{[ promptMessage ]}

Bookmark it

{[ promptMessage ]}

HPLC - CHROMATOGRAPHY CHEM3001(C30J Chemical Analysis II...

Info iconThis preview shows pages 1–12. Sign up to view the full content.

View Full Document Right Arrow Icon
CHROMATOGRAPHY CHEM3001- (C30J) Chemical Analysis II Donna A Minott-Kates
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
DMK: CHEM3001 - Chromatography (2009-10) CHROMATOGRAPHY LIQUID GAS MOBILE PHASE PLANAR COLUMN STATIONARY PHASE PAPER, SILICA, ALUMINA PACKED CAPILLARY SFC
Background image of page 2
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)
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
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)
Background image of page 4
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)
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
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)
Background image of page 6
Gradient vs Isocratic Elution DMK: CHEM3001 - Chromatography (2009-10) Source : http://www.forumsci.co.il/HPLC (2009-03-24)
Background image of page 7

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
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
Background image of page 8
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)
Background image of page 9

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
DMK: CHEM3001 - Chromatography (2009-10) CHROMATOGRAPHY LIQUID GAS MOBILE PHASE PLANAR COLUMN STATIONARY PHASE PAPER, SILICA, ALUMINA PACKED CAPILLARY SFC
Background image of page 10
DMK: CHEM3001 - Chromatography (2009-10) PACKED COLUMN PRINCIPLE OF SEPARATION ADSORPTION NORMAL PHASE PARTITION REVERSED PHASE SOLUTES: LIPOPHILIC: Oils, lipids HYDROPHILIC: Biomedical CONDITIONS: ORGANIC SOLVENTS: n-hexane, chloroform, alcohols AQUEOUS MIXTURES: Buffers, ion pairs, MeOH, CH 3 CN, THF, with additives
Background image of page 11

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full Document Right Arrow Icon
Image of page 12
This is the end of the preview. Sign up to access the rest of the document.

{[ snackBarMessage ]}