Midazolam MRM Method

Measured Analytes

  1. Midazolam


Atmospheric Pressure Ionization Liquid Chromatography tandem mass spectrometry (APCI-LC-MS) was carried out in the positive polarity with a Shimadzu Prominence HPLC system and SCIEX 4000 mass spectrometer. An aliquot of 10 µl of each sample was injected onto an Phenyl-hexyl, 3 µm 100 x 2.1 mm column for gradient separation. Mobile phases were: A, ddH2O with 0.1% Formic Acid; and B, Acetonitrile with 0.1% Formic Acid. Gradient schedule starts at 10% B and is linearly increased to 100% B at 5 minutes, then held at 100% B until 7 minutes, and returned to 10% B at 8 minutes and re-equilibration until 10 minutes. Time per sample injection was 10 minutes. The flow rate was 300µL/min.

Extraction Protocols:

Solvent precipitation was used to extract midazolam from human serum sample. An aliquot of 200µL of plasma was mixed with 800 µL of 1% (v/v) acetic acid in MeOH and 200 µL of triazolam (600 nM) as internal standard, then vortexed and centrifuged at 1,800 ×g for 10 minutes.

Method derived from the following reference


[1] Hemodynamic characteristics of midazolam, propofol, and dexmedetomidine in healthy volunteers
Michael A. Frölich, Ali Arabshahi, Charles Katholi, Jeevan Prasain, Stephen Barnes
J Clin Anesth. Author manuscript; available in PMC 2013 Nov 14.
Published in final edited form as: J Clin Anesth. 2011 May; 23(3): 10.1016/j.jclinane.2010.09.006. doi: 10.1016/j.jclinane.2010.09.006 PMCID: PMC3828053


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