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Determination of S-adenosylmethionine and S-adenosylhomocysteine in Blood Plasma by UPLC With Fluorescence Detection

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, etc.

J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Aug 15;1124:366-374.

PMID: 31295723

Abstract:

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP168618426	2-(Methylthio)phenylboronic acid		168618-42-6	Price

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