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<article article-type="research-article" dtd-version="1.3" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xml:lang="ru"><front><journal-meta><journal-id journal-id-type="publisher-id">actabiomedica</journal-id><journal-title-group><journal-title xml:lang="ru">Acta Biomedica Scientifica</journal-title><trans-title-group xml:lang="en"><trans-title>Acta Biomedica Scientifica</trans-title></trans-title-group></journal-title-group><issn pub-type="ppub">2541-9420</issn><issn pub-type="epub">2587-9596</issn><publisher><publisher-name>Scientific Centre for Family Health and Human Reproduction Problems</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.29413/ABS.2024-9.1.18</article-id><article-id custom-type="elpub" pub-id-type="custom">actabiomedica-4612</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="ru"><subject>ФАРМАКОЛОГИЯ И ФАРМАЦИЯ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>PHARMACOLOGY AND PHARMACEUTICS</subject></subj-group></article-categories><title-group><article-title>Разработка и валидация методики количественного определения моноаминовых нейромедиаторов и их метаболитов в тканях мозга крыс с помощью ВЭЖХ-МС/МС</article-title><trans-title-group xml:lang="en"><trans-title>Development and validation of a method for the quantitative determination of monoamine neurotransmitters and their metabolites in rat brain tissue using HPLC-MS/MS</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-0032-0341</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Хохлов</surname><given-names>А. Л.</given-names></name><name name-style="western" xml:lang="en"><surname>Khokhlov</surname><given-names>A. L.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Хохлов Александр Леонидович – доктор медицинских наук, профессор, академик РАН, заместитель директора по научной работе Центра трансфера фармацевтических технологий им. М.В. Дорогова, 150010, г. Ярославль, ул. Технопарковая, 11/2;</p><p>заведующий кафедры фармакологии и клинической фармакологии, ректор, 150000, г. Ярославль, ул. Революционная, 5</p></bio><bio xml:lang="en"><p>Alexander L. Khokhlov – Dr. Sc. (Med.), Professor, Member of RAS, Deputy Director for Science of the Center of Pharmaceutical Technologies Transfer named after M.V. Dorogov, Tekhnoparkovaya str. 11/2, Yaroslavl 150010;</p><p>Head of the Department of Pharmacology and Clinical Pharmacology, Rector, Revolutsionnaya str. 5, Yaroslavl 150000</p></bio><email xlink:type="simple">al460935@yandex.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-0066-7388</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Яичков</surname><given-names>И. И.</given-names></name><name name-style="western" xml:lang="en"><surname>Yaichkov</surname><given-names>I. I.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Яичков Илья Игоревич – кандидат фармацевтических наук, научный сотрудник Центра трансфера фармацевтических технологий им. М.В. Дорогова, 150010, г. Ярославль, ул. Технопарковая, 11/2;</p><p>научный сотрудник Института фармации, 150000, г. Ярославль, ул. Революционная, 5</p></bio><bio xml:lang="en"><p>Ilya I. Yaichkov – Cand. Sc. (Med.), Research Officer at the Center of Pharmaceutical Technologies Transfer named after M.V. Dorogov, Tekhnoparkovaya str. 11/2, Yaroslavl 150010;</p><p>Research Officer at the Institute of Pharmacy, Revolutsionnaya str. 5, Yaroslavl 150000</p></bio><email xlink:type="simple">ilya_1993_08@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-0913-2571</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Корсаков</surname><given-names>М. К.</given-names></name><name name-style="western" xml:lang="en"><surname>Korsakov</surname><given-names>M. K.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Корсаков Михаил Константинович – доктор химических наук, профессор кафедры химии, теории и методики преподавания химии, директор Центра трансфера фармацевтических технологий им. М.В. Дорогова, </p><p>150010, г. Ярославль, ул. Технопарковая, 11/2</p></bio><bio xml:lang="en"><p>Mikhail K. Korsakov – Dr. Sc. (Chem.), Professor at the Department of Chemistry, Theory and Methods for Teaching of Chemistry, Director of the Center of Pharmaceutical Technologies Transfer named after M.V. Dorogov, </p><p>Tekhnoparkovaya str. 11/2, Yaroslavl 150010</p></bio><email xlink:type="simple">m.korsakov@yspu.org</email><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-2139-6847</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Каграманян</surname><given-names>И. Н.</given-names></name><name name-style="western" xml:lang="en"><surname>Kagramanyan</surname><given-names>I. N.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Каграманян Игорь Николаевич – доктор медицинских наук, профессор Института лидерства и управления здравоохранением, </p><p>119991, г. Москва, ул. Большая Пироговская, 2, стр. 4</p></bio><bio xml:lang="en"><p>Igor N. Kagramanyan – Dr. Sc. (Med.), Professor at the Institute of Leadership and Management of Healthcare, </p><p>Bolshaya Pirogovskaya str. 2, build. 