HPLC - February 25 - CHEM 4128 Lab #3 Separation by HPLC...

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CHEM 4128 Lab #3 Separation by HPLC and the Impact of Dead-Volume Objective In this experiment, HPLC will be used to perform a evaluation of the role of dead-volume in determining column efficiency. Chemical separation in HPLC is a process in which analyte species are transported through a column by a pumped solvent system, the mobile phase. The column also contains a solid material called the stationary phase. The degree of affinity of analyte molecules for the mobile and stationary phases dictates the transit time of molecules through the column. Desired separation is achieved by the selection of an optimal stationary/mobile phase through the column. Dead volume will then be added before and after the column and its affects will be evaluated. Dead volume is volume within the chromatographic system that does not contain stationary phase. Experiment Instrument: SpectraSystem P4000, SpectraSystem AS3000, UV1000 Flow Rate: 1 mL/min Column length: 15 cm Column: Silica gel, coated with octadectane Wavelength: 275 nm Trail Time: 6 min Mobile Phase: 5 M 80% KHPO 4 , 16% NaCn, 4% MeOH
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HPLC - February 25 - CHEM 4128 Lab #3 Separation by HPLC...

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