Selected article for: "chromatography ultra performance and liquid chromatography"
Author: Patton, John B.; Bennuru, Sasisekhar; Eberhard, Mark L.; Hess, Jessica A.; Torigian, April; Lustigman, Sara; Nutman, Thomas B.; Abraham, David
Title: Development of Onchocerca volvulus in humanized NSG mice and detection of parasite biomarkers in urine and serum
  • Document date: 2018_12_12
    • ID: 0yh5k6jk_31
    Snippet: Each digestion mixture was analyzed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). LC was performed using an Easy-nLC 1000 UHPLC system (Thermo Fisher Scientific, Waltham, MA). Mobile phase A was 97.5% MilliQ water, 2% acetonitrile, 0.5% formic acid. Mobile phase B was 99.5% acetonitrile, 0.5% formic acid. The 240 min LC gradient ran from 0% B to 35% B over 210 min, then to 80% B for the remaining 30 min. .....
    Document: Each digestion mixture was analyzed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). LC was performed using an Easy-nLC 1000 UHPLC system (Thermo Fisher Scientific, Waltham, MA). Mobile phase A was 97.5% MilliQ water, 2% acetonitrile, 0.5% formic acid. Mobile phase B was 99.5% acetonitrile, 0.5% formic acid. The 240 min LC gradient ran from 0% B to 35% B over 210 min, then to 80% B for the remaining 30 min. Samples were loaded directly to the column. The column was 50 cm x 75 um I.D. and packed with 2 μm C18 media (Thermo Easy Spray PepMap). The LC was interfaced to a quadrupole-Orbitrap mass spectrometer (Q-Exactive, Thermo Fisher Scientific, Waltham, MA) via nano-electrospray ionization using a source with an integrated column heater (Thermo Easy Spray, Thermo Fisher Scientific, Waltham, MA). The column was heated to 50˚C. An electrospray voltage of 2.2 kV was applied. The mass spectrometer was programmed to acquire, by data-dependent acquisition, tandem mass spectra from the top 10 ions in the full scan from 400-1200 m/z. Dynamic exclusion was set to 15 s, singly-charged ions were excluded, isolation width was set to 1.6 Da, full MS resolution to 70,000 and MS/MS resolution to 17,500. Normalized collision energy was set to 25, automatic gain control to 2e5, max fill MS to 20 ms, max fill MS/MS to 60 ms and the underfill ratio to 0.1%.

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