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Rapid Separation and Mass Spectrometry Detection of Intact Protein Samples Using Monolithic Capillary Columns
Shane Bechler and Kelly Flook, Thermo Fisher Scientific
This application note demonstrates the rapid separation of a wide mass range of peptides and proteins using a reversed-phase monolithic column with both UV and MS detection. The separation is conducted on Thermo Scientific™ ProSwift™ RP-4H columns using conventional water/acetonitrile-based eluents. These simple, straightforward methods can be used to optimize chromatographic separations for analysis time, resolution, and MS detection especially for top-down proteomics. Read More |
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Separation of Biochemical Buffering Agents Using Multi-Mode Liquid Chromatography with Charged Aerosol Detection
Mark Tracy and Xiaodong Liu, Thermo Fisher Scientific
Good’s buffers are a series of zwitterionic buffering agents popularly used in biochemical experiments and HPLC mobile phases. We present analytical conditions using the Thermo Scientific™ Acclaim™ Trinity™ P2 column to separate 22 of these and other common buffer ingredients by simultaneous anion exchange (AEX), cation exchange (CEX), and hydrophilic interaction chromatography (HILIC) with simple isocratic conditions. Since by design these buffer ingredients have low UV absorption, we use the Thermo Scientific™ Dionex™ Corona™ Veo™ charged aerosol detector. Read More |
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Improved Determination of Antioxidants in Edible Oils Using Solid Core LC Columns
Mark Tracy, Thermo Fisher Scientific
Phenolic antioxidants are commonly used as preservatives in foods containing edible oils and fats to prevent development of rancidity. AOAC Method 983.15 is used to assay levels of these compounds in finished food products. The method uses a C18 column but can suffer from matrix interference effects. In this work, we demonstrate the suitability of both Thermo Scientific™ Accucore™ C18 and Thermo Scientific™ Accucore™ Polar Premium columns for this application. Read More |
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Rapid Screening Method for Veterinary Antibiotics Using an Advanced Solid Core UHPLC Column and UHPLC System with MS-MS Detection
Gordon Wilson and Derek Hillbeck, Thermo Fisher Scientific
This application shows the advantages of using the Thermo Scientific™ Accucore™ Vanquish™ C18 UHPLC 1.5 µm column and Vanquish UHPLC system for the analysis of 36 antibiotics. Advanced capabilities of the Vanquish UHPLC system allow the Accucore Vanquish UHPLC columns to be operated at high flow rates that enable development of rapid analytical methods while maintaining performance. The need for chromatographic separation of isobaric compounds prior to MS detection is also highlighted. Read More |
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Rapid Separation of 18 Herbicides Using Advanced UHPLC and Sub–2 µm Solid Core Column Technologies
Kristina Kirkham, Luisa Pereira, and Derek Hillbeck, Thermo Fisher Scientific
This application shows the advantages of using the Thermo Scientific™ Accucore™ Vanquish™ C18 1.5 µm UHPLC column and the Vanquish UHPLC system for the separation of 18 herbicides. The advanced capabilities of the Vanquish UHPLC system allow the Accucore Vanquish columns to be operated at higher flow rates that enable development of rapid, high-performance analytical methods. Comparisons are made with other commercially available sub-2-µm solid core columns, demonstrating improved peak capacity and resolution. Read More |
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Determination of Polyacrylic Acid in Boiler Water Using Size-Exclusion Chromatography with Charged Aerosol Detection
Mark Tracy, Xiaodong Liu, and Ian Acworth, Thermo Fisher Scientific
Polyacrylic acid (PAA) is widely used as an antiscaling additive for boiler water in conventional steam generators and, more recently, in nuclear steam generators. This work demonstrates the application of Thermo Scientific™ Acclaim™ SEC columns and the Thermo Scientific™ Dionex™ Corona Veo™ charged aerosol detector to the measurement of PAA in boiler water. No sample extraction or pre-concentration is needed to attain detection limits below 10 µg/L. The linear range exceeds 200 µg/L, with a useful dynamic range exceeding 400 µg/L, and recoveries are 60-120% over the range of 3-400 µg/L in a boiler water matrix. Read More |
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High-Resolution Separation of Oligonucleotides on a Pellicular Anion-Exchange Column
J.R. Thayer, Thermo Fisher Scientific
Multiple oligonucleotide (ON)-based therapeutic models have been developed to date including antisense oligonucleotides (ASOs), aptamer oligonucleotides (AOs), and short interfering RNA oligonucleotides (siRNA). In addition, modifications to oligonucleotide base-, backbone-, and ribosemoieties have all been introduced to limit nuclease-mediated degradation in therapeutic ONs. Mechanisms to resolve oligonucleotides from their modified forms, synthetic failure fragments, and their metabolic products are therefore necessary, although these can be technically challenging to develop. The Thermo Scientific™ DNAPac™ PA200 column offers industry leading selectivity and resolution for the separation of therapeutic and diagnostic oligonucleotides. Read More |
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High-Resolution Separation of Intact Monoclonal Antibody Isoforms
Dai Zhenyu, Xu Qun, and Jeffrey Rohrer, Thermo Fisher Scientific
Learn to develop a high-resolution, fast separation of intact protein sialylation isoforms using the Thermo Scientific™ MAbPac™ SCX-10 column and CX-1 pH gradient buffer on a Thermo Scientific™ Dionex™ UltiMate™ 3000 Dual Biocompatible Analytical LC system and an UltiMate 3000 BioRS LC system. Read More |
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Rapid Screening Method for Illicit Drugs, Using an Advanced Solid Core UHPLC Column and UHPLC System with MS-MS Detection
Derek Hillbeck, Thermo Fisher Scientific
This application shows the advantages of using the Thermo Scientific™ Accucore™ Vanquish™ C18 UHPLC, 1.5 µm column and Vanquish UHPLC system for the analysis of 47 illicit drugs. Advanced capabilities of the Vanquish UHPLC system allow the Accucore Vanquish UHPLC columns to be operated at high flow rates that enable development of rapid analytical methods while maintaining performance. The need for chromatographic separation of isobaric compounds prior to MS detection is also highlighted. Read More |
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