1-Decene, 1-ethoxy-

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Between 12 October 2001 and 15 November 2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted in compliance with agreed protocols and under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2001

Report date:

2002

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

yes

Remarks:

Five animals w'ere above the weight range specified in the study protocol (450g).

GLP compliance:

yes (incl. QA statement)

Type of study:

guinea pig maximisation test

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

440-020-0

EC Name:

-

Cas Number:

61668-40-4

Molecular formula:

C12H24O

IUPAC Name:

Dec-1-en-1-yl ethyl ether

Test material form:

other: liquid

Details on test material:

Sponsor's identification: ARCTICAL
Chemical name: 1-decene, 1-ethoxy
CAS number: 61668-40-4
Purity: 97.8%
Stability: stable under normal temperatures and pressure
Description: clear colourless liquid
Purity: 97.8%
Batch number: ES 7565L12
Date received: 23 July 2001
Storage conditions: room temperature in the dark

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

male

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Supplied by a reputable supplier. For full details please see full study report
- Age at study initiation: Approximately eight to twelve weeks old
- Weight at study initiation: 384 to 499g
- Housing: The animals were housed singly or in pairs in solid-floor polypropylene cages furnished with woodflakes.
- Diet: Free access to food (Certified Guinea Pig Diet (Code 5026
- Water: Free access to mains tap water


ENVIRONMENTAL CONDITIONS
- Temperature: 17 to 23°
- Humidity: 30 to 70%
- Air changes (per hr): At least fifteen changes per hour
- Photoperiod (hrs dark / hrs light): Twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

arachis oil

Induction 2

Concentration / amount:

INDUCTION
Intradermal: 5% v/v
Topical: 100%

CHALLENGE CONCENTRATIONS: 100% and 75% v/v

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

arachis oil

Challenge 2

Concentration / amount:

INDUCTION
Intradermal: 5% v/v
Topical: 100%

CHALLENGE CONCENTRATIONS: 100% and 75% v/v

No. of animals per dose:

A group of fifteen guinea pigs was used for the main study, ten test and five control

Details on study design:

Selection of Concentrations for Main Study (Sighting Tests)

The concentrations of test material to be used at each stage of the main study were determined by 'sighting tests' in which groups of guinea pigs were treated with various concentrations of test material. The procedures were as follows:
Selection of Concentration for Intradermal Induction
Intradermal injections (0.1 ml/injection site) were made on the clipped shoulder of two guinea pigs, using concentrations of 1 % and 5% v/v in arachis oil BP. The degree of erythema at the injection sites was assessed approximately 24, 48, 72 hours and 7 days after injection. The degree of oedema was not evaluated. Any evidence of systemic toxicity was also recorded. The highest concentration that caused only mild to moderate skin irritation, and which was well tolerated systemically, was selected for the intradermal induction stage of the main study.
Selection of Concentration for Topical Induction
Two guinea pigs (intradermally injected with Freund's Complete Adjuvant fourteen days earlier) were treated with the undiluted test material and three preparations of the test material (75%, 50% and 25% v/v in arachis oil BP). Applications were made to the clipped flanks under occlusive dressings for an exposure period of 48 hours. The degree of erythema and oedema was evaluated approximately 1, 24 and 48 hours after dressing removal. The highest concentration producing only mild to moderate dermaI irritation was selected for the topical induction stage of the main study.
Selection of Concentration for Topical Challenge
The undiluted test material and three preparations of the test material (75%, 50% and 25% v/v in arachis oil BP) were applied to the clipped flanks of two guinea pigs under occlusive dressings for an exposure period of 24 hours. These guinea pigs did not form part of the main study but had been treated identically to the control animals of the main study, up to Day 14. The degree of erythema and oedema was evaluated approximately 1, 24 and 48 hours after dressing removal. The highest non-irritant concentration of the test material and one lower concentration were selected for the topical challenge stage of the main study.

