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

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 microg/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.

Quantification of vancomycin in human serum by LC-MS/MS

“…Indeed, several LC-MS/MS methods for the quantification of serum vancomycin concentrations have been described previously [7][8][9]. These methods, however, rely on internal standard compounds with are structurally not related to the target analyze (teicoplanin, atenolol).…”

Analytical methods for vancomycin determination in biological fluids and in pharmaceuticals

“…Comparing HPLC methods with the immunologic, it is observed that the former presents better accuracy and precision but higher analytical delay and involve pre extraction of the analyte. To avoid the necessity of the pre treatment some HPLC with direct injection, using pre extraction columns, have been suggested 54,66 .…”

“…When the HPLC technique is used for the determination of vancomycin in biological fluids, the extraction procedure of the analyte is done based on chemical processes, involving protein precipitation and use of organic solvents. In some methods, however, the extraction process is associated to the chromatography as continuous systems 56,59,66 . Recently analytical techniques, where the samples are injected without treatment, have been developed.…”

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“…In the clinical laboratory the use of this technique has been efficient for determination of analytes in several biological fluids. Therefore, HPLC has been developed for the analysis of vancomycin in medicines, as well as for the determination in biological fluids 17,24,[26][27][28][29][30][31][32][33][34] . Kitahashi and Furuta 35 developed an assay for measuring vancomycin concentration by micellar electrokinetic capillary chromatography with detection at 210 nm using direct serum injection.…”