Project description:The concentrations of twenty kinds of hormones in the follicular fluid were detected by high-performance liquid chromatography–mass spectrometry (HPLC-MS/MS). An Agilent 1200 series high-performance liquid chromatography (HPLC) instrument (Agilent, USA) was utilized. A PAL autosampler (CTC, Swiss) and a Gemini-NX-C18 column (2.0 mm×50 mm, 3 μm, Waters, USA) were used. The ion source was an API-4000 quadrupole electrostatic field orbit trap high-resolution mass spectrometer (Applied Biosystems, USA). The scanning mode was multiple reaction monitoring (MRM) (Agilent-1200 LC system coupled to an API400 mass spectrometer).
Project description:We established a spectral library for data-independent analysis of human plasma samples, using a pooled plasma sample from Danish adults. The samples were prepared for proteome analysis using an in-house developed protocol consuming one microliter of plasma per sample, and the tryptic digest and subsequent C18 purification was performed in 96-well plates. The spectral library consists of plasma which was analyzed neat (i.e. non-treated), and top 14 depleted. The samples were subjected to high pH reversed-phase peptide fractionated using C18 tips (8 fraction) and a HPLC system (20 fraction). The resulting peptides were analyzed on a Q Exactive with a capflow reversed-phase C18 LC-MS/MS setup in data-dependent mode. The final spectral library contains 21788 precursors and 1000 proteins (both at FDR<0.01).