A brief review on: Separation techniques chromatography
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Chromatography is an important biophysical process, allowing the components of a mixture to be isolated, classified and extracted for qualitative and quantitative analysis. Proteins can be purified based on features such as size and shape, total charge, surface hydrophobic groups and stationary phaset binding ability. Four separation strategies based on molecular characteristics and form of interaction Four separation techniques based on molecular characteristics and form of interaction use ion exchange mechanisms, surface adsorption, partition, and exclusion of the size. Column chromatography is one of the most common methods of protein purification Chromatography is based on the theory of separation of fluid stationary phase (stable phase) when traveling with the support of a mobile phase. The variables that are involved in this separation process include adsorption (liquid-solid), partition (liquid-solid), and affinity or variations between their molecular weights. Due of these variations, certain components of the mixture stay in the stationary phase longer, and travel gradually in the chromatography system, while others move rapidly through the mobile phase, leaving the system faster. Based on this method, three components form the basis of the chromatography technique. Stationary phase: is a phase consisting of a "solid" phase or a "solid surface adsorbed layer" Mobile phase: is a phase consisting of a "liquid" or a "gaseous part." Separate molecules which form the interaction between stationary phase, mobile phase and substances contained in the mixture are the basic component that is successful in separating molecules from the partition-based chromatography methods are very effective in separating small molecules and distinguishing them as amino acids, carbohydrates and fatty acids. In the separation of macromolecules as nucleic acids and proteins, however, affinity chromatography’s (i.e. ion-exchange chromatography) are more efficient. Paper chromatography is used in protein separation and protein synthesis studies; gas-liquid chromatography is used to isolate alcohol, ethanol, lipid, and amino groups, and to analyze enzymatic interactions, while molecular-sieve chromatography is used in particular to evaluate molecular protein weights. Agarose gel chromatography is used to purify RNA, DNA particles, and viruses. The chromatography stationary phase is a solid phase or liquid phase that is sprayed on a solid phase surface. A gaseous or liquid phase is a mobile phase which flows through the stationary phase. If the mobile phase is liquid, it is called liquid chromatography, and if it is gas, it is called gas chromatography. of gases and mixtures of volatile liquids, and solid material, gas chromatography is used. Liquid chromatography is used in particular for thermal unstable and non-volatile materials. The purpose of applying chromatography, which is used as a quantitative analysis tool apart from its separation, is to record a sufficient separation within an acceptable time interval. Various chromatography methods have been developed to that end. Some of these have chromatography on columns, thin layer chromatography (TLC), paper chromatography, gas chromatography, ion exchange chromatography, gel permeation chromatography, liquid high pressure chromatography, and affinity chromatography.
HPLC, Mobile Phase, Stationary Phase, Detector, Chromatography, Instrumentation.