chemicallytechFine Chemical TechnologiesТонкие химические технологии2410-65932686-7575MIREA – Russian Technological University (RTU MIREA).10.32362/2410-6593-2020-15-3-70-77chemicallytech-1625Research ArticleANALYTICAL METHODS IN CHEMISTRY AND CHEMICAL TECHNOLOGYАНАЛИТИЧЕСКИЕ МЕТОДЫ В ХИМИИ И ХИМИЧЕСКОЙ ТЕХНОЛОГИИQuantification of polysorbate 80 in biopharmaceutical formulations implementing an optimized colorimetric approachОпределение полисорбата 80 в биофармацевтических препаратах с помощью оптимизированной колориметрической методикиhttps://orcid.org/0000-0002-5842-7344СерегинА. С.SereginA. S.

Cерегин Александр Сергеевич, аспирант

117545, Москва, 1-й Дорожный проезд, д. 1

Aleksandr S. Seregin, Postgraduate Student

1, 1st Dorozhnyi pr., Moscow, 117545

penpney@mail.ruhttps://orcid.org/0000-0003-4161-5880ОрловаН. В.OrlovaN. V.

Орлова Наталья Владимировна, кандидат биологических наук, заведующая испытательной лабораторией

117545, Москва, 1-й Д рожный проезд, д. 1

Natalya V. Orlova, Cand. of Sci. (Biology), Head of Research Laboratory

1, 1st Dorozhnyi pr., Moscow, 117545

orlova.chemist@gmail.comhttps://orcid.org/0000-0003-0110-8323АскретковА. Д.AskretkovA. D.

Аскретков Александр Дмитриевич, старший химик-аналитик

117246, Москва, Научный проезд, д. 8, стр. 1

Aleksandr D. Askretkov, Chief Chemist-Analyst

8, b. 1, Nauchnyi pr., Moscow, 117246

askretkov.a.d@gmail.comhttps://orcid.org/0000-0002-5542-982XЗыбинД. И.ZybinD. I.

Зыбин Дмитрий Игоревич, старший химик-аналитик. ResearсherID P-8049-2016

117246, Москва, Научный проезд, д. 8, стр. 1

Dmitry I. Zybin, Chief Chemist-Analyst. ResearсherID P-8049-2016

8, b. 1, Nauchnyi pr., Moscow, 117246

zybin@pharmapark.ruСерегинД. И.SereginYu. A.

Серегин Юрий Александрович, кандидат биологических наук, заместитель директора. ResearсherID I-1874-2017

117545, Москва, 1-й Дорожный проезд, д. 1

Yuri A. Seregin, Cand. of Sci. (Biology), Deputy Director. ResearсherID I-1874-2017

1, 1st Dorozhnyi pr., Moscow, 117545

yuriy.seregin@pharmapark.ruНациональный исследовательский центр «Курчатовский институт» – ГосНИИгенетикаРоссияScientific Center “Kurchatov Institute,” Research Institute for Genetics and Selection of Industrial MicroorganismsRussian FederationООО «ФАРМАПАРК»РоссияPHARMAPARKRussian Federation2020060720201537077Copyright © Seregin A.S., Orlova N.V., Askretkov A.D., Zybin D.I., Seregin Y.A., 20202020Серегин А.С., Орлова Н.В., Аскретков А.Д., Зыбин Д.И., Серегин Д.И.Seregin A.S., Orlova N.V., Askretkov A.D., Zybin D.I., Seregin Y.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.finechem-mirea.ru/jour/article/view/1625Objectives

Objectives. We hereby describe an improvement of a previously developed quantification technique for polysorbate 80 in biopharmaceutical formulations (darbepoetin alfa and eculizumab) and report the validation of the new approach.

Methods

Methods. Polysorbate was isolated from analyte samples by protein precipitation using an organic solvent, followed by supernatant evaporation in vacuum. Polysorbate was derivatized using a ferric thiocyanate reagent and extracted into an organic phase; the relevant optical density measurements were performed.

Results

Results. We established the optimal conditions for each step of the analysis procedure. The accuracy was 97–102% in the tested analytical range, the relative standard deviation did not exceed 5%, and the limit of quantification was 0.01 mg/mL.

Conclusions

Conclusions. The reported approach is highly sensitive; polysorbate isolation and quantification do not depend on the matrix or, most importantly, the protein.

Цели

Цели. В данной работе была усовершенствована ранее разработанная методика определения полисорбата 80 в биотехнологических препаратах (дарбэпоэтин альфа, экулизумаб), а также проведена ее валидация.

Методы

Методы. Полисорбат извлекали из пробы осаждением белка органическим растворителем, затем выпаривали супернатант в вакууме. Полисорбат дериватизировали оптимизированным железо–тиоцианатным реагентом; дериват экстрагировали в слой органического растворителя и измеряли оптическую плотность.

Результаты

Результаты. Были установлены оптимальные условия для каждой стадии методики. Правильность находится в диапазоне степени извлечения 97–102%, относительное стандартное отклонение составляет не более 5%, предел количественного определения методики 0.01 мг/мл.

Выводы

Выводы. Представленная методика имеет высокую чувствительность. Извлечение и определение полисорбата не зависят от матрикса пробы – прежде всего, от присутствующего белка.

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