Octadecene, reaction products with hexadecene, hydrogenated

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

08 April 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Version / remarks:

OECD Guidelines for the Testing of Chemicals No. 438 (2018)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU method B.48 (Isolated chicken eye test method for identifying occular corrosives and severe irritants)

Version / remarks:

Commission Regulation (EU) 2017/735 of 14 February 2017 amending, the Annex to Regulation (EC) No 440/2008

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Oligomerisation products of alpha-alkenes C16-18 (even numbered), hydrogenated, hydroisomerised

EC Number:

832-827-5

Cas Number:

2241366-04-9

Molecular formula:

Variable - UVCB

IUPAC Name:

Oligomerisation products of alpha-alkenes C16-18 (even numbered), hydrogenated, hydroisomerised

Test material form:

liquid

Details on test material:

Name: SynNova Base Oil
CAS number: 2241366-04-9
Batch/Lot number: TS20371/TS21270
Description: Liquid - water white colorless oil.
Purity: 100%
Expiry date: 25 February 2021/31 July 2021
Storage conditions: Controlled room temperature (15-25°C, ≤70% relative humidity)
Safety precautions: Routine safety precautions (gloves, goggles, face mask, lab coat) for unknown materials were applied to ensure personnel health and safety.

Specific details on test material used for the study:

No further details specified in the study report.

Test animals / tissue source

Species:

chicken

Strain:

other: ROSS 308

Details on test animals or tissues and environmental conditions:

Strain of chicken: ROSS 308
Source: TARAVIS KFT. (Address: 9600 Sárvár, Rábasömjéni út 129., Hungary)
Chicken heads were collected after slaughter in a commercial abattoir from chickens (approximately 7 weeks old) which are used for human consumption. Heads were collected by a slaughter house technician and heads transported to Citoxlab Hungary Ltd. at ambient temperature at the earliest convenience.
After collection, the heads were inspected for appropriate quality and wrapped with tissue paper moistened with saline, then placed in a plastic box which was closed (4-5 heads per box). The heads were received at Citoxlab Hungary Ltd. and processed within 2 hours of collection.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

30 μL of the test item was applied onto the entire surface of the cornea.
One negative control eye was treated with 30 μL of physiological saline
Three positive control eyes were treated with 30 μL 5% (w/v) Benzalkonium chloride solution.

Duration of treatment / exposure:

exposure period of 10 seconds from the end of the application

Duration of post- treatment incubation (in vitro):

The control eyes and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse.

Number of animals or in vitro replicates:

7 replicates (3 test item/3 positive control/ 1 negative control)

Details on study design:

SELECTION AND PREPARATION OF EYES FOR THE TEST
Eyes selection
After removing the head from the plastic box, it was put on soft paper. The eyelids were carefully cut away with scissors, avoiding damaging the cornea. One small drop of 2% (w/v) fluorescein solution was applied onto the cornea surface for a few seconds and subsequently rinsed off with 20 mL physiological saline. Then the fluorescein-treated cornea was examined with a hand-held slit lamp or slit lamp microscope, with the eye in the head, to ensure that the cornea was not damaged. If the cornea was in good condition, the eyeball was carefully removed from the orbit.

Preparation of eyes
The eye ball was carefully removed from the orbit by holding the nictitating membrane with a surgical forceps, while cutting the eye muscles with bent scissors. Care was taken to remove the eyeball from the orbit without cutting off the optical nerve too short. The procedure avoided pressure on the eye while removing the eyeball from the orbit, in order to prevent distortion of the cornea and subsequent corneal opacity. Once removed from the orbit, the eye was placed onto damp paper and the nictitating membrane was cut away with other connective tissue. The prepared eyes were kept on the wet papers in a closed box so that the appropriate humidity was maintained.

