Targeted Metabolic Profiling Using IDQ p400 HR

The Absolute IDQ p400 HR kit (test plates in the 96-well format; Biocrates Life Sciences AG, Innsbruck, Austria) was used to perform targeted quantitative analysis of the metabolites, according to the manufacturer’s protocol. The kit allows the quantitative measurement of up to 408 compounds (or their isomer groups) covering 11 classes of metabolites (including amino acids, biogenic amines, hexoses, acylcarnitines, di- and triglycerides, (lyso)phosphatidylcholines, sphingolipids, and cholesteryl esters) in 10 µl of sample thanks to the combination of direct flow injection and liquid chromatography (LC) with mass spectrometry (MS). The MS analyses were performed using Orbitrap Q Exactive Plus spectrometer (Thermo Fisher Scientific, Waltham, MA, USA), which was equipped with a 1290 Infinity UHPLC (Agilent, Santa Clara, CA, USA) system; Xcalibur 4.1 software (Thermo Fisher Scientific, Waltham, MA, USA) controlled the whole system. The concentrations of metabolites in µM were established by processing the registered spectra and chromatograms using Xcalibur 4.1. and MetIDQ DB110-2976 software (Biocrates Life Sciences AG) (19 (link)).