Analysis of Metabolites

Authored by: Kathleen R. Noon , Ara Kirakosyan , Maureen McKenzie , Peter B. Kaufman

Handbook of Molecular and Cellular Methods in Biology and Medicine

Print publication date:  December  2011
Online publication date:  April  2016

Print ISBN: 9781420069389
eBook ISBN: 9781439881958
Adobe ISBN:

10.1201/b11351-26

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Abstract

Chromatography is a method of separation that uses two phases: a stationary phase and a mobile phase. A mixture of components called analytes is introduced into the system by either applying it directly to the stationary phase or indirectly through the use of mobile phase. As the mobile phase passes through the stationary phase, the components of the mixture will distribute between the two phases, resulting in differential rates of migration through the system. The migration rates of the analytes are determined by their relative tendencies to interact with the stationary phase and mobile phase. The molecular mechanisms for separation of analytes include adsorption, hydrophobic interactions, partitioning, electrostatic interactions, hydrodynamic movement, and migration in an electric field. The amount of time it takes for a compound to travel through the stationary phase and reach the detector is called the retention time (Rt). These techniques are very powerful because they allow one to separate compounds of similar physical and chemical properties within a complex sample mixture that may be structurally very similar or even isomers of each other. It is also fast, easy, and economical. 1 10

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