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

作者信息

Chen Mu, Lu Wenzhe, Lu Yang, Kang Lijuan, Zhao Harry, Lin Zhongping John, Wang Weimin, Fraier Daniela, Ottaviani Giorgio

机构信息

Frontage Laboratories Inc., 700 Pennsylvania Drive, Exton, PA 19341, USA.

Roche Innovation Center Shanghai, 720 Cai Lun Road, Building 5/917 Ha Lei Road, Building 6, Pudong, Shanghai 201203, China.

出版信息

Bioanalysis. 2017 Feb;9(3):297-312. doi: 10.4155/bio-2016-0191. Epub 2017 Jan 23.

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.