Two-stage bottom-up tiered approach combining several alternatives for identification of eye irritation potential of chemicals including insoluble or volatile substances.

For the assessment of eye irritation, one alternative test may not completely replace the rabbit Draize test. In the present study, we examined the predictive potential of a tiered approach analyzing the results from several alternatives (i.e., the Short Time Exposure (STE) test, the EpiOcular assay, the Hen's Egg Test-Chorioallantoic Membrane (HET-CAM) assay and the Bovine Corneal Opacity and Permeability (BCOP) assay) for assessing Globally Harmonized System (GHS) eye irritation categories. Fifty-six chemicals including alcohols, surfactants, and esters were selected with a balanced GHS category and a wide range of chemical classes. From a standpoint of both assessable sample numbers and predictive accuracy, the more favorable tiered approach was considered to be the two-stage bottom-up tiered approach combining the STE test, the EpiOcular assay followed by the BCOP assay (accuracy 69.6%, under prediction rate 8.9%). Moreover, a more favorable predictive capacity (accuracy 71.4%, under prediction rate 3.6%) was obtained when high volatile alcohols/esters with vapor pressures >6 kilopascal (kPa) at 25°C were evaluated with EpiOcular assay instead of the STE test. From these results, the two-stage bottom-up tiered approach combining the STE test, the EpiOcular assay followed by the BCOP assay might be a promising method for the classification of GHS eye irritation category (Not classified (NC), Category 2 (Cat. 2), and Category 1 (Cat. 1)) for a wide range of test chemicals regardless of solubility.

[1]  Yutaka Takahashi,et al.  A tiered approach combining the short time exposure (STE) test and the bovine corneal opacity and permeability (BCOP) assay for predicting eye irritation potential of chemicals. , 2012, The Journal of toxicological sciences.

[2]  E Borenfreund,et al.  Toxicity determined in vitro by morphological alterations and neutral red absorption. , 1985, Toxicology letters.

[3]  S Kato,et al.  An in vitro alternative to the Draize eye-irritation test: Evaluation of the crystal violet staining method. , 1991, Toxicology in vitro : an international journal published in association with BIBRA.

[4]  Robert Landsiedel,et al.  In-house Validation of the EpiOcular™ Eye Irritation Test and its Combination with the Bovine Corneal Opacity and Permeability Test for the Assessment of Ocular Irritation , 2011, Alternatives to laboratory animals : ATLA.

[5]  H Babich,et al.  Comparisons of two in vitro cytotoxicity assays-The neutral red (NR) and tetrazolium MTT tests. , 1988, Toxicology in vitro : an international journal published in association with BIBRA.

[6]  M. Umeda,et al.  Cytotoxicity test with simplified crystal violet staining method using microtitre plates and its application to injection drugs. , 1989, Toxicology in vitro : an international journal published in association with BIBRA.

[7]  M Liebsch,et al.  Eye Irritation Testing: The Way Forward. The Report and Recommendations of ECVAM Workshop 34. , 1999, Alternatives to laboratory animals : ATLA.

[8]  P. Botham,et al.  Eye irritation: Reference chemicals data bank , 1992 .

[9]  Y Ohno,et al.  Interlaboratory validation of the in vitro eye irritation tests for cosmetic ingredients. (1) Overview of the validation study and Draize scores for the evaluation of the tests. , 1999, Toxicology in vitro : an international journal published in association with BIBRA.

[10]  H. Itagaki,et al.  An In Vitro Tier Evaluation for the Identification of Cosmetic Ingredients Which are Not Ocular Irritants , 2008, Alternatives to laboratory animals : ATLA.

[11]  Yutaka Takahashi,et al.  Validation study of the Short Time Exposure (STE) test to assess the eye irritation potential of chemicals. , 2011, Toxicology in vitro : an international journal published in association with BIBRA.

[12]  Y. Ohno,et al.  Interlaboratory validation of in vitro eye irritation tests for cosmetic ingredients. (2) Chorioallantoic membrane (CAM) test. , 1999, Toxicology in vitro : an international journal published in association with BIBRA.

[13]  J. Sina,et al.  Bovine corneal opacity and permeability test : an in vitro assay of ocular irritancy , 1992 .

[14]  Nicola J Hewitt,et al.  A tiered approach to the use of alternatives to animal testing for the safety assessment of cosmetics: skin irritation. , 2009, Regulatory toxicology and pharmacology : RTP.

[15]  Valérie Zuang,et al.  Alternative (non-animal) methods for cosmetics testing: current status and future prospects—2010 , 2011, Archives of Toxicology.

[16]  Valérie Zuang,et al.  A proposed eye irritation testing strategy to reduce and replace in vivo studies using Bottom-Up and Top-Down approaches. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[17]  Hiroshi Honda,et al.  Development of the short time exposure (STE) test: an in vitro eye irritation test using SIRC cells. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[18]  S Kalweit,et al.  Validation project of alternatives for the Draize eye test. , 1987, Molecular toxicology.

[19]  K. Cooper,et al.  Accuracy of blood cytological screening techniques for the diagnosis of a possible hematopoietic neoplasm in the bivalve mollusc, Mya arenaria , 1982 .

[20]  John H. Draize,et al.  METHODS FOR THE STUDY OF IRRITATION AND TOXICITY OF SUBSTANCES APPLIED TOPICALLY TO THE SKIN AND MUCOUS MEMBRANES , 1944 .

[21]  Yutaka Takahashi,et al.  Inter-laboratory study of short time exposure (STE) test for predicting eye irritation potential of chemicals and correspondence to globally harmonized system (GHS) classification. , 2009, The Journal of toxicological sciences.