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

Autor: Ivanov AV; Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia. Electronic address: ivanov_av82@mail.ru., Dubchenko EA; City Clinical Hospital V.V. Veresaeva, Moscow, Russia., Kruglova MP; Institute Translational Medicine and Biotechnology, Sechenov First Moscow State Medical University, Moscow, Russia., Virus ED; Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Bulgakova PO; Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Alexandrin VV; Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia., Fedoseev AN; State Budgetary Institution of the city of Moscow 'City clinical hospital №24', Moscow Healthcare Department, Moscow, Russia., Boyko AN; Pirogov Russian National Research Medical University, Moscow, Russia., Grachev SV; City Clinical Hospital V.V. Veresaeva, Moscow, Russia., Kubatiev AA; Federal State Budgetary Scientific Institution 'Institute of General Pathology and Pathophysiology', Moscow, Russia; Russian Medical Academy of Postdoctoral Education, Moscow, Russia.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2019 Aug 15; Vol. 1124, pp. 366-374. Date of Electronic Publication: 2019 Jun 27.
DOI: 10.1016/j.jchromb.2019.06.032
Abstrakt: 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 KH 2 PO 4 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.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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