5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-aminium chloride

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 15, 2013 - March 21, 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

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 Ocular Corrosives and Severe Irritants)

Version / remarks:

2009

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)

Version / remarks:

2010

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

(2R)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-amine hydrochloride

Cas Number:

93601-85-5

Molecular formula:

C14H21NO.HCl

IUPAC Name:

(2R)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-amine hydrochloride

Constituent 2

Reference substance name:

(2S)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-amine hydrochloride (1:1)

EC Number:

695-797-0

Cas Number:

93601-86-6

Molecular formula:

C14H21NO.HCl

IUPAC Name:

(2S)-5-methoxy-N-propyl-1,2,3,4-tetrahydronaphthalen-2-amine hydrochloride (1:1)

Test material form:

solid: particulate/powder

Details on test material:

- Appearance: Mauve powder
- Storage condition of test material: At room temperature in the dark

Test system

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

TEST MATERIAL
- Since no workable suspension in physiological saline could be obtained, the test substance was added as such on top of the corneas
- Amount applied: 315 to 353 mg per cornea

NEGATIVE CONTROL
- Amount applied: 750 µL of physiological saline per cornea

POSITIVE CONTROL
- Amount applied: 750 µL of a 20% (w/v) Imidazole solution in physiological saline per cornea

Duration of treatment / exposure:

4 hours

Details on study design:

TEST SITE
- Isolated bovine cornea
- Corneas that had an initial opacity reading higher than 7 were not used
- Three corneas were used for each treatment group, at random
- Uniform distribution of the control or the test substance over the entire cornea was ensured by horizontal position of the corneas

REMOVAL OF TEST SUBSTANCE
- Washing: yes, with MEM+phenol red
- Time after start of exposure: 4 hours (at 32°C)

SCORING SYSTEM
- After exposure, the cornea is thoroughly rinsed to remove the test substance followed by immediate opacity measurement
- Following the final opacity measurement, permeability of the cornea to Na-fluorescein was evaluated, after another incubation period of 85 minutes at 32°C. OD490 values of less than 1500 were used in the calculation.
- The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate the mean in vitro score:
Mean in vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).
- Opacity and permeability values were also evaluated individually

TOOL USED TO ASSESS SCORE:
- Opacitometer (OP-KIT, MC2, Clermont, France) and microplate reader (TECAN Infinite® M200 Pro Plate Reader)

DATA EVALUATION:
- A test substance that induces a mean IVIS >55 is classified with Eye Irr. Cat. 1 in accordance with the CLP Regulation
- A test substance that induces a mean IVIS ≤ 3 is not classified for Eye irritation/corrosion in accordance with the CLP Regulation
- For a test substance that induces a mean IVIS of >3 - ≤55, no prediction on the classification can be made. More information is needed.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

Mean of 3

Value:

0.1

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Summary of opacity, permeability and in vitro scores:

Treatment	Mean opacity	Mean permeability	Mean in vitro irritation score
Negative control	0	0.000	0.0
Positive control	98	1.614	122
Test substance	0	0.005	0.1

 

Individual in vitro irritancy scores:

Eye	In vitro Irritancy Score
Negative control
1	0.1
2	0.0
3	0.9
Positive control
4	108
5	116
6	142
Test substance
13	2.0
14	-0.9
15	-0.9

No pH effect of the test substance was observed on the rinsing medium.

Applicant's summary and conclusion

Interpretation of results:

other: the substance does not need to be classified for eye irritation/corrosion according to GHS and CLP

Conclusions:

Based on a mean in vitro irritancy score of 0.1 in a Bovine Corneal Opacity and Permeability test, it is concluded that the substance did not induce ocular irritation and subsequently does not need to be classified for Eye irritation/corrosion in accordance with the CLP Regulation.

Executive summary:

Using the Bovine Corneal Opacity and Permeability test (BCOP test) the substance was screened for its eye irritancy potential in accordance with OECD 437 and according to GLP principles. Since no workable suspension in physiological saline could be obtained, the substance was applied as such directly on top of the corneas in such a way that the cornea was completely covered (315 to 353 mg per cornea). Adequate negative and positive controls were included. The substance did not induce ocular irritation, resulting in a mean in vitro irritancy score of 0.1 after 240 minutes of treatment. Since the substance induced a mean IVIS ≤ 3, the substance does not need to be classified for Eye irritation/corrosion in accordance with the CLP Regulation.