pharmjournalРазработка и регистрация лекарственных средствDrug development & registration2305-20662658-5049LLC «CPHA»pharmjournal-469Research ArticleАНАЛИТИЧЕСКИЕ МЕТОДИКИ И МЕТОДЫ КОНТРОЛЯANALYTICAL AND QUALITY CONTROL METHODSОПРЕДЕЛЕНИЕ ПОЛИСОРБАТОВ СПЕКТРОФОТОМЕТРИЧЕСКИМ МЕТОДОМ В ЛЕКАРСТВЕННЫХ ПРЕПАРАТАХ НА ОСНОВЕ РЕКОМБИНАНТНЫХ БЕЛКОВDETERMINATION OF POLYSORBATES BY SPECTROPHOTOMETRY IN DRUGS BASED ON RECOMBINANT PROTEINSАскретковА. Д.AskretkovA. D.noemail@neicon.ruИсайкинаП. М.IsaykinaP. M.noemail@neicon.ruЧерепушкинС. А.CherepushkinS. A.noemail@neicon.ruОрловаН. В.OrlovaN. V.noemail@neicon.ruФГБОУ ВО «Московский технологический университет» (Институт тонких химических технологий)Moscow Technological University (MITHT)ФГБУ «Государственный научно-исследовательский институт генетики и селекции промышленных микроорганизмов» («ГосНИИгенетика»)Scientific Center of Russian Federation «Research Institute for Genetics and Selection of Industrial Microorganisms»20170901201903124129Copyright © Аскретков А.Д., Исайкина П.М., Черепушкин С.А., Орлова Н.В., 20192019Аскретков А.Д., Исайкина П.М., Черепушкин С.А., Орлова Н.В.Askretkov A.D., Isaykina P.M., Cherepushkin S.A., Orlova N.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.pharmjournal.ru/jour/article/view/469

Разработана методика определения полисорбата-20 и полисорбата-80 в лекарственных препаратах на основе рекомбинантных белков методом спек трофотометрии. Выделение полисорбата из раствора проводится с помощью твердофазной экстракции с последующим элюированием полисорбата ацетонитрилом. Ацетонитрильный раствор упаривается, и полисорбат дериватизуется железотиоцианатным реагентом, полученный дериватизат извлекается в органическую фазу, и измеряется оптическая плотность. Подобраны наиболее оптимальные условия дериватизации, экстракции в органическую фазу и измерения полисорбата. Проведена валидация методики: определены её специфичность, линейность, правильность, прецизионность, устойчивость, предел количественного определения.

Method of determination of polysorbate 20 and polysorbate 80 in recombinant protein drugs based on spectrophotometry has been designed. Isolation of polysorbate from solution is carried out by solid phase extraction followed by elution of polysorbate with acetonitrile. Acetonitrile solution is evaporated and then polysorbate is derivatized with ferrous-thiocyanate reagent. Obtained derivative is extracted into organic phase and optical density is measured. The most optimal conditions of derivatization, extraction into organic phase and measurment was chosen. The validation was performed: specificity, linearity, accuracy, precision, robustness, limit of quantification.

