HPLC - CHROMATOGRAPHY CHEM3001(C30J Chemical Analysis II Donna A Minott-Kates DMK CHEM3001 Chromatography(2009-10 CHROMATOGRAPHY LIQUID GAS MOBILE

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Unformatted text preview: CHROMATOGRAPHY CHEM3001- (C30J) Chemical Analysis II Donna A Minott-Kates DMK: CHEM3001 - Chromatography (2009-10) CHROMATOGRAPHY LIQUID GAS MOBILE PHASE PLANAR COLUMN STATIONARY PHASE PAPER, SILICA, ALUMINA PACKED CAPILLARY SFC DMK: CHEM3001 - Chromatography (2009-10) High Performance Liquid Chromatography Useful HPLC links http://www.forumsci.co.il/HPLC/program.html (2009-03-17) http://www.studyhplc.com/hplclinks.php (2009-02-04) Royal Society of Chemistry educational video http://www.youtube.com/watch?v=kz_egMtdnL4 (2009-03-26) Source : http://www.pharmainfo.net/files/images/stories/article_images/ INTRODUCTION_ANALYTICALMETHODfig4.JPG (2009-03-17) HPLC Schematic Diagram 1) Solvent reservoirs, (2) Solvent degasser, (3) Gradient valve, (4) Mixing vessel for delivery of the mobile phase, (5) High-pressure pump, (6) Switching valve in "inject position", (6') Switching valve in "load position", (7) Sample injection loop, (8) Pre-column, (9) Analytical column, (10) Detector (i.e. IR, UV), (11) Data acquisition, (12) Waste or fraction collector. DMK: CHEM3001 - Chromatography (2009-10) Source : http://commons.wikimedia.org/wiki/File:HPLC_apparatus.svg (2009-03-17) HPLC Mobile Phase Pre-filtered Degassed Removal of dissolved air & other gases Sparged with inert gas eg N 2 , Ar Under applied vacuum - “Evacuated” Pump Pulse-free (prevents detector interference) High pressure (up to 6000 psi), reproducible flow Gradient device Optional Allows combination of more than one solvent over the chromatographic run DMK: CHEM3001 - Chromatography (2009-10) LC - Gradient Profiles Initial and final mobile phase composition programmed eg Initial: 5% A, 95% B where A and B may be single solvent of solvent mixture Gradient profile selected Affords ability to manage rate of gradient change for more effective separations DMK: CHEM3001 - Chromatography (2009-10) Source : http://www.forumsci.co.il/HPLC/ gradient_curves.gif (2009-03-24) Gradient vs Isocratic Elution DMK: CHEM3001 - Chromatography (2009-10) Source : http://www.forumsci.co.il/HPLC (2009-03-24) DMK: CHEM3001 - Chromatography (2009-10) Injection System 6-Port injection valve consists of injection port, switching valve and sample loop Animation : http://www.restek.com/hplcsixport/6_Port_Injector_content.html (2010-01-20) Source : Harris, D.C. 2007. Quantitative Chemical Analysis , 7th Edition. W.H. Freeman and Company, New York. p571 Injection port Columns Guard Short (1 – 2 cm) pre-column Contains same stationary phase as separation column Provides buffer zone - prevents contamination Separation High performance analytical or preparative Stationary phase typically packed in stainless steel column (10 - 30 cm) May be jacketed for temperature control DMK: CHEM3001 - Chromatography (2009-10) DMK: CHEM3001 - Chromatography (2009-10) CHROMATOGRAPHY LIQUID GAS MOBILE PHASE PLANAR COLUMN STATIONARY PHASE PAPER, SILICA, ALUMINA PACKED CAPILLARY SFC DMK: CHEM3001 - Chromatography (2009-10)...
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This note was uploaded on 02/23/2011 for the course CHEMISTRY C30J taught by Professor Vaugnrattray during the Spring '11 term at University of the West Indies at Mona.

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HPLC - CHROMATOGRAPHY CHEM3001(C30J Chemical Analysis II Donna A Minott-Kates DMK CHEM3001 Chromatography(2009-10 CHROMATOGRAPHY LIQUID GAS MOBILE

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