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Toxicological information

Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Remarks:
in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2002-10-03 to 2002-11-18.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: No certificate of analysis; only few animals are tested: 5 instead of 10.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2003
Report Date:
2003

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Principles of method if other than guideline:
Comparable to OECD 406. However, only few animals are tested (Simplified study).
GLP compliance:
yes
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
Study performed before the ECHA recommendations.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Vanillic acid
- Expiration date of the lot/batch: no data
- Purity test date: no data

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: ambient temperature
- Stability under test conditions: yes

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Elm Hill Breeding Labs, Chelmsford, MA.
- Age at study initiation: at least 4 weeks old
- Weight at study initiation: 377.5 to 402.6 grams.
- Housing: the animals were housed individually in group in suspended stainless steel.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: Minimum 5 days under the same conditions as for the actual test.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 +/- 5 °C
- Humidity (%): 30 – 70 %
- Air changes (per hr): 10 to 15 air exchanges/hour
- Photoperiod (hrs dark / hrs light): 12hr/12hr

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal
Vehicle:
cotton seed oil
Concentration / amount:
100% TS +/- FCA (treated group) or cottonseed oil +/- Freund Complete Adjuvant (control group) or FCA (both groups) in scapular region.
Day(s)/duration:
Day 1
Adequacy of induction:
highest concentration used causing mild-to-moderate skin irritation and well-tolerated systemically
Route:
intradermal
Vehicle:
cotton seed oil
Concentration / amount:
treatment with 10% SLS in petrolatum
Day(s)/duration:
Day 7
Route:
epicutaneous, occlusive
Vehicle:
cotton seed oil
Concentration / amount:
100% TS occlusive dressing for 48h (treated) or cottonseed oil (control)
Day(s)/duration:
Day 8
Challenge
No.:
#1
Route:
epicutaneous, occlusive
Vehicle:
cotton seed oil
Concentration / amount:
100% TS under occlusive dressing for 24 h (all group) on the right flank and cottonseed oil on the left flank.
Day(s)/duration:
Day 22
Adequacy of challenge:
highest non-irritant concentration
No. of animals per dose:
5 animals and a control group of 3 animals.
Details on study design:
RANGE FINDING TESTS:
1- Intradermal injection tolerance test:
Six concentrations of the test substance were administered in Cottonseed Oil (CSO). The hair was removed from an area overlying the scapulae of a single animal. One of 6 concentrations of the test substance was administered by intradermal injection (0.1 mL) to each of 6 test sites. A single examination was undertaken 5 to 7 days after injection for evidence of skin reaction.
2- Dermal application tolerance test:
Four concentrations of the test substance were investigated. At each site, a 0.2 mL aliquot of the substance was spread over a 20 mm square of gauze or filter paper. This was then placed onto the skin and covered by an occlusive dressing for 24 hours. The treated sites on each animal were examined 24 and 48 hours after removal of the dressings for evidence of skin reaction which was scored according to the following scheme:
Reaction to treatment:
No visible change (score: 0)
Discrete or patchy erythema (score: 1)
Moderate and confluent erythema (score: 2)
Intense erythema and swelling (score: 3)
3- Selection of dose-levels for the sensitization test:
* Induction by intradermal injection:
the test substance was used at a concentration (100%) that is systemically tolerated and is judged not to cause excessive reaction at the injection site.
* Induction by topical application:
The test substance was used at the concentration (100%) that is systemically tolerated and is judged not to cause excessive reaction at the injection site.
* Induction by topical application:
The test substance was used at the concentration 100% that is found to be systemically tolerated and provokes a weak inflammatory response.
* Challenge:
The test substance was used at the maximum non-irritant concentration.
MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2
- Exposure period: 24 hours (Day 1: Induction by intradermal injection. Day 2: Day 2: Assessment of reaction), 48 hours (Day 8: Induction by dermal application. Day 10: removal of dressings).
- Site:
* Anterior sites: Freund's complete adjuvant (FCA) in emulsified form
* Middle sites: Test substance in selected vehicle (or selected vehicle alone)
* Posterior sites: Test substance (or selected vehicle) in FCA.
- Duration: Day 1 and Day 8
- Concentrations: 100%

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: 22
- Exposure period: 24 hours
- Test groups: yes
- Control group: yes
- Concentrations: 100%
- Evaluation (hr after challenge): 24 and 48 hours
Positive control substance(s):
no

Results and discussion

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
100%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
3
Remarks on result:
no indication of skin sensitisation
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
0%
No. with + reactions:
0
Total no. in group:
3
Remarks on result:
no indication of skin sensitisation
Key result
Group:
positive control
Remarks on result:
not measured/tested

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
Under the test conditions of this study, Vanillic acid is not considered as sensitizing to skin. Therefore, no classification is required according to EU criteria.
Executive summary:

In a dermal sensitization study using the method of Maximization test within Vanillic acid, Dunkin-Hartley guinea pigs (11 females) were tested under test conditions comparable to the guideline OECD 406 (Simplified study, Reliability 2, GLP). On day 1, three intradermal injections were performed in a row on each side of the shoulder. 24 hours before the topical sensitization (day 7), all the animals were treated on the shoulder with a preparation consisting of sodium lauryl sulphate (SLS) 10% in petrolatum. On day 8, topical sensitization on the same skin area was then performed with a solution of the test substance. The patch was covered by an occlusive dressing for 48 hours. On day 22 (= Challenge ), a 24 -hour occluded patch was performed on the right flank with 0.2 mL aliquot of the test substance. On day 24, assessment of challenge reaction was performed. On day 25, assessment of challenge reaction was again assessed. The concentration used for induction phase (intradermal and topical) and for challenge phase was 100%.

Under the test conditions of this study, 0/5 treated animals showed no indication of skin sensitization. Based on these results, no classification is required according to EU criteria.