New Generation of Chromatographic Packings and Columns for Determination of Biologically Active Compounds

Abstract

Analysis of biologically active substances is particularly important in the pharmaceutical and biomedical area. For separation of polar compounds or complex mixtures by normal (NP) or reversed phase liquid chromatography (RP-HPLC) and/or electromigration techniques, it is necessary to apply a new generation of packings and columns with strictly defined properties. It is connected to the definition of chromatographic behavior and determination of compounds that are described by its structure, as well as chemical and physical properties. One of the factors playing a predominant role in the separation process is the interaction between analyte and stationary phases. During recent years a large variety of stationary phases have become available and have been also applied in routine and research chromatographic separations. Bonded phases are in widespread use and popular because of the great number of available packing materials, allowing the solution of a scale of different separation problems. At the present time analytical methods cannot be restricted only to the so-called “black box.” To meet the requirements of modern analytical techniques, strong demands are put on further, deeper understanding of the essence of the separation process, among other things, on the basis of conclusions about interactions between solute and stationary phase surfaces. For surface characterization different physicochemical methods such as CP/MAS, NMR, FT IR, DCS, chromatography, etc., have been described. The resolution, as reflected by efficiency, selectivity, and retention patterns on these materials, has been demonstrated. The effect of structure of stationary phases on retention of a model series of test analytes has been proved and numerically expressed by means of the Quantitative Structure-Retention Relationship (QSSR). The main aim of this paper is to present possibilities of determining different biologically active compounds (e.g., vitamins, steroids, nucleosides, peptides) in complex chromatographic methods (sample preparation, final analysis, validation) using new generation of stationary phases, columns, and chips divides. Special attention is dedicated to the advantage of packing materials imitating natural membranes because of the possible examination of the interaction between drug membrane. They can permit to the design of new pharmaceuticals and observation of processes taking place on the border of phases without interfering in natural systems.