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Prolonged deficiency allows more extensive damage to accumulate, particularly to the nervous system, which requires longer to repair

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This method involves four main steps: isolation by strong cation-exchange SPE, mild derivatization with pentafluorophenyl trifluoromethanesulfonate, chromatographic separation using sequential ion-exchange/reversed-phase (ultra) high-performance liquid chromatography [(U)HPLC], and detection by electrospray ionization MRM-MS