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Simultaneous determination of midazolam and 1′‐hydroxymidazolam in human plasma by liquid chromatography with tandem mass spectrometry
Author(s) -
Li Wenkui,
Luo Suyi,
Smith Harold T.,
Tse Francis L. S.
Publication year - 2007
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.829
Subject(s) - chemistry , chromatography , mass spectrometry , ammonium acetate , extraction (chemistry) , electrospray ionization , tandem mass spectrometry , liquid chromatography–mass spectrometry , electrospray , selected reaction monitoring , hydrophilic interaction chromatography , liquid–liquid extraction , analytical chemistry (journal) , high performance liquid chromatography
A sensitive and simple liquid chromatography–tandem mass spectrometry method for the determination of midazolam and 1′‐hydroxymidazolam in human plasma has been developed and validated with a dynamic range of 0.1–250 ng/mL. The analysis was based on semi‐automated liquid–liquid extraction followed by evaporation of the extraction solvent, reconstitution and chromatography on a reversed‐phase C 18 column. The mobile phase consists of 5 m m ammonium acetate and methanol and runs in gradient at a flow rate of 0.25 mL/min with column temperature of ∼20°C. The entire column effluent was transferred into the LC‐MS/MS interface operated in positive electrospray ionization mode. The chromatographic run time was 4.3 min per injection, with retention times for midazolam, 1′‐hydroxymidazolaml and the internal standard, triazolam, of 2.5, 2.3 and 2.1 min, respectively. The intra‐day and inter‐day precision (RSD %) and accuracy (bias %) of the quality control samples were <15.0% and within ±13%, respectively. The current method has been applied to a clinical drug–drug interaction study in human. Copyright © 2007 John Wiley & Sons, Ltd.