Reaction mass of (E)-3-methylcyclotetradec-5-en-1-one and (Z)-3-methylcyclotetradec-5-en-1-one

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

27 jan 2004 to 30 mar 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

conducted under GLP conditions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

2002

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.4 (Acute Toxicity: Dermal Irritation / Corrosion)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(E)-3-methylcyclotetradec-5-en-1-one

Cas Number:

259854-70-1

Molecular formula:

C15H26O

IUPAC Name:

(E)-3-methylcyclotetradec-5-en-1-one

Constituent 2

Reference substance name:

(Z)-3-methylcyclotetradec-5-en-1-one

Cas Number:

259854-71-2

Molecular formula:

C15H26O

IUPAC Name:

(Z)-3-methylcyclotetradec-5-en-1-one

Test material form:

liquid

Specific details on test material used for the study:

Identification: KARMALONE
Description: Colourless to pale yellow liquid
Batch number: TOT0300497
Purity: 90.9%
Stability of test item: Stable under storage conditions
Expiry date: 28-NOV-2004
Storage conditions: At room temperature (range of 20 ± 3 °C), light protected.
Safety precautions: Routine hygienic procedures were used to ensure the health and satety of the personnel.

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Source: Elevage scientifique des Dombes F-01400 Chatillon sur Chalaronne / France
Number of animals per test : 3 (animals of both sexes were used)
Age at treatment: 12 weeks (male and females)
Identification: By unique cage number and corresponding ear number
Acclimatization: Under laboratory conditions after health examination. Only animals without any visual signs of illness were used for the study.
Allocation: Male No. 53 and Females Nos 54 and 55
Room number: 106 / RCC Ltd, Fullinsdorf
Conditions:
Standard Laboratory Conditions
Air-conditioned with target ranges for room temperature 17-23 °C, relative humidity 30-70 % and approximately 10-15 air changes per hour. Room temperature and humidity were monitored continuously and values outside of these ranges may have occasionally occurred, usually following room cleaning. These transient variations are considered not to have any influence on the study and, therefore, these data are not reported but are retained at RCC. The animals were provided with an automatically controlled light cycle of 12 hours light and 12 hours dark. Music was played during the daytime light period.
Accommodation: Individually in stainless steel cages equipped with feed hoppers and drinking water bowls. Wood blocks (RCC Ltd, Fullinsdorf) and haysticks 4646 (batch no. 0403, Provimi Kliba AG) were provided for gnawing.
Diet: Pelleted standard Provimi Kliba 3418 rabbit maintenance diet ad libitum (batch no. 86/03) provided by Provimi Kliba AG, CH-4303 Kaiseraugst. Results of analysis for contami­ nants are archived at RCC Ltd, ltingen.
Water: Community tap water from Fullinsdorf, ad libitum. Results of bacteriological, chemical and contaminant analyses are ar­ chived at RCC Ltd, ltingen.

Test system

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

o.5 mL of Karmalone undiluted

Duration of treatment / exposure:

4 hours

Observation period:

Approximately 1, 24, 48 and 72 hours, as well as 7 and 10 days after the removal of the dressing, gauze patch and test item.

Number of animals:

3(one male and two females)

Details on study design:

Four days before treatment, the left flank was clipped with an electric clipper, exposing an area of approximately 100 cm2 (1O cm x 1O cm). The skin of the animals was examined one day before treatment, and regrown fur of all animals was clipped again.
Animals with overt signs of skin injury or marked irritation which may have interfered with the interpretation of the results were not used in the test.
On the day of treatment, 0.5 ml of KARMAlONE was placed on a surgical gauze patch (ca. 4 cm x 4 cm). This gauze patch was applied to the intact skin of the clipped area. The patch was covered with a semi-occlusive dressing. The dressing was wrapped around the abdomen and anchored with tape.
The duration of treatment was 4 hours. Then the dressing was removed and the skin was flushed with lukewarm tap water to clean the application site so that any reactions (erythema) were clearly visible at that time.

Observations:
Viability/Mortality: Daily from acclimatization of the animals to the termination of test.
Clinical signs: Daily from acclimatization of the animals to the termination of test.
Body weights: At start of acclimatization, on the day of application and at termination of observation.

Irritation scores: The skin reaction was assessed according to the numerical scoring system listed in the EEC Commission Directive 92/69/EEC, July 31, 1992 approximately 1, 24, 48 and 72 hours, as well as 7 and 10 days after the removal of the dressing, gauze patch and test item.
To allow further examination of the test sites animal Nos. 53 and 55 were re-clipped on completion of the 24-hour examination as well as No. 53 again on completion of the 72-hour reading.

Treatment of results:
Data are summarized in tabular form, showing the irritation scores for erythema and oedema for each individual animal at all observation intervals after patch removal, any lesions, a description of the degree and nature of irritation, corrosion or reversibility, and any other toxic effects observed.
To evaluate the irritation of the test item the mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. An animal is positive when the mean score is 2 or greater. The test is positive for irritation when at least 2 animals are positive for the same endpoint (erythema/eschar or oedema).