Eyes examination and acclimatization time
The prepared eye was placed in a steel clamp with the cornea positioned vertically with the eye in the correct relative position (same position as in the chicken head). Again avoiding too much pressure on the eye by the clamp. Because of the relatively firm sclera of the chicken eyeball, only slight pressure was needed to fix the eye properly. The clamp with the eyeball was transferred to a chamber of the superfusion apparatus. The clamp holding the eye was positioned in such a way that the entire cornea was supplied with physiological saline solution dripping from a stainless steel tube, at a rate of approximately 3-4 drops/minute or 0.1 to 0.15 mL/minutes. The door of the chamber was closed except for manipulations and examinations, to maintain temperature and humidity.
The appropriate number of eyes was selected and were placed in the superfusion apparatus. There they were examined again with the slit lamp microscope to ensure that they were in good condition. The focus was adjusted to see clearly the physiological saline which was flowing on the cornea surface. Eyes with a high baseline fluorescein staining (i.e., > 0.5) or corneal opacity score (i.e., > 0.5) were rejected. The cornea thickness was measured, any eye with cornea thickness deviating more than 10 % from the mean value for all eyes, or eyes that showed any other signs of damage, were rejected and replaced. If the selected eyes were appropriate for the test, acclimatization started and it was conducted for approximately 45 to 60 minutes. The chambers of the superfusion apparatus were at controlled temperature (32±1.5°C) during the acclimatization and treatment periods.

Identification
The eyes were identified by chamber number, marked on the door of the chamber.

THE BASELINE ASSESSMENTS
At the end of the acclimatization period, a zero reference measurement was recorded for cornea thickness and opacity to serve as a baseline (t=0) for each individual eye. The cornea thickness of the eyes should not change during acclimatization by more than 5% between the -45 minute and the zero time. No changes in thickness (0.0%) were observed in the eyes used in the study. Following the equilibration period, the fluorescein retention was measured. Baseline values were required to evaluate any potential test item related effect after treatment. All eyes were considered to be suitable for the assay.

TEST PROCEDURE
Treatment
After the zero reference measurements, the eye in its retainer was taken out of the chamber and placed on a layer of tissue with the cornea facing upwards. The eye was held in horizontal position, while the test material was applied onto the centre of the cornea. In the study, 30 μL of the test item was applied onto the entire surface of the cornea attempting to cover the cornea surface uniformly with the test item, taking care not to damage or touch the cornea. Three test item treated eyes were examined in the study.
One negative control eye was treated with 30 μL of physiological saline; three positive control eyes were treated with 30 μL 5% (w/v) Benzalkonium chloride solution.

Test item removal
The time of application was observed, then after an exposure period of 10 seconds from the end of the application, the cornea surface was rinsed thoroughly with 20 mL physiological saline* at ambient temperature, taking care not to damage the cornea but attempting to remove all residual of the test item if possible.
*Note: Physiological saline (Manufacturer: B. Braun Pharmaceuticals SA, Lot number: 84222Y05-1, Expiry date: 30 September 2021) was used for rinsing.

OBSERVATION
Observation and assessment of corneal effects
The control eyes and test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Minor variations within approximately ±5 minutes were considered acceptable.
Corneal thickness and corneal opacity were measured at all time points. Fluorescein retention was measured on two occasions, at baseline (t=0) and approximately 30 minutes after the post-treatment rinse. Haag-Streit BP 900® slit-lamp microscope was used for the measurements.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

percent corneal swelling

Run / experiment:

up to 75 min - Test item

Value:

0.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

percent corneal swelling

Run / experiment:

up to 240 min - Test item

Value:

1.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

MORPHOLOGICAL EFFECTS
Severe loosening of epithelium was observed on one eye at 120 minutes and on one eye at 180 minutes after the post-treatment rinse in case of the positive control material.

VALIDITY OF THE TEST
The results from all eyes used met the quality control standards. The negative control and positive control results were within the historical data range in the experiment. This study was considered to be valid.

