Simultaneous determination of L-693,612, a topical carbonic anhydrase inhibitor, and two potential metabolites in human whole blood by ion-pair high-performance liquid chromatography |
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Authors: | E.J. Woolf T. Au M. Kasper M. Constanzer B. Matuszewski |
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Affiliation: | Merck Research Laboratories, Department of Drug Metabolism, West Point, PA 19486 USA |
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Abstract: | A method for the simultaneous determination of a topical carbonic anhydrase inhibitor, L-693,612, and two of its potential metabolites in human whole blood is described. The analytes are isolated from the matrix via liquid-liquid extraction with a mixture of toluene, ethyl acetate and isopropanol (49:50:1, v/v/v). The analytes are then back extracted into dilute phosphoric acid prior to injection into the HPLC system. A cyano column (Zorbax SB-CN, 150 × 4.6 mm) with a mobile phase of phosphoric acid(0.085%)-acetonitrile (73.5:26.5) containing 10 mM sodium decane sulfonate and adjusted to pH 3 is used for the analysis. Detection is based on UV absorbance at 252 nm. The assay was found to be linear in the concentration range of 5–500 ng/ml for each analyte when 1-ml aliquots of whole blood were extracted. |
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