1-(2,4,6-trichlorophenyl)acetone O-methyloxime

Eye irritation

Administrative data

Endpoint:

eye irritation

Remarks:

in vitro

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-12-02 until 2015-06-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: The study was carried out in accordance with OECD guideline 438 and in cimpliance with GLP. The validity criteria have been fulfilled.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2015

Report date:

2015

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 438 (Isolated Chicken Eye Test Method for Identifying Ocular Corrosives and Severe Irritants)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EPA OPPTS 870.2400 (Acute Eye Irritation)

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

1-(2,4,6-trichlorophenyl)acetone O-methyloxime

EC Number:

807-534-0

Cas Number:

1228284-89-6

Molecular formula:

C10H10Cl3NO

IUPAC Name:

1-(2,4,6-trichlorophenyl)acetone O-methyloxime

Test material form:

other: liquid

Details on test material:

- Physical state: Brown liquid
- Purity test date: 25 September 2013
- Expiration date of the lot/batch: End September 2015
- Stability under test conditions: Stable until end September 2015
- Storage condition of test material: Room temperature (<30°C), protected from light and humidity

Test animals / tissue source

Species:

other: Isolated chicken eye

Strain:

other: ROSS 308

Details on test animals or tissues and environmental conditions:

EYES
- Source: Commercial abattoir

EXPERIMENTAL DATES: 2 December 2013

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: Yes (0.9% sodium chloride solution)

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 30 μL

Duration of treatment / exposure:

10 seconds

Observation period (in vivo):

240 minutes. The negative and positive control eyes and all test eyes were evaluated pre-treatment and at approximately 30, 75, 120, 180 and 240 minutes after the post-treatment rinse. Corneal thickness and corneal opacity were measured at all time points. Fluorescein retention was measured on two occasions, at base line (t=0) and approximately 30 minutes after the post-treatment rinse.

Number of animals or in vitro replicates:

7 isolated eyes (3 test item, 3 positive control, 1 negative control)

Details on study design:

METHOD
- After the zero reference measurements, the eye was held in a horizontal position and 30 μL of test item was applied onto the centre of the cornea such that the entire surface of the cornea was covered. After 10 seconds, the surface was rinsed with saline. The positive control eyes were treated in a similar way with 30 μL benzalkonium chloride 5 % (w/v). The negative control eye was treated with 30 μL of sodium chloride (0.9% solution). Corneal thickness, corneal opacity and fluorescein retention were measured and any morphological effects (pitting or loosening of the epithelium) evaluated.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): After the 10 second exposure period, the cornea surface was rinsed thoroughly with 20 mL isotonic saline at ambient temperature.

SCORING SYSTEM:
- Corneal thickness or swelling. Measurements made with an optical pachymeter on a slit-lamp microscope and expressed as a percentage calculated from corneal thickness measurements. Corneal swelling was classified by the following scale: Mean cornea swelling (%) of 0-5, >5-12, >12-18 (>75 mins after treatment), >12-18 (<75 mins after treatment), >18-26, >26-32 (>75 mins after treatment), >26-32 (<75 mins after treatment), >32 classified as ICE Class I, II, II, III, III, III, IV or IV, respectively. i.e. No swelling = Class I, slight swelling = Class II, moderate swelling = Class III, severe swelling = Class IV.
- Corneal opacity. Corneal opacity was scored (on a 0-4 scale) using the area of the cornea that was most densely opacified and the mean ΔCOmax (mean value of the highest opacity scores) calculated. Mean maximum opacity scores of 0-0.5, 0.6-1.5, 1.6-2.5 and 2.6-4.0 were classified as ICE Class I, II, III and IV, respectively. i.e. No opacity = Class I, Slight opacity = Class II, Moderate opacity = Class III, Severe or total opacity = Class 4.
- Fluorescein retention: Fluorescein retention change was calculated and scored using a 0-3 scale. Mean fluorescein retention scores of 0-0.5, 0.6-1.5, 1.6-2.5 and 2.6-3.0 were classified as ICE Class I, II, III and IV, respectively. i.e. No fluorescein retention = Class I, slight fluorescein retention = Class II, Moderate fluorescein retention = Class III, Severe fluorescein retention = Class IV.

TOOL USED TO ASSESS SCORE: Hand-slit lamp / fluorescein

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: Mean maximum corneal swelling

Basis:

mean

Time point:

other: up to 75 minutes

Score:

0

Remarks on result:

other: ICE Class I

Results 2

Irritation parameter:

other: Mean maximum corneal swelling

Basis:

mean

Time point:

other: up to 240 minutes

Score:

0

Remarks on result:

other: ICE Class I

Results 3

Irritation parameter:

other: Mean maximum corneal opacity change

Basis:

mean

Score:

0.67

Remarks on result:

other: ICE Class II

Results 4

Irritation parameter:

other: Mean fluorescein retention change

Basis:

mean

Score:

0

Remarks on result:

other: ICE Class I

Results 5

Irritation parameter:

other: Overall ICE Class

Remarks on result:

other: 2xI, 1xII

Irritant / corrosive response data:

No corneal swelling or fluorescein retention was observed during the four hour observation period. Corneal opacity (severity 0.5 or 1) was noted in all eyes. No other corneal effect was observed. The negative and positive control group results demonstrate that the study was valid.

The test item is non-irritating, GHS Classification: Non-classified.

The positive control benzalkonium chloride 5% (w/v) was classed as severely irritating, GHS Classification: Category 1.

The negative control Saline (NaCl 0.9% w/v) was classed as non-irritating, GHS Classification: Non-classified.

Any other information on results incl. tables

Table 1: Mean values of the treated eyes for maximum corneal thickness change, corneal opacity change and fluorescein retention change – Run 1

Observation	Test item		Positive control		Negative control
	Value	ICE Class	Value	ICE Class	Value	ICE Class
Mean maximum corneal swelling at up to 75 min	0%	I	0%	I	0%	I
Mean maximum corneal swelling at up to 240 min	0%	I	-5.3%	II	0%	I
Mean maximum corneal opacity change	0.67	II	4.00	IV	0	I
Mean fluorescein retention change	0	I	2.83	IV	0	I
Other Observations	None		Loosening of epithelium observed in all eyes (3/3) 75 mins after post-treatment rinse.		None
Overall ICE Class	2xI 1xII		1xII 2xIV		3xI

 

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Remarks:

Migrated information: non-irritant

Conclusions:

Based on this in vitro eye irritation study in isolated chicken eyes, the test item is non-irritating, GHS Classification: Non-classified.

Executive summary:

An in vitro eye irritation study was performed in isolated chicken’s eyes. The irritation effects of the test item were evaluated according to OECD 438 (2013). After the zero reference measurements, the eye was held in a horizontal position and 30 μL of test item was applied onto the centre of the cornea such that the entire surface of the cornea was covered. After 10 seconds, the surface was rinsed with saline. The positive control eyes were treated in a similar way with 30 μL benzalkonium chloride 5 % (w/v). The negative control eye was treated with 30 μL of sodium chloride (0.9% solution). Corneal thickness, corneal opacity and fluorescein retention were measured and any morphological effects (pitting or loosening of the epithelium) evaluated.

No corneal swelling or fluorescein retention was observed during the four hour observation period. Corneal opacity (severity 0.5 or 1) was noted in all eyes. No other corneal effect was observed. The negative and positive control group results demonstrated that the study was valid.

Based on this in vitro eye irritation study in isolated chicken eyes, the test item is non-irritating, GHS Classification: Non-classified.