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. 2011 Jun 1;55(3):487-93.
doi: 10.1016/j.jpba.2011.01.040. Epub 2011 Mar 5.

Simultaneous determination of alfentanil and midazolam in human plasma using liquid chromatography and tandem mass spectrometry

Thomas Kim  1 , Amy London, Evan D Kharasch
Affiliations

Simultaneous determination of alfentanil and midazolam in human plasma using liquid chromatography and tandem mass spectrometry

Thomas Kim et al. J Pharm Biomed Anal. .

Abstract

A fast, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of alfentanil and midazolam in human plasma has been developed and validated. Alfentanil and midazolam were extracted from plasma using a mixed-mode cation exchange solid phase extraction method, with recoveries of both compounds greater than 80% at 3 different concentrations (1, 10 and 100ng/ml). Compounds were analyzed on a C(18) column with a water and methanol mobile phase gradient with acetic acid as an additive, at a flow rate of 0.3ml/min. The working assay range was linear from 0.25 to 100ng/ml for each compound. The signal to noise ratio was 80 and 40 for alfentanil and midazolam, respectively, at the lowest concentration calibration standard, with less than 10% matrix suppression by human plasma at this concentration. Alfentanil and midazolam were stable in human plasma during storage at -80°C, processing, and analysis. The procedure was validated and applied to the analysis of plasma samples from healthy human subjects administered oral and intravenous alfentanil and midazolam.

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Conflict of interest statement

Conflict of interest: No author has any conflict of interest

Figures

Figure 1
Figure 1
Representative chromatograms of the lowest calibration standard (0.25 ng/ml) of (A) alfentanil and (B) midazolam in human plasma.
Figure 2
Figure 2
Representative chromatograms of blank human plasma to assess potential interfering peaks at the retention times and MRM transitions for (A) midazolam, (B) midazolam d4, (C) alfentanil, and (D) alfentanil d5.
Figure 3
Figure 3
Representative LC-MS/MS chromatograms of human plasma from a subject administered alfentanil and midazolam. Shown are (A) midazolam, (B) midazolam d4, (C) alfentanil, and (D) alfentanil-d5.
Figure 4
Figure 4
Plasma concentration vs time profiles of alfentanil and midazolam after administering (A) intravenous midazolam, (B) intravenous alfentanil, (C) oral midazolam, and (D) oral alfentanil.
See this image and copyright information in PMC

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