Chromatography Methods and Their Application

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.