Protein hydrolyzates, milk

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

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)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Protein hydrolyzates, milk

EC Number:

296-575-2

EC Name:

Protein hydrolyzates, milk

Cas Number:

92797-39-2

IUPAC Name:

Protein hydrolyzates, milk

Test material form:

solid

Test animals / tissue source

Species:

cattle

Details on test animals or tissues and environmental conditions:

Test System: Bovine eyes were used as soon as possible after slaughter.
Rationale: In the interest of sound science and animal welfare, a sequential testing strategy is recommended to minimize the need of in vivo testing (1-6). As a consequence a validated and accepted in vitro test for eye irritation should be performed before in vivo tests are conducted. One of the proposed validated in vitro eye irritation tests is the Bovine Corneal Opacity and Permeability (BCOP) test.
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.
Transport: Eyes were collected and transported in physiological saline in a suitable container under cooled conditions.

Test system

Vehicle:

other: saline

Remarks:

The test item was tested neat.

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

Prior to the application 651.94 mg of the test item were suspended in 3.26 mL saline to
reach a concentration of 20% (w/v).

Duration of treatment / exposure:

topical application for 10 minutes

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

2.33

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Executive summary:

This in vitro study was performed to assess the corneal irritation and damage potential of

YOGURTENE 20 CQ (PFR) U/A - 6099943 by means of the BCOP assay using fresh bovine

corneae.

After a first opacity measurement of the fresh bovine corneae (t0), the 20% (w/v) suspension

in saline (0.9% (w/v) NaCl in deionised water of the test item YOGURTENE 20 CQ (PFR)

U/A - 6099943, the positive, and the negative controls were applied to corneae and

incubated for 240 minutes at 32 ± 1 °C. The posterior chamber contained MEM medium

supplemented with sodium bicarbonate and L-glutamine and 1% fetal calf serum (FCS)

(complete medium = cMEM). After the incubation phase the test item, the positive, and the

negative controls were each rinsed from the corneae and opacity was measured again (t240).

After the opacity measurements permeability of the corneae was determined by measuring

spectrophotometrically the transfer of sodium fluorescein after incubation in a horizontal

position for 90 minutes at 32 ± 1 °C.

With the negative control (saline) neither an increase of opacity nor permeability of the

corneae could be observed.

The positive control (10% (w/v) Benzalkonium chloride in saline) induced clear opacity on the

corneae corresponding to a classification as corrosive / severe irritant to the eye

(CLP/EPA/GHS (Cat 1)).

Relative to the negative control, the test item YOGURTENE 20 CQ (PFR) U/A - 6099943 did

not cause any increase of the corneal opacity or permeability. The calculated mean in vitro

irritation score was 2.33. According to OECD 437 the test item is classified as not corrosive /

not severe irritant to the eye.

In conclusion, according to the current study and under the experimental conditions

reported, the test item YOGURTENE 20 CQ (PFR) U/A - 6099943 is not corrosive / not

severe irritant to the eye (CLP/EPA/GHS (Cat 1)).