No erythema, no edema observable.
Key result
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
No erythema, edema observable.
Remarks on result:
no indication of skin sensitisation

Any other information on results incl. tables

Pretest intradermal: All concentrations of the test substance, as well as the vehicle, caused well-defined erythema and edema 24 hours after injection. Therefore the highest concentration, 10 % w/w, was chosen for the intradermal induction in the main study. Pretest epicutaneous: None of the concentrations of the test substance, including the undiluted substance itself, caused any skin irritation upon application in an occlusive patch (24 hours).

Main study, intradermal induction: The injection of Freund¿s Complete Adjuvans (FCA, diluted 1:1 with saline) caused well-defined to severe erythema and edema. 10 animals of the test group and 12 of the control groups developed necrosis. The injection of the test substance (10 % in corn oil) caused very slight erythema and edema, which developed into well-defined erythema and edema in half of the animals 24 hours after the injection. Reactions of the same grade were observed in the control animals after the injection of the vehicle corn oil. Injection of the test substance in FCA (diluted 1:1 with corn oil) caused very slight to well-defined erythema and edema, which developed into moderate edema and erythema after 24 hours; one animal developed necrosis.

Main study, epicutaneous induction: All injection sites of FCA (50% in saline, mixed with corn oil, or mixed with 10% of the test substance and corn oil) were affected by severe erythema, eschar formation, bleeding, and strong edema. Injection sites of the test substance in corn oil, or the pure vehickle in the controls, showed only scale formation.

Main study, challenge: No signs of irritation in the test and control group.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met