Fast HPLC analysis, macromolecules

For this reason, the performance of these instruments is less satisfactory in applications with high speed columns which allow the analysis of biopolymers to be carried out on a time scale of a minute or less. In order to expand the scope of HPLC to such applications, some of the considerations described below should be taken in to account in the design of fast HPLC analyzers for biological macromolecules. 

The analysis of lipids is dominated by chromatographic methods such as gas chromatography (GC) and high-pressure liquid chromatography (HPLC) and some other physical methods such as thermo analysis. Nuclear magnetic resonance (NMR) spectroscopy covers only a small part of lipid analysis. In most cases it is used for qualitative analysis such as structure elucidation of lipids. Unlike the other methods NMR spectroscopy is coupled with computer technology, and its development is as fast as that of computer software and hardware. In 1991 the Nobel Prize for chemistry was given to Professor Ernst for the development of Fourier Transform NMR spectroscopy (FT-NMR). The capabilities of these methods even for the elucidation of the three-dimensional structures of small molecules up to macromolecules are impressive. 

Nowadays, interfacing separation methods such as high-performance hquid chromatography (HPLC) or CE [54] to MS is already a routine technique. MS methods based on so-called soft ionization techniques, which include fast atom bombardment, laser desorption and electrospray ionization (ESI), have allowed the analysis of biological macromolecules that in the past could have been analyzed only by extensive cleavage and derivatization. Of these methods, the two most preferred for biomolecules are ESI [55] and matrix-assisted laser desorption/ionization(MALDI) [56,57], for which time-of-flight (TOE) and ion trap mass analyzers are the most frequentiy used mass analysis methods. However, these methods are equally suitable for small molecules, such as metabolites or the products of organochemical reactions. 

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