Chromatography KSU色谱 KSU.ppt

Chromatography History Mikhail Tswett, Russian, 1872-1919 Botanist In 1906 Tswett used to chromatography to separate plant pigments He called the new technique chromatography because the result of the analysis was written in color along the length of the adsorbent column Chroma means “color” and graphein means to “write” Importance Chromatography has application in every branch of the physical and biological sciences 12 Nobel prizes were awarded between 1937 and 1972 alone for work in which chromatography played a vital role Chromatography is a physical method of separation in which the components to be separated are distributed between two phases one of which is stationary (stationary phase) while the other (the mobile phase) moves through it in a definite direction. The chromatographic process occurs due to differences in the distribution constant of the individual sample components. Chromatography Classification of chromatography according to mobile phase: 1- Liquid chromatography: mobile phase is a liquid. (LLC, LSC). 2- Gas chromatography : mobile phase is a gas. (GSC, GLC). Classification according to the packing of the stationary phase: 1- Thin layer chromatography (TLC): the stationary phase is a thin layer supported on glass, plastic or aluminium plates. 2- Paper chromatography (PC): the stationary phase is a thin film of liquid supported on an inert support. 3- Column chromatography (CC): stationary phase is packed in a glass column. Classification according to the force of separation: 1- Adsorption chromatography. 2- Partition chromatography. 3- Ion exchange chromatography. 4- Gel filtration chromatography. 5- Affinity chromatography. Thin layer chromatography (TLC) is a method for identifying substances and testing the purity of compounds. TLC is a useful technique because it is relatively quick and requires small quantities of material. Separations in TLC involve distributing a mixture of two or more substances between a stationary phase and a