Featured Application Note:
Relative Quantitation of TMT-Labeled Proteomes – Focus on Sensitivity and Precision
R. Viner, M. Scigelova, M. Zeller, M. Oppermann, T. Moehring, V. Zabrouskov, Thermo Fisher Scientific
This note we evaluated the impact of the technological advances of hybrid instruments on overall experimental outcome in terms of protein/peptide identification and quantitation. The focus was on assessing the percentage of quantifiable peptides, the quantitative precision, and the quantitative accuracy. |
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Fast Screening of Mobile and Stationary Phases with the Agilent 1290 Infinity LC and Seamless Method Transfer to an Agilent 1200 Series LC Using ISET
Angelika Gratzfeld-Huesgen, Agilent Technologies
This application note describes how the Agilent 1290 Infinity LC System Method Development Solution was used to find optimum separation conditions for degradation products of a pharmaceutical preparation. Having evaluated the optimum combination, a transfer to conventional conditions was performed. Finally, the Agilent Intelligent System Emulation Technology (ISET) of the 1290 Infinity LC was used to simulate the target Agilent 1200 Infinity Series LC. The method was then transferred to an Agilent 1200 Series LC to prove the functionality of the ISET tool. |
Label-free Phosphopeptide Quantification and Occupancy Determination
Oliver Drews, Stephanie Kaspar, Carsten Baessmann, Detlev Suckau, Bruker Daltonik GmbH, Bremen, Germany
The identification of post-translational modifications, such as phosphorylation, is a key task in proteomics research. Most approaches focus on the pure detection of phosphorylation sites in peptides in complex mixtures, but miss the quantitative determination of the occupancy particularly in case of multiple phosphorylated proteins. In this study, a straight-forward label-free strategy for quantitative assessment of phosphorylation status at various target sites was established for a highly phosphorylated protein using nanoflow LC-separation on Bruker’s nano-Advance UHPLC and ultrafleXtreme MALDI-TOF/TOF system and amazon speed ETD ion trap respectively. |
Analysis of Isoflavones with UHPLC
Njies Pedjie, PerkinElmer, Inc.
Clinical studies suggest consumption of isoflavones can exert positive physiological effects. Yet a study done in by the University of Missouri showed that injections of genistein in mice caused a decrease of thymocyte by 86% and a decrease of humoral immunity, thus raising the possibility that a high concentration of genistein in food could cause thymic and immune abnormalities in infants. Due to potential health benefit of isoflavones, many soy products and isoflavone supplements are available to consumers. The health benefits and concerns including labeling requirements have created the need for analytical
techniques to determine the type and amount of isoflavone in nutraceutical products. |
Characterization of Monoclonal Antibodies
Wyatt Technology
Multiangle light scattering (MALS) represents a uniquely powerful technique to determine the absolute molar masses and sizes of proteins in solution. MALS detectors cover a wide detection range of sizes and masses and enable the characterization of protein molecules as monomers, dimers, or higher order aggregates. |
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Automated Solid Phase Extraction and Concentration of Semi-Volatile Organic Compounds in Water
Phil Bassignani, FMS, Inc.
The following note was developed to demonstrate the benefits of automating EPA Method 525.2 using SPE cartridges, the FMS PowerPrep™ SPE, and the FMS PowerVap™ Direct-to-Vial Concentration system. This one step method delivers rapid, consistent, and reproducible extraction and concentration of semi volatile organic compounds in water samples for analysis by GC–MS. |
Semivolatile Organic Compounds In Drinking Water By Solid-Phase Extraction And Capillary Column (GC/MS) EPA Method 525.3
Xiaoyan Wang and Don Shelly, UCT
A1-liter water sample is fortified with surrogates and then extracted using a solid phase extraction (SPE) cartridge (ECUNI525). Analytes are eluted from the solid phase with a small amount of organic solvents. The extract is dried using sodium sulfate and concentrated using N2. IS are added and the volume adjusted to 1 mL with ethyl acetate. A splitless injection is made into a GC/MS with either scan, SIM or SIS detection. The GC/MS may be calibrated using standards prepared in solvent or using matrix-matched standards. |
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Extraction of Amphetamines from Urine with GC-MS Analysis
Rhys Jones, Lee Williams, Adam Senior, Gavin Jones, Biotage
This application note describes the supported liquid extraction clean-up of a range of amphetamines and metabolites from urine prior to quantitative GC-MS analysis. Analyte recoveries achieved using this method ranged from 99-104% with RSDsbelow 10% for all analytes. |
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