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Polar metabolites were methoximated and trimethylsilylated to improve chromatographic behavior.
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120µL of 15mg/mL methoxyamine hydrochloride in pyridine was added, incubated at 50°C, and sonicated until crystallized metabolites went into solution.
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120µL MSTFA + 1%TMCS was subsequently added and the solution incubated for 30 min at 50°C.
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1.0 µL of the solution was injected at 15:1 split ratio onto a HP 6890 GC equipped with a 60M DB-5-MS column coupled to a HP 5973 MS.
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The injection port and transfer arm were held at 280°C, and the oven ramped from 80°C (2 min) to 315°C (6 min) at 5°C/min.
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The MS source was held at 250°C and the quadrupole at 150°C and scanned from 50-650.