Raw data from the manuscript entitled "The impact of normothermic and hypothermic preservation methods on kidney lipidome – comparative study using chemical biopsy with microextraction probes."
Authors: Natalia Warmuzińska1, Kamil Łuczykowski1, Iga Stryjak1, Hernando Rosales-Solano2, Peter Urbanellis3, Janusz Pawliszyn2, Markus Selzner3,4, Barbara Bojko1
1Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Nicolaus Copernicus University in Torun, Collegium Medicum in Bydgoszcz, Bydgoszcz, Poland
2Department of Chemistry, University of Waterloo, Waterloo, Ontario N2L 3G1, Canada
3Ajmera Transplant Center, Department of Surgery, Toronto General Hospital, University Health Network, Toronto, ON, Canada
4Department of Medicine, Toronto General Hospital, Toronto, ON, Canada
This study was funded by National Science Centre (Poland), grant Opus No. 2017/27/B/NZ5/01013
Contact: Barbara Bojko bbojko@cm.umk.pl; Natalia Warmuzińska n.warmuzinska@cm.umk.pl
SPME fibers coated with a mixed-mode extraction phase (coating length: 7 mm) were applied for direct kidney sampling in three porcine models of renal DCD autotransplantation using different preservation methods: an 8-hour SCS group (n=3), an 8-hour NEVKP group (n=3), and an 8-hour HMP group (n=2). SPME sampling was performed in vivo prior to kidney procurement; after 1 h and 2 h of warm ischemia; after 1 h, 3 h, 5 h, and 7 h of perfusion; in vivo immediately after revascularization (reperfusion), and in vivo under deep anesthesia at the time of sacrifice on postoperative day 3 (POD3). The extractions were performed by inserting the SPME fiber into the kidney cortex for 30 min at each time point. All fibers were desorbed immediately before instrumental analysis. For desorption, the fibers were inserted into 200μl of isopropanol:methanol (1:1 v/v) solution with the use of silanized inserts and agitated (1200 rpm) for 120 min. Untargeted lipidomics analysis was performed using an LC-HRMS procedure based on the coupling of an ultra-high performance liquid chromatograph and a Q-Exactive Focus Orbitrap mass spectrometer. During analysis, the samples were randomized and pooled quality control (QC) samples containing 10 μL of each sample were run every 8-10 injections to monitor instrument performance and analyte stability. Chromatographic separation was carried out on a hydrophilic stationary phase (HILIC) column (SeQuant ZIC-cHILIC, 3μm 100x2.1 mm) and in reversed-phase (RP) using a C18 column (Waters, XSelect CSH C18, 3.5µm, 2.1x75mm). The analyses were performed in positive and negative electrospray ionization mode. Four separate analyses were performed, and four sets of raw data were obtained: RP pos, RP neg, HILIC pos, and HILIC neg. Additionally, for each analysis, the putative identification of compounds was confirmed in Full MS/dd-MS2 mode using pooled QC samples. Detailed information about LC-MS parameters has been described in the Materials and Methods section of the manuscript.
All animals received humane care in compliance with the "Principles of Laboratory Animal Care" formulated by the National Society for Medical Research and the "Guide for the Care of Laboratory Animals" published by the National Institutes of Health and the ARRIVE guidelines 2.0. The study protocol was approved by the Animal Care Committee at the Toronto General Research Institute, Ontario, Canada.
Each sample was named according to the following scheme: type of chromatographic separation_scan polarity_animal identification number_sampling time point
Abbreviations: HMP- hypothermic machine perfusion; NEVKP- normothermic ex vivo kidney perfusion; SCS- static cold storage; WIT- warm ischemia time
License: CC BY - Creative Commons Attribution 4.0
