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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:
1996
Report Date:
1996

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Qualifier:
according to
Guideline:
EU Method B.6 (Skin Sensitisation)
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
The study was conducted 1996 before the adoptance of LLNA.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid: viscous
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- lot/batch No.of test material: 6602
- Purity test date: 95.2 % (Total Tocopherols: 98.4 %)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: 2-10 °C in the dark

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Remarks:
Crl:(HA)BR
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River, Germany
- Females (if applicable) nulliparous and non-pregnant: yes
- Microbiological status of animals, when known: SPF-bred albino guinea pigs
- Age at study initiation: 4 weeks old
- Weight at study initiation: 249 - 274 g
- Housing: in a mobile battery, containing 6 cages; one cage per group of at most 10 animals.
- Diet (e.g. ad libitum): Standard laboratory diet ad libitum.
- Water (e.g. ad libitum): Tap water (N. V. Waterleidingbedrijf Midden-Nederland) ad libitum.
- Acclimation period: 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 +/- 3°C (from 15/11 - 18/11: 18-19°C)
- Humidity (%): 50-75% (upper limit higher than the intended 70%, because of meteorological circumstances and/or because of wet clean­ing of the animal room; the 75% peak occurred for one hour only).
- Air changes (per hr): ca 10 air changes/hour
- Photoperiod (hrs dark / hrs light): 12/12

Study design: in vivo (non-LLNA)

Induction
Route:
intradermal and epicutaneous
Vehicle:
maize oil
Concentration / amount:
Amounts of 0.1 ml of the selected concentrations:
30, 10, 3%
Day(s)/duration:
1. Intradermal injections and secondly, one week later, topical application over the injection sites.
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
maize oil
Concentration / amount:
30% and vehicle alone
Day(s)/duration:
Skin readings were made at 24 and 48 hours after removal of the patches.
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
Test group of 5 males and 5 females and one control group of 3 males and 3 females.
Details on study design:
RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures:
- 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):

OTHER:
Positive control substance(s):
yes
Remarks:
Benzocaine (BZ)

Results and discussion

Positive control results:
The sensitivity of this test system was checked by means of a positive control study with benzocaine (BZ). The challenge treatment with a 30% dilution of BZ in vaseline induced positive reactions in 50% of the test animals, whereas no skin reactions were observed in the controls. These results clearly show sensitization by BZ. Therefore, it can be concluded that the experimental design and the strain of guinea pigs used are suitable to detect possible sensitizing potential of lest materials.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
30%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
All animals remained in good health during the experimental period
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
30%
No. with + reactions:
0
Total no. in group:
5
Clinical observations:
All animals remained in good health during the experimental period.
Remarks on result:
no indication of skin sensitisation

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Because upon challenge none of the test animals reacted it can be concluded that according to the EC-standards (mentioned in EC Directive 93/21/EC and published in the Official Journal of the European Communities, L 110 A, Volume 36, 4 May 1993), d-delta-rich tocopherols concentrate (EMD) is not sensitizing.
Executive summary:

1. The test material EMD was examined for possible sensitizing properties by a maximization test in guinea pigs using 10 test animals and 6 controls.

2. The test comprised:

Test animals

- induction treatment by intradermal injections of Freund's Complete Adjuvant (FCA) 1:1 diluted with isotonic saline, a 10% dilution of EMD in maize oil, and a 10% test dilution in FCA/diluent 1:1, followed one week later by topical application of undiluted EMD (test site pretreated with sodium Iauryl sulfate),

- challenge treatment, 14 days after the last induction, by topical application of a 30% dilution of EMD in maize oil and with the vehicle alone

Controls

- induction treatment by intradermal injections of FCA/saline 1:1, maize oil alone, and FCA/diluent 1:1, followed one week Iater by topical appli­cation of empty patches (test site pretreated with sodium lauryl sulfate), and

- challenge treatment, 14 days after the last induction, by topical application of a 10% dilution of EMD in maize oil and with the vehicle alone.

3. The challenge treatment with EMD did not cause any skin reaction in the ten test animals. On the basis of the results obtained under the conditions of this study and according to the EC-standards (mentioned in EC Directive 93/21/EC and published in the Official Journal of the European Communities, L 110 A, Volume 36, 4 May 1993), it was concluded that EMD is not sensitizing.