A total of 50 μL of protein solution was thoroughly mixed with 500 μL of acetone following centrifugation at 13,000 rpm for 10 min. The protein residues were evaporated to dryness after discarding the supernatant. Protein residues were redissolved with 25 mM of ammonium bicarbonate aqueous solution and digested with sequence-grade trypsin (Promega) at 37 °C for 16 h. Subsequently, 2 μL of tryptic peptide solution was injected into the nanoACQUITY UPLC system (Waters, Milford, MA, USA) containing a desalting column (Symmetry C18, 5 μm, 180 μm × 20 mm) and an analytical column (BEH C18, 1.7 μm, 75 μm × 100 mm) and was detected by an LTQ Orbitrap Discovery Hybrid fourier transform mass spectrometer (Thermo Fisher Scientific Inc., Bremen, Germany) at a resolution of 30000 coupled with a nanospray source in a positive ion mode. Individual raw data were processed using Mascot Distiller software (Version 2.2, Matrix Science Inc., Boston, MA) and uploaded to the in-house Mascot server for protein identification.
Ltq orbitrap discovery hybrid fourier transform mass spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The LTQ Orbitrap Discovery Hybrid Fourier Transform Mass Spectrometer is an analytical instrument designed to perform high-resolution, accurate-mass measurements of molecular ions. It combines the features of a linear ion trap (LTQ) and an Orbitrap mass analyzer, providing enhanced sensitivity and mass accuracy for a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Nanoacquity uplc system, by Waters Corporation (2 mentions)
Symmetry c18, by Thermo Fisher Scientific (1 mentions)
Sequence grade trypsin, by Promega (1 mentions)
Mascot distiller software, by Matrix Science (1 mentions)
Nanoacquity ultra, by Waters Corporation (1 mentions)
Symmetry c18, by Waters Corporation (1 mentions)
Beh c18, by Waters Corporation (1 mentions)
3 protocols using ltq orbitrap discovery hybrid fourier transform mass spectrometer
1
Mass Spectrometry Proteomics Workflow
2
Proteomic Analysis by Nanoflow LC-MS/MS
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MS/MS analysis was essentially performed according to the procedure described in [21 (link)]. In brief, peptide solution was injected into the nanoACQUITY ultra performance liquid chromatography (UPLC) system (Waters, Milford, MA, USA) and detected by linear trap quadropole (LTQ) Orbitrap Discovery hybrid Fourier Transform Mass Spectrometer (Thermo Fisher Scientific Inc., Bremen, Germany) in the positive ion mode at a resolution of 30,000. Mobile phase A (0.1% formic acid) and mobile phase B (100% ACN with 0.1% formic acid) were used to elute the desalting column (Waters Symmetry C18, 5 μm, 180 μm × 20 mm) and analytical column (Waters BEH C18, 1.7 μm, 75 μm × 150 mm).
3
Nano-UPLC-MS Analysis of Quetiapine Metabolites
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A nanoACQUITY UPLC system (Waters, MA, USA) was coupled with an LTQ orbitrap discovery hybrid Fourier transform mass spectrometer (Thermo Fisher Scientific, Inc. Bremen, Germany) equipped with a nanoelectrospray positive ionization (nano ESI) interface. The desalting column was equipped with a symmetric μm, 180 μm × 20 mm), and the analytical column was equipped with a BEH C18 column (1.7 μm, 75 μm × 150 mm). The mobile phase included 0.1% formic acid as solvent A and acetonitrile containing 0.1% formic acid as solvent B. The sample loading volume was 2 μL. The initial gradient elution conditions were solvent B at 1%, then conditions were adjusted to 100% after 5 min and maintained in isocratic mode for 45 min. Finally, solvent B was adjusted to 1% after 60 min. The flow rate was set at 300 nL min−1. Each plasma sample was pretreated and then analyzed with nano-UPLC-MS in positive ionization mode with MRM mode to identify the QTP at 384.17 (m/z), NorQTP at 296.12 (m/z) and 7-QTP at 400.16 (m/z).
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