Oct-7-enal

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-08-09

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

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)

Version / remarks:

2013

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Oct-7-enal

EC Number:

244-449-2

EC Name:

Oct-7-enal

Cas Number:

21573-31-9

Molecular formula:

C8H14O

IUPAC Name:

oct-7-enal

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Chemical name: Oct-7-enal (OEL)
- CAS no.: 21573-31-9
- EC-no.: not assigned
- Source and lot/batch No.of test material: Kuraray / 82725
- Expiration date of the lot/batch: 24 April 2018
- Molecular weight: 126.198 g/mol
- Purity: 95.4%

Test animals / tissue source

Species:

cattle

Strain:

not specified

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

TEST MATERIAL
- Amount applied (volume or weight with unit): 750 uL
- Concentration (if solution): as recieved

VEHICLE
- Amount applied (volume or weight with unit): 750 uL
- Concentration (if solution): 0.9% NaCl

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

10 minutes

Number of animals or in vitro replicates:

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%

Details on study design:

SELECTION AND PREPARATION OF CORNEAS

QUALITY CHECK OF THE ISOLATED CORNEAS

NUMBER OF REPLICATES: 3 corneas/dose

NEGATIVE CONTROL USED: yes, physiological saline 0.9% NaCl

POSITIVE CONTROL USED: yes, ethanol 100%

APPLICATION DOSE AND EXPOSURE TIME: Single 750 uL application for 10 minutes at 32°C

TREATMENT METHOD: open chamber

POST-INCUBATION PERIOD: no

REMOVAL OF TEST SUBSTANCE
- Number of washing steps after exposure period: washed three times with Minimum essential medium (MEM) with phenol red (used for the rinsing of test substances only). Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red).

- POST-EXPOSURE INCUBATION: 2 hours.

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: opacitometer
- Corneal permeability: 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 °C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490) was determined, using a spectrophotometer (Jenway 6405 UV/VIS).
- Others (e.g, pertinent visual observations, histopathology): each cornea was observed visually and pertinent observations were recorded

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: The IVIS cut-off values for identifying test substances as inducing serious eye damage (UN GHS Category 1) and test substances not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given below:
IVIS UN GHS
= 3 No Category
> 3; = 55 No prediction can be made
> 55 Category 1

An identification of test substances that should be classified as irritating to eyes (UN GHS Category 2 or Category 2A) or test substances that should be classified as mildly irritating to eyes (UN GHS Category 2B) cannot be made

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

ca. 59.17

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

positive indication of irritation

Other effects / acceptance of results:

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control:
- Acceptance criteria met for positive control:

Any other information on results incl. tables

Individual Data

 

Table CA 7.3.2/02-1:Opacity

Test item	Cornea no.	Initial opacity	Final opacity	Change of opacity value	Corrected opacity value
Negative control (saline, 0.9% NaCl)	1	1.65	1.08	-0.57	n/a
	2	1.65	1.54	-0.11
	3	1.58	1.04	-0.53
	Mean	1.63	1.22	-0.40
Test item (undiluted)	1	0.02	13.12	13.10	13.51
	2	1.15	13.24	12.09	12.49
	3	2.54	16.72	14.19	14.59
	Mean	1.23	14.36	13.13	13.53
Positive control (ethanol, 100%)	1	3.11	40.60	37.49	37.90
	2	2.88	30.21	27.34	27.74
	3	3.03	33.25	30.22	30.62
	Mean	3.01	34369	31.68	32.09

 

Table CA 7.3.2/02-2: Permeability

Test item	Cornea no.	OD490	Corrected OD490 value
Negative control (saline, 0.9% NaCl)	1	0.009	n/a
	2	0.013
	3	0.004
	Mean	0.009
Test item (undiluted)	1	2.645	2.636
	2	2.995	2.986
	3	3.515	3.506
	Mean	3.052	3.043
Positive control (ethanol, 100%)	1	1.1013	1.004
	2	1.147	1.138
	3	2.120	2.111
	Mean	1.427	1.418

 

Table CA 7.3.2/02-3: In Vitro Irritation Score

Test item	Cornea no.	Corrected opacity	Corrected OD490 value	IVIS
Negative control (saline, 0.9% NaCl)	1	-0.57	0.009	-0.27
	2	-0.11	0.013
	3	-0.53	0.004
	Mean	-0.40	0.009
Test item (undiluted)	1	13.51	2.636	59.17
	2	12.49	2.986
	3	14.59	3.506
	Mean	13.53	3.043
Positive control (ethanol, 100%)	1	37.90	1.004	53.36
	2	27.74	1.138
	3	30.62	2.111
	Mean	32.09	1.418

 

Table CA 7.3.2/02-4: Historical Mean In Vitro Irritation Score of the Positive Control

	IVIS positive control – ethanol 100%
Mean value:	48.47
Standard deviation (SD):	10.08
Mean value - 2 SD:	28.31
Mean value + 2 SD:	68.64
No. of replicates providing historical mean: 43

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

Under the conditions of this study the Oct-7-enal showed irreversible effects on the eye using the BCOP permeability assay. The mean in vitro irritancy score (IVIS) was 59.17 after a 10 minute exposure (+ 2 hour post-incubation). Therefore, according to Annex I for Regulation (EC) 1272/2008 the active ingredient, Oct-7-enal is classified as Category 1: Irreversible effects on the eye, H318: Causes serious eye damage.

Executive summary:

The BCOP test method was used to identify if Oct-7-enal induced serious eye damage as defined by UN GHS, i.e. chemicals to be classified as UN GHS Category. In the present study Oct-7-enal was applied topically to the isolated bovine corneas for 10 minutes followed by a 2 hour post-incubation period. Negative (0.9% NaCl) and positive controls (ethanol, 100% were included). Irritant potential of the test article was predicted using an opacitometer to assess corneal opacity, with corneal permeability assessed with the sodium fluorescein solution, with optical density at 490 nm ready using a spectrophotometer. These values were corrected for background opacity and the negative control permeability. The mean opacity and permeability OD490 values for each treatment group were combined in an empirically-derived formula to calculate an in vitro irritancy score (IVIS) for each treatment group. An IVIS score of 53.36 was obtained for the positive control (ethanol, 100%), thereby confirming that serious eye damage could be detected in this test system. The test item induced slight opacity, with an IVIS scored of 59.17 calculated after a 10 minute exposure (+ 2 hour post-incubation). Therefore, according to Annex I for Regulation (EC) 1272/2008 the active ingredient, Oct-7-enal is classified as Category 1: Irreversible effects on the eye, H318: Causes serious eye damage.