Any other information on results incl. tables

TEST ITEM

Observation	Value	ICE Class
Mean maximum corneal swelling up to 75 min	0.6%	I
Mean maximum corneal swelling up to 240 min	1.7%	I
Mean maximum corneal opacity change	0.00	I
Mean fluorescein retention change	0.33	I
Other Observations	None
Overall ICE Class	3xI

Based on this in vitro eye irritation assay in isolated chicken eyes with SynNova Base Oil, the test item is non-irritant, UN GHS Classification: No Category

 

POSITIVE CONTROL

Observation	Value	ICE Class
Mean maximum corneal swelling up to 75 min	11.5%	II
Mean maximum corneal swelling up to 240 min	28.4%	III
Mean maximum corneal opacity change	4.00	IV
Mean fluorescein retention change	3.00	IV
Other Observations	Severe loosening of epithelium was observed on one eye at 120 minutes and on one eye at 180 minutes after post-treatment rinse.
Overall ICE Class	1xIII 2xIV

The positive control (5% (w/v) Benzalkonium chloride solution) was classified as severely irritating, UN GHS Classification: Category 1.

 

NEGATIVE CONTROL

Observation	Value	ICE Class
Mean maximum corneal swelling up to 75 min	0.0%	I
Mean maximum corneal swelling up to 240 min	0.0%	I
Mean maximum corneal opacity change	0.00	I
Mean fluorescein retention change	0.00	I
Other Observations	None
Overall ICE Class	3xI

The negative control Physiological saline was classified as non-irritating, UN GHS Classification: No Category.

 

SUMMARY TABLE FOR UN GHS CLASSIFICATION

Criteria for “No category” (all true)
3 endpoints classed as I or 2 endpoints classed as I and 1 endpoint classed as II, or 2 endpoints classed as II and 1 endpoint classed as I:	True
No severe corneal morphological changes:	True
Test item was not stuck to the cornea at 240 minutes after the post-treatment rinse:	True
Criteria for “Category 1” (one or more true)
2 or more endpoints classed as IV:	False
Corneal opacity = 3 at 30 min (in at least 2 eyes):	False
Corneal opacity = 4 at any time point (in at least 2 eyes):	False
Severe loosening of epithelium (in at least 1 eye):	False
Criteria for “No prediction can be made” (one or two true)
Based on the endpoints not classifiable for No Category, or for Category 1:	False
Particles of test item were stuck to the cornea and could not be washed off during the study:	False

 

Historical Control Data (updated on 07 November 2018):

 

Negative Control: Physiological Saline

Observation	Min. Value	Max. Value
Maximum corneal swelling at up to 75 min	-3.2%	3.4%
Maximum corneal swelling at up to 240 min	-4.8%	3.4%
Maximum corneal opacity change	0.00	0.50
Fluorescein retention	0.00	0.50
Number of cases	416

 

Positive Control: 5% (w/v) Benzalkonium chloride solution

Observation	Min. Value	Max. Value
Maximum corneal swelling at up to 75 min	-8.5%	27.0%
Maximum corneal swelling at up to 240 min	-10.7%	38.3%
Maximum corneal opacity change	2.50	4.00
Fluorescein retention	1.50	3.00
Number of cases	234

 

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on this in vitro eye irritation assay in isolated chicken eyes with SynNova Base Oil, the test item is non-irritant, UN GHS Classification: No Category.

Executive summary:

An in vitro eye irritation study of the test item was performed in isolated chicken’s eyes. The irritation effects of the test item were evaluated according to the OECD No. 438 (25 June 2018).

 

After the zero time reference measurements, the eyes were held in horizontal position and 30 μL of the test item was applied onto the centre of the corneas such that the entire surface of the corneas were covered. After 10 seconds, the surface was rinsed with physiological saline. The positive control eyes were treated with 30 μL of 5% (w/v) Benzalkonium chloride solution. The negative control eye was treated with 30 μL of physiological saline (0.9% (w/v) NaCl solution). In the study, three treated eyes per test item, three positive control treated eyes and one negative control treated eye were examined.

 

The results from all eyes used in the study met the quality control standards. The negative control and positive control results were in good correlation with the historical control data. Thus, the experiment was considered to be valid.

 

No significant corneal swelling (mean <5.0%) was observed during the four-hour observation period on the test item treated eyes. No corneal opacity change (severity 0.0) was noted on all eyes. No significant fluorescein retention change (severity 0.0 on one eye and severity 0.5 on two eyes) was noted on all eyes. No other corneal effects were observed.

 

Based on this in vitro eye irritation assay in isolated chicken eyes with SynNova Base Oil, the test item is non-irritant, UN GHS Classification: No Category.