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Lecture 9 Introdution to Chromatography “The Science of 9讲介绍色谱学”
PHCMt561 Instrumental Analysis - Lecture #7 RN, 2008-12-01 * The majority of HPLC columns range from 10-30 cm, the inside diameter is often 4-10 mm and the most common particle size is 5-10 μm. Guard columns are connected to the analytical column to increase its life by removing not only particulate matter and contaminants form the solvents but also from sample components that bind irreversibly to the stat. phase. The composition of the guard column should be closely similar to that of the analytical column. When the guard column has been contaminated, it is discarded and replaced with a new one. Thus the guard column is sacrificed to protect the more expensive analytical column. We normally carry out chromatography for analytical purposes (to separate and to quali- and quantitative determination) or for preparative purposes to purify a significant quantity of a component in a mixture. While analytical chromatography is usually performed in thin columns that provide good separation, in preparative chromatography, fatter columns are used that can handle more load. Preparative chromatography is especially important in the pharmaceutical industry in case of separating optical isomers of drugs. PHCMt561 Instrumental Analysis - Lecture #7 RN, 2008-12-01 * Packed columns contain fine particles of solid support coated with non volatile liquid stationary phase or the solid phase itself may be the stationary phase. Compared with packed columns, open tubular columns offer: higher resolution, shorter analysis time, greater sensitivity, lower sample capacity. The wall coated column has a 0.1 to 0.5 μm thick film of stat. liquid phase on the inner wall of the column. The support coated column has solid particles coated with stationary phase and attached to the inner walls of the column. In the porous layer column, solid particles are the active stationary phase. With their high surface area, SCOT can handle larger capacity samples than wall coated. The open tubular columns ar
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