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A general approach for the quantitative analysis of bisphosphonates in human serum and urine by high‐performance liquid chromatography/tandem mass spectrometry
Author(s) -
Zhu Lee S.,
Lapko Veniamin N.,
Lee Jean W.,
Basir Yousef J.,
Kafonek Chris,
Olsen Richard,
Briscoe Chad
Publication year - 2006
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.2755
Subject(s) - chemistry , chromatography , derivatization , diazomethane , solid phase extraction , bioanalysis , extraction (chemistry) , mass spectrometry , urine , tandem mass spectrometry , detection limit , liquid chromatography–mass spectrometry , sample preparation , organic chemistry , biochemistry
Bisphosphonates are extremely hydrophilic and structurally similar to many endogenous phosphorylated compounds, making their selective extraction from serum or urine very challenging. Many bisphosphonates lack strong chromophores for sensitive UV or fluorescence detection. We report here the first general approach to enable sensitive and selective quantitation of N‐containing bisphosphonates by liquid chromatography/tandem mass spectrometry (LC/MS/MS) following derivatization with diazomethane. The novelty of the strategy lies in performing the derivatization on silica‐based anion‐exchange sorbents as an integrated step in the sample purification by solid‐phase extraction (SPE). The ‘on‐cartridge’ reaction with diazomethane not only led to higher efficiency of derivatization, but also enabled a more discriminatory recovery of the drug's derivatives. The derivatized bisphosphonates demonstrated improved chromatographic separation and increased sensitivity of the detection. The general applicability of the approach was demonstrated by validation of bioanalytical methods for risedronate and alendronate in human serum and urine. Sensitivity was achieved at the pg/mL level with merely 100–200 µL of sample. Copyright © 2006 John Wiley & Sons, Ltd.