High-performance liquid chromatography (HPLC) is an analytical chemistry technique used to separate compounds in a chemical mixture. These separations utilize the pressure-driven flow of a liquid mobile phase through a column packed with a stationary phase.
An HPLC instrument generally has four major hardware components: a pump, an autosampler, a column compartment, and a detector. Many factors, including mobile phase composition, stationary phase chemistry, and temperature, influence HPLC separations. Successful separation only occurs if the analytes have differing affinities for the stationary phase, so selecting the appropriate stationary phase for your compounds is crucial.
The applications of HPLC encompass a broad range of fields. They include testing the quality of products you use every day. From impurity analysis in medicine, characterizing the quality of ingredients in foods and drinks, or monitoring the levels of pollutants in water and soil, HPLC helps keep us as consumers safe.
This whiteboard video provides a brief history of chromatography, and explains the fundamental principles behind HPLC and UHPLC. It also describes the essential components, or modules, of a chromatography system.
Do you have questions about how HPLC instruments work? How to develop or transfer an HPLC method? Or the typical applications of HPLC? Take a closer, in-depth look into each of these topics.
Explore the basic principles of how HPLC works along with the most common types of techniques and when to apply each.
Learn how each component of an HPLC system works, the various column chemistries, and detectors.
Read expert tips for developing an HPLC method plus best practices for sample preparation.
Discover best practices for transferring and optimizing HPLC methods from one system to another with the same or different brand of instruments.
Learn about the applications of HPLC — from the analysis of small molecules to large complex compounds like proteins and antibodies.
Find important technical information, tips for working with an HPLC system, and answers to your frequently asked questions.
High-performance liquid chromatography (HPLC) is an analytical chemistry technique used to separate, identify, and quantify compounds in a liquid mixture.
UHPLC columns contain sub-2 µm particles or less while in HPLC the columns are 3-5 µm particles. UHPLC separations enable higher peak resolution, improved analyte sensitivity, shorter analysis times and less solvent consumption. UHPLC instruments also require much higher operating pressure and performance compared to HPLC systems.
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