Related Experiment Video
Updated: Mar 8, 2026
Author Spotlight: An Efficient Methodology to Confidently Differentiate and Characterize Fentanyl Analogs
Published on: November 8, 2024
An ultra-sensitive LC-MS/MS method to determine midazolam levels in human plasma: development, validation and
Mu Chen1, Wenzhe Lu2, Yang Lu1
1Frontage Laboratories Inc., 700 Pennsylvania Drive, Exton, PA 19341, USA.
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.
