2-Butenoic acid, 1-ethyl-2-methylpropyl ester, (2E)-

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was conducted between 20 May 2014 and 03 June 2014

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study is considered to be a reliability 1 as it has been conducted according to OECD Test Guideline 404 using the in vivo Acute Dermal Irritation/Corrosion method and in compliance with GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

2-methylpentan-3-yl (2E)-but-2-enoate

EC Number:

806-544-2

Cas Number:

1370711-06-0

Molecular formula:

C10H18O2

IUPAC Name:

2-methylpentan-3-yl (2E)-but-2-enoate

Test material form:

other: liquid

Details on test material:

Identification: IFF TM 12-209
Storage Conditions: approximately 4 °C in the dark
Physical state / Appearance: extremely pale yellow liquid

Test animals

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Three New Zealand White (Hsdlf:NZW) strain rabbits were supplied by Harlan Laboratories UK Ltd., Leicestershire, UK. At the start of the study the animals weighed 2.92 to 3.10 kg and were twelve to twenty weeks old. After an acclimatization period of at least five days each animal was given a number unique within the study which was written with a black indelible marker pen on the inner surface of the ear and on the cage label.

The animals were individually housed in suspended cages. Free access to mains drinking water and food (2930C Teklad Global Rabbit diet supplied by Harlan Laboratories UK Ltd., Oxon, UK) was allowed throughout the study. The diet and drinking water were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

The temperature and relative humidity were set to achieve limits of 17 to 23 °C and 30 to 70% respectively. The rate of air exchange was at least fifteen changes per hour and the lighting was controlled by a time switch to give twelve hours continuous light (06:00 to 18:00) and twelve hours darkness.

The animals were provided with environmental enrichment items which were considered not to contain any contaminant of a level that might have affected the purpose or integrity of the study.

Test system

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

0.5 mL of the test item was applied directly to the skin.

Test Item Formulation
For the purpose of the study the test item was used as supplied. The absorption of the test item was not determined.

Duration of treatment / exposure:

Four hours

Observation period:

14 days

Number of animals:

Three

Details on study design:

On the day before the test each of a group of three rabbits was clipped free of fur from the dorsal/flank area using veterinary clippers. Only animals with a healthy intact epidermis by gross observation were selected for the study.

On the day of the test a suitable test site was selected on the back of each rabbit. A quantity of 0.5 mL of the undiluted test item was applied directly to the skin under a 2.5 cm x 2.5 cm cotton gauze patch. The patch was secured in position with a strip of surgical adhesive tape. To prevent the animals interfering with the patches, the trunk of each rabbit was wrapped in an elasticated corset and the animals were returned to their cages for the duration of the exposure period.

Four hours after application the corset and patches were removed from each animal and any residual test item removed by gentle swabbing with cotton wool soaked in distilled water.

Immediately following removal of the patches and approximately 1, 24, 48 and 72 hours later, the test sites were examined for evidence of primary irritation and scored.

Any other skin reactions and clinical signs of toxicity, if present, were also recorded.

Additional observations were made on Days 7 and 14 to assess the reversibility of skin reactions.

Individual body weights were recorded on Day 0 (the day of dosing) and at the end of the observation period.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: Erythema/Eschar Formation

Basis:

animal #1

Remarks:

74312 Male

Time point:

other: 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Erythema extended approximately 10 mm around the treated skin site. At the 7 day timepoint a score of 1 was given with slight desquamation noted.

Results 2

Irritation parameter:

other: Erythema/Eschar Formation

Basis:

animal #2

Remarks:

74313 Male

Time point:

other: 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Erythema extended approximately 10 mm around the treated skin site. At the 7 day timepoint a score of 1 was given with slight desquamation noted.

Results 3

Irritation parameter:

other: Erythema/Eschar Formation

Basis:

animal #3

Remarks:

74314 Male

Time point:

other: 24, 48 and 72 hours

Score:

1.67

Max. score:

4

Reversibility:

fully reversible within: 14 days

Remarks on result:

other: Erythema extended approximately 10 mm around the treated skin site. At the 7 day timepoint a score of 1 was given with slight desquamation noted.

Results 4

Irritation parameter:

edema score

Basis:

animal #1

Remarks:

74312 Male

Time point:

other: 24, 48 and 72 hours

Score:

2

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 5

Irritation parameter:

edema score

Basis:

animal #2

Remarks:

74313 Male

Time point:

other: 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Results 6

Irritation parameter:

edema score

Basis:

animal #3

Remarks:

74314 Male

Time point:

other: 24, 48 and 72 hours

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 7 days

Irritant / corrosive response data:

Well-defined erythema and very slight or slight edema were noted at all treated skin sites at the 24-Hour observation. Well defined erythema and very slight or slight edema were noted at two treated skin sites with very slight erythema and very slight edema noted at one treated skin site at the 48 Hour observation. Well defined erythema and very slight or slight edema were noted at all treated skin sites at the 72 Hour observation. The erythema extended approximately 10 mm around all treated skin sites at the 24 and 48 Hour observations. Very slight erythema and slight desquamation were noted at all treated skin sites at the 7 Day observation. All treated skin sites appeared normal at the 14 Day observation.

Other effects:

Body Weight
All animals showed expected gain in body weight during the study.

Any other information on results incl. tables

Individual Skin Reactions

Skin Reaction	Observation Time (following patch removal)	Individual Scores – Rabbit Number and Sex			Total
		74312 Male	74313 Male	74314 Male
Erythema/Eschar Formation	Immediately	0	0	0	(0 )
	1 Hour	0	0	0	( 0 )
	24 Hours	2R	2R	2R	6
	48 Hours	2R	2R	1R	( 5 )
	72 Hours	2	2	2	6
	7 Days	1D	1D	1D	( 3 )
	14 Days	0	0	0	( 0 )
Edema Formation	Immediately	0	0	0	( 0 )
	1 Hour	0	0	0	( 0 )
	24 Hours	2	1	1	4
	48 Hours	2	1	1	( 4 )
	72 Hours	2	1	1	4
	7 Days	0	0	0	( 0 )
	14 Days	0	0	0	( 0 )
Sum of 24 and 72-Hour Readings (S)           :          20
Primary Irritation Index (S/6)            :          20/6 = 3.3
Classification                                     :          MODERATE IRRITANT

(   ) = Total values not used for calculation of primary irritation index

R = Erythema extended approximately 10 mm around the treated skin site

D = Slight desquamation

Individual Body Weights and Body Weight Change

Rabbit Number and Sex	Individual Body Weight (kg)		Body Weight Change (kg)
	Day 0	Day 14
74312 Male	3.04	3.38	0.34
74313 Male	3.10	3.47	0.37
74314 Male	2.92	3.40	0.48

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

The test item produced a primary irritation index of 3.3 and was classified as a moderate irritant to rabbit skin according to the Draize classification scheme. No corrosive effects were noted.

The test item meets the criteria for classification as a Mild irritant (Category 3) according to the Globally Harmonized System of Classification and Labelling of Chemicals.

The test item does not meet the criteria for classification according to Regulation (EC) No 1272/2008, relating to the Classification, Labelling and Packaging of Substances and Mixtures.

Executive summary:

The dermal irritation potential of the test substance, TM 12-209, was assessed as not irritating according to OECD Test Guideline 404 using the in vivo Acute Dermal Irritation/Corrosion method.