Necropsy:
No necropsy was performed on the animals sacrificed at termination of observation.
All rabbits were sacrificed by an intravenous injection of Vetanarcol into the ear vein at a dose of at least 1 mUkg body weight (equivalent to 162 mg sodium pentobarbitone/kg body weight) and discarded.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

erythema score

Basis:

animal: 53

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 2

Irritation parameter:

erythema score

Basis:

animal: 54

Time point:

24/48/72 h

Score:

0.67

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 3

Irritation parameter:

erythema score

Basis:

animal: 55

Time point:

24/48/72 h

Score:

1.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 4

Irritation parameter:

erythema score

Basis:

animal: 53, 54 and 55

Time point:

other: 1 hour

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 5

Irritation parameter:

erythema score

Basis:

animal: 53, 54 and 55

Time point:

24 h

Score:

>= 1 - <= 2

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 6

Irritation parameter:

erythema score

Basis:

animal: 53, 54 and 55

Time point:

48 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 7

Irritation parameter:

erythema score

Basis:

animal: 53, 54 and 55

Time point:

72 h

Score:

>= 0 - <= 1

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 8

Irritation parameter:

erythema score

Basis:

other: 53, 54 and 55

Time point:

7 d

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 9

Irritation parameter:

erythema score

Basis:

animal: 53, 54 and 55

Time point:

10 d

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 10

Irritation parameter:

edema score

Basis:

animal: 53

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 11

Irritation parameter:

edema score

Basis:

animal: 54

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 12

Irritation parameter:

edema score

Basis:

animal: 55

Time point:

24/48/72 h

Score:

0.33

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 13

Irritation parameter:

edema score

Basis:

animal: 53, 54 and 55

Time point:

other: 1 hour

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 14

Irritation parameter:

edema score

Basis:

animal: 53, 54 and 55

Time point:

24 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 15

Irritation parameter:

edema score

Basis:

animal: 53, 54 and 55

Time point:

48 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 16

Irritation parameter:

edema score

Basis:

animal: 53, 54 and 55

Time point:

72 h

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 17

Irritation parameter:

edema score

Basis:

animal: 53, 54 and 55

Time point:

7 d

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 days

Results 18

Irritation parameter:

edema score

Basis:

animal: 53, 54 and 55

Time point:

10 d

Score:

0

Max. score:

4

Reversibility:

fully reversible within: 10 days

Irritant / corrosive response data:

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score of the three animals was 0.67, 0.67 and 1.33, respectively and the mean oedema score was 0.33 for all three animals.
Well-defined erythema was observed in all animals at the 1-hour reading and very slight to well-defined erythema was noted in all animals 24 hours after treatment. Very slight erythema was present in all animals at the 48-hour examination and persisted in one animal up to 72 hours after treatment.
Very slight swelling (oedema) was present in all animals at the 1- and 24-hour reading. Scaling was observed in one animal 72 hours and 7 days after treatment.
No abnormal findings were observed on the treated skin of any animal 10 days after treatment, the end of the observation time.

Other effects:

No clinical signs of systemic toxicity were observed in the animals during the study and no mortality occurred.
No staining produced by the test item of the treated skin was observed.
The body weights of all rabbits were considered to be within the normal range of variability.

Any other information on results incl. tables

Body weight in grams:

Body weight in grams
AnimalNo.	Sex	First Day of Acclimatization	Day of Treatment	Last Day ofObservation
53	male	2225	2476	2723
54	female	2127	2306	2718
55	female	2098	2385	2635

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), KARMALONE is considered to be "not irritating" to rabbit skin.

Executive summary:

The primary skin irritation potential of KARMALONE was investigated according to OECD test guideline no. 404. The test item was applied by topical semi-occlusive application of 0.5 ml to the intact left flank of each of three young adult New Zealand White rabbits. The duration of treatment was four hours. The scoring of skin reactions was performed 1, 24, 48 and 72 hours, as well as 7 and 10 days after removal of the dressing.

The mean score was calculated across 3 scoring times (24, 48 and 72 hours after patch removal) for each animal for erythema/eschar grades and for oedema grades, separately. The mean erythema/eschar score of the three animals was 0.67, 0.67 and 1.33, respectively and the mean oedema score was 0.33 for all three animals.

The application of KARMALONE to the skin resulted in slight, early-onset and transient signs of irritation such as erythema, oedema and scaling. These effects were reversible and were no longer evident 10 days after treatment, the end of the observation period for all animals. The test item caused no staining of the treated skin. No corrosive effects were noted on the treated skin of any animal at any of the measuring intervals and no other clinical signs of substance related effects were observed.

Thus, the test item did not induce significant or irreversible damage to the skin.

Based upon the referred classification criteria (Commission Directive 2001/59/EC of August 2001), KARMALONE is considered to be "not irritating" to rabbit skin.