Analytes in tissue extract supernatant were quantified using liquid chromatography/mass spectrometry (LC/MS) following derivatization with benzoyl chloride (BZC). Five microliters of supernatant was then mixed with 10 μL each of 500 mM NaCO3 (aq) and 2% BZC in acetonitrile in an LC/MS vial. After 2 min, the reaction was stopped by adding a 10 μL internal standard solution.
LC was performed on a 2.1 × 100 mm, 1.6 μm particle CORTECS Phenyl column (Waters Corporation, Milford, MA, USA) using a Waters Acquity UPLC. Mobile phase A was 0.1% aqueous formic acid, and mobile phase B was acetonitrile with 0.1% formic acid. MS analysis was performed using a Waters Xevo TQ‐XS triple quadrupole tandem mass spectrometer. The source temperature was 150°C, and the desolvation temperature was 400°C. The LC gradient is shown in TableS2 . Metabolite measurements were completed in the Vanderbilt University Neurochemistry Core.
LC was performed on a 2.1 × 100 mm, 1.6 μm particle CORTECS Phenyl column (Waters Corporation, Milford, MA, USA) using a Waters Acquity UPLC. Mobile phase A was 0.1% aqueous formic acid, and mobile phase B was acetonitrile with 0.1% formic acid. MS analysis was performed using a Waters Xevo TQ‐XS triple quadrupole tandem mass spectrometer. The source temperature was 150°C, and the desolvation temperature was 400°C. The LC gradient is shown in Table
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