Schuyler Mincemoyer
High Performance Liquid Chromatography
To Waste
Block Diagram:
Pulse Damper
Regulated
Helium Source
Output
Check Valve
Pump
Drain
Valve
Inlet Check
Valve
Solvent
Reservoirs
Sparger
Priming
Syringe
Solvent
Proportioning
Valve
Column
Inlet
Filter
Pressure
Transducer
Injector Valve
Back-Pressure
Regulator
Filter
To Detector
Components
Solvent Reservoir: Holds 500mL or more of a solvent.
Sparger: dissolves gases that are swept out of solution by fine bubbles of an inert gas that is not soluble in the
mobile phase.
Pump: There are generally two types of pumps (reciprocating and displacement), but are used to pump the
liquids. Needs to generate pressure up to 6000 psi, have pulse free output, flow rates ranging from 0.110mL/min and resistance to corrosion by a variety of solvents.
Pulse Damper: Used to remove any pulses that may be in the solvent as a result of pumping.
Back-Pressure Regulator: Balances inlet and outlet pressures going to injector.
Injector valve: Inject sample into a loop and then move the handle and the entire loop of sample is inserted
between the pump and the column so the mobile phase sweeps the sample onto the column.
Column: Can be made from heavy-walled glass tubing, polymer tubing, or smooth-bore stainless steel
tubing. There are different kinds of columns including analytical columns and guard columns.
Detectors: The sample is then sent to some kind of detector such as a UV-Vis, Infrared Absorption,
Fluorescence, or Refractive-Index detectors.
Vendors:
Agilent Technologies, Beckman Coulter INC. Cecil Instruments, Dion Corp, Perkin Elmer, Shimadzu, Theromo
Fisher, and Varian
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The most expensive HPLC I could find was selling for $24,000. It was a USED Waters Acquity 2007
HPLC system on
http://www.dotmed.com/listing/740676?utm_source=base&utm_medium=search&utm_campaign=Base
The cheapest HPLC I could find was selling for $499.00 on Ebay. It was a Walter 717 Autosampler.
http://cgi.ebay.com/Waters-717-Plus-HPLCAutosampler_W0QQitemZ110490475944QQcmdZViewItemQQptZLH_DefaultDomain_0?hash=item1
9b9bedda8
Articles:
Analysis of Pharmaceuticals in Water by Isotope Dilution Liquid Chromatography/Tandem
Spectrometry
Brett J. Vanderford and, Shane A. Snyder
Environmental Science & Technology 2006 40 (23), 7312-7320
Mass
Selective Analysis of Secondary Amino Acids in Gelatin Using Pulsed Electrochemical Detection
Jason D. Russell, John M. Dolphin, and Miles D. Koppang
Analytical. Chemistry., 2007, 79 (17), pp 6615–6621
Facile Identification and Quantitation of Protein Phosphorylation via β-Elimination and Michael
Addition with Natural Abundance and Stable Isotope Labeled Thiocholine
Meng Chen, Xiong Su, Jingyue Yang, Christopher M. Jenkins, Ari M. Cedars and Richard W. Gross
Analytical Chemistry., 2010, 82 (1), pp 163–171
Everyday Uses For HPLC
1) The government uses HPLC to test confiscated narcotics
2) It can also be used to help test environmental samples such as water from a drain area or stream.