Liquid chromatography-mass biopolymers

Electrospray mass spectrometry has developed into a well-established method of wide scope and potential over the past 15 years. The softness of electrospray ionization has made this technique an indispensable tool for biochemical and biomedical research. Electrospray ionization has revolutionized the analysis of labile biopolymers, with applications ranging from the analysis of DNA, RNA, oligonucleotides, proteins as well as glycoproteins to carbohydrates, lipids, gly-colipids, and lipopolysaccharides, often in combination with state-of-the-art separation techniques like liquid chromatography or capillary electrophoresis [1,2]. Beyond mere analytical applications, electrospray ionization mass spectrometry (ESMS) has proven to be a powerful tool for collision-induced dissociation (CID) and multiple-stage mass spectrometric (MSn) analysis, and - beyond the elucidation of primary structures - even for the study of noncovalent macromolecular complexes [3]. 

A new on-line approach to SPE is the use of so-called turbulent flow chromatography which combines rapid mobile phase linear velocities with larger particle sizes this approach was discussed in Section 3.5.9 in the context of breakdown of the conventional (van Deemter) rate theory. Whether or not the flow can truly be described as turbulent (Ayrton 1998), there is no question that eddies are formed that enhance the interactions of smaller molecules with the stationary phase. In contrast the large proteins and other biopolymers have rates of mass transfer from the liquid to stationary phase (C term in the van Deemter equation) that are too... 

Countercurrent chromatography is based on the distribution of substances in two liquid phases [128,129]. The liquid is fed into a coiled tube that is moved along an orbital trajectory. Due to centrifugal power, the liquids move in a counter-current. For proteins and many other biomolecules, this method is not practical because of denaturation in a nonaqueous phase. In aqueous two-phase systems, at least one phase exhibits high viscosity and, therefore, mass transfer between the two phases is limited. Similar problems occur with reversed micelle extraction as were observed with the aqueous two-phase extraction [130]. CCC has not been used for large-scale purification of proteins and other biopolymers. 

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