zldmЗаводская лаборатория. Диагностика материаловIndustrial laboratory. Diagnostics of materials1028-68612588-0187ООО «Издательство «ТЕСТ-ЗЛ»10.26896/1028-6861-2026-92-4-5-14zldm-2790Research ArticleАНАЛИЗ ВЕЩЕСТВАSUBSTANCES ANALYSISПростое и экспрессное определение ключевых интермедиатов трансметилирования в плазме крови спортсменов методом высокоэффективной жидкостной хроматографии в сочетании с тандемной масс-спектрометриейRapid and simple determination of key transmethylation intermediates in athletes’ blood plasma by high-performance liquid chromatography combined with tandem mass spectrometryДикунецМ. А.DikunetsM. A.

Марина Александровна Дикунец

105005, Москва, Елизаветинский пер., д. 10, стр. 1

Marina A. Dikunets

10 bld. 1, Elizavetinskii per., Moscow, 105005

ДудкоГ. А.DudkoG. A.

Григорий Алексеевич Дудко

105005, Москва, Елизаветинский пер., д. 10, стр. 1

Grigory A. Dudko

10 bld. 1, Elizavetinskii per., Moscow, 105005

ВирюсЭ. Д.VirusE. D.

Эдуард Даниэлевич Вирюс

105005, Москва, Елизаветинский пер., д. 10, стр. 1

Edward D. Virus

10 bld. 1, Elizavetinskii per., Moscow, 105005

edwardvirus@yandex.ruФедеральный научный центр физической культуры и спортаРоссияFederal Science Center of Physical Culture and SportRussian Federation202628042026924514Copyright © Дикунец М.А., Дудко Г.А., Вирюс Э.Д., 20262026Дикунец М.А., Дудко Г.А., Вирюс Э.Д.Dikunets M.A., Dudko G.A., Virus E.D.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.zldm.ru/jour/article/view/2790

Разработка простых и экспрессных методов определения различных биомаркеров в плазме крови необходима как для обеспечения оперативного мониторинга функционального состояния спортсменов, так и для валидации новых биомаркеров. В частности, ключевые интермедиаты трансметилирования S-аденозилметионин (SAM) и S-аденозилгомоцистеин (SAH) рассматривают в качестве потенциальных ранних маркеров нарушения долговременной адаптации спортсменов к физическим нагрузкам. Разработана методика хромато-масс-спектрометрического определения этих соединений в плазме крови: найдены приемлемые условия, обеспечивающие баланс чувствительности, селективности и экспрессности анализа. Для хроматографического разделения определяемых метаболитов в условиях градиентного элюирования использовали колонку с обращенной фазой Zorbax Eclipse XDB-C18 и смесь вода – метанол в качестве подвижной фазы. Оптимизированы значения потенциала декластеризации, изучена фрагментация ионов-предшественников SAM и SAH, исследовано влияние энергии соударений на интенсивность сигналов характеристичных фрагментных ионов. Для определения SAM и SAH выбраны ионы-продукты с m/z 250 и 136 соответственно. Оценены метрологические характеристики разработанной методики и матричный эффект при определении аналитов: предел определения SAM и SAH составляет 1 нг/мл, линейный динамический диапазон — 1 – 500 и 1 – 1000 нг/мл соответственно. Правильность методики внутри отдельной серии определений и между сериями подтверждена путем анализа образцов известного состава. Продолжительность анализа не превышает 4 мин.

The development of simple and rapid methods for determining various biomarkers in blood plasma is necessary both to ensure prompt monitoring of the functional state of athletes and to validate new biomarkers. In particular, the key transmethylation intermediates S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) are considered to be potential early markers of impaired long-term adaptation of athletes to physical exertion. A technique for the determination of these compounds in blood plasma by liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been developed: acceptable conditions have been found to ensure a balance of sensitivity, selectivity and expressiveness of the analysis. For chromatographic separation of the determined metabolites under gradient elution conditions, a Zorbax Eclipse XDB-C18 reversed-phase column and a water-methanol mixture as the mobile phase were used. The values of the declusterization potential have been optimized, fragmentation of SAM and SAH precursor ions has been studied, and the effect of collision energy on the signal intensity of characteristic fragment ions has been investigated. Product ions with m/z 250 and 136, respectively, were selected to determine SAM and SAH. The metrological characteristics of the developed technique and the matrix effect in the determination of analytes are evaluated: the limit of quantification of SAM and SAH is 1 ng/mL, the linear dynamic range is 1 – 500 and 1 – 1000 ng/mL, respectively. The correctness of the technique within a separate series of analysis and between series was confirmed by analyzing samples of known composition. The duration of the analysis does not exceed 4 min.

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The authors declare that there are no conflicts of interest present.