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Size-Exclusion Ultra Performance Liquid Chromatography for the Analysis of Covalent High Molecular Weight Insulin

Waters
8 Apr 2013

Presented in this application note are the advantages that may be realized using a 125Å pore size, sub-2-µm ethylene-bridged hybrid (BEH) silica packing material and Waters UltraPerformance Liquid Chromatography (UPLC) instrumentation for this traditional analysis. Among these advantages are faster run times, higher sensitivity, and higher resolving separations of insulin and covalent insulin HMW, while at the same time greatly reducing acetonitrile containing waste-stream volumes.

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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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BEH (Ethylene Bridged Hybrid) Particles for ACQUITY UPLC

Waters

The 1.7 µm Ethylene Bridged Hybrid [BEH] particle is one of the key enablers behind UPLC® technology.It is available in three different pore sizes [130Å, 200Å and 300Å] and several bonded phases for reversed-phase and hydrophilic interaction chromatography and is applicable from small molecule to large biopharmaceutical analysis. Due to the intrinsic chemical stability of hybrid particle technology, a wider usable pH range [pH 1-12] can be employed, enabling a versatile and robust separation technology for method development. BEH particle technology is available in HPLC particle sizes [2.5, 3.5, 5 and 10 µm] in the XBridge® family of HPLC columns, enabling seamless transfer between HPLC and UPLC technology platforms.

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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.DiabetesDiabetes is a metabolic disorder characterized by high blood sugar levels, either due to insufficient insulin production or resistance to its effects. Ongoing diabetes innovation focuses on developing better diagnostic tools, treatments, and preventive measures. Browse our peer-reviewed product directory to find the best diabetes diagnostic tools, compare products, check reviews, and get pricing directly from manufacturers.AggregationProtein 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.Chromatography Method DevelopmentChromatography method development is the process of designing and optimizing chromatographic techniques to separate and analyze complex mixtures. This includes selecting appropriate stationary and mobile phases, determining flow rates, and optimizing detection methods. Successful method development is essential for high-quality results in fields like pharmaceuticals, food safety, and environmental analysis. Explore chromatography method development tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Molecular Weight AnalysisMolecular weight analysis determines the size and structure of biomolecules, essential in protein characterization, polymer studies, and drug development. Techniques like mass spectrometry ensure precise results. Compare solutions for molecular weight analysis in our peer-reviewed product directory.

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