Direct determination of diazepam and its glucuronide metabolites in human whole blood by μElution solid-phase extraction and liquid chromatography-tandem mass spectrometry

Xin Wang, Rong Wang, Yurong Zhang

17 Citations (Scopus)

Abstract

A μElution solid-phase extraction (SPE) liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of diazepam, nordiazepam, oxazepam, oxazepam glucuronide, temazepam and temazepam glucuronide in human whole blood is presented. 200. μL of whole blood samples were loaded onto a Waters Oasis HLB 96-well μElution SPE plate using 75. μL of methanol as the elution solvent, and the eluents were injected into an Eclipse XDB C18 column. No hydrolysis, solvent transfer, evaporation or reconstitution was involved in the sample preparation procedures. Tandem mass spectrometric detection with Turbo Ion Spray was conducted via multiple reaction monitoring (MRM) under positive ionization mode. The method was validated and proved to be accurate (accuracy within 93-108%), precise (intra-day RSD. <. 9.9% and inter-day RSD. <. 7.2%) and sensitive with limits of detection (LOD) in the range of 0.05-0.25. ng/mL for all the compounds. Extraction recoveries were in the range of 31-80% for all the analytes. This method demonstrated to be reproducible and reliable. The applicability of the method was demonstrated by analysis of several forensic cases involving diazepam and its metabolites.

Original languageEnglish
JournalForensic Science International
Volume233
Issue numberISSN: 0379-0738
Pages (from-to)304-311
Number of pages8
ISSN0379-0738
Publication statusPublished - 10 Dec 2013

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