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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:
24 October 1994 - 26 November 1994
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1996
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The Murine Local Lymph Node Assay (LLNA) is the first-choice method for in vivo testing according to the REACH Regulation. However, this reliable GPMT test was performed before entry into force of the REACH Regulation.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
Name: Ethanol, 2,2'-oxybis-, reaction products with ammonia, morpholine derivs. residues
Physical state: liquid
Appearance: brown liquid
Specific details on test material used for the study:
- Name of test material (as cited in study report): AMINE C-8
- Substance type: dark brown slightly viscous liquid with crystalline sediment
- Physical state: liquid
- Analytical purity: no data
- Composition of test material, percentage of components: no data
- Lot/batch No.: no data
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: at room temperature

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: David Hall Ltd., Burton-on-Trent, Staffordshire, UK
- Age at study initiation: 8-12 weeks
- Weight at study initiation: 307-399 g
- Housing: singly or in pairs in solid floor polypropylene cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-22 °C
- Humidity (%): 50-63%
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
water
Concentration / amount:
Intradermal induction: 0.5% (w/v) in distilled water
Topical induction: undiluted
Topical challenge: 50% and 75% (v/v) in distilled water
Challenge
Route:
epicutaneous, occlusive
Vehicle:
water
Concentration / amount:
Intradermal induction: 0.5% (w/v) in distilled water
Topical induction: undiluted
Topical challenge: 50% and 75% (v/v) in distilled water
No. of animals per dose:
10 test animals and 5 control animals
Details on study design:
RANGE FINDING TESTS:
For the intradermal injections, 4 animals were injected with 0.1%, 0.5%, 1% or 5% test material and erythema was assessed 24, 48 and 72 hours and 7 days after injection. For the topical induction and challenge, 2 animals were treated with 25%, 50%, 75% and 100% test material under occlusion. The highest concentration without severe irritation after intradermal injection was used in the main test. The highest concentration producing mild-moderate irritation after topical induction was used in the main study. The highest non-irritating dose and one lower dose after topical challenge were used in the main test.

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 injections and one occlusive topical application
- Exposure period: one week between injection and topical application; 48 hours topical application
- Test groups: 0.1 ml injection of 1) FCA + water (1:1); 2) 0.5% test material; 3) 0.5% test material + FCA (1:1)
- Control group: 0.1 ml injection of 1) FCA + water (1:1); 2) water; 3) 50% formulation of FCA + water (1:1)
- Site: 40 mm x 60 mm on shoulder region
- Frequency of applications: one series of 3 injections followed by one topical induction 1 week later
- Duration: 7 days between intradermal and topical induction, 48 hours topical exposure, 24 hours observation after topical exposure
- Concentrations: undiluted test material for topical induction

B. CHALLENGE EXPOSURE
- No. of exposures: 3 per animal
- Day(s) of challenge: 1 day
- Exposure period: 1 day
- Test groups: 50% and 75% of test material (right flank)
- Control group: vehicle (left flank)
- Site: flanks
- Concentrations: 0, 50% and 75%
- Evaluation (hr after challenge): 24 and 48 h
Challenge controls:
Distilled water (vehicle) was used as the negative control.
Positive control substance(s):
yes
Remarks:
2-mercaptobenzothiazole

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
75%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
yellow-coloured staining
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
yellow-coloured staining
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
yellow-coloured staining
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
75%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
yellow-coloured staining
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
50%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
yellow-coloured staining
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
10
Clinical observations:
yellow-coloured staining
Reading:
1st reading
Group:
positive control
Dose level:
2-mercaptobenzothiazole at 5% in arachis oil B.P.
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Reading:
2nd reading
Group:
positive control
Dose level:
2-mercaptobenzothiazole at 5% in arachis oil B.P.
No. with + reactions:
8
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation

Any other information on results incl. tables

After intradermal injection, erythema was noted at the exposure sites, mainly in the test group animals (well-defined after 24h and very slight to well-defined after 48h). After topical induction, all test group animals showed yellow-coloured staining and very slight to well-defined erythema. The control group animals did not show skin reactions. After topical challenge, all animals, including control group animals showed yellow-coloured staining. No other skin reactions were observed.

No adverse effects in body weight (gain) was observed when comparing test group animals and control group animals.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In the guinea pig maximisation test, the test material was negative for skin sensitisation.