Method of processing a biological sample

A method of quantitative metabolic profiling of a biological sample containing multiple metabolites representing a plurality of different metabolite classes, the method comprising the steps of:

• pre-treating the biological sample with a metabolite extraction solvent to provide a pre-treated sample;
• separating a first aliquot of the pre-treated sample by reverse phase liquid chromatography (RPLC) to provide a first eluent containing resolved hydrophobic metabolites;
• separating a second aliquot of the pre-treated sample by hydrophilic interaction liquid interaction chromatography (HILIC) to provide a second eluent containing resolved hydrophilic metabolites;
• and
• assaying the first and second eluents using targeted tandem mass spectroscopy operated in multiple reaction monitoring mode to quantitatively profile the metabolites representing the plurality of different metabolite classes,
• including quantitative profiling of Dilinoleoyl-glycerol:
• 1,3-Dilinoleoyl-glycerol:
• 1,2-Dilinoleoyl-glycerol (isomer mixture) (DLG).

A method of quantitative metabolic profiling of a biological sample containing multiple metabolites representing a plurality of different metabolite classes, the method comprising the steps of:

• pre-treating the biological sample with a metabolite extraction solvent to provide a pre-treated sample;
• separating a first aliquot of the pre-treated sample by reverse phase liquid chromatography (RPLC) to provide a first eluent containing resolved hydrophobic metabolites;
• separating a second aliquot of the pre-treated sample by hydrophilic interaction liquid interaction chromatography (HILIC) to provide a second eluent containing resolved hydrophilic metabolites;
• and
• assaying the first and second eluents using targeted tandem mass spectroscopy operated in multiple reaction monitoring mode to quantitatively profile the metabolites representing the plurality of different metabolite classes,
• including quantitative profiling of L-leucine (L-LEU);
• Citrulline (CR);
• Dilinoleoyl-glycerol:
• 1,3-Dilinoleoyl-glycerol:
• 1,2-Dilinoleoyl-glycerol (isomer mixture) (DLG);
• L-isoleucine (L-ISO);
• L-methionine (L-MET);
• NG-Monomethyl-L-arginine (NGM);
• Asymmetric dimethylarginine (ADMA);
• 1-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (1-HD);
• or Biliverdin (BV).