Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

21-12-2018 - 23-01-.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:

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

Version / remarks:

February 2017

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants)

Version / remarks:

October 2007

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

EC Number:

810-418-2

Cas Number:

1703784-30-8

IUPAC Name:

Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

Test material form:

liquid

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 2298695
- Expiration date of the lot/batch: 3 February 2019

Test animals / tissue source

Species:

other: Bovine Corneas

Details on test animals or tissues and environmental conditions:

Bovine eyes from cattle in the age range of 6 to 12 months were obtained from a slaughterhouse.

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

Each 0.75 mL of the undiluted test item were applied to three bovine corneas.

Duration of treatment / exposure:

The corneas were exposed to the test item solution for 10 minutes.

Duration of post- treatment incubation (in vitro):

two hours

Number of animals or in vitro replicates:

3 corneas per group (test item, negative control, positive control)

Details on study design:

Three corneas were epxosed to each 0.75 mL of the undiluted test item in physiological saline. After treatment the test item was rinsed off the corneas and they were incubated for further 2 hours. After the 2 hours incubation the cornea's opacity was determined. In a second step the permeability of the corneas was determined phototometrically after 90 minutes treatment with fluorescein solution.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Value:

3.479

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

other: Permeability score

Value:

0.486

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

in vitro irritation score

Value:

10.769

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Results after 10 Minutes Incubation Time:

	Cornea No.	Opacity	Permeability	IVIS
				Per Cornea	Per Group
					Mean	SD
0.9% NaCl	1	-0.836	0.001	-0.821	-0.411	0.357
	2	-0.319	0.010	-0.169
	3	-0.319	0.005	-0.244
Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated	4	1.168	0.423	7.513	10.769	3.027
	5	4.276	0.468	11.296
	6	4.993	0.567	13.498
1% NaOH	7	55.829	1.819	83.114	83.027	3.781
	8	54.794	1.627	79.199
	9	64.874	1.459	86.759

SD: Standard deviation

Applicant's summary and conclusion

Interpretation of results:

other: no prediction concerning irritant or severely irritant potential of the test item can be made

Remarks:

Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

Under the present test conditions Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated tested in the in vitro BCOP test method, had an IVIS value of 10.769, which is above the cut-off value of 3 (UN GHS no category) and below the cut-off value of 55, identifying test substances as inducing serious eye damage (UN GHS Category 1). Consequently no prediction concerning irritant or severely irritant potential of the test item can be made.

Executive summary:

The purpose of this study was to determine the ocular irritancy potential of Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

in an in vitro system. The Bovine Corneal Opacity and Permeability Assay (BCOP) test method is an organotypic model that provides short-term

maintenance of normal physiological and biochemical function of the bovine cornea in vitro. In this test method, possible damage by the test item was assessed by quantitative measurements of changes in corneal opacity and permeability in

isolated corneas from bovine eyes.

Three corneas were used for each treatment group (test item, negative control and positive control). The liquid test item Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

was used undiluted as recommended in the test guideline OECD 437. 0.9% NaCl solution was used as the

negative control and 1% NaOH in water (highly purified water) as the positive control item.

The test and control items were applied to the epithelial surface of the cornea by addition to the anterior chamber of the corneal holder. The exposure time for the test item and the controls was 10 minutes. The optical density (OD) was

measured at a wavelength of 490 nm.

The acceptance criteria of validity were fulfilled in this test.

Following treatment with Reaction products of acrylonitrile and 2,2’-Iminodi(ethylamine), hydrogenated

a mean opacity of 3.479 ± 2.033 and a mean permeability value of 0.486 ± 0.074 compared to the negative control were determined. The calculated IVIS of 10.769 ± 3.027 is above the cut-off value of 3

(UN GHS no category) and below the cut-off value of 55, identifying test substances as inducing serious eye damage (UN GHS Category 1). Consequently no prediction concerning irritant or severely irritant potential of the test item can be made.