High-Performance Liquid Chromatographic Determination of Pyrazinamide In Human Plasma, Bronchoalveolar Lavage Fluid, and Alveolar Cells

Published: Journal of Chromatographic Science, ISSN 0021-9665 Volume 38, Number 1, January 2000, pp. 33-37

High-Performance Liquid Chromatographic Determination of Pyrazinamide In Human Plasma, Bronchoalveolar Lavage Fluid, and Alveolar Cells
John E. Conte, Jr.1,2,3,*, Emil Lin4, and Elisabeth Zurlinden1
1-Department of Epidemiology & Biostatistics, Infectious Diseases Research Laboratory, 2-Department of Medicine, 3-Department of Microbiology and Immunology, and 4-Department of Biopharmaceutical Sciences, University of California San Francisco, 350 Parnassus Avenue, Suite 210, San Francisco, CA 94117

A technique is presented for the measurement of pyrazinamide in human plasma, bronchoalveolar lavage, and alveolar cells by reversed-phase column chromatography. The assay utilizes acetazolamide as an internal standard, ultraviolet detection at 268 nm, and an acetonitrile-based mobile phase. Preparation of plasma samples requires a simple deproteinization step, resulting in the development of sharp peaks with retention times of 8.4 and 17 minutes for pyrazinamide and acetazolamide, respectively. Bronchoalveolar lavage and alveolar cell suspensions require an acid extraction with ethyl acetate, evaporation to dryness, and reconstitution. This method provides specific, rapid, and reliable determinations of drug concentrations and therefore is suitable for pharmacological studies, particularly those that are designed to quantitate the intrapulmonary concentrations of pyrazinamide.

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