4, Moscow 119991</p></bio><email xlink:type="simple">info@hsha.ru</email><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-4275-9037</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Вольхин</surname><given-names>Н. Н.</given-names></name><name name-style="western" xml:lang="en"><surname>Volkhin</surname><given-names>N. N.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Вольхин Никита Николаевич – младший научный сотрудник Центра трансфера фармацевтических технологий им. М.В. Дорогова, </p><p>150010, г. Ярославль, ул. Технопарковая, 11/2</p></bio><bio xml:lang="en"><p>Nikita N. Volkhin – Junior Research Officer at the Center of Pharmaceutical Technologies Transfer named after M.V. Dorogov,</p><p>Tekhnoparkovaya str. 11/2, Yaroslavl 150010</p></bio><email xlink:type="simple">nnvolkhin@ysmu.ru</email><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0009-0007-8435-7689</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Петухов</surname><given-names>С. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Petukhov</surname><given-names>S. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Петухов Сергей Станиславович – инженер Центра трансфера фармацевтических технологий им.  М.В.  Дорогова, 150010, г. Ярославль, ул. Технопарковая, 11/2;</p><p>младший научный сотрудник Института фармации, 150000, г. Ярославль, ул. Революционная, 5</p></bio><bio xml:lang="en"><p>Sergey S. Petukhov – Engineer at the Center of Pharmaceutical Technologies Transfer named after M.V. Dorogov, Tekhnoparkovaya str. 11/2, Yaroslavl 150010;</p><p>Junior Research Officer at the Institute of Pharmacy, Revolutsionnaya str. 5, Yaroslavl 150000</p></bio><email xlink:type="simple">sspp465@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0009-0008-9431-1980</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Зайкова</surname><given-names>В. Е.</given-names></name><name name-style="western" xml:lang="en"><surname>Zaikova</surname><given-names>V. E.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Зайкова Валерия Евгеньевна – лаборант Центра трансфера фармацевтических технологий им. М.В. Дорогова, 150010, г. Ярославль, ул. Технопарковая, 11/2;</p><p>специалист Института фармации, 150000, г. Ярославль, ул. Революционная, 5</p></bio><bio xml:lang="en"><p>Valeria E. Zaykova – Clinical Research Assistant at the Center of Pharmaceutical  Technologies Transfer named after M.V. Dorogov, Tekhnoparkovaya str. 11/2, Yaroslavl 150010;</p><p>Specialist at the Institute of Pharmacy, Revolutsionnaya str. 5, Yaroslavl 150000</p></bio><email xlink:type="simple">valeria.zaykova@bk.ru</email><xref ref-type="aff" rid="aff-1"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru"><institution>ФГБОУ ВО «Ярославский государственный педагогический университет им. К.Д. Ушинского»;&#13;
ФГБОУ ВО «Ярославский государственные медицинский университет» Минздрава России</institution></aff><aff xml:lang="en"><institution>Yaroslavl State Pedagogical University named after K.D. Ushinsky;&#13;
Yaroslavl State Medical University</institution></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru"><institution>ФГБОУ ВО «Ярославский государственный педагогический университет им. К.Д. Ушинского»</institution></aff><aff xml:lang="en"><institution>Yaroslavl State Pedagogical University named after K.D. Ushinsky</institution></aff></aff-alternatives><aff-alternatives id="aff-3"><aff xml:lang="ru"><institution>ФГАОУ ВО Первый Московский государственный медицинский университет имени И.М. Сеченова Минздрава России (Сеченовский Университет)</institution></aff><aff xml:lang="en"><institution>I.M. Sechenov First Moscow State Medical University (Sechenov University)</institution></aff></aff-alternatives><pub-date pub-type="collection"><year>2024</year></pub-date><pub-date pub-type="epub"><day>27</day><month>03</month><year>2024</year></pub-date><volume>9</volume><issue>1</issue><fpage>177</fpage><lpage>191</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Хохлов А.Л., Яичков И.И., Корсаков М.К., Каграманян И.Н., Вольхин Н.Н., Петухов С.С., Зайкова В.Е., 2024</copyright-statement><copyright-year>2024</copyright-year><copyright-holder xml:lang="ru">Хохлов А.Л., Яичков И.И., Корсаков М.К., Каграманян И.Н., Вольхин Н.Н., Петухов С.С., Зайкова В.Е.</copyright-holder><copyright-holder xml:lang="en">Khokhlov A.L., Yaichkov I.I., Korsakov M.K., Kagramanyan I.N., Volkhin N.N., Petukhov S.S., Zaikova V.E.</copyright-holder><license xml:lang="ru" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>Данная работа распространяется под лицензией Creative Commons Attribution 4.0.</license-p></license><license xml:lang="en" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>This work is licensed under a Creative Commons Attribution 4.0 License.</license-p></license></permissions><self-uri xlink:href="https://www.actabiomedica.ru/jour/article/view/4612">https://www.actabiomedica.ru/jour/article/view/4612</self-uri><abstract><sec><title>Обоснование</title><p>Обоснование. Определение изменения содержания моноаминовых нейромедиаторов и  их  метаболитов в  структурах головного мозга является необходимой частью изучения фармакодинамики противопаркинсонических лекарственных средств. Методика совместного определения норадреналина, адреналина, допамина, серотонина, 5-гидроксииндол-3-уксусной кислоты, 3,4-дигидроксифенилуксусной кислоты, гомованилиновой кислоты, ванилилминдальной кислоты в тканях мозга крыс ранее не была разработана.