High-performance liquid chromatography, an invaluable component in the analytical chemistry toolbox, is used to separate, identify, and quantitate compounds in liquid samples. HPLC systems are made up of a mobile phase, pump, injector, column, and detector.
HPLC was originally termed high-pressure chromatography, due to the pressure required to force the mobile phase and sample through the packed columns. With HPLC, a pump is used to move the mobile phase and sample through the column, as opposed to classical liquid chromatography, which relies on gravity. Concentrations down to ppt level can easily be identified.
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How do I choose an HPLC system?
Both integrated, all-in-one systems and the modular components approach have their benefits: With an integrated system, there is no need to choose components (which may not work well together) and these systems are often less expensive, but mixing separate components (sometimes from different vendors) ups your flexibility and upgradeability—it’s your call. Since HPLCs tend to be long-lasting instruments, system upgradeability is often a primary concern.
Identifying your compound of interest will lead to the correct column chemistry, including reversed phase, the most popular type, ion exchange, affinity, or gel filtration. The most widely used detector is UV-VIS, but fluorescence, electrochemical, conductivity, and refractive index are additional options. Are you performing analytical or preparative chromatography? The flow rate of your pump (from 0.4 to 150 mL/min) should match those needs.
New developments in HPLC
Ultra-high-performance LC (UHPLC) continues to provide advances in resolution, sensitivity, and speed, as well as reduced eluent consumption. Improvements have often focused on the columns themselves, with a reduction in particle size below 2 µm, upgraded packing material design and column construction, and extended pH range.
Vendors have focused on upgrades for older instruments, greater diagnostic sensitivity and specificity, more precise solvent delivery, upgrades in intelligent software, and always welcome system price reduction.
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