pmedpharmФармация и фармакологияPharmacy & Pharmacology2307-92662413-2241Pyatigorsk Medical and Pharmaceutical Institute - branch of Volgograd State Medical Univer10.19163/2307-9266-2018-6-3-229-240pmedpharm-304Research ArticleФАРМАЦЕВТИЧЕСКАЯ И ТОКСИКОЛОГИЧЕСКАЯ ХИМИЯPHARMACEUTICAL AND TOXICOLOGICAL CHEMISTRYРАЗРАБОТКА МЕТОДИКИ ПРЕДКОЛОНОЧНОЙ ДЕРИВАТИЗАЦИИ ГЛУТАТИОНА ВОССТАНОВЛЕННОГО 4-МЕТОКСИ-2-НИТРОФЕНИЛ- ИЗОТИОЦИОНАТОМ ДЛЯ ОПРЕДЕЛЕНИЯ МЕТОДОМ ВЫСОКОЭФФЕКТИВНОЙ ЖИДКОСТНОЙ ХРОМАТОГРАФИИDEVELOPMENT OF METHODS OF PRE-COLUMNAR DERIVATIZATION OF GLUTATHIONES RECOVERED BY 4-METHOXY-2-NITROPHENYLISOTHIOTOCIONATE FOR DETERMINATION BY METHOD OF HIGH-EFFECTIVE LIQUID CHROMATOGRAPHYhttps://orcid.org/0000-0002-0711-3505АлексееваК. А.AlexeevaK. A.

аспирант кафедры фармацевтической химии фармакогнозии

a post-graduate student of Pharmaceutical Chemistry and PharmacognosyDepartment

740890@bsu.edu.ruhttps://orcid.org/0000-0002-2996-7712ПисаревД. И.PisarevD. I.

доктор фармацевтических наук, доцент, профессор кафедры фармацевтической химии и фармакогнозии

PhD (Pharmacy), Associate Professor, Professor of Pharmaceutical Chemistryand Pharmacognosy Department

pisarev@bsu.edu.ruhttps://orcid.org/0000-0003-3145-6783НовиковО. О.NovikovO. O.

доктор фармацевтических наук, профессор, заведующий кафедройфармацевтической химии и фармакогнозии

PhD (Pharmacy), Professor, Head of Pharmaceutical Chemistry and Pharmacognosy Department

novikov@bsu.edu.ruhttps://orcid.org/0000-0001-6170-2151МалютинаА. Ю.MalyutinaA. Yu.

кандидат фармацевтических наук, доцент кафедры фармацевтической химии и фармакогнозии

PhD (Pharmacy), Associate Professor of Pharmaceutical Chemistryand Pharmacognosy Department

malyutina_a@bsu.edu.ruФГАОУ ВО НИУ «БелГУ» Минобрнауки РоссииРоссияFSAEI HE Belgorod National State Research University of the Russian FederationRussian Federation20181607201863229240Copyright © Алексеева К.А., Писарев Д.И., Новиков О.О., Малютина А.Ю., 20182018Алексеева К.А., Писарев Д.И., Новиков О.О., Малютина А.Ю.Alexeeva K.A., Pisarev D.I., Novikov O.O., Malyutina A.Y.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.pharmpharm.ru/jour/article/view/304

В настоящее время активно исследуется фармакологическая роль глутатиона в терапии канцерогенеза, нервно-дегенеративных и глазных болезней, заболеваний сердца, иммунной системы и старения организма. Поэтому для разработки фармацевтических объектов на его основе необходимо создание оптимальнойаналитической базы. Целью настоящего исследования является разработка методики анализа глутатиона восстановленного с помощью предколоночной дериватизации 4-метокси-2-нитрофенил-изотиоцианатом. Материалы и методы. Поскольку глутатион не имеет необходимых спектральных характеристик для непосредственного анализа, то исходя из этого, разработана методика определения глутатиона с помощью предколоночной дериватизации 4-метокси-2-нитрофенил-изотиоцианатом методом обращенно-фазной высокоэффективной хроматографии (ОФ ВЭЖХ). Детекцию образовавшегося деривата проводили по поглощению в УФ-свете с помощью диодно-матричного детектора. Результаты и обсуждение. В ходе описанного эксперимента, были получены хроматограммы деривата глулатиона с 4-метокси-2-нитрофенил-изотиоцианатом. Данную методику также оценивали на возможность количественного определения глутатиона. Чувствительность методики составила 0,01% или 3,1*10-1 моль. Прямолинейная зависимость между аналитическим сигналом (площадь пика) и концентрацией наблюдалась в диапазоне 0,01–0,08%, коэффициент корреляции – 0,995. Заключение. В ходе проведённых исследований разработана методика определения глутатиона с помощью предколоночной дериватизации 4-метокси-2-нитрофенил-изотиоцианатом методом ОФ ВЭЖХ. При этом образуется дериват со временем удерживания 22,3 мин и максимумом поглощения 398 нм. Также данная методика позволяет оценить количественное содержание исследуемого объекта.

Nowadays the pharmacological role of glutathione in the therapy of carcinogenesis, neurodegenerative and ocular diseases, heart diseases, the immune system and aging of the organism is being actively investigated. Therefore, for the development of pharmaceutical medical forms on its basis, it is necessary to create an optimal analytical base. The aim of this study is to develop a methodology for the analysis of glutathione recovered by pre-columnar derivatization of 4-methoxy-2-nitrophenyl isothiocyanate. Materials and methods. Since glutathione does not have the necessary spectral characteristics for its direct analysis, a methodology for the determination of glutathione with the use of pre-columnar derivatization of 4-methoxy-2-nitrophenyl-isothiocyanate by reversed-phase high-performance chromatography (RP HPLC) has been developed on that basis. Detection of the resulting derivative has been carried out by absorption in UV light using a diode array detector. Results and discussion. In the course of the experiment described, chromatograms of a glulathione derivative with 4-methoxy-2-nitrophenyl isothiocyanate were obtained. This technique was also evaluated for the possibility of quantitative determination of glutathione. The sensitivity of the methods was 0.01% or 3.1*10-1 mol. The linear relationship between the analytical signal (peak area) and concentration was observed within the range of 0.01–0.08% and the correlation coefficient of 0.995. Conclusion. In the course of the studies, a methodology for the determination of glutathione has been developed with the use of pre-columnarderivatization of 4-methoxy-2-nitrophenyl-isothiocyanate by RP HPLC. In this case, the derivative is formed with the retention time of 22.3 minutes and the absorption maximum of 398 nm. This method also allows estimating the quantitative content of the object under study.

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