Chromatography Methods and Their Application
Sofija Sasnauskienė, Regina Firantienė, Valerija Jablonskienė
Summary
Chromatography is the collective term for a set of laboratory techniques for the separation of mixtures and analysis. Chromatografic systems consist of the stacionarry phase and the mobile phase. The mixture is dissolved in a fluid called the “mobile phase”, which carries it through a structure holding another adsorbent material called the “stationary phase” or matrice. The various constituents of the mixture travel at different speeds, causing them to separate. The separation is based on differential parti- tion-ng between the mobile and stationary phases. The choice of stationary phase is made so that the compounds to be separated have differences in a compound’s structure result in differential retention on the stationary phase and thus changing the separation. Chromatography may be preparative or analytical. The purpose of preparative chromatography is to separate the components of a mixture for further use (and is thus a form of purification). Analytical chromatography is done normally with smaller amounts of material and is for measuring the relative proportions of compounds in a mixture. Chromatography - is one of the most important and rapidly developing laboratory techniques in chemistry, biochemistry, molecular biology and experimental medicine. It is applicable in fields of life and medical sciences, laboratory diagnostics and clinical purposes. Chromatography was first employed by Russian botanist M. Tsvet in the first decade of the 20th century, primarily for the separation of plant pigments using a column of calcium carbonate. In process became the first scientist to recognise that chlorophyll was not a single plant pigment, separate chlorophyll and carotenes. After chromatographic separation methods developed intensively and chromatographic technique has advanced rapi dly. The main principles of chromatography could be applied in many different ways. Chromatography can separate complex mixtures with great precision. Chromatography can purify basically any substance if the right adsorbent material, carrier fluid, and operating conditions are employed. Chromatography can be used to separate delicate, labile, compounds, enzymes, nucleic acids, antibodies, is quite well suited to a variety of uses in the field of biotechnology, biochemistry, experimental and practical medicine.
Keywords: chromatography, HPLC, affinity, ion exchange, MARS, diagnostic.
