Kaumil Hplc

Basics of - HPLC ,[object Object],[object Object]

Index ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

What is HPLC ? ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Applications of HPLC ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Types of HPLC ,[object Object],[object Object],[object Object],[object Object],[object Object]

Separation based on selection of column material

HPLC instrumentation ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Detector Mobile phase Pump Injection Valve Separation column => => => =>

Solvent system ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Pumping system ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Reciprocating pump : Single piston pump

Reciprocating pump : Double piston pump

Sample injection system ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Column ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Detectors ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Types of HPLC detectors Ability to ionize analyte Low LOD; analyte identification Mass Spec Non-specific; high LOD Commercially available Electrochemical Non-specific; interference from solvent Uniform response; 5ng/25  L LOD Scattering Temperature sensitive; high LOD Works w/ nearly all molecules Refractive Index Many solvents IR active Works w/all molecules IR Not everything fluoresces Very specific; low LOD Fluorescence High LOD Works for all wavelengths DAD Non-specific; complex samples; absorption wavelength Works w/all molecules UV-Vis Name Advantage Disadvantage

UV-VIS detector. ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

ELSD (Evaporative Light Scattering Detector)

P9 RP- HPLC Salient features. ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Typical P9 RP-HPLC peak (Agilent-1200).

Kaumil Hplc

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