Main Study

A group of fifteen guinea pigs was used for the main study, ten test and five control. The bodyweight of each animal was recorded at the start and end of the study.
Two phases were involved in the main study: (a) an induction of a response and (b) a challenge of that response.
Induction
Induction of the Test Animals: Shortly before treatment on Day 0 the hair was removed from an area approximately 40 mm x 60 mm on the shoulder region of each animal with veterinary clippers. A row of three injections (0.1 m) each) was made on each side of the mid-line into a 20 mm x 40 mm area. The injections were:
a) Freund's Complete Adjuvant plus distilled water in the ratio 1:1
b) a 5% v/v formulation of the test material in arachis oil BP
c) a 5% v/v formulation of the test material In a 1 :1 preparation of Freund's Complete Adjuvant plus distilled water.
Approximately 24 and 48 hours after intradermal injection the degree of erythema at the test material injection sites (ie. injection site b) was evaluated.
On Day 7 the same area on the shoulder region used previously for intradermal injections was clipped again and treated with a topical application of the undiluted test material. A filter paper patch (WHATMAN No. 4: approximate size 40 mm x 20 mm), saturated with the undiluted test material was applied to the prepared skin and held in place with a strip of surgical adhesive tape covered with an overlapping length of aluminium foil. The patch and foil were further secured with a strip of elastic adhesive bandage wound in a double layer around the torso of each animal. This occlusive dressing was kept in place for 48 hours.
The degree of erythema and oedema was quantified one and twenty-four hours following removal of the patches.
Any other reactions were also recorded.
Induction of the Control Animals: The intradermal induction was performed using an identical procedure to that used for the test animals except that the test material was omitted from the intradennal injections. Injection b) was therefore the vehicle alone, injection c) was a 50% formulation of the vehicle in a 1:1 preparation of Freund's Complete Adjuvant plus distilled waler. Similarly, the topical induction procedure was identical to that used for the test animals except that the test material was omitted.
Challenge
Shortly before treatment on Day 21, an area of approximately 50 mm x 70 mm on both flanks of each animal, was clipped tree of hair with veterinary clippers.
A square filter paper patch (WHATMAN No. 4: approximate size 20 mm x 20 mm), saturated with the undiluted test material was applied to the shorn right flank of each animal and was held in place with a strip of surgical adhesive tape. To ensure that the maximum non-irritant concentration was used at challenge, the test material at a concentration of 75% v/v in arachis oil BP was similarly applied to a skin site on the left shorn flank. The patches were occluded with an overlapping length of aluminium foil and secured with a strip of elastic adhesive bandage wound in a double layer around the torso of each animal.
After 24 hours, the dressing was carefully removed and discarded. The challenge sites were swabbed with cotton wool soaked in diethyl ether to remove residual material. The position of the treatment sites was identified by using a black indelible marker-pen.
Prior to the 24-hour obsenration the flanks were clipped using veterinary clippers 10 remove regrown hair.
Approximately 24 and 48 hours after challenge dressing removal, the degree of exythema and oedema was quantified.
Any other reactions were also recorded.

Positive control substance(s):

no

Results and discussion

In vivo (non-LLNA)

Results

Results 1

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

75% v/v in arachis oil

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: 75% v/v in arachis oil. No with. + reactions: 0.0. Total no. in groups: 10.0.

Results 2

Reading:

2nd reading

Hours after challenge:

47

Group:

test chemical

Dose level:

75% v/v in arachis oil

No. with + reactions:

0

Total no. in group:

10

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 47.0. Group: test group. Dose level: 75% v/v in arachis oil. No with. + reactions: 0.0. Total no. in groups: 10.0.

Results 3

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

100%

No. with + reactions:

1

Total no. in group:

10

Clinical observations:

1 animal with score [1]. 1 = Discrete or patchy erythema

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 100%. No with. + reactions: 1.0. Total no. in groups: 10.0. Clinical observations: 1 animal with score [1]. 1 = Discrete or patchy erythema.

Results 4

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

100%

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: 100%. No with. + reactions: 0.0. Total no. in groups: 10.0.

Results 5

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

75% v/v in arachis oil

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: 75% v/v in arachis oil. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 6

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

75% v/v in arachis oil

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: 75% v/v in arachis oil. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 7

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

100%

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: 100%. No with. + reactions: 0.0. Total no. in groups: 5.0.

Results 8

Reading:

2nd reading

Hours after challenge:

48

Group:

negative control

Dose level:

100%

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: 100%. No with. + reactions: 0.0. Total no. in groups: 5.0.

Any other information on results incl. tables

Skin Reactions Observed After Intradermal Induction Discrete or patchy to moderate and confluent erythema was noted at the intradermal induction sites of test group animals. Discrete or patchy to moderate and confluent erythema was noted at the intradermal induction sites of control group animals.Skin Reactions Observed After Topical Induction Discrete or patchy to moderate and confluent erythema was noted at the topical induction sites of test group animals. No skin reactions were noted at the topical induction sites of control group animals. Bleeding from the intradermal injection sites was noted in one test group animal. Desquamation was noted at the intradermal injection site of one test group animal at the 24 -hour observation. Skin Reactions Observed After Topical Challenge Undiluted as Supplied: Discrete or patchy erythema was noted at the challenge sites of two test group animals at the 24 -hour observarion. These reactions were not apparent at the 48-hour observation and were therefore not attributed to contact sensitisation. No skin reactions were noted at the challenge sites of the control group animals at the 24 or 48 -hour observations. 75% v/v in Arachis Oil BP: No skin reactions were noted at the challenge sites of the test or control group animals at the 24 or 48-hour observations.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test material produced a 0% (0/10) sensitisation rate and was classified as a NON-SENSITISER to guinea pig skin under the conditions of the test.
The test material did not meet the criteria for classification as a sensitiser according to CLP Regulation (EC) 1272/2008

Executive summary:

A GPMT study according to OECD 406 was performed to assess the contact sensitisation potential of the test material in the albino guinea pig. Ten test and five control animals were used for the study. Two phases were involved in the main study; an induction of a response by intradermal injection and topical application and a topical challenge of that response. Based on the results of sighting tests, the concentrations of test material for the induction and challenge phases were selected as: 5% v/v in arachis oil BP for Intradermal Induction, undiluted as supplied for Topical Induction and undiluted as supplied and 75% v/v in arachis oil BP for Topical Challenge. Under the conditions of the test, the test material produced a 0% (0/10) sensitisation rate and was classified as a non-sensitiser to guinea pig skin. The test material did not meet the criteria for classification as a sensitiser according to CLP Regulation (EC) 1272/2008.