Lipidome profiles of postnatal day 2 vaginal swabs reflect fat composition of gilt’s postnatal diet

KaLynn Harlow; Christina R. Ferreira; Tiago J.P. Sobreira; Theresa Casey; Kara Stewart

Samples were centrifuged, the organic phase (bottom phase) was separated from aqueous phase, 163 divided into four aliquots, and dried in a centrifugal evaporator (Savant SpeedVac AES2010, 164 ThermoFisher Scientific, San Jose, CA). Dried lipid extracts were stored at -20ËšC until mass 165 spectrometry analysis. Supplemental Table S1 for classes scanned). Initial data processing 190 of the profiles obtained was carried out by converting each set of MRM-profiling method data 191 into mzML format using MSConvert20. Next, the signal intensity for the ions present in NL and 192 Prec mass spectra was obtained using an in-house script. Ions with values of counts >1000 were 193 selected as parent ions and the product ion or neutral loss information was used for selecting ion Table S2 ). TAG 198 have no polar head or diagnostic functional group fragment, precluding them from the discovery 199 step of analysis. During screening phase, TAG were profiled using parent ions and a product ion The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/593392 doi: bioRxiv preprint 206 swabs, prior to screening lipid extract amount was recorded across samples by scanning for the 207 precursor ion of m/z 184, which profiles the most abundant lipids in cells, namely PC and SM 208 lipids. In this way, intensities for PC lipids were measured, and samples were diluted using the 209 same flow injection solvent to achieve a similar concentration for comparative relative analysis. Table S3 ) and 197 MRMs 218 (Supplemental Table S4 ) were organized into two distinct methods (Method 1 and Method 2, 219 respectively) for Screening Phase II analysis. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/593392 doi: bioRxiv preprint