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Separation of the (1) isocitrate, (2) citrate (m/z 191.019), (3) 3-hydroxybutyrate, (4) 3-hydroxyisobutyrate, and (5) 2-hydroxybutyrate (m/z 103.040) isomers using an ACQUITY UPLC HSS T3 column (50 mm × 2.1 mm i.d.; 1.8 μm particle size) coupled to an ACQUITY UPLC HSS T3 VanGuard pre-column (5 mm × 2.1 mm i.d.; 1.8 μm particle size). (a) Gradient water/0.2% formic acid and methanol/0.1% formic acid; (b) gradient water/0.2% formic acid and acetonitrile/0.1% formic acid.
