DOI: 10.5061/dryad.1vhhmgqz0
title: Metabolome data from bovine preovulatory follicular fluid

description: The intrafollicular milieu influences mammalian fertility by providing the
 microenvironment for oocyte growth and maturation. A number of studies
 have linked abundance of follicular fluid metabolites to oocyte
 developmental competence and pregnancy outcome. Few studies have
 interrogated the preovulatory follicular fluid metabolome in cattle. This
 dataset includes preovulatory follicular fluid metabolome profiles from
 non-lactating Jersey cows. Ultra-High-Performance Liquid
 Chromatography-High Resolution Mass Spectrometry was performed on
 preovulatory follicular fluid samples and Xcalibur (RAW) files were
 converted to an open-source mzML format (msconvert software; ProteoWizard
 package). The converted files were processed using the Metabolomic
 Analysis and Visualization Engine (MAVEN; mzroll software, Princeton
 University) to complete an untargeted analysis of the liquid
 chromatography mass spectrometry data. The pre-processed peak data tables
 generated by MAVEN are provided in this dataset. 
description: Ultra-High-Performance Liquid
 Chromatography - High Resolution Mass Spectrometry (UHPLC-HRMS) was
 performed on follicular fluid samples at the Biological and Small Molecule
 Mass Spectrometry Core (RRID: SCR_021368) at the University of Tennessee,
 Knoxville. Sixty μL aliquots of follicular fluid samples (n=29) were
 thawed at room temperature, and metabolites within each sample were
 extracted using a 40:40:20 methanol/acetonitrile/water solution with 0.1 M
 formic acid following a previously reported procedure. An extraction
 solvent was added to the samples which underwent agitation and vortexing
 before samples were then chilled at -20°C for 20 min. Once samples were
 properly chilled, the tubes were centrifuged to form pellets and remove
 debris from the sample. A second set of 2 mL microcentrifuge tubes were
 used to collect the supernatant from the first set of tubes. The
 extraction solvent was again dispensed into the first set of tubes, which
 contained the pellet, and the mixture was pipetted to resuspend the
 pellet. The tubes were then re-submitted to agitation and vortexing,
 chilling, and centrifugation as described above. The supernatant from the
 tube was added to the second set of tubes that contained the previous
 supernatant. The first tube set with the final pellet was then discarded.
 The second set of tubes containing the supernatant underwent drying using
 nitrogen gas. Once dried, tubes were filled with MilliQ water to resuspend
 samples and moved to new autosampler vials. Metabolomic samples were
 separated using columns (Synergi Hydro RP, 2.5 μm, 100 mm x 2.0 mm column;
 Phenomenex, Torrance, CA, USA) which were maintained at 25°C. Solvents to
 elute metabolites during the mobile phase were (i) 97:3 methanol/water
 with 15 mM acetic acid and 11 mM tributylamine and (ii) 100% methanol. At
 a flow rate of 0.2 μL/min, the solvent gradient was (i) 100% and (ii) 0%
 from 0 to 5 min, (i) 80% and (ii) 20% from 5 to 13 min, (i) 45% and (ii)
 55% from 13 to 15.5 min, (i) 5% and (ii) 95% from 15.5 to 19 min, and (i)
 100% and (ii) 0% from 19 to 25 min. An Exactive Plus Orbitrap mass
 spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) with an
 electrospray ionization probe attached was used, operating in negative
 mode with a scan range between 72–1000 m/z, a resolution of 140,000, and
 an acquisition gain control of 3x10<sup>6</sup>.
 The mass spectrometry generated
 Xcalibur (RAW) files were converted to an open-source mzML format
 (msconvert software; ProteoWizard package). The converted files were
 processed using the Metabolomic Analysis and Visualization Engine (MAVEN;
 mzroll software, Princeton University) to complete an untargeted analysis
 of the liquid chromatography mass spectrometry data. MAVEN identifies
 metabolites using a variety of factors including peak shape, retention
 time, and signal-to-noise-ratio. The program then generated pre-processed
 peak data tables which were used for statistical analysis.
description: Microsoft Excel is required to open the dataset.