Synapt g2 s high definition mass spectrometer

The processing and analysis of LT muscle’s protein data from 105 of the animals used in this study were previously performed by Poleti [43 (link)], using an integrated transcriptome-assisted label-free quantitative proteomic approach by High Definition Mass Spectrometry. In summary, peptide samples were separated using the nanoACQUITY UPLC 2D Technology system [44 (link)] and identified by Synapt G2-S High Definition mass spectrometer (Waters, Manchester, UK). For protein identification and quantification, the raw data were searched against a Nelore transcriptome database built from the RNA-sequencing data of LT muscle. Label-free protein quantification values were generated based on the Hi3 method [45 (link)]. Only proteins identified with at least two peptides present in at least 80% of the animals were considered. Raw data of 938 proteins were used for genetic analysis.

High-resolution mass spectra of aliquots of all fractions and the isolated compound 1 and its isotopically labeled analog (1-IS, 0.1 mg/mL) were generated using a Synapt G2-S high-definition mass spectrometer (HDMS) (Waters) combined with an Acquity UPLC core system (Waters). A 150 × 2.1 mm, 130 Å, 1.7 µm BEH C18 column with a corresponding guard column (Waters) was used for chromatic separation. A mixture of water (solvent A) and acetonitrile (solvent B), each with an additive of 0.1% formic acid and a gradient starting with 5% B, increasing within 4.0 min to 100% B, holding for 0.5 min, and decreasing within 0.5 min to 5% B, was used. All instrument parameters were applied according to [32 (link)]. Data acquisition and processing were carried out using MassLynx 4.1 SCN 8.5.1 (Waters).