N'-(ethylcarbonimidoyl)-N,N-dimethylpropane-1,3-diamine monohydrochloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

17 July 2017 to 18 July 2017

Reliability:

1 (reliable without restriction)

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)

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

N'-(ethylcarbonimidoyl)-N,N-dimethylpropane-1,3-diamine monohydrochloride

EC Number:

247-361-2

EC Name:

N'-(ethylcarbonimidoyl)-N,N-dimethylpropane-1,3-diamine monohydrochloride

Cas Number:

25952-53-8

Molecular formula:

C8H17N3.ClH

IUPAC Name:

N-[3-(dimethylamino)propyl]-N'-ethylcarbodiimide hydrochloride

Test material form:

solid

Specific details on test material used for the study:

Appearance: White crystalline powder
Batch: AAN0680
Purity/Composition: 99.9 %
Test item storage: At room temperature

Test animals / tissue source

Species:

cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

Bovine eyes were used as soon as possible after slaughter. Bovine eyes from young cattle were obtained from the slaughterhouse (Vitelco, -'s Hertogenbosch, The Netherlands), where the eyes were excised by a slaughterhouse employee as soon as possible after slaughter. Eyes were collected and transported in physiological saline in a suitable container under cooled conditions

Test system

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750ul of a 20% (w/v) solution of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride prepared in physiological saline was administered

Duration of treatment / exposure:

Corneas were incubated in a horizontal position for 240 +/-10 minutes at 32 +/-1 degreesC

Duration of post- treatment incubation (in vitro):

After the incubation the solutions were removed and the epithelium was washed at least three times with MEM with phenol red (Earle’s Minimum Essential Medium Life Technologies). Possible pH effects of the test item on the corneas were recorded

Number of animals or in vitro replicates:

3 replicates

Details on study design:

The eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded.
The isolated corneas were stored in a petri dish with cMEM (Earle’s Minimum Essential Medium (Life Technologies, Bleiswijk, The Netherlands) containing 1% (v/v) L-glutamine
(Life Technologies) and 1% (v/v) Fetal Bovine Serum (Life Technologies)). The isolated corneas were mounted in a corneal holder (one cornea per holder) of BASF (Ludwigshafen,
Germany) with the endothelial side against the O-ring of the posterior half of the holder. The anterior half of the holder was positioned on top of the cornea and tightened with screws. The compartments of the corneal holder were filled with cMEM of 32  1C. The corneas were incubated for the minimum of 1 hour at 32 +/- 1 deg C.

Results and discussion

In vitro

Results

Results 1

Irritation parameter:

cornea opacity score

Value:

24

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation parameter:

in vitro irritation score

Value:

24

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 3

Irritation parameter:

other: Permeability

Value:

0.007

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Applicant's summary and conclusion

Interpretation of results:

study cannot be used for classification

Conclusions:

The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 123 and within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly.
1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride induced ocular irritation through one endpoint (opacity) only, resulting in a mean in vitro irritancy score of 24 after 240 minutes of treatment.
1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride induced an IVIS > 3 ≤ 55, no prediction on the classification can be made.

Executive summary:

The objective of this study was to evaluate the eye hazard potential of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride as measured by its ability to induce opacity and increase permeability in an isolated bovine cornea using the Bovine Corneal Opacity and Permeability test (BCOP test). This report describes the potency of chemicals to induce serious eye damage using isolated bovine corneas. The eye damage of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride was tested through topical application for approximately 240 minutes. The study procedures described in this report were based on the most recent OECD guideline. Batch AAN0680 of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride was a white crystalline powder with a purity of 99.9%. The test item was applied as a 20% (w/v) solution (750 µl) directly on top of the corneas. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (20% (w/v) Imidazole) was 123 and within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride induced ocular irritation through one endpoint (opacity) only, resulting in a mean in vitro irritancy score of 24 after 4 hours of treatment. In conclusion, since 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride induced an IVIS > 3 ≤ 55, no prediction on the classification can be made.