zldmЗаводская лаборатория. Диагностика материаловIndustrial laboratory. Diagnostics of materials1028-68612588-0187ООО «Издательство «ТЕСТ-ЗЛ»10.26896/1028-6861-2021-87-10-18-25zldm-1503Research ArticleАНАЛИЗ ВЕЩЕСТВАSUBSTANCES ANALYSISОпределение метаболитов азотистых ипритов в моче с использованием жидкостной хромато-масс-спектрометрии высокого разрешенияDetermination of nitrogen mustard metabolites in urine using high-performance liquid chromatography and high-resolution mass spectrometryБайгильдиевТ. М.BaygildievT. M.

Тимур Муратович Байгильдиев

119991, Москва, Ленинские горы, д. 1, стр. 3

Timur M. Baygildiev

1-3, Leninskie gory, Moscow, 119991

timurbaychem@yandex.ruБраунА. В.BraunA. V.

Аркадий Владимирович Браун

105005, Москва, Бригадирский переулок, д. 13

Arkady V. Braun

13, Brigadirsky per., Moscow, 105005

ВокуевМ. Ф.VokuevM. F.

Михаил Федорович Вокуев

119991, Москва, Ленинские горы, д. 1, стр. 3

Mikhail F. Vokuev

1-3, Leninskie gory, Moscow, 119991

РыбальченкоИ. В.RybalchenkoI. V.

Игорь Владимирович Рыбальченко

105005, Москва, Бригадирский переулок, д. 13

Igor V. Rybalchenko

13, Brigadirsky per., Moscow, 105005

РодинИ. А.RodinI. A.

Игорь Александрович Родин

119991, Москва, Ленинские горы, д. 1, стр. 3

Igor A. Rodin

1-3, Leninskie gory, Moscow, 119991

Московский государственный университет имени М. В. Ломоносова, химический факультетРоссияDepartment of Chemistry, M. V. Lomonosov Moscow State UniversityRussian Federation27 Научный Центр Министерства Обороны РФРоссияLaboratory for the Chemical and Analytical Control of the Military Research CentreRussian Federation20211810202187101825Copyright © Байгильдиев Т.М., Браун А.В., Вокуев М.Ф., Рыбальченко И.В., Родин И.А., 20212021Байгильдиев Т.М., Браун А.В., Вокуев М.Ф., Рыбальченко И.В., Родин И.А.Baygildiev T.M., Braun A.V., Vokuev M.F., Rybalchenko I.V., Rodin I.A.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/1503

Для обнаружения биологически активных компонентов в смесях сложного состава, к которым относятся биологические жидкости (кровь, моча и т.д.), широко используют метод жидкостной хромато-масс-спектрометрии высокого разрешения, сочетающий в себе возможности высокоселективного разделения исследуемых смесей, достоверного обнаружения неизвестных веществ и высокую чувствительность. Предложен способ одновременного извлечения из мочи N-триэтаноламина (ТЭА), N-этилдиэтаноламина (ЭДЭА) и N-метилдиэтаноламина (МДЭА) — высокополярных биомаркеров азотистых ипритов — с последующим определением методом ВЭЖХ в сочетании с тандемной масс-спектрометрией высокого разрешения. Исследованы масс-спектры фрагментации протонированных молекулярных ионов ТЭА, ЭДЭА и МДЭА, приведены возможные структурные формулы фрагментных ионов. Оптимизированы условия пробоподготовки образцов мочи и масс-спектрометрического детектирования в режиме мониторинга выбранных реакций. В качестве способа подготовки проб мочи к анализу выбран вариант разбавления деионизированной водой в 5 раз. Разделение компонентов проводили в варианте обращенно-фазовой хроматографии, при этом времена удерживания ТЭА, ЭДЭА и МДЭА составили 2,00, 2,05 и 1,92 мин соответственно. Время, необходимое для выполнения всех стадий анализа проб мочи, не превышает 25 мин. Пределы обнаружения биомаркеров в моче составили: 1 нг/мл ТЭА и 2 нг/мл ЭДЭА и МДЭА. Разработанный подход позволяет установить факт применения специфических азотистых ипритов в случаях расследования возможного попадания кожно-нарывных отравляющих веществ в живой организм.

Detection of biologically active components in mixtures of complex composition which include biological fluids (blood, urine, etc.) is the most difficult problem of the analytical chemistry and requires the use of modern highly informative research methods. One of the most effective and universal ways to study the structure of unknown substances is the method of liquid chromatography- high resolution mass spectrometry that combines the possibility of highly selective separation of the mixtures under study, the reliable detection of unknown substances and high sensitivity of the procedure. We propose a method for the simultaneous extraction of highly polar biomarkers of nitrogen mustard — N-triethanolamine (TEA), N-ethyl diethanolamine (EDEA) and N-methyldiethanolamine (MDEA) — from urine with subsequent determination by HPLC and detection by high resolution tandem mass spectrometry. The mass spectra of fragmentation of protonated molecular ions of TEA, EDEA, and MDEA have been studied and possible structural formulas of the fragment ions are given. The sample preparation of urine and mass spectrometric detection in the multiple reaction monitoring mode were optimized. The five-fold dilution with deionized water was chosen as a method of urine sample preparation for analysis. Separation of the components was performed in the reversed-phase chromatography mode with retention times for TEA, EDEA, and MDEA of 2.00, 2.05, and 1.92 min, respectively. The time required to complete all steps of the urine sample analysis did not exceed 25 min. The detection limits in urine were 1 ng/ml for TEA and 2 ng/ml for EDEA and MDEA. The developed approach provides determination of the fact of application of specific nitrogen mustard in enquiry of possible exposure of living organism to the blister agents.

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