Methyl 2-[(2-methylundecylidene)amino]benzoate

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

24/05/2018 - 5/12/2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Test guideline 1

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)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Methyl 2-[(2-methylundecylidene)amino]benzoate

EC Number:

267-104-8

EC Name:

Methyl 2-[(2-methylundecylidene)amino]benzoate

Cas Number:

67800-80-0

Molecular formula:

C20H31NO2

IUPAC Name:

methyl 2-[(2-methylundecylidene)amino]benzoate

Test material form:

liquid

Specific details on test material used for the study:

CAS No. 67800-80-0
EINECS-No. 267-104-8
Solubility H2O: <0.1g/L; EtOH: >1g/L; acetone: >1g/L; CH3CN:
>1g/L

Test animals / tissue source

Species:

cattle

Strain:

other: Bos primigenius Taurus

Details on test animals or tissues and environmental conditions:

Fresh bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2-4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months old. The eyes were transported to the test facility in Hanks’ Balanced Salt Solution with 1% Penicillin-Streptomycin solution (Penicillin 100 U/mL, Streptomycin 100 µg/mL) in a suitable cooled container within 1 hour.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 µL , undiluted

Duration of treatment / exposure:

10 min

Duration of post- treatment incubation (in vitro):

2 hours
Permeation: + 90 min

Number of animals or in vitro replicates:

3

Details on study design:

EVALUATION

- Calculation of Opacity
The change of opacity value of each treated cornea with test item, positive control and negative control was calculated by subtracting the initial basal opacity from the post treat- ment opacity reading for each cornea.
The average change in opacity of the negative control cornea was calculated and this val- ue was subtracted from the change in opacity of each treated cornea with test item and positive control to obtain a corrected opacity.

Opacity = [(I0/I)-b]/a

a = 0.0251 and b = 0.9894 being Opacitometer-specific empirically determined variables I0 = the empirically determined illuminance through a cornea holder with windows and Medium, here: Io= 1096.80
I = the measured illuminance (unit: LUX)


- Calculation of Permeability
The corrected OD492 value of each cornea treated with test item and positive control was calculated by subtracting the mean negative control cornea value from the original perme- ability value for each cornea.
The mean OD492 value for each treatment group (test item, positive control and negative control) was determined by averaging the final corrected OD492 values of the treated cor- neas for one treatment group.


- Calculation of IVIS (In Vitro Irritancy Score)
The IVIS of each replicate of the negative control was calculated from the following equa- tion:
IVIS = opacity difference + (15 x corrected OD492 value)

The IVIS of each replicate of the positive control and of the test item were calculated from the following equation:
IVIS = (opacity difference – mean opacity difference of the negative control) + [15 x (OD492
– mean OD492 of the negative control)]

Note: All calculations are performed with unrounded values. Therefore, re-calculation with rounded values may lead to slightly different results.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Main

Value:

ca. -0.18

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

1.34

Positive controls validity:

valid

Remarks:

120.36

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

According to OECD Guideline no. 437 (Oct. 2017), Methyl 2-[(2- methylundecylidene)amino]benzoate requires no classification for eye irritation or serious eye damage under GHS.

Executive summary:

This in vitrostudy was performed to assess corneal damage potential of Methyl 2-[(2- methylundecylidene)amino]benzoate by quantitative measurements of changes in opacity and permeability in a bovine cornea.

The test item Methyl 2-[(2-methylundecylidene)amino]benzoate was brought onto the cornea of a bovine eye which previously had been incubated with cMEM without phenol red at 32 ± 1 °C for 1 hour and whose opacity had been determined. The test item was incubated on the cornea for 10 minutes at 32 ± 1 °C. After removal of the test item and 2 hours post- incubation, opacity and permeability values were measured.

 

The test item was tested neat. Under the conditions of this test, the test item Methyl 2-[(2 - methylundecylidene)amino]benzoate showed no effects on the cornea of the bovine eye. The calculated IVIS is -0.18.

According to OECD Guideline no. 437 (Oct. 2017), a substance with an IVIS ≤ 3 requires no classification for eye irritation or serious eye damage.

 

The negative control (HBSS) and the positive control (undiluted dimethylformamide) have met the validity criteria.

 

No observations were made which might cause doubts concerning the validity of the study outcome. The test is considered valid.