Phosphoric acid, mono- and di-isotridecyl esters, compd. with 2,2'-iminobis[ethanol]

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Remarks:

Bovine Corneal Opacity and Permeability (BCOP) test

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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 i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Deviations:

yes

Remarks:

but the study integrity was not affected by this deviation

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Phosphoric acid, mono- and di-isotridecyl esters, compd. with 2,2'-iminobis [ethanol]

Cas Number:

85409-75-2

IUPAC Name:

Phosphoric acid, mono- and di-isotridecyl esters, compd. with 2,2'-iminobis [ethanol]

Constituent 2

Reference substance name:

Bis(2-hydroxyethyl)ammonium dihydrogen phosphate

EC Number:

249-913-8

EC Name:

Bis(2-hydroxyethyl)ammonium dihydrogen phosphate

Cas Number:

29870-32-4

Molecular formula:

C4H11NO2.xH3O4P

IUPAC Name:

bis(2-hydroxyethyl)ammonium dihydrogen phosphate

Constituent 3

Reference substance name:

bis(2-hydroxyethyl)ammonium diphosphate

IUPAC Name:

bis(2-hydroxyethyl)ammonium diphosphate

Constituent 4

Reference substance name:

Water

EC Number:

231-791-2

EC Name:

Water

Cas Number:

7732-18-5

Molecular formula:

H2O

IUPAC Name:

Dihydrogen oxide

Test material form:

other: highly viscous liquid

Details on test material:

- Name of test material (as cited in study report): Phosphoric acid, mono- and di-isotridecyl esters, compd. with 2,2'-iminobis [ethanol]
- ZS name: PHOSPHORSÄUREPARTIALESTER, DEASALZ I-C13
- ZS number: 1577
- Batch: RE 10-7
- Appearance, Color: Colorless, highly viscous liquid

Specific details on test material used for the study:

Batch no.: RE 10-7
Purity/composition: 10% dry matter
Appearance: whitish liquid

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

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.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750 µL

Duration of treatment / exposure:

10 min

Duration of post- treatment incubation (in vitro):

After exposure the cornea was thoroughly rinsed to remove the test substance and incubated for 2 hours with fresh medium followed by opacity measurement and the permeability of the corneas was determined after a 90 minutes incubation period with sodium fluorescein.

Number of animals or in vitro replicates:

3

Details on study design:

The Bovine Corneal Opacity and Permeability (BCOP) test is an organic model that provides short-term maintenance of normal physiological and biological function of the bovine cornea in an isolated system. In this test method, damage by the test substance is assessed by quantitative measurements of changes in corneal opacity and permeability with an opacitymeter and an ultraviolet/visible spectrophotometer, respectively.

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

corneal opacity and permeability

Value:

34

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

not determinable

Remarks:

no prediction could be made

Other effects / acceptance of results:

The test substance was applied as it is (750 µL) directly on top of the corneas. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (ethanol) was 51 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. Since the test substance induced an IVIS > 3 ≤ 55, no prediction can be made.

Any other information on results incl. tables

Interpretation of the results:

- In vitro irritancy score

The mean opacity and mean permeability values (OD490) were used for each treatment group to calculate an in vitro score: in vitro irritancy score (IVIS) = mean opacity value + (15 x mean OD490 value).

Additionally the opacity and permeability values were evaluated independently to determine whether the test substance induced irritation through only one of the two endpoints. The IVIS cut-off values for identifying the test substances as inducing serious eye damage (UN GHS Category 1) and test substances not requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given hereafter:

< or = 3: no category,

>3 and < or = to 55: no prediction can be made,

> 55: category 1.

Applicant's summary and conclusion

Conclusions:

Under the study conditions, no prediction for eye irritation could be made as the substance induced an in vitro irritancy score of 34 (IVIS > 3 ≤ 55).

Executive summary:

A study was conducted to determine the in vitro eye irritation potential of the substance according to OECD Guideline 437, in compliance with GLP. Freshly collected bovine corneas were exposed to 750 µL test substance for 10 min. After exposure, the corneas were thoroughly rinsed to remove the test substance and incubated for 2 h with fresh medium followed by opacity measurement and the permeability of the corneas was determined after a 90 min incubation period with sodium fluorescein. An in vitro irritancy score was then established. Physiological saline and ethanol were used as negative and positive controls, respectively. The negative control responses for opacity and permeability were less than the upper limits of the laboratory historical range indicating that the negative control did not induce irritancy on the corneas. The mean in vitro irritancy score of the positive control (ethanol) was 51 and was within two standard deviations of the current historical positive control mean. It was therefore concluded that the test conditions were adequate and that the test system functioned properly. Under the study conditions, no prediction for eye irritation could be made as the substance induced an in vitro irritancy score of 34 (IVIS > 3 ≤ 55) (Verbaan, 2016).