BCOP Testing - Bovine Corneal Opacity/Permeability Assay

(OECD TG 437) Alternative Ocular Irritation Testing

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What is the BCOP?

The Bovine Cornea Opacity/Permeability test (BCOP) is an alternative ocular irritation assay designed to replace the rabbit eye test. It is commonly utilized to test for eye irritation and corrosive potential of test materials that may be used in our around the eye. The BCOP uses excised bovine corneas, normally discarded as waste in the production of beef and does not utilize live animals for research.

Ocular Irritation Testing Experts

MB Research has pioneered the use and development of alternative and in vitro ocular toxicity models used mainly for cosmetics and consumer product safety evaluations. We have performed over 10,000 alternative ocular irritancy tests, which include over 3,000 materials assayed with the BCOP. The BCOP can also be used in combination with other alternatives such as the CAMVA or HET-CAM to further determined ocular irritancy classification. Also see: ROBatt

BCOP General Information:

  • Draize Rabbit Eye Test Alternative
  • Uses excised bovine corneas normally discarded
  • Measures two endpoints; 1) changes in opacity, reflecting protein denaturation and corneal injury, and 2) fluorescent dye permeability reflecting damage to corneal epithelium
  • Optional histological endpoint is available (H & E staining)
  • Measurement of pro-inflammatory mediators in liberated corneal cells (by flow cytometry or ELISA)
  • See BCOP-EIT- Dual Assay for Identifying Eye Irritants (GHS Category 2) - OECD TG 405 -Draize Rabbit Eye Test Replacement.
BCOP Cornea in Holder -Normal and Opacity

Bovine corneas have been used since the '50s. Current BCOP protocols were developed by Pierre Gautheron and Joseph Sina* as an alternative to the Draize Rabbit Eye Test to evaluate the response of an excised cornea (a waste byproduct of food production) by measuring light transmission and fluorescent dye permeability.

* “Bovine Corneal Opacity and Permeability Test: An In Vitro Assay of Ocular Irritancy”, Gautheron P., Dukic, M., Alix, D. and Sina, J. Fundamental and Applied Toxicology 18, 442-449.

Current BCOP Testing Protocols

OECD Test Guideline 437 - MB Research Labs offer GLP compliant protocols for the BCOP based on regulatory agency requirements.

Current OECD Protocol 437

Validation Studies


In Vitro Toxicol 1994;7(3):283-90.

The author's task was to develop an approach to assessing ocular irritation, with minimal use of animals, so that it could be applied inhouse for worker safety testing.

The centerpiece of the multifaceted approach was the bovine corneal opacity and permeability (BCOP) assay, a test that measures opacity and cell sheet integrity in a target tissue: the cornea. Initial studies indicated that the BCOP gave an overall concordance with in vivo data of 85%. Two specific findings were that although suspensions could be tested, hydroscopic compds. tended to float on the exposure medium and adequate exposures could not be obtained; and that some compounds caused irritation in animals only after a delay of 24-72 hrs, and these could not be detected in vitro.

The BCOP method proved to be easily transferable, with good reproducibility in a small interlaboratory study. Overall concordance was approximately 83% among the six participants. Some slight modifications were made to the assay based on the results of this study, and a larger interlaboratory study was begun in order to broaden further experience and to attempt to define other areas of potential difficulty. Twelve labs participated in evaluating 52 diverse chemicals, and interlaboratory variability was quite low. Concordance was approximately 80%, with the false-negative and false-positive rate dependent upon the in vivo classification system generally in use at a particular lab.

In a comparison study in which 36 pharmaceutical manufacturer intermediates from 11 companies were all tested in the BCOP and 6 other assays, the BCOP was found to be among the most accurate tests. A tiered approach, combining results from other assays with the BCOP data, did not appear to provide an enhanced predictive value. Based on this body of data, it was considered that the BCOP assay is valid for the specific purposes and for the types of chemicals included in the studies (i.e., mainly pharmaceutical intermediates and raw materials).