</p></sec><sec><title>Цель исследования</title><p>Цель исследования. Разработка и  валидация методики количественного определения норадреналина, адреналина, допамина, серотонина, 5-гидроксииндол-3-уксусной кислоты, 3,4-дигидроксифенилуксусной кислоты, гомованилиновой кислоты, ванилилминдальной кислоты в  тканях мозга крыс с помощью высокоэффективной жидкостной хроматографии в сочетании с тандемной масс-спектрометрией (ВЭЖХ-МС/МС).</p></sec><sec><title>Методы</title><p>Методы. Методика определения моноаминовых медиаторов и их метаболитов разработана c  применением метода ВЭЖХ-МС/МС. Гомогенаты тканей мозга готовились с  помощью механического ручного гомогенизатора. Изучено влияние различных антиоксидантов на стабильность норадреналина, адреналина, допамина и 3,4-дигидроксифенилуксусной кислоты в испытуемых образцах.</p></sec><sec><title>Результаты</title><p>Результаты. Хроматографическое разделение компонентов пробы осуществлялось с помощью двух хроматографических колонок Synergi Max RP (20 × 2,0 мм, 2,5 мкм) и Synergi Fusion RP 80Å (250 × 4,6 мм, 4 мкм). Элюирование проводили в градиентном режиме с применением подвижной фазы на основе метанола и 0,1%-го раствора муравьиной кислоты в воде. Для подготовки проб гомогенатов использовалось разведение образцов раствором внутренних стандартов в метаноле. В качестве стабилизатора-антиоксиданта был выбран 5%-й водный раствор аскорбиновой кислоты.</p></sec><sec><title>Заключение</title><p>Заключение. Разработанная методика прошла полную валидацию и соответствует требованиям российских и международных руководств. Выбранный способ стабилизации позволяет хранить образцы гомогенатов мозга в течение 30 дней после отбора.</p></sec></abstract><trans-abstract xml:lang="en"><sec><title>Background</title><p>Background. Determining changes in the content of monoamine neurotransmitters and their metabolites in brain structures is a necessary part of studying the pharmacodynamics of  antiparkinsonian drugs. A  method for  the  joint determination of norepinephrine, adrenaline, dopamine, serotonin, 5-hydroxyindole-3-acetic acid, 3,4-dihydroxyphenylacetic acid, homovanillic acid, vanillylmandelic acid in rat brain tissue has not previously been developed.</p></sec><sec><title>The aim of the study</title><p>The aim of the study. To develop and to validate a method for the quantitative determination of norepinephrine, adrenaline, dopamine, serotonin, 5-hydroxyindole3-acetic acid, 3,4-dihydroxyphenylacetic acid, homovanillic acid, vanillylmandelic acid in rat brain tissue using high-performance liquid chromatography in combination with tandem mass spectrometry (HPLC-MS/MS).</p></sec><sec><title>Materials and  methods</title><p>Materials and  methods. A method for  determining monoamine mediators and their metabolites was developed using the HPLC-MS/MS method. Brain tissue homogenates were prepared using a  mechanical hand-operated homogenizer. The  effect of  various antioxidants on  the  stability of  norepinephrine, adrenaline, dopamine and 3,4-dihydroxyphenylacetic acid in the test samples was studied.</p></sec><sec><title>Results</title><p>Results. Chromatographic separation of sample components was carried out using two Synergi Max RP (20 × 2.0 mm, 2.5 µm) and Synergi Fusion RP 80Å (250 × 4.6 mm, 4 µm) chromatographic columns. Elution was carried out in a gradient mode using a  mobile phase based on  methanol and  a  0.1% solution of  formic acid in  water. To prepare homogenate batches, the samples were diluted with a solution of internal standards in methanol. A 5% aqueous solution of ascorbic acid was chosen as an antioxidant stabilizer.</p></sec><sec><title>Conclusion</title><p>Conclusion. The  developed methodology has been fully validated and meets the requirements of Russian and international guidelines. The chosen stabilization method allows samples of brain homogenates to be stored for 30 days after collection.</p></sec></trans-abstract><kwd-group xml:lang="ru"><kwd>ВЭЖХ-МС/МС</kwd><kwd>моноаминовые нейромедиаторы</kwd><kwd>ткани мозга</kwd><kwd>стабилизация образцов</kwd></kwd-group><kwd-group xml:lang="en"><kwd>HPLC-MS/MS</kwd><kwd>monoamine neurotransmitters</kwd><kwd>brain tissue</kwd><kwd>sample stabilization</kwd></kwd-group><funding-group><funding-statement xml:lang="ru">Исследование выполнено за счёт средств гранта Российского научного фонда «Разработка новых фармакологических агентов для терапии нейродегенеративных заболеваний» № 22-13-20085.</funding-statement></funding-group></article-meta></front><back><ref-list><title>References</title><ref id="cit1"><label>1</label><citation-alternatives><mixed-citation xml:lang="ru">Руководство по проведению доклинических исследований лекарственных средств. Часть первая. 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