p-mentha-1,4-diene

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:

2017

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-mentha-1,4-diene

EC Number:

202-794-6

EC Name:

p-mentha-1,4-diene

Cas Number:

99-85-4

Molecular formula:

C10H16

IUPAC Name:

p-mentha-1,4-diene

Specific details on test material used for the study:

Designation in Test Facility: 17022802G
Date of Receipt: 28. Feb. 2017
Condition at Receipt Room temperature, in proper conditions
6.1.1 Specification
The following information concerning identity and composition of the test item was provid-ed by the sponsor.
Name Gamma Terpinene
Batch no. 161215
Appearance Clear liquid
Composition p-mentha-1,4-diene
Purity 98%
Homogeneity homogeneous
Expiry date 14. Dec. 2018
Storage Room Temperature (20 ± 5 °C), keep away from light

The following additional information was relevant to the conduct of the study, according to
OECD 437:

CAS No. 99-85-4
EINECS-No. 202-794-6
Chemical Class not stated
Volatility 0.9 hPa at 20 °C
pH-value unknown
Stability H2O: unknown; EtOH: unknown; acetone: unknown; CH3CN: unknown; DMSO: unknown
Solubility H2O: < 0.1 g/L; EtOH: unknown; acetone: unknown; CH3CN: unknown; DMSO: unknown

It was provided by the sponsor as well.

Test animals / tissue source

Species:

cattle

Strain:

other: Bovine corneas were used. They were collected from slaughtered cattle which were be-tween 12 and 60 months old.

Details on test animals or tissues and environmental conditions:

Specification
Species:Bos primigenius Taurus (fresh bovine corneas)
Origin
Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hank’s balanced salt solu-tion (supplemented with 0.01% streptomycin and 0.01% penicillin). Then, the corneas were dissected and incubated in medium at 32 ± 1 °C in an incubation chamber for 1 h.

Test system

Vehicle:

Hank's balanced salt solution

Controls:

yes, concurrent positive control

yes, concurrent negative control

Duration of treatment / exposure:

1 hour

Duration of post- treatment incubation (in vitro):

2 hours

Number of animals or in vitro replicates:

3

Details on study design:

One valid experiment was performed.
Bovine corneas were used. They were collected from slaughtered cattle which were be-tween 12 and 60 months old.
The test item Gamma Terpinene was brought onto the cornea of a bovine eye which had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been measured.
The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of the test item and 2 hours post-incubation, opacity and permeability values were measured.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

in vitro irritation score

Run / experiment:

Negative Control HBSS-solution

Value:

1.28

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Run / experiment:

Positive Control-DMF undiluted

Value:

120.29

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Run / experiment:

Test Item-Gamma Terpinene

Value:

6.55

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

This in vitro study was performed to assess corneal damage potential of Gamma Terpinene by quantitative measurements of changes in opacity and permeability in a bo-vine cornea.
The test item Gamma Terpinene was brought onto the cornea of a bovine eye which pre-viously had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been determined. The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of the test item and 2 hours post-incubation, opacity and permeability values were measured.

The test item was tested pure.
Under the conditions of this test, the test item Gamma Terpinene showed effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 6.55.
According to OECD Guideline no. 437 (Jul. 2013), a substance with an IVIS > 3 and ≤ 55 induces effects on the cornea, that cannot be classified in an UN GHS Category.

The negative control (HBSS-solution) and the positive control (undiluted dimethylforma-mide) have met the validity criteria.

No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.