Borate(1-), bis[ethanedioato(2-)-.kappa.O1,.kappa.O2]-, potassium (1:1), (T-4)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2014-01-15 to 2014-01-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP and guideline compliant study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2014

Report date:

2014

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)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

potassium bisoxalatoborate KBOB

IUPAC Name:

potassium bisoxalatoborate KBOB

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

Stored at room temperature, protected from light.

Test animals / tissue source

Species:

other: cow

Strain:

not specified

Test system

Vehicle:

physiological saline

Controls:

other: not applicable; but in vitro controls

Amount / concentration applied:

20 % in physiological saline (0.9 % NaCl).

Duration of treatment / exposure:

4 hours ± 5 minutes

Observation period (in vivo):

90 minutes

Number of animals or in vitro replicates:

not applicable

Details on study design:

The assay uses isolated corneas from animals freshly slaughtered at an abattoir. Fresh eyes were collected from the abattoir on the test day and transported in HBSS containing Pen/Strep on ice to the laboratory, where cornea preparation was initiated immediately. The eyes were carefully examined for defects and any defective eyes discarded. The tissue surrounding the eyeball was carefully pulled away and the cornea excised, leaving 2-3 mm of sclera. The isolated corneas were stored in a petri dish containing HBSS and again examined for defects before use in the test.

Negative control: physiological saline (0.9 % NaCl)
Positive control: imidazole 20 % in physiological saline (0.9 % NaCl)

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Value:

102.43

In vivo

Irritant / corrosive response data:

As the mean in vitro irritation score for the test item was determined to be > 55 (102.43), the test item is identified as inducing serious eye damage.

Applicant's summary and conclusion

Interpretation of results:

Category 1 (irreversible effects on the eye)

Remarks:

Criteria used for interpretation of results: other: UN GHS

Conclusions:

Based on the in vitro eye corrosion/irritation study with bovine corneas, KBOB is considered a severe eye irritant.

Executive summary:

They eye corrosion potential of KBOB was investigated in the in vitro bovine corneal opacity and permeability assay according to OECD Technical Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants).

The test item was suspended in physiological saline (0.9 %) as a concentration of 20 %. Isolated bovine corneas were mounted in corneal holders, and after an initial incubation in RPMI medium (in both anterior and posterior chamber), their opacity was measured with an opacitometer. Three corneas were used as negative controls, positive controls and for the test item, each. 750 μL of the test item or the control substance was introduced into the anterior chamber, and after 4 hours ± 5 minutes of incubation at 32 ± 1 °C, the substances were removed and the epithelium washed with MEM media containing phenol red until free of test substance, then with RPMI, then an opacity measurement was performed. After a 90-minute incubation (32 ± 1 °C) in fresh RPMI with 1 mL of a 5 mg/mL sodium fluorescein solution added to the anterior chamber, the medium from the posterior chamber was removed and its optical density (OD490) at 490 nm was determined, using a spectrophotometer. The initial opacity reading of each cornea was subtracted from the final opacity reading to obtain the change in opacity, corrected by subtracting the average change in opacity observed for the negative controls, and a mean opacity value was calculated for each treatment. The OD490 values were similarly corrected by subtracting a mean blank-well OD490 from each OD490 for a test article, then the average corrected negative-control OD490 from each test-item or positive-control OD490, and a mean OD490 was then calculated for each treatment. The in vitro irritation score (IVIS) was then calculated as the mean opacity value – (15 x mean OC490 value).

The resulting mean in vitro irritation score for the test item was 102.43. According to the evaluation criteria, KBOB is classified into UN GHS Category 1.