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Happy Healthy Period Course

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Mass spectrometry data was obtained in positive and negative mode on an Orbitrap Fusion Tribrid mass spectrometer (ThermoFisher Scientific, Bremen, Germany) operating in full scan mode (400-1,400 m/z). The Orbitrap Fusion was operated in FTMS, HCD, and IT MS/MS mode using a resolution of 120,000 with ion injection time set to 30 ms for the FTMS scan, and 15 for HCD. The target value was set to 1E6 for the intact precursor masses, 1E5 for the MS/MS spectra, and 1E4 for the MS/MS spectra (top 10). Peak lists were obtained from raw data using DataAnalysis version 5.0 (Bruker Daltonics). Lipid compounds were identified using the Lipid Search from SimLipid and Lipidomics Gateway databases. All detected peaks were manually inspected and peak picking was performed using LipidBlast version 3.0. The elemental composition and isotope pattern were characterized using the CAMERA algorithm for isotope patterns and the elemental composition search of apLCMS version 2.9.3. For lipid identification, required match factors were set to 5 or 10, respectively, for intact and MS/MS masses, and 10 for MS/MS spectra. LIPID MAPS classifications were given by manually checking the spectra against the LIPID MAPS database. Identification of oxidized lipids was performed by setting the following elemental composition search parameters: (M+NH4)+ ion and a match factor of 3.0. All mass spectra were searched against in silico reconstructed database using ProteoWizard from version and MPDL2 version 2.2.2 (Bruker Daltonics, Germany). Peptide identifications were done in MaxQuant version (Bruker Daltonics, Germany). An initial precursor ion mass tolerance of 6 ppm and a maximum of two missed cleavages for peptides were used. Carbamidomethylation was set as fixed modification. Oxidation of methionine and proline, acetylation of the N-terminus and the alkylation of cysteine were set as variable modifications. MS2 data were searched with a precursor ion tolerance of 6ppm and a fragment ion tolerance of 20ppm. The false discovery rate (FDR) for peptide and protein identification was set to 0.01. All other settings were default.




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