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High‐throughput approaches to the quantitative analysis of ketoconazole, a potent inhibitor of cytochrome P450 3A4, in human plasma
Author(s) -
Ramos L.,
Brignol N.,
Bakhtiar R.,
Ray T.,
Mc Mahon L. M.,
Tse F. L. S.
Publication year - 2000
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/1097-0231(20001215)14:23<2282::aid-rcm164>3.0.co;2-v
Subject(s) - chemistry , ketoconazole , cyp3a4 , cytochrome p450 , pharmacology , human plasma , pharmacokinetics , chromatography , biochemistry , enzyme , antifungal , medicine , dermatology
Abstract Ketoconazole, an imidazole‐piperazine compound, is an orally active antimycotic agent. In addition, ketoconazole is a specific inhibitor of cytochrome P450 3A4. As about 60% of oxidized drugs are biotransformed by this isoform, the potential effect of a concomitant administration of ketoconazole on drug disposition may be of interest during drug development. The present paper describes three different approaches (methods A, B, and C) to attain high‐throughput sample preparation and analysis in the quantification of ketoconazole in human plasma. Method A consisted of acetonitrile precipitation in a 96‐well plate, transfer of the supernatant via a Tomtec Quadra 96 Model 320, and subsequent injection onto a 50 × 4.6 mm (i.d.) Develosil Combi‐RP‐5 column (packed with C30 bonded silica particles). Method B consisted of an identical sample preparation to method A with the exception that a Michrom Magic Bullet ™ column, 2.0 → 0.50 mm (i.d., tapered bore) ×25 mm length, was used. Lastly, in method C, a turbulent‐flow chromatography (TurboFlow LC/APCI‐MS/MS) module was used for the direct analysis of ketoconazole in human plasma. A Sciex API 3000 was used in methods A and B, while a Micromass Quattro LC was employed in method C. Based on the values obtained for the calibrator (standard) and quality control samples, all three protocols yielded satisfactory accuracy, precision, and reduced manual sample preparation time. Copyright © 2000 John Wiley & Sons, Ltd.