(3Z)-hex-3-en-1-yl 2-methylprop-2-en-1-yl ether

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 06 December 2000 and 17 January 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in accordance with OECD guidelines and in compliance with GLP.

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

the temperature recorded in the animal room was sometimes outside of the target ranges specified in the protocol.

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

yes

Remarks:

the temperature recorded in the animal room was sometimes outside of the target ranges specified in the protocol.

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Species:

guinea pig

Strain:

Hartley

Sex:

male/female

Details on test animals and environmental conditions:

Animals
Species and sex: male and nulliparous and non-pregnant female guinea pigs.
Strain and sanitary status: Hartley CrI: (HA) BR, Caesarian obtained, Barrier sustained - Virus Antibody Free (COBS - VAF)
Reason for this choice: species generally accepted by regulatory authorities for this type of study.
The strain used has been shown to produce a satisfactory sensitization response using known sensitizers.
Breeder: Charles River France, 76410 Saint-Aubin-les-Elbeuf, France.
Number: three males and three females for the preliminary test, 30 animals (15 males and 15 females) for the main test.
Allocation of the animals to the groups: on day -1, the animals were weighed and randomly allocated to two groups: a control group of ten animals (five males and five females) and a treated group of 20 animals (ten males and ten females).
Age/weight: on day 1, the animals of the main test were 1-3 months old and had a mean body weight ± standard deviation of 367 ± 18 g for the males and 365 ± 19 g for the females.
Acclimation: at least 5 days before the beginning of the study.
Identification of the animals: ear-tattoo.

Environmental conditions
The conditions in the animal room were set as follows:
temperature: 21 ± 2°C
relative humidity: 30 to 70%
light/dark cycle: 12 h / 12 h
ventilation: approximately 12 cycles/hour of filtered, non-recycled air.
The temperature and relative humidity were under continuous control and recording. The records were checked daily and filed. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular intervals.
During the acclimation period and throughout the study, the animals were housed individually in polycarbonate cages with stainless steel lid (48 cm x 27 cm x 20 cm) equipped with a po!ypropylene bottle.
Dust-free sawdust was provided as litter (SICSA, 94142 AlfortvilIe, France).
Sawdust is analysed by the supplier for composition and contaminant levels.

Food and water
During the study, the animals had free access to "106 pelleted diet" (UAR, 91360 Villemoisson-sur-Orge, France).
Food is analysed regularly by the supplier for composition and contaminant levels.
Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum.
Bacteriological and chemical analyses of water are performed regularly by external laboratories.
These analyses include the detection of possible contaminants (pesticides, heavy metals and nitrosamines).
No contaminants were known to have been present in the diet, drinking water or bedding material at levels which may be expected to have interfered with or prejudiced the outcome of the study.

Route:

intradermal and epicutaneous

Vehicle:

other: intradermal injections: corn oil, topical application: ethanol:water or acetone

Concentration / amount:

Intradermal: 10% (w/w) in corn oil
Topical: a mixture of ethanol/water (80/20; w/w) for the cutaneous application on day 8 at the maximum concentration of 50% (w/w),
acetone for the cutaneous application on day 22 at the maximum concentration of 50% (w/w).

Route:

epicutaneous, occlusive

Vehicle:

other: intradermal injections: corn oil, topical application: ethanol:water or acetone

Concentration / amount:

Intradermal: 10% (w/w) in corn oil
Topical: a mixture of ethanol/water (80/20; w/w) for the cutaneous application on day 8 at the maximum concentration of 50% (w/w),
acetone for the cutaneous application on day 22 at the maximum concentration of 50% (w/w).

No. of animals per dose:

Three males and three females for the preliminary test.
30 animals (15 males and 15 females) for the main test.

Details on study design:

Preliminary test
A preliminary test was conducted in order to determine the concentrations to be tested in the main study.
By intradermal route (tested concentrations: 25%, 10%,5% and 1 % (w/w))
24 hours before treatment, the dorsal region of the animals was clipped, intradermal injections of the dosage form preparations (0.1 ml) were performed in the interscapular region, cutaneous reactions were evaluated approximately 24, 48 hours and 6 days after the injections.
By cutaneous route for the challenge phase (tested concentrations: 100%, 50% and 25% (w/w))
24 hours before treatment, both flank regions of the animals were clipped and shaved, the filter paper of a chamber (Finn Chamber®) was fully-loaded with one dosage form preparation. The chamber was then applied to the clipped area of the skin (one concentration per flank). The chamber was held in place by means of an occlusive dressing for 24 hours, cutaneous reactions were evaluated approximately 24 and 48 hours after removal of the
dressings.
By cutaneous route for the induction phase (tested concentrations: 50%, 25% (w/w))
24 hours before treatment, the interscapular region of the animals was clipped, a pad of filter paper (approximately 8 cm2) was fully-loaded with one dosage form preparation. The filter paper was then applied to the clipped area of the skin. The filter paper was held in place by means of an occlusive dressing for 48 hours, cutaneous reactions were evaluated approximately 24 and 48 hours after removal of dressings.
Criteria for selection of concentrations
The following criteria were used:
the concentrations should be well-tolerated systemically and locally, intradermal injections should cause moderate irritant effects (no necrosis or ulceration of the skin), cutaneous application for the induction should cause at most weak or moderate skin reactions or be the maximal practicable concentration, cutaneous application for the challenge phase should be the highest concentration which does not cause irritant effects.