валидациярекомбинантные белкиполисорбаттвердофазная экстракциятвинтиоцианат железаvalidationrecombinant proteinspolysorbatesolid phase extractiontweeniron thiocyanateReferencesE.Y. Chi, S. Krishnan, T.W. Randolph, J.F. Carpenter. Physical Stability of Proteins in Aqueous Solution: Mechanism and Driving Forces in Nonnative Protein Aggregation // Pharm. Res. 2003. V. 20(9). P. 1325–1336.E.Y. Chi, S. Krishnan, T.W. Randolph, J.F. Carpenter. Physical Stability of Proteins in Aqueous Solution: Mechanism and Driving Forces in Nonnative Protein Aggregation // Pharm. Res. 2003. V. 20(9). P. 1325–1336.B.A. Kerwin. Polysorbates 20 and 80 used in the formulation of protein biotherapeutics: Structure and degradation pathway // J. Pharm. Sci. 2008. V. 97(8). P. 2924–2935.B.A. Kerwin. Polysorbates 20 and 80 used in the formulation of protein biotherapeutics: Structure and degradation pathway // J. Pharm. Sci. 2008. V. 97(8). P. 2924–2935.M. Agarkhed, C. O’Dell, M.C. Hsieh et al. Effect of Polysorbate 80 Concentration on Thermal and Photostability of a Monoclonal Antibody // AAPS Pharm. Sci. Tech. 2013. V. 14(1). P. 1–9.M. Agarkhed, C. O’Dell, M.C. Hsieh et al. Effect of Polysorbate 80 Concentration on Thermal and Photostability of a Monoclonal Antibody // AAPS Pharm. Sci. Tech. 2013. V. 14(1). P. 1–9.W. Norde. Adsorption of proteins at solid-liquid interfaces (Review) // Cells and Materials. 1995. V. 5(1). P. 97–112.W. Norde. Adsorption of proteins at solid-liquid interfaces (Review) // Cells and Materials. 1995. V. 5(1). P. 97–112.S. Kiese, A. Pappenberger., W. Friess, H.C. Mahler. Shaken, not stirred: mechanical stress testing of an IgG1 antibody // J. Pharm. Sci. 2008. V. 97(10). P. 4347–4366.S. Kiese, A. Pappenberger., W. Friess, H.C. Mahler. Shaken, not stirred: mechanical stress testing of an IgG1 antibody // J. Pharm. Sci. 2008. V. 97(10). P. 4347–4366.L. Kreilgaard, L.S. Jones, T.W. Randolph et al. Effect of tween 20 on freeze-thawing- and agitation-induced aggregation of recombinant human factor XIII // J. Pharm. Sci. 1998. V. 87(12). P. 1597–1603.L. Kreilgaard, L.S. Jones, T.W. Randolph et al. Effect of tween 20 on freeze-thawing- and agitation-induced aggregation of recombinant human factor XIII // J. Pharm. Sci. 1998. V. 87(12). P. 1597–1603.L.S. Jones, T.W. Randolph, U. Kohnert. et al. The effects of Tween 20 and sucrose on the stability of anti-L-selectin during lyophilization and reconstitution // J. Pharm. Sci. 2001. V. 90(10). P. 1466–1477.L.S. Jones, T.W. Randolph, U. Kohnert. et al. The effects of Tween 20 and sucrose on the stability of anti-L-selectin during lyophilization and reconstitution // J. Pharm. Sci. 2001. V. 90(10). P. 1466–1477.J.X. Zhou, J. Qiua, G. Jiangb et al. Non-specific binding and saturation of Polysorbate-20 with aseptic filter membranes for drug substance and drug product during mAb production // J. Membrane Sci. 2008. V. 325(2). P. 735–741.J.X. Zhou, J. Qiua, G. Jiangb et al. Non-specific binding and saturation of Polysorbate-20 with aseptic filter membranes for drug substance and drug product during mAb production // J. Membrane Sci. 2008. V. 325(2). P. 735–741.T.H. Tani, J.M. Moore, T.W. Patapoff. Single step method for the accurate concentration determination of polysorbate 80 // J. Chromatogr. A. 1997. V. 786(1). P. 99–106.T.H. Tani, J.M. Moore, T.W. Patapoff. Single step method for the accurate concentration determination of polysorbate 80 // J. Chromatogr. A. 1997. V. 786(1). P. 99–106.D. Ilko, A. Braun, O. Germershaus et al. Fatty acid composition analysis in polysorbate 80 with high performance liquid chromatography coupled to charged aerosol detection // Eur. J. Pharm. Biopharm. 