Liquid Chromatography HPLC HPLC: High Performance Liquid Chromatography

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Liquid Chromatography HPLC HPLC: High Performance Liquid Chromatography ( liquid-chromatography-hplc-hplc-high-performance-liquid-chro )

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However these systems require a high pressure pump to push the mobile phase through the column. P=f uηL πr2d2 p f depends on particle shape and packing Stationary Phase Column support: small, pure, spherical, micro-porous particles of silica or polymer particles. Often these particles are coated with s.p. or the s.p is chemically bonded to the surface. Stationary phase coatings on silica particles are not very durable. Sometimes silica particles are used (as is) for LC (adsorption mechanism, and normal mode). Typical molecular size ~ 0.1*pore size Stationary phase coated or bonded onto the m size silica/polymer particles (base ‘inert’ particles). Partition chromatography on bonded phases: Chemically bonding the hydroxy groups of silica particles would generate a rugged s.p. and most importantly the thinnest possible (monolayer) coating, tiny df. Bonded carbon chains forms the primary s.p. material C4  C18 (RP). Derivatization of the terminal carbons allows us to ‘tailor’ s.p.’s with different polarities. Such derivatized s.p. are used in the reverse phase mode, generally. Highly polar Adsorption chromatography, NP Si OH pH 2-3, Si-OH pH>3 de-protonation may occur Si-O-, resulting in tailing of positively charged analyte species pH > 8, silica dissolves use polymeric supports pH 2-8 optimum range (silica)

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