Main study
Preparation of the animals
For all animals, the application sites were:
clipped on days -1 and 7 (interscapular region 4 cm x 2 cm), clipped and shaved on day 21 (each flank 2 cm x 2 cm).
Induction phase by intradermal and cutaneous routes
Intradermal route
On day 1, six injections were made deep into the dermis of a 4 cm x 2 cm clipped interscapular area, using a needle (diameter: 0.50 x 16 mm) mounted on a 1 ml plastic syringe (0.01 ml graduations).
Three injections of 0.1 ml were made into each side of this interscapular region (i.e. three pairs of sites), as follows:

Injection Site Treated group Control Group
1 Anterior FCA at 50% (v/v) in 0.9% NaCl FCA at 50% (v/v) in 0.9% NaCI
2 Middle test substance at 10% (w/w)
in corn oil corn oil
3 Posterior* test substance at 10% (w/w) in the vehicle at 50% (w/v) in a mixture
mixture FCA/0.9% NaCI (50/50) FCA/0.9% NaCI (50/50)

FCA: Freund's complete adjuvant
* The test substance was suspended in FCA prior to combining with the aqueous phase.
The final concentration of the test substance was equal to that used in injection 2.

The anterior and middle pairs of injections were performed close to each other and nearest the head, while the posterior pair was performed towards the caudal part of the test area.
Cutaneous route
On day 7, the interscapular area was clipped.
As the test substance was shown to be non-irritant during the preliminary test, the animals were treated with 0.5 ml of sodium lauryl sulfate at the concentration of 10% (w/w) in vaseline, in order to induce local irritation.
On day 8, a pad of fIlter paper (approximately 8 cm2) was fully-loaded with the test substance at the concentration of 50% (w/w) and was then applied to the interscapular region of the animals of the treated group.
The animals of the control group received an application of the vehicle alone under the same experimental conditions.
The pad was held in place for 48 hours by means of an adhesive hypo allergenic dressing and an adhesive anallergenic waterproo f plaster.

Challenge phase
On day 22, the animals of treated and control groups received an application of the test substance and vehicle. The filter paper of a chamber (Finn Chamber®) was fully-loaded with the test substance at the concentration of 50% (w/w) and was then applied to a clipped area of the skin of the posterior right flank of all animals.
The vehicle was applied under the same experimental conditions to the skin of the posterior left flank.
The chambers were held in contact with the skin for 24 hours by means of an adhesive anallergenic waterproof plaster.

Challenge controls:

On day 22, the animals of the control group received an application of the test substance and vehicle. The filter paper of a chamber (Finn Chamber®) was fully-loaded with the test substance at the concentration of 50% (w/w) and was then applied to a clipped area of the skin of the posterior right flank of all animals.
The vehicle was applied under the same experimental conditions to the skin of the posterior left flank.
The chambers were held in contact with the skin for 24 hours by means of an adhesive anallergenic waterproof plaster.

Positive control substance(s):

no

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

vehicle (acetone)

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: vehicle (acetone). No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

vehicle (acetone)

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: vehicle (acetone). No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

negative control

Dose level:

vehicle (acetone)

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: vehicle (acetone). No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

Vivaldie at 50% w/w in acetone

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: Vivaldie at 50% w/w in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

2nd reading

Hours after challenge:

4

Group:

negative control

Dose level:

Vivaldie at 50% w/w in acetone

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 4.0. Group: negative control. Dose level: Vivaldie at 50% w/w in acetone. No with. + reactions: 0.0. Total no. in groups: 10.0.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

negative control

Dose level:

Vivaldie at 50% w/w in acetone

No. with + reactions:

1

Total no. in group:

10

Clinical observations:

1 animal with score of Grade 1/dryness of skin

Remarks on result:

other: see Remark

Remarks:

Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: negative control. Dose level: Vivaldie at 50% w/w in acetone. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: 1 animal with score of Grade 1/dryness of skin.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

vehicle (acetone)

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: vehicle (acetone). No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

vehicle (acetone)

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 72.0. Group: test group. Dose level: vehicle (acetone). No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

vehicle (acetone)

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: vehicle (acetone). No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

Vivaldie at 50% w/w in acetone

No. with + reactions:

0

Total no. in group:

20

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: Vivaldie at 50% w/w in acetone. No with. + reactions: 0.0. Total no. in groups: 20.0.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

Vivaldie at 50% w/w in acetone

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

2 animals with score of Grade 1

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: Vivaldie at 50% w/w in acetone. No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: 2 animals with score of Grade 1.