2015. V. 94. P. 569–574.D. Ilko, A. Braun, O. Germershaus et al. Fatty acid composition analysis in polysorbate 80 with high performance liquid chromatography coupled to charged aerosol detection // Eur. J. Pharm. Biopharm. 2015. V. 94. P. 569–574.D. Lee, S. Rumbelow, S.K.R. Williams. Identification and Quantitation of Trace Impurities in Fatty Alcohol Ethoxylates Using HPLC and MALDI-TOF Mass Spectrometry // Anal. Chim. Acta. 2009. V. 654(1). P. 59–63.D. Lee, S. Rumbelow, S.K.R. Williams. Identification and Quantitation of Trace Impurities in Fatty Alcohol Ethoxylates Using HPLC and MALDI-TOF Mass Spectrometry // Anal. Chim. Acta. 2009. V. 654(1). P. 59–63.S.D. Baker, M. Zhao, P. He et al. Simultaneous analysis of docetaxel and the formulation vehicle polysorbate 80 in human plasma by liquid chromatography/tandem mass spectrometry // Anal. Biochem. 2004. V. 324(2). P. 276–284.S.D. Baker, M. Zhao, P. He et al. Simultaneous analysis of docetaxel and the formulation vehicle polysorbate 80 in human plasma by liquid chromatography/tandem mass spectrometry // Anal. Biochem. 2004. V. 324(2). P. 276–284.M. Liu, C. Xie, W. Xu, W. Lu. Separation of polyethylene glycols and their amino-substituted derivatives by high-performance gel filtration chromatography at low ionic strength with refractive index detection // J. Chromatogr. A. 2004. V. 1046(1-2). P. 121–126.M. Liu, C. Xie, W. Xu, W. Lu. Separation of polyethylene glycols and their amino-substituted derivatives by high-performance gel filtration chromatography at low ionic strength with refractive index detection // J. Chromatogr. A. 2004. V. 1046(1-2). P. 121–126.M.D. Wenger, A.M. Bowman, M.V. Thorsteinsson et al. An automated homogeneous method for quantifying polysorbate using fluorescence polarization // Anal. Biochem. 2005. V. 337(1). P. 48–54.M.D. Wenger, A.M. Bowman, M.V. Thorsteinsson et al. An automated homogeneous method for quantifying polysorbate using fluorescence polarization // Anal. Biochem. 2005. V. 337(1). P. 48–54.Z. Wei, S. Bilbulian, J. Li et al. Universal method for the determination of nonionic surfactant content in the presence of protein // J. Sep. Sci. 2015. V. 38(8). P. 1318–1325.Z. Wei, S. Bilbulian, J. Li et al. Universal method for the determination of nonionic surfactant content in the presence of protein // J. Sep. Sci. 2015. V. 38(8). P. 1318–1325.N.B. Cucakovich. Determination of Tween 80 in tissue culture media, vaccines, and related products // Anal. Biochem. 1971. V. 40(1). P. 183–186.N.B. Cucakovich. Determination of Tween 80 in tissue culture media, vaccines, and related products // Anal. Biochem. 1971. V. 40(1). P. 183–186.N. Savjani, E. Babcock, H.K. Khor, A. Raghani. Use of ferric thiocyanate derivatization for quantification of polysorbate 80 in high concentration protein formulations // Talanta. 2014. V. 130. P. 542–546.N. Savjani, E. Babcock, H.K. Khor, A. Raghani. Use of ferric thiocyanate derivatization for quantification of polysorbate 80 in high concentration protein formulations // Talanta. 2014. V. 130. P. 542–546.H. Yang, B.W. Glickman, G. de Boer Johan. Sex-specific induction of mutations by PhIP in the kidney of male and female rats and its modulation by conjugated linoleic acid // Environ. Mol. Mutagen. 2002. V. 40(2). P. 116–121.H. Yang, B.W. Glickman, G. de Boer Johan. Sex-specific induction of mutations by PhIP in the kidney of male and female rats and its modulation by conjugated linoleic acid // Environ. Mol. Mutagen. 2002. V. 40(2). P. 116–121.J. Kim, J. Qiu. Quantitation of low concentrations of polