1-Propanaminium, N,N,N-trimethyl-3-[(2-methyl-1-oxo-2-propen-1-yl)amino]-, 4-C10-13-sec-alkylbenzenesulfonates

Eye irritation

Administrative data

Endpoint:

eye irritation

Remarks:

other: ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

The study was performed on 13 August 2011

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guideline and the study was conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD 437

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-(methacryloylamino)propyl trimethylammonium sec-C10-13-alkylbenzene sulfonate

IUPAC Name:

3-(methacryloylamino)propyl trimethylammonium sec-C10-13-alkylbenzene sulfonate

Constituent 2

Reference substance name:

N,N,N-trimethyl-3-[(2-methylacryloyl)amino]propan-1-aminium 4-(C10-13-sec-alkyl)benzenesulfonate

EC Number:

688-159-8

Cas Number:

1024699-81-7

Molecular formula:

UVCB substance - not available

IUPAC Name:

N,N,N-trimethyl-3-[(2-methylacryloyl)amino]propan-1-aminium 4-(C10-13-sec-alkyl)benzenesulfonate

Constituent 3

Reference substance name:

MAPTA-ABS salt

IUPAC Name:

MAPTA-ABS salt

Details on test material:

Sponsor's identification : 1-Propanaminium, N,N,N-trimethyl-3-[(2-methyl-1-oxo-2-propen-1-yl)amino]-, 4-C10-13-sec-alkylbenzenesulfonates
Description : amber coloured slightly viscous liquid
Batch number : P1118
Purity : 78.6 w/w%
Date received : 01 July 2011
Expiry date : 01 June 2012
Storage conditions : room temperature in the dark under nitrogen

Test animals / tissue source

Species:

other: Excised Bovine Cornea

Strain:

other: Not Applicable

Details on test animals or tissues and environmental conditions:

Not applicable

Test system

Vehicle:

unchanged (no vehicle)

Controls:

no

Amount / concentration applied:

TEST MATERIAL

-Amounts(s) applied (volume or weight with unit):
0.75 mL of the test material was applied to triplicate corneas.

-Concentration (if solution):
The test material was used as supplied.

VEHICLE
No vehicle used

Duration of treatment / exposure:

The undiluted test material was applied for 10 minutes followed by an incubation period of 120 minutes.

Observation period (in vivo):

Not applicable

Number of animals or in vitro replicates:

Not applicable

Details on study design:

TEST SITE
-Area of exposure
0.75 mL of the test material was applied to triplicate corneas.

-% coverage:
The test material was topically applied to the cornea. The holders were gently tilted back and forth to ensure a uniform application of the material over the entire cornea.

-Type of wrap used:
None used

REMOVAL OF TEST SUBSTANCE
-Washing (if done):
At the end of the exposure period the test material was removed from the anterior chamber and each cornea was rinsed three times with fresh complete MEM containing phenol red before a final rinse with complete MEM.

-Time after start of exposure:
10 minutes post exposure

SCORING SYSTEM:
Results from the two test method endpoints, opacity and permeability, were combined in an empirically derived formula to generate an In Vitro Irritancy Score.
Opacity Measurement-
The change in opacity for each cornea (including the negative control) was calculated by subtracting the initial opacity reading from the final opacity reading. These values were then corrected by subtracting from each the average change in opacity observed for the negative control corneas. The mean opacity value of each treatment group was then calculated by averaging the corrected opacity values of each cornea for that treatment group.

Permeability Measurement
The corrected OD492 was calculated by subtracting the mean OD492 of the negative control corneas from the OD492 value of each treated cornea. The OD492 value of each treatment group was calculated by averaging the corrected OD492 values of the treated corneas for the treatment group.

The following formula was used to determine the in vitro score:
In Vitro Irritancy Score = mean opacity value + (15 x mean OD492 value)

DATA INTERPRETATION
A test material that induces an in vitro irritancy score > than or equal to 55.1 is defined as an ocular corrosive or severe irritant.

Additionally, the opacity and permeability values were evaluated independently to determine whether the test material induced a response through only one of the two endpoints.

Visual Observation
The condition of the cornea was visually assessed immediately after rinsing and at the final opacity measurement.

Results and discussion

In vivo

Results

Irritation parameter:

cornea opacity score

Basis:

other: Mean Score of Opacity & Permeability

Time point:

other: 120 Minutes Post Rinsing

Max. score:

36.3

Reversibility:

other:

Remarks on result:

other: See Below

Irritant / corrosive response data:

The test material induced an in vitro irritancy score of 36.3.

Other effects:

The corneas treated with the test item were clear post treatment and cloudy post incubation.

Any other information on results incl. tables

RESULTS

Corneal Opacity and Permeability Measurement

Individual and mean corneal opacity measurements and individual and mean corneal permeability measurements are given in Table 1.

Corneal Epithelium Condition

The condition of the cornea immediately after rinsing and at the final opacity measurement is given in Table 2.

The corneas treated with the test item were clear post treatment and cloudy post incubation. The corneas treated with the negative control item were clear post treatment and post incubation. The corneas treated with the positive control item were cloudy post treatment and post incubation.

In Vitro Irritancy Score

The results are summarised as follows:

Treatment	In Vitro Irritancy Score
Test Material	36.3
Negative Control	1.6
Positive Control	31.8

Criteria for an Acceptable Test

The positive control In Vitro irritancy Score was within the range of 30.9 to 67.7. The positive control acceptance criterion was therefore satisfied.

Table 1          Individual and Mean Corneal Opacity and Permeability Measurements

Treatment	Cornea Number	Opacity					Permeability (OD)		In vitroIrritancy Score
		Pre-Treatment	Post-Treatment	Post-Incubation	Post-Incubation-Pre‑Treatment	Corrected Value		Corrected Value
Negative Control	1	2	3	4	2		0.020
	2	3	3	4	1		0.014
	3	3	3	4	1		0.013
					1.3*		0.016#		1.6
Positive Control	4	4	33	29	25	23.7	0.729	0.713
	5	3	21	20	17	15.7	0.793	0.777
	6	3	31	25	22	20.7	0.881	0.865
						20.0·		0.785·	31.8
Test Item	10	2	12	22	20	18.7	1.000	0.984
	11	2	9	21	19	17.7	1.037	1.021
	12	3	13	27	24	22.7	1.338	1.322
						19.7·		1.109·	36.3

OD= Optical density                 * = Mean of the post treatment-pre‑treatment values      # = Mean permeability              · = Mean corrected value

Table 2          Corneal Epithelium Condition

Treatment	Cornea Number	Observation
		Post Treatment	Post Incubation
Negative Control	1	clear	clear
	2	clear	clear
	3	clear	clear
Positive Control	4	cloudy	cloudy
	5	cloudy	cloudy
	6	cloudy	cloudy
Test Item	7	clear	cloudy
	8	clear	cloudy
	9	clear	cloudy

Applicant's summary and conclusion

Interpretation of results:

other: Non-Corrosive

Remarks:

Criteria used for interpretation of results: expert judgment

Conclusions:

The test item was considered not to be an ocular corrosive or severe irritant.

Executive summary:

Introduction. 

A study was performed to assess the ocular irritancy potential of the test material to the isolated bovine cornea. The method was designed to meet the requirements of the following:

OECD Guidelines for the Testing of Chemicals No. 437 (2009) “Bovine Corneal Opacity and Permeability Assay”

Method. 

The undiluted test material was applied for 10 minutes followed by an incubation period of 120 minutes. Negative and positive control materials were tested concurrently. The two endpoints, decreased light transmission through the cornea (opacity) and increased passage of sodium fluorescein dye through the cornea (permeability) were combined in an empirically derived formula to generate anIn VitroIrritancy Score (IVIS).

Results. 

Thein vitroIrritancy scores are summarised as follows:

Treatment	In VitroIrritancy Score
Test Material	36.3
Negative Control	1.6
Positive Control	31.8

The corneas treated with the test item were clear post treatment and cloudy post incubation.

 

Conclusion. 

The test item was considered not to be an ocular corrosive or severe irritant.