Acetic acid, esters with turpentine-oil myrcene fraction terpene alcs.

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

p-menth-1-en-8-yl acetate

EC Number:

201-265-7

EC Name:

p-menth-1-en-8-yl acetate

Cas Number:

80-26-2

Molecular formula:

C12H20O2

IUPAC Name:

1-methyl-1-(4-methylcyclohex-3-en-1-yl)ethyl acetate

Constituent 2

Reference substance name:

Myrcenyl acetate

EC Number:

214-262-0

EC Name:

Myrcenyl acetate

Cas Number:

1118-39-4

Molecular formula:

C12H20O2

IUPAC Name:

1,1-dimethyl-5-methylenehept-6-en-1-yl acetate

Constituent 3

Reference substance name:

(E)-2-(3,3-dimethylcyclohexylidene)ethyl acetate

EC Number:

308-183-1

EC Name:

(E)-2-(3,3-dimethylcyclohexylidene)ethyl acetate

Cas Number:

97890-04-5

Molecular formula:

C12H20O2

IUPAC Name:

(E)-2-(3,3-dimethylcyclohexylidene)ethyl acetate

Constituent 4

Reference substance name:

(Z)-2-(3,3-dimethylcyclohexylidene)ethyl acetate

EC Number:

308-184-7

EC Name:

(Z)-2-(3,3-dimethylcyclohexylidene)ethyl acetate

Cas Number:

97890-05-6

Molecular formula:

C12H20O2

IUPAC Name:

(Z)-2-(3,3-dimethylcyclohexylidene)ethyl acetate

Constituent 5

Reference substance name:

Geranyl acetate

EC Number:

203-341-5

EC Name:

Geranyl acetate

Cas Number:

105-87-3

Molecular formula:

C12H20O2

IUPAC Name:

(E)-3,7-dimethylocta-2,6-dien-1-yl acetate

Constituent 6

Reference substance name:

1-methyl-4-(1-methylethylidene)cyclohexyl acetate

EC Number:

233-564-3

EC Name:

1-methyl-4-(1-methylethylidene)cyclohexyl acetate

Cas Number:

10235-63-9

Molecular formula:

C12H20O2

IUPAC Name:

4-isopropylidene-1-methylcyclohexyl acetate

Constituent 7

Reference substance name:

4-(5-methyl-1-methylene-4-hexenyl)-1-(4-methylpent-3-enyl)cyclohexene

EC Number:

208-546-3

EC Name:

4-(5-methyl-1-methylene-4-hexenyl)-1-(4-methylpent-3-enyl)cyclohexene

Cas Number:

532-87-6

Molecular formula:

C20H32

IUPAC Name:

4-(5-methyl-1-methylenehex-4-en-1-yl)-1-(4-methylpent-3-en-1-yl)cyclohexene

Constituent 8

Reference substance name:

cis-1-methyl-4-(1-methylvinyl)cyclohexyl acetate

EC Number:

244-028-3

EC Name:

cis-1-methyl-4-(1-methylvinyl)cyclohexyl acetate

Cas Number:

20777-47-3

Molecular formula:

C12H20O2

IUPAC Name:

4-isopropenyl-1-methylcyclohexyl acetate

Constituent 9

Reference substance name:

p-menth-1-en-8-ol

EC Number:

202-680-6

EC Name:

p-menth-1-en-8-ol

Cas Number:

98-55-5

Molecular formula:

C10H16O

IUPAC Name:

2-(4-methylcyclohex-3-en-1-yl)propan-2-ol

Constituent 10

Reference substance name:

Neryl acetate

EC Number:

205-459-2

EC Name:

Neryl acetate

Cas Number:

141-12-8

Molecular formula:

C12H20O2

IUPAC Name:

(Z)-3,7-dimethylocta-2,6-dien-1-yl acetate

Constituent 11

Reference substance name:

1-(3,3-dimethylcyclohex-1-en-1-yl)ethyl acetate

Cas Number:

150461-96-4

Molecular formula:

C12H20O2

IUPAC Name:

1-(3,3-dimethylcyclohex-1-en-1-yl)ethyl acetate

Constituent 12

Reference substance name:

Tentative monoterpene diacetate

IUPAC Name:

Tentative monoterpene diacetate

Constituent 13

Reference substance name:

Tentative oxygenated monoterpene acetate

IUPAC Name:

Tentative oxygenated monoterpene acetate

Constituent 14

Reference substance name:

Tentative Diterpene hydrocarbon

IUPAC Name:

Tentative Diterpene hydrocarbon

Constituent 15

Reference substance name:

Unresolved/non-elucidated mixed terpene/diterpene/oxygenated terpene/terpene acetate derivatives

IUPAC Name:

Unresolved/non-elucidated mixed terpene/diterpene/oxygenated terpene/terpene acetate derivatives

Test material form:

liquid

Specific details on test material used for the study:

Identification: Neobergamate Forte
Appearance: Pale yellow liquid
Batch: SC00015234
Purity/Composition: See Certificate of Analysis
Test item storage: At room temperature protected from light
Stable under storage conditions until: 07 November 2016 (expiry date)

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

Test System: Bovine eyes were used as soon as possible after slaughter.
Rationale: In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimise the need of in vivo testing (1-6). As a consequence a validated and accepted in vitro test for eye irritation should be performed before in vivo tests are conducted. One of the proposed validated in vitro eye irritation tests is the Bovine Corneal Opacity and Permeability (BCOP) test.
Source: Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter.
Transport: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:

unchanged (no vehicle)

Remarks:

The test item was tested neat. To protect the test item from light, glassware was wrapped in tinfoil.

Controls:

yes, concurrent positive control

yes, concurrent negative control

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

>= 1 - <= 2

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

In vitro irritancy score

The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score:
In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value)

Additionally the opacity and permeability values were evaluated independently to determine whether the test item induced irritation through only one of the two endpoints.

The IVIS cut-off values for identifying the test items as inducing serious eye damage (UN GHS Category 1) and test items not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given hereafter:

In vitro score range UN GHS
≤ 3 No Category
> 3; ≤ 55 No prediction can be made
>55 Category 1

Acceptability of the assay
The assay is considered acceptable if:
- The positive control gives an in vitro irritancy score that falls within two standard deviations of the current historical mean.
- The negative control responses should result in opacity and permeability values that are less than the upper limits of the laboratory historical range

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (10% (w/v) Benzalkonium Chloride) was 141 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.

Neobergamate Forte did not induce ocular irritation through both endpoints, resulting in a mean in vitro irritancy score of 1.4 after 10 minutes of treatment.
Since Neobergamate Forte induced an IVIS ≤ 3, no classification is required for eye irritation or serious eye damage.