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Metabolomic Profiling of <em>Neurospora crassa</em> Fungus Using HILIC and Reversed-Phase LC-MS

14 Jul 2014

Much is known regarding the genome of Neurospora crassa, specifically in the determination of circadian rhythms. However, little is known regarding how the metabolome changes over the course of the day under control of the circadian clock. Profiling metabolic changes in biological samples can be a complex and tedious task. The use of the high-resolution Ascentis® Express HPLC columns greatly aids in the resolution of components in even the most complex sample matrix.

Ascentis® Express F5 HPLC Columns

Sigma-Aldrich Supelco

The pentafluorophenylpropyl (PFP) stationary phase of Ascentis® Express F5 provides a stable, reversed-phase packing with electron-deficient phenyl rings due to the presence of electronegative fluorines. In addition to forming p-p and mildly steric interactions, F5 phases also retain compounds by polar interactions. As a result of having both polar and non-polar character, F5 phases can show dual-mode retention behavior, sometimes producing a “U-shaped” retention as a function of acetonitrile content of the mobile phase, with retention increasing at both low and high concentrations of ACN (reversed-phase and HILIC retention modes). Ascentis Express F5 can be used for basic, acidic, or neutral compounds with alternate selectivity from C18. Column Details: Chemistry: Reversed Phase, Hydrophilic Interaction Particle Size: 2.7, 5.0 μm Pore Size: 90 Å Length: 20, 30, 50, 75, 100, 150, 250 mm ID: 2.1, 3.0, 4.6 mm

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Ascentis® Express RP-Amide HPLC Columns

Sigma-Aldrich Supelco

Ascentis Express RP-Amide HPLC columns are the most recent product additions to the Supelco HPLC product line. Combining an embedded polar group (EPG) stationary phase with the Fused-Core particles, Ascentis Express RP-Amide provides a host of useful benefits to the HPLC chromatographer.   Ascentis Express RP-Amide is a blend of modern phase technology and innovative particle technology. The Fused-Core particle provides benefits in terms of speed, resolution, sensitivity, and ruggedness. The one-step RP-Amide bonding chemistry provides benefits in terms of selectivity, aqueous stability, and improved peak shape for bases. Read about how this product was used to analyze Riboflavin and impurities here.     Benefits: Alternative reversed-phase selectivity to C18 Improved peak shape for bases 100% aqueous compatible reversed-phase column Column Details: Chemistry: RP-Amide Particle Size: 2.7, 5.0 μm Pore Size: 90 Å Length: 20, 30, 50, 75, 100, 150, 250 mm ID: 2.1, 3.0, 4.6 mm

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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.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.MetabolitesMetabolites are the intermediate and end products of metabolism. Analyzing metabolites offers insight into cellular processes and disease mechanisms and is critical in fields like pharmacology, toxicology, and diagnostics. Explore our peer-reviewed product directory to find the best tools for metabolite analysis, compare products, check reviews, and get pricing directly from manufacturers.MetabolomicsMetabolomics is the study of small metabolites (the intermediates and products of metabolism). It involves the identification and quantification of cellular metabolites using analytical technologies such as GC, HPLC, NMR, and LC/MS.Reversed-Phase LC