HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)

1 high PERFORMANCELIQUID CHROMATOGRAPHY (HPLC)Chapter 28 NOTE: HPLC came about because not allcompounds can be vaporizedand analyzed on a GCSource: Harris, Exploring Chemical Analysis, 2nded. (2001) 2 Summary of MethodMobile phase: LiquidHigh PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) An analytical separation technique that involves the high -pressure flow of a LIQUID through a column that contains the stationary phase: Can be a solid (LSC) or a LIQUID (LLC) A mixture of compounds injected at one end of the column separates as the compounds pass through. Separated compounds are detected electronicallyas they elute at the other end of the with GCGCHPLCB asis of separationInteractionof solutes with the ; solute vapor pressureInteractionof solutes with boththe and timeFast(a few minutes for simple mixtures)Slower than GC (several minutes for a simple mixtureTemperature for separationUsually requires a high temperature(>40 0C)Usually a room temperature techniqueApplicationsSeparation of volatileand thermally stablecompounds - cannot be used for high MW and highly polar compoundsSeparation of a wider range of compounds -- high MW, polar, and ioniccompoundsParameter3 InstrumentationInstrumentation(Cont.))

High Performance Liquid Chromatography (HPLC) An analytical separation technique that involves the high-pressure flow of a liquid through a column that contains the stationary phase. Stationary phase: Can be a solid (LSC) or a liquid (LLC) A mixture of compounds injected at one end of the column separates as the compounds pass through.

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