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Multidimensional Chromatography - Essay Example

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Biochemical separation techniques have long depended on chromatographic modes that evolved into newer and more advanced forms with complicated procedures for separation and analysis. In fact, chromatography as a whole has found its application in all fields of natural and biological science, with extensive use in biomedical and biopharmaceutical research…
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Multidimensional Chromatography
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Download file to see previous pages Hence, essentially speaking, one dimensional chromatographic separation is definitely not the last word for isolation and characterization of biomolecules, chiefly proteins. The limitation is due to its low resolving power in presenting a homogenous protein sample from a complex mixture of several other unwanted proteins, associated matrix components and metabolites that can otherwise be segregated in a much more efficient way by various types of matrix loaded multiple columns utilizing their differential properties and affinities. Moreover, combination of two or more analytical separation steps involve perfectly standardized pre-treatment processes that are to be coordinated in an automated way for a complete segregation of the test samples. (Mondello, P. 251)
Multidimensional separation techniques were first devised with the implementation of two dimensional separation beds, in which two different parameters for migration corresponding to two distribution coefficients help to identify the separated components on the bed with more certainty and a high peak capacity than that achieved by one dimensional separation procedures. (Cortes, P. 14 - 15) A high value of peak density is an important requisite for effective separation process and it can easily be achieved by combining two or more chromatographic separation techniques. The greater the number of peaks that can be resolved effectively in a given multidimensional separation procedure within a fixed time frame greater is the overall efficiency of the process. Likewise with effective resolution of a higher number of peaks more and more complex samples can be duly segregated, characterized and analyzed for further investigation. In a simple two dimensional chromatographic process that is carried out by a combination of two different columns, the maximum peak capacity will be the product of the peak capacities of the individual columns, thereby generating a higher capacity of resolution for the overall analytical process. As for example, the peak capacities for each separation process being 100 and 200, the combined value of the maximum peak capacity of the two dimensional separation procedure becomes a product of the two, that is, 20000, thereby ensuring high resolution and larger separation between the collected fractions. (Chromatography, 1998 - 2008)
Multidimensional chromatography, therefore, finds a maximum usage in biomedical research and especially in proteomics, where investigation of biological samples of protein mixtures requires prior separation and characterization in order to tag the target protein. In protein analysis research several novel methods of multidimensional chromatographic separations are being currently used in complementation with the 2-D polyacrylamide gel electrophoretic separation (2D-PAGE), for a much better resolution and greater separation space between analyzable eluents. Multidimensional approach involving multidimensional liquid chromatography (MDCL) used in unison with bio-mass ...Download file to see next pagesRead More
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