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level: CHROMATOGRAPHIC METHODS

Questions and Answers List

level questions: CHROMATOGRAPHIC METHODS

Question	Answer
STATIONARY PHASE: solid in which sample components are adsorbed MOBILE PHASE: liquid (liquid – solid chrom) or gas (gas – solid chrom)	ADSORPTION CHROMATOGRAPHY:
ADSORPTION CHROMATOGRAPHY: types	1 Column chrom (CC) - SP: solid packed in tubular column - MP: allow to flow thru the solid 2 Thin Layer Chrom (TLC) - SP: plane
STATIONARY PHASE: liquid supported on an inert solid MOBILE PHASE: liquid (liq-liq chrom) gas (gas- liq chrom)	PARTITION CHROMATOGRAPHY
PARTITION CHROMATOGRAPHY: types	Paper chromatography SP: layer of water adsorbed on a sheet of paper
STATIONARY PHASE: polar solvents (water or methanol) MOBILE PHASE: non polar (hexane) This favors retention of polar compounds and elution of non polar compounds	NORMAL PHASE CHROMATOGRAPHY
STATIONARY PHASE: non polar (octadylsilane -ODS) MOBILE PHASE: polar (mixture of water, methanol or acetonitrile) In this phase, non polar solutes are retained favoring elution of polar solute	REVERSE PHASE CHROMATOGRAPHY
a process that allows the separation of ions and polar molecules based on their charge. Use an ion exchange resin as stationary phase - a polymeric matrix w/ the surface of w/c ionic functional groups such as carboxylic acids or quarternary amines have been chemically bonded For analysis of inorganic ions; ionized in solution	ION EXCHANGE CHROMATOGRAPHY Also known as ion chromatography
STATIONARY PHASE: polymeric subst. containing numerous pores of molecular dimensions MOBILE PHASE: contains analytes that are separated according to their size by their ability to penetrate a sieve like structure (stationary phase) Large molecules remain MP and are not retained Use for separation of macromolecules of biological origin (proteins, triglycerides)	SIZE EXCLUSION CHROMATOGRAPHY:
__ separate on the basis of molecular size. Individual gel beads have pores of set size, that restrict entry to molecules of a minimum size.	Size exclusion gels
a method of separating biochemical mixtures and based on a highly specific interaction such as that between antigen and antibody When crude mixture containing protein passed thru column, it reacts w/ antibody and is bound Protein is dislodge from antibody by changing pH Use for antibiotic and enzyme	AFFINITY CHROMATOGRAPHY:
MOBILE PHASE: electro-osmotically driven thru chromatographic bed For chiral analytes in biological fluids	CAPILLARY ELECTRO-CHROMATOGRAPHY (CEC)