Propyl oleate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017

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

Test guideline 1

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)

Deviations:

yes

Remarks:

SD of positive control slightly increased in comparison to historical mean.

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

Deviations:

yes

Remarks:

SD of positive control slightly increased in comparison to historical mean.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Propyl (9Z,12Z)-octadeca-9,12-dienoate

Molecular formula:

C21H38O2

IUPAC Name:

Propyl (9Z,12Z)-octadeca-9,12-dienoate

Constituent 2

Reference substance name:

Propyl oleate

EC Number:

203-885-3

EC Name:

Propyl oleate

Cas Number:

111-59-1

Molecular formula:

C21H40O2

IUPAC Name:

propyl oleate

Constituent 3

Reference substance name:

Propyl stearate

EC Number:

222-855-0

EC Name:

Propyl stearate

Cas Number:

3634-92-2

Molecular formula:

C21H42O2

IUPAC Name:

propyl stearate

Constituent 4

Reference substance name:

Heptadecanoic acid, propyl ester

Cas Number:

propyl heptadecanoate

Molecular formula:

C20H40O2

IUPAC Name:

Heptadecanoic acid, propyl ester

Constituent 5

Reference substance name:

Propyl (9Z)-9-hexadecenoate

Molecular formula:

C19H36O2

IUPAC Name:

Propyl (9Z)-9-hexadecenoate

Constituent 6

Reference substance name:

Propyl palmitate

EC Number:

218-803-1

EC Name:

Propyl palmitate

Cas Number:

2239-78-3

Molecular formula:

C19H38O2

IUPAC Name:

propyl palmitate

Constituent 7

Reference substance name:

Pentadecanoic acid, propyl ester

Molecular formula:

C18H36O2

IUPAC Name:

Pentadecanoic acid, propyl ester

Constituent 8

Reference substance name:

Propyl myristate

EC Number:

238-241-0

EC Name:

Propyl myristate

Cas Number:

14303-70-9

Molecular formula:

C17H34O2

IUPAC Name:

propyl myristate

Constituent 9

Reference substance name:

Propyl laurate

EC Number:

222-961-7

EC Name:

Propyl laurate

Cas Number:

3681-78-5

Molecular formula:

C15H30O2

IUPAC Name:

propyl laurate

Test material form:

liquid

Details on test material:

Name: 9-Octadecenoic acid (9Z)-, propyl ester
Product Description: Propyl oleate
CAS No.: 111-59-1
Physical state: colourless to yellow liquid at 20 °C
Batch No.: 37584
Re-certification date of batch: 08 March 2018
Purity: > 95 % (mono constituent substance, water content 0.33 % (w/w))
Free Fatty Acid, % Oleic: 0,1
Color, Gardner: 3
Iodine Value, cg I2/g 75
Cloud Point, degrees C 0
Saponification Number, mg KOH/g 177
Moisture, % 0,07
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

Specific details on test material used for the study:

Name: 9-Octadecenoic acid (9Z)-, propyl ester
Product Description: Propyl oleate
CAS No.: 111-59-1
Physical state: pale yellow to yellow liquid at 20 °C
Batch No.: 37584
Re-certification date of batch: 08 March 2018
Purity: > 99 % (fatty acid propyl esters, max water content 0.3 % (w/w))
Free Fatty Acid, % Oleic: 0.1
Color, Gardner: 3
Iodine Value, cg I2/g 75
Cloud Point, degrees C 0
Saponification Number, mg KOH/g 177
Moisture, % 0,07
Stability: stable under test conditions
Storage condition of test material: Room temperature, protected from light

Test animals / tissue source

Details on test animals or tissues and environmental conditions:

Preparation of the Corneas

The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered at the abattoir A. Moksel AG, Buchloe, Germany. On the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the laboratories. Immediately after arrival of the eyes, cornea preparation was initiated. The eyes were carefully examined for defects and any defective eyes were discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded. The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first. The corneas were incubated for one hour at 32 +- 1 °C.

Test Groups:

- 3 corneas for the test item
- 3 corneas as negative controls treated with physiological saline 0.9% NaCl
- 3 corneas as positive controls treated with ethanol 100%

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 µL of the test substance or the control substance was introduced into the anterior chamber. As the viscosity of the test item was relatively high, it was applied directly onto the cornea by removing the window-locking ring and glass window prior to treatment. After 10 minutes incubation at 32 +- 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red).

Duration of treatment / exposure:

After the equilibration period, the medium was removed from both chambers and replaced with fresh complete RPMI. An initial measurement was performed on each of the corneas using the opacitometer. Three corneas with illuminance readings approximately equivalent to the median illuminance of all corneas were selected as negative-control corneas. The illuminance of each cornea was read and recorded. Only corneas that had an initial illuminance reading I > I0/1.1651 lux were used for the assay. The medium was removed from the anterior chamber and replaced with the test item or control. 750 µL of the test substance or the control substance was introduced into the anterior chamber. As the viscosity of the test item was relatively high, it was applied directly onto the cornea by removing the window-locking ring and glass window prior to treatment. After 10 minutes incubation at 32 +- 1 °C either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber expect of cornea no. 8 was refilled with complete RPMI and an illuminance measurement was performed after 2 hours incubation at 32 +- 1 °C. Also, each cornea was observed visually and pertinent observations were recorded.

Duration of post- treatment incubation (in vitro):

After the illuminance measurement was performed, the medium was removed from both chambers of the holder. The posterior chamber was refilled with fresh complete RPMI. 1 mL of a 4 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were incubated for 90 minutes at 32 +- 1 °C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer.

Number of animals or in vitro replicates:

Test Groups:

- 3 corneas for the test item
- 3 corneas as negative controls treated with physiological saline 0.9% NaCl
- 3 corneas as positive controls treated with ethanol 100%

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

mean

Value:

0.93

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

physiological saline 0.9% NaCl; IVIS: 1.18

Positive controls validity:

valid

Remarks:

ethanol 100%; IVIS: 74.52

Remarks on result:

no indication of irritation

Other effects / acceptance of results:

Validity

The in vitro irritation score obtained with the positive control is slightly increased (deviation of 5.47 SD) in comparison to the two standard deviations of the current historical mean. The deviation of the positive control did not influence the quality or integrity of the present study and therefore the assay is considerd to be valid.
The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

The eye irritancy potential of the test item was investigated in the bovine corneal opacity and permeability assay. The following mean in vitro irritation score was calculated to 0.93. Therefore the test item was classified into UN GHS No Category.

Executive summary:

The eye irritancy potential of the test item was investigated in the bovine corneal opacity and permeability assay. The following mean in vitro irritation score was calculated to 0.93. Therefore the test item was classified into UN GHS No Category.