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Determination of S-adenosylmethionine and S-adenosylhomocysteine in blood plasma by UPLC with fluorescence detection

Alexander Vladimirovich Ivanov, Ekaterina Alexandrovna Dubchenko, Maria Petrovna Kruglova, Edward Danielevich Virus, Polina Olegovna Bulgakova, Valery Vasil'evich Alexandrin, Anatolij Nikolaevich Fedoseev, Alexey Nikolaevich Boyko, Sergej Vitalievich Grachev, Aslan Amirkhanovich Kubatiev
Journal of chromatography 2019 v.1124 pp. 366-374
S-adenosylmethionine, acetic acid, acetonitrile, blood plasma, chemical species, derivatization, detection limit, fluorescence, metabolites, phenylboronic acids, potassium dihydrogen phosphate, solid phase extraction, ultra-performance liquid chromatography, volunteers
A validated approach to determine various methionine cycle metabolites (S-adenosylmethionine, S-adenosylhomocysteine, and methylthioadenosine) in human blood plasma is offered. The approach is based on solid-phase extraction (with grafted phenylboronic acid) and derivatization with chloroacetaldehyde followed by ultra-performance liquid chromatography with fluorescence detection. We used a 100 × 2.1 mm × 1.8 μm C18 column for the selective separation of analytes. Chromatographic separation was achieved with gradient elution of acetonitrile (flow rate 0.2 mL/min) from 2 to 20%. The eluent was initially composed of 10 mM KH2PO4 with 10 mM acetic acid and 25 μM heptafluorobutyric acid. The total analysis time was 11 min. Validation of the method included detection of the limit of detection (2 nM), limit of quantification (5 nM), accuracy (97.2–101%), and intra- and interday precision (2.2–9.0%). Analysis of plasma samples from healthy volunteers revealed that the average levels of S-adenosylmethionine, S-adenosylhomocysteine, and methylthioadenosine were 93.6, 20.9 and 14.8 nM, respectively.