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Successful Transfer of Size-Exclusion Separations between HPLC and UPLC

Waters
24 Nov 2014

Waters’ recent development of HPLC compatible 200Å and 450Å, 3.5 µm particles based on the same diol-coated BEH-particle chemistry as that used in ACQUITY UPLC Protein BEH SEC Columns provides the capability to readily transfer between SE-UPLC and SE-HPLC for the first time. This application note details and provides examples of the methodology that can successfully employed for this method transfer.

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ACQUITY UPLC H-Class PLUS Bio System

Waters

The ACQUITY UPLC H-Class PLUS Bio System delivers the benefits of UPLC's resolution, sensitivity, and throughput in a system purpose-built for the analysis of proteins, peptides, nucleic acids, and glycans.

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ACQUITY UPLC BEH Columns

Waters

In 2004, Waters revolutionized liquid chromatography with the introduction of the ACQUITY UPLC System. At the heart of the system was Waters second generation hybrid, the Ethylene Bridged Hybrid (BEH) particle. In 2005, BEH Technology preparative particle sizes (2.5, 3.5, 5 and 10 μm) with the introduction of XBridge between HPLC and UPLC Technology platforms. BEH Technology particles offer: Excellent peak shape and efficiency for basic analytes A rational array of chromatographic selectivity Improvements in chemical stability at mobile phase extremes, particularly at elevated pH

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Size Exclusion ChromatographySize exclusion chromatography (SEC) is a liquid chromatographic technique used to separate and identify molecules based on their size, including polymers, proteins, nanoparticles & nucleic acid. SEC is further subdivided into gel filtration chromatography (GFC) and gel permeation chromatography (GPC) when an aqueous or organic mobile phase is used respectively. HPLC-based SEC systems and columns are available for analytical separation alongside SEC spin columns or filters to prep samples for many protocols. Find the best size exclusion chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein BiologyThe analysis of protein expression, identity and function is vital for many areas of life science research and drug discovery. Some of the most commonly used techniques in protein analysis include Western blotting, electrophoresis and mass spectrometry.

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