• Application Note

Diazepam in Serum by LC-MS

Diazepam in Serum by LC-MS
  • Waters Corporation
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This is an Application Brief and does not contain a detailed Experimental section.

Abstract

This application brief demonstrates the analysis of diazepam by LC-MS. 

Introduction

The compound analyzed in this study is diazepam. 

Experimental

HPLC Method

Column:

Symmetry C18, 2.1 x 100 mm, 3.5 μm

Part number:

WAT058965

Mobile phase:

2 mM Ammonium Acetate/Acetonitrile/Formic Acid 65:35:0.1

Flow rate:

200 μL/min

Injection volume:

10 μL

MS:

Micromass Quattro LC

Ion Mode:

ES+

Cone Voltage:

45 V

Collision Energy:

25 eV

Results and Discussion

Background-subtracted electrospray mass spectrum of pure diazepam standard (5μg), under optimum conditions
Figure 1. Background-subtracted electrospray mass spectrum of pure diazepam standard (5μg), under optimum conditions. Chromatographed as described above to remove contaminants.
Background-subtracted electrospray product tion spectrum of pure diazepam standard (5μg), under optimum conditions
Figure 2. Background-subtracted electrospray product tion spectrum of pure diazepam standard (5μg), under optimum conditions. Chromatographed as described above to remove contaminants.
Calibration curve with triplicate injections for each point and demonstrating LOQ at 0.2ng/mL
Figure 3. Calibration curve with triplicate injections for each point and demonstrating LOQ at 0.2ng/mL.
MRM Chromatograms under optimum conditions of pure diazepam
Figure 4. MRM Chromatograms under optimum conditions of pure diazepam standard at (A) 0.2ng/mL (LOD) and (B) 5.0ng/mL and (C) a processed human plasma sample with a low concentration of diazepam (calculated as 0.75ng/mL).

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WA31763.72, June 2003

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