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Separation and Quantitation of R and S Enantiomers using Normal Phase Chromatography on an API 3000 LC/MS/MS System

Purpose

Normal phase chromatography can be used for the analysis of chiral compounds using the API 3000 LC/MS/MS System.

Overview

In recent years, pharmaceutical companies have placed greater importance on examining the stereoisomeric composition of drugs having a chiral center. Isomerspecific pharmaceuticals often exhibit increased potency, higher bioavailability and reduced side effects when compared to racemic pharmaceutical compounds. Previously, chiral compounds could not be separated by conventional HPLC techniques without derivatizing enantiomers into diastereomers. Todays improved chiral stationary phases allow for separation and quantitation of these enantiomers. These chiral HPLC columns are amenable to normal phase chromatography. This technique was not often utilized in LC/MS in the past as there is a serious non-polar mobile phase incompatibility with atmospheric pressure ionization mass spectrometry. Mobile phases primarily consisting of hexane present an explosion hazard when paired with the extreme temperatures of atmospheric pressure chemical ionization (APCI).

The experiment outlined here demonstrates that it is possible to safely and effectively perform normal phase chromatography by APCI LC/MS/MS using an API 3000 triple quadrupole mass spectrometer.

Key Features

Excellent APCI performance across a wide range of flow rates and mobile phase compositions

Simplified MS/MS tuning

Analyst quantitation software for fast and easy data acquisition and reduction

Experiment

The goal was to separate and quantitate the precise amount of R and S enantiomers of three commonly used drugs found in biological fluids of animals dosed with racemic mixtures. These drugs include the an
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