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Simple and Fast Quantitation of Nicotinic Acid and Nicotinamide in Human Plasma by Applying Impact™ Protein Precipitation Plate Technology with Gemini® 3 μm C18 HPLC Columns

Phenomenex Inc
9 Dec 2014

In this application note, nicotinic acid and nicotinamide were extracted from human plasma by performing a rapid protein precipitation using Impact Protein Precipitation Plates followed by HPLC analysis using a Gemini 3 μm C18 100 x 4.6 mm HPLC column and positive polarity ESI LC/MS/MS system. Impact technology offers easy, fast protein removal while providing maximized recovery of the target analytes. The Gemini 3 μm C18 HPLC column produced excellent chromatographic resolution, sensitivity, and high peak capacities.

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Impact™ Protein Precipitation Plates

Phenomenex Inc

Impact™ protein precipitation plates with Solvent Shielding Technology™ offer a rapid and convenient way to remove proteins from plasma and tissue homogenate samples prior to analysis.The Solvent Shielding Technology design will withhold organic solvents above the filter membranes for up to 25 minutes, allowing for direct in-well precipitation upon sample addition. The precipitate is then filtered out via vacuum, centrifuge or positive pressure resulting in a clean, protein depleted extract.

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Gemini pH Flexible LC Columns

Phenomenex Inc

Setting the Standard for pH Method DevelopmentRugged reversed phase HPLC columns that offer extended lifetime under extreme pH conditions and excellent stability for reproducible, high efficiency separations. Take full advantage of high and low pH conditions (pH 1-12) to manipulate selectivity Expect longer column lifetime with patented TWIN-NX™ technology Ideal for analytical and preparative separations of basic and acidic compounds Who needs Phenomenex's Gemini HPLC columns? Method Developers: Extreme pH method flexibility Reproducible performance Purification Chemists: Purify bases at high pH Increased loading Increased retention Increased resolution Lab Managers: Extended column lifetime Consistent results.

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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.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Robotic InstrumentsRobotic instruments can be used for high-throughput automation of many lab processes. Such processes use instruments for assays like cell based assays and ELISA, for sample preparation like shakers, centrifuges and incubators, and for analysis such as sequencing and western blot analyzers. Useful features of robotic instruments include speed, reproducibility, barcode readers, software and automation.HPLC Method DevelopmentIt is important to develop and validate HPLC methods in order to identify, quantify or purify compounds of interest in the most efficient way. Effective method development ensures optimization, quality control and cost effectiveness.Human PlasmaHuman plasma is the liquid component of blood, which carries blood components - nutrients, hormones, proteins, and waste - throughout the body. Plasma-based studies provide insights into biomarkers, immune responses, and disease mechanisms. Compare plasma handling tools and systems in our peer-reviewed product directory.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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Simple and Fast Quantitation of Nicotinic Acid and Nicotinamide in Human Plasma by Applying Impact™ Protein Precipitation Plate Technology with Gemini® 3 μm C18 HPLC Columns