An ultra-sensitive LC–MS/MS method to determine midazolam levels in human plasma: development, validation and application to a clinical study. (February 2017)

• Record Type: Journal Article
• Authors: Chen, Mu; Lu, Wenzhe; Lu, Yang; Kang, Lijuan; Zhao, Harry; Lin, Zhongping John; Wang, Weimin; Fraier, Daniela; Ottaviani, Giorgio
• Is Part Of: Bioanalysis. Volume 9:Number 3(2017)
• Page Start: 297
• Page End: 312
• Publication Date: 2017-02
• Subjects: CYP3A4 -- LC–MS/MS -- microdosing -- midazolam -- Phase 0 clinical study -- SPE -- ultra sensitive; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2016-0191
• Languages: English
• ISSNs: 1757-6180

Abstract

Aim: Midazolam is a commonly used marker substrate for the in vivo assessment of CYP3A activity. Reliable pharmacokinetic assessment at sub-pharmacological doses of midazolam requires an ultra-sensitive analytical method.Methods: A new, ultra-sensitive LC–MS/MS method for the determination of midazolam in human plasma using SPE was developed and fully validated. The lowest limit of quantitation is 0.1 pg/ml with a sample volume of 500 μl.Results/conclusion: The following parameters were validated: sensitivity, assay accuracy and precision, linearity, selectivity, and stability of midazolam at pertinent analytical and storage conditions. The validated method was utilized successfully for the sample assay during a midazolam microdosing study for the evaluation of CYP3A4 activity of a clinical candidate.