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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:
2006-01-16 to 2006-02-23
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
GLP followed OECD, well conducted and documented study, substance details and certificate of analysis. However there are deviations: slight higher temperature, longer rest phase between induction and challenge, and insufficient number of test animals.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2006
Report Date:
2006

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
slight higher temperature, longer rest phase between induction and challenge and insufficient number of test animals
Principles of method if other than guideline:
Not applicable
GLP compliance:
yes (incl. certificate)
Type of study:
guinea pig maximisation test
Justification for non-LLNA method:
A study conducted according to Guideline OECD 406 before 2008 and is available on the registered substance.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Type:
Constituent
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Type:
impurity
Test material form:
liquid
Details on test material:
Batch No.: 049807
Purity: 67.2% (sum of the three main constituents)
Name of test material (as cited in study report): TERPINEOL MULTICONSTITUENT
Physical state: colourless liquid
Storage conditions: +2°C to +8°C, under nitrogen and protected from light
Expiry date: 30 November 2017

In vivo test system

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Centre de Production Animale, Olivet, France
- Weight at study initiation: between 359 g and 500 g
-housing: housed in groups of 2 or 3 in makrolon containers
-diet(ad libitum): pelleted guinea pig breeding diet, ad libitum
- Water (e.g. ad libitum): tap water from public distribution system, ad libitum
- Acclimatation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): between 20 and 25
- Humidity (%): between 30 and 49
-air changes: at least 10 cycles per hour
-photoperiod: 12 hours dark / 12 hours light

IN-LIFE DATES: From: 2006-01-16 To: 2006-02-23

Study design: in vivo (non-LLNA)

Inductionopen allclose all
Route:
intradermal and epicutaneous
Vehicle:
olive oil
Concentration / amount:
For maximal non necrotising concentration test (through intradermal injections): 100, 50, 25, 12.5, 6.25 and 3.125%
For maximal non irritant concentration (topical application): - Induction phase 100, 50,25, and 12.5% - Challenge 25, 12.5, 6.25 and 3.125%
Challengeopen allclose all
Route:
epicutaneous, occlusive
Vehicle:
olive oil
Concentration / amount:
For maximal non necrotising concentration test (through intradermal injections): 100, 50, 25, 12.5, 6.25 and 3.125%
For maximal non irritant concentration (topical application): - Induction phase 100, 50,25, and 12.5% - Challenge 25, 12.5, 6.25 and 3.125%
No. of animals per dose:
For maximal non necrotising concentration test: 2 males
For maximal non irritant concentration: - Induction phase: 2 males
- Challenge phase: 1 male and 2 females
Details on study design:
RANGE FINDING TESTS: For maximal non necrotising concentration test (through intradermal injections): 100, 50, 25, 12.5, 6.25 and 3.125% (2 males)
For maximal non irritant concentration (topical application): - Induction phase 100, 50,25, and 12.5% (2 males) - Challenge 25, 12.5, 6.25 and 3.125% (1 male and 2 females)

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 pairs of intradermal injections in treated group and 3 pairs of intradermal injections in control group on day 0 and 2nd induction topical application in both treated and control groups on 7th day
- Exposure period: 9 days
- Test groups: 10 males
- Control group: 5 males
- Site: inter scapular zone
- Frequency of applications: only once
- Concentrations:
- Intradermal injections: 2 intradermal injections of the test item diluted at 6.25 % in olive oil. 2 intradermal injections of Freund's Complete Adjuvant diluted at 50 % in a physiological saline solution. 2 intradermal injections of a mixture with equal volumes of Freund's Complete Adjuvant at 50% and the test item diluted at 12.5% in olive oil.
- Topical application: on the same zone, with the test item at 100%

B. CHALLENGE EXPOSURE
- No. of exposures: one time
- Day(s) of challenge: 2
- Exposure period: 24 h
- Test groups: 10 males
- Control group: 5 males
- Site: dorso-lombar zone
- Concentrations: for group 1 and 2 topical application under occlusive dressing at the following concentrations: 25 and 12.5 %
- Evaluation (hr after challenge): skin reactions at 24 and 48 h
Challenge controls:
5 males, topical application under occlusive dressing at the following concentrations: 25 and 12.5%
Positive control substance(s):
yes
Remarks:
alpha-hexylcinnamaldehyde (CAS No 101-86-0) and 2-mercaptobenzothiazole (CAS No 149-30-4)

Results and discussion

Positive control results:
The test substances Hexylcinnamaldehyde (50, 25 and 12.5%) and 2-Mercaptobenzothiazole (50 and 25%) were used as a positive control and induce positive sensitization response

In vivo (non-LLNA)

Resultsopen allclose all
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 25% . No with. + reactions: 0.0. Total no. in groups: 5.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
25%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
12.5%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 12.5%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
12.5%
No. with + reactions:
0
Total no. in group:
5
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 12.5%. No with. + reactions: 0.0. Total no. in groups: 5.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
25%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 25%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
12.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 12.5%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
12.5%
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 12.5%. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
12.5%
No. with + reactions:
45
Total no. in group:
11
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
Dose level:
12.5%
No. with + reactions:
18
Total no. in group:
11
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
24
Group:
positive control
Dose level:
25%
No. with + reactions:
100
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation
Reading:
1st reading
Hours after challenge:
48
Group:
positive control
Dose level:
25%
No. with + reactions:
60
Total no. in group:
10
Remarks on result:
positive indication of skin sensitisation

Any other information on results incl. tables

No remarks

Applicant's summary and conclusion

Interpretation of results:
not sensitising
Conclusions:
Terpineol multiconstituent should not be classified according to CLP Regulation (EC) No 1272/2008.
Executive summary:

In a skin sensitization study performed according to the OECD guideline 406 and conducted in compliance with GLP, Terpineol multiconstituent was tested in male albino guinea pigs using the Guinea pig Maximisation test method (10 treated animals and 5 control animals).

Preliminary studies were conducted to dertermine the maximal non necrotising concentration through intradermal injections and the maximal non irritant concentration by topical application. Six concentrations were tested for intradermal injections and for topical application: 100, 50, 25, 12.5, 6.25 and 3.125% in olive oil.

In the main experiment, the induction phase for the test group consisted of:

one pair of intradermal injections of test substance at 6.25% in olive oil

one pair of intradermal injections of Freund's complete adjuvant diluted 50% in physiological saline

one pair of intradermal injections of mixture of equal volumes of Freund's complete adjuvant at 50% and test substance at 12.5% in olive oil.

For control group, test substance was replaced by isotonic solution of NaCl.

Seven days later, undiluted test substance was applied topically for 48 h, followed by a rest phase of 17 days.

For the challenge phase, all animals were exposed to 25 and 12.5% of the test susbtance by topical application under occlusive conditions for 24 h. Cutaneous reactions were recorded 24 and 48 h after the removal of the test substance.

Hexylcinnamicaldehyde and mercaptobenzothiazole were used as positive controls and induced positive sensitization responses in test animals.

No cutaneous reactions following the removal of the occlusive dressing from all animals were observed at 24 and 48 h readings.

Terpineol multiconstituent can be considered as a non sensitiser and should not be classified according to CLP Regulation (EC) No 1272/2008.