1290 infinity 2 lc

Metabolic profiling of the samples was performed at the Fraunhofer Institute of Toxicology and Experimental Medicine (ITEM, Hanover, Lower-Saxony, Germany) using a Biocrates AbsoluteIDQ p180 Kit (Biocrates Life Sciences AG, Innsbruck, Tyrol, Austria). This assay includes up to 188 metabolites related to glycolysis, oxidative processes, lipid degradation and inflammatory signalling. Amino acids and biogenic amines were measured by liquid chromatography (Agilent 1290 Infinity II LC, Santa Clara, CA, USA) tandem mass spectrometry (AB SCIEX 5500 QTrap mass spectrometer; AB SCIEX, Darmstadt, Germany), while acylcarnitines, hexoses, glycerophospholipids (PC and lysophosphatidylcholines), and sphingomyelins were quantified using flow injection analysis-tandem mass spectrometry.

For Metabolite profiling, all plasma samples were shipped on dry ice and analyzed at the Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM), Hannover, Germany, using a targeted metabolomics kit (MxP® Quant 500 kit: BIOCRATES Life Sciences AG, Innsbruck, Austria). This approach allows simultaneous absolute quantification of up to 630 metabolites covering 26 compound classes including 14 small molecule and 12 lipid classes using a combination of liquid chromatography (Agilent 1290 Infinity II LC, Santa Clara, CA, USA) and mass spectrometry (AB SCIEX 5500 QTrap™ mass spectrometer; AB SCIEX, Darmstadt, Germany). After normalization and pre-processing of the data, using MetIDQ™ software (Biocrates, Innsbruck, Austria) for peak integration and calculation of metabolite concentrations, 15 sphingolipids, distinct acylcarnitines and bile acids were employed for further investigation in the present study, whereas the unmentioned metabolites are documented in the supplemental Table S3.
Fast performance liquid chromatography (FPLC) was used for lipoprotein separation by means of two Superose 6 columns connected in series.