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Rapid bioanalysis of vancomycin in serum and urine by high‐performance liquid chromatography tandem mass spectrometry using on‐line sample extraction and parallel analytical columns
Author(s) -
Cass Robert T.,
Villa Josephine S.,
Karr Dane E.,
Schmidt Donald E.
Publication year - 2001
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/rcm.246
Subject(s) - chemistry , chromatography , bioanalysis , extraction (chemistry) , urine , tandem mass spectrometry , liquid chromatography–mass spectrometry , sample preparation , mass spectrometry , sample (material) , biochemistry
Abstract A novel high‐performance liquid chromatography tandem mass spectrometry (LC/MS/MS) method is described for the determination of vancomycin in serum and urine. After the addition of internal standard (teicoplanin), serum and urine samples were directly injected onto an HPLC system consisting of an extraction column and dual analytical columns. The columns are plumbed through two switching valves. A six‐port valve directs extraction column effluent either to waste or to an analytical column. A ten‐port valve simultaneously permits equilibration of one analytical column while the other is used for sample analysis. Thus, off‐line analytical column equilibration time does not require mass spectrometer time, freeing the detector for increased sample throughput. The on‐line sample extraction step takes 15 seconds followed by gradient chromatography taking another 90 seconds. Having minimal sample pretreatment the method is both simple and fast. This system has been used to successfully develop a validated positive‐ion electrospray bioanalytical method for the quantitation of vancomycin. Detection of vancomycin was accurate and precise, with a limit of detection of 1 ng/mL in serum and urine. The calibration curves for vancomycin in rat, dog and primate were linear in a concentration range of 0.001–10 µg/mL for serum and urine. This method has been successfully applied to determine the concentration of vancomycin in rat, dog and primate serum and urine samples from pharmacokinetic and urinary excretion studies. Copyright © 2001 John Wiley & Sons, Ltd.