2.2.4. Targeted Metabolomic Determinations
AC18OH
A targeted metabolome analysis was carried out, including glucose, free carnitine, acylcarnitines (AC2, AC3, AC4, AC5, AC5:1, AC6, AC8, AC8:1, AC10, AC10:1, AC10:2, AC12, AC12:1, AC14, AC14:1, AC14:2, AC14OH, AC16, AC16:1, AC16:1OH, AC16OH, AC18, AC18:1, AC18:1OH, AC18:2, and AC18OH), arginosuccinate (ASA), and 12 L-amino acids (alanine, glycine, arginine, methionine, proline, arginine, valine, leucine, phenylalanine, tyrosine, citrulline, and ornithine). The above was achieved by using a Quattro Micro API (MicroMass, Cary, NC, USA) tandem mass spectrometer (MS-MS). All procedures for sample preparation and MS-MS analysis were performed using a NeoBase Non-derivatized kit (PerkinElmer, Waltham, MA, USA) according to the manufacturer’s protocol. The serum was dried in filter papers, and single disks were punched from each spot using a 3 mm punch. One disk was used per well. Using a multichannel pipette, 190 μL of extraction solution containing a mixture of the respective stable isotope-labeled internal standards was added to each well. The plate was covered with aluminum foil, shaken at 650× 