Reading:

other: 3rd reading

Hours after challenge:

72

Group:

test chemical

Dose level:

Vivaldie at 50% w/w in acetone

No. with + reactions:

2

Total no. in group:

20

Clinical observations:

2 animals with score of Grade 2, dryness of skin noted in 8 animals

Remarks on result:

other: see Remark

Remarks:

Reading: other: 3rd reading. . Hours after challenge: 72.0. Group: test group. Dose level: Vivaldie at 50% w/w in acetone. No with. + reactions: 2.0. Total no. in groups: 20.0. Clinical observations: 2 animals with score of Grade 2, dryness of skin noted in 8 animals.

CHOICE OF THE VEHICLE

The test substance was not soluble in 0.9% NaCl: two phases were observed.For the intradermal injections, the vehicle chosen was corn oil: a homogeneous dosage form preparation was obtained whatever the proportion.For the topical applications, the vehicles chosen were as follows: a mixture of ethanol/water (80/20; w/w) for the cutaneous application on day 8: a homogeneous dosage form preparation was obtained at the maximum concentration of 50% (w/w), acetone for the cutaneous application on day 22: a homogeneous dosage form preparation was obtained at the maximum concentration of 50% (w/w).

PRELIMINARY STUDY

Administration by intra-dermal route Results were as follows: see attachment 3. In order to respect the criteria for the selection of concentrations (the concentration should be well-tolerated systemically and locally), intra-dermal injections should cause moderate irritant effect but no necrosis or ulceration of the skin), concentration chosen for the main study was 10% (w/w), because this concentration caused moderate irritation. Application by cutaneous route Results were as follows: see attachment 4. On removal of the dressing, no residual test substance was observed. In order to respect the criteria for the selection of concentrations (the concentrations should be well-tolerated systemically and locally), cutaneous application for the induction should cause at most weak or moderate skin reactions or be the maximal practicable concentration. At 100 % moderate irritation was seen (grade 1). Therefore in the inducation and challenge phase the maximum concentration of 50% (w/w) was used.

MAIN STUDY

Clinical examinations:

 No clinical signs and no deaths were observed during the study.

Body weight The body weight gain of the treated animals was similar to that of the control animals . Challenge phase - Scoring of cutaneous reactions On removal of the dressing, no residual test substance was observed. Scoring of skin reactions was as follows: No cutaneous reactions were observed in the animals of the control group at the 24 and 48-hour readings. At the 72-hour reading, a discrete erythema (grade 1) and dryness of the skin were noted in 1/10 animals. In the treated group, no cutaneous reactions were observed at the 24-hour reading. At the 48-hour reading, a discrete erythema (grade 1) was noted in 2/20 animals. At the 72-hour reading, a moderate erythema (grade 2) and dryness of the skin were recorded in these same animals; dryness of the skin was also noted in 6 other animals. As the cutaneous reactions recorded in animals of the treated group were of low incidence and as a discrete skin reaction was recorded in the control group with similar incidence, these cutaneous reactions were most probably not attributable to a sensitizing process.

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Under our experimental conditions and according to the maximization method of Magnusson and Kligman, the test substance Vivaldie should not be considered as a skin sensitizer.

Executive summary:

The potential of the test substance Vivaldie to induce delayed contact hypersensitivity was evaluated in guinea pigs according to the maximization method of Magnusson and Kligman and to OECD (No. 406, 17th July 1992) and EC (96/54/EEC, B.6, 30 July 1996) guidelines. Thirty guinea pigs were allocated to two groups: a control group of five males and five females and a treated group of ten males and ten females. During the induction phase the treated group received intradermal injections of Vivaldie at a concentration of 10% (w/w) in corn oil and topical application of Vivaldie at the concentration of 50% (w/w) in ethanol/water (80/20). During the Challenge phase all groups received a topical application of Vivaldie at a concentration of 50% (w/w) in acetone. No clinical signs and no deaths were noted during the study. After the challenge application, no cutaneous reactions were observed in the animals of the control group at the 24 and 48-hour readings. At the 72-hour reading, a discrete erythema and dryness of the skin were noted in 1/ 10 animals.

In the treated group, no cutaneous reactions were observed at the 24-hour reading. At the 48-hour reading, a discrete erythema was noted in 2/20 animals. At the 72-hour reading, a moderate erythema and dryness of the skin were recorded in these same animals; dryness of the skin was also noted in 6 other animals .

As the cutaneous reactions recorded in animals of the treated group were of low incidence and as a discrete skin reaction was recorded in the control group with similar incidence, these cutaneous reactions were most probably not attributable to a sensitizing process. It can be concluded that under the experimental conditions of this study and according to the maxirnization method of Magnusson and Kligman, the test substance Vivaldie should not be considered as a skin sensitizer.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

Two studies are available a GPMT test and an HRIPT study. The GPMT study is the key study because up to 50% test substance concentration has been used. In the HRIPT with sufficient number of subjects (>100) 0.5% test concentration has been used.

Migrated from Short description of key information:
A GPMT, according to OECD TG 406 has been performed up to 50%. It can be concluded that Vivaldie is not sensitising.

Justification for selection of skin sensitisation endpoint:
Key study at reliability 1

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no study available

Justification for classification or non-classification

Based on the results available Vivaldie does not need to be classified for sensitisation according to the DSD 67/548/EC and the CLP Regulation EC 1272/2008.