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Toxicological information

Eye irritation

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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:
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

Reference
Name:
Unnamed
Type:
Constituent
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

Resultsopen allclose all
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
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)
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 Opacity1

Mean Permeability1

Mean In vitro Irritation Score1, 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 Opacity1

Negative control corrected Final Opacity2, 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 11

Negative control corrected OD490 21

Negative control corrected OD490 31

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