C8-18 (even numbered) and C18 (unsaturated) sarcosinates, 2-hydroxypropylammonium salt

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

26 Jan 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study without restrictions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

(adopted July 26, 2013)

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

2-(N-methyldodecanamido)acetic acid 2-hydroxypropan-1-aminium 2-(N-methyldodecanamido)acetate

EC Number:

943-406-1

Molecular formula:

Not applicable. This substance is UVCB.

IUPAC Name:

2-(N-methyldodecanamido)acetic acid 2-hydroxypropan-1-aminium 2-(N-methyldodecanamido)acetate

Test material form:

liquid: viscous

Details on test material:

- Name of test material (as cited in study report): MIPA-Cocoyl-Sarkosinat
- Substance type: organic (UVCB)
- Physical state: clear yellow-brown viscous liquid
- Analytical purity: 100%, according to definition of UVCB
- Impurities (identity and concentrations): no impurities, UVCB
- Composition of test material, percentage of components: 100% MIPA-Cocoyl-Sarkosinat
- Lot/batch No.: 070715
- Expiration date of the lot/batch: 31 Dec 2016
- Stability under test conditions: stable in vehicle water
- Storage condition of test material: at room temperature
- Other: pH 5.7 - 6.0 (1% in water)

Test animals / tissue source

Species:

other: cattle

Strain:

not specified

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
Not applicable, in vitro test performed with bovine eyes as soon as possible after slaughter.
- Source: 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. The eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:

physiological saline

Controls:

other: yes, 3 corneas concurrently treated with vehicle

Amount / concentration applied:

TEST MATERIAL
- Concentration (if solution): 750 µL of a 10% dilution (since test item is a surfactant)

VEHICLE
- Amount(s) applied (volume or weight with unit): 750 µL physiological saline

Duration of treatment / exposure:

10 ± 1 minutes

Observation period (in vivo):

Opacity determination after 120 ± 10 minutes incubation time, followed by permeability measurement

Number of animals or in vitro replicates:

3 isolated corneas per treatment group

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with MEM with phenol red (Eagle's Minimum Essential Medium, Life Technologies), thereafter with phenol red free cMEM
- Time after start of exposure: 10 ± 1 min

SCORING SYSTEM: not applicable; opacity measurement, followed by permeability determination

TOOL USED TO ASSESS SCORE: opacity measured with device OP-KIT; permeability determined by measurement of optical density of medium of posterior compartment of cornea holder at 490 nm (OD490) in a microplate reader (TECAN Infinite M200 Pro Plate Reader) after incubation of cornea for 90 ± 5 minutes at 32 ± 1 °C to 1 mL of 4mg sodium-fluorescein/mL phenol red free cMEM in anterior compartment.

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

other: Mean In vitro irritancy score (IVIS)

Basis:

mean

Remarks:

of 3 corneas

Time point:

other: 10 ± 1 min

Score:

-0.6

Reversibility:

other: not applicable

Remarks on result:

other: Negative control

Results 2

Irritation parameter:

other: Mean In vitro irritancy score (IVIS)

Basis:

mean

Remarks:

of 3 corneas

Time point:

other: 10 ± 1 min

Score:

140.5

Reversibility:

other: not applicable

Remarks on result:

other: Positive control (10% Benzalkonium chloride)

Results 3

Irritation parameter:

other: Mean In vitro irritancy score (IVIS)

Basis:

mean

Remarks:

of 3 corneas

Time point:

other: 10 ± 1 min

Score:

19.3

Reversibility:

other: not applicable

Remarks on result:

other: 10% Test substance

Irritant / corrosive response data:

The corneas treated with the test substance showed opacity values ranging from 5.5 to 6.3 and permeability values ranging from 0.807 to 0.981. The corneas were slightly translucent after the 10 minutes of treatment with test substance. A pH effect of the test item was observed on the rinsing medium, the corneas were rinsed until no colour change of the medium was observed. Hence, the in vitro irritancy scores ranged from 18 to 20 after 10 minutes of treatment with test substance.

Any other information on results incl. tables

The test substance induced ocular irritation through both endpoints, opacity and permeability, resulting in a mean in vitro irritancy score of 19 after 10 minutes of treatment.

Since the test substance induced an IVIS > 3 ≤ 55, no prediction on the classification can be made.

Summary of opacity, permeability and in vitro scores

Treatment	Mean Opacity^1	Mean Permeability^1	Mean In vitro Irritation Score^1, 2
Negative control	-0.4	-0.014	-0.6
Positive control (Benzalkonium Chloride)	93.1	3.159	140.5
Test substance	5.9	0.896	19.3

¹             Calculated by subtracting the negative control mean opacity and mean permeability values from those for the positive control and test item.

²             Mean In vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD 490 value).

Opacity score

Treatment	Opacity before treatment	Opacity after treatment	Final Opacity^1	Negative control corrected Final Opacity^2, 3	Mean Final Opacity
Negative control	3.0	2.5	-0.6		-0.4
	2.9	3.6	0.8
	3.5	2.0	-1.5
Positive control	3.6	105.3	101.7	101.7	93.1
	3.3	93.5	90.2	90.2
	2.5	89.9	87.5	87.5
Test substance	3.4	8.9	5.5	5.5	5.9
	3.0	9.4	6.3	6.3
	3.1	8.9	5.8	5.8

¹             Final Opacity = Opacity after treatment – Opacity before treatment.

²             Negative control corrected Final Opacity = Final Opacity – Mean final opacity negative control.

³             In case the mean final opacity of the negative control is below zero, no correction was made.

 

Permeability score individual values (corrected)

Treatment	Dilution factor	Negative control corrected OD490 1^1	Negative control corrected OD490 2^1	Negative control corrected OD490 3^1	Negative control corrected OD490 Average	Negative control corrected final OD490	Average OD
Positive control	6	0.604	0.595	0.599	0.599	3.596	3.159
	6	0.483	0.481	0.481	0.482	2.890
	6	0.503	0.494	0.498	0.498	2.990
Test substance	1	0.973	0.980	0.989	0.981	0.981	0.896
	1	0.896	0.899	0.909	0.901	0.901
	1	0.810	0.798	0.814	0.807	0.807

¹             OD490 values corrected for the mean final negative control permeability (-0.014).

Applicant's summary and conclusion

Interpretation of results:

other: no classification possible

Remarks:

Criteria used for interpretation of results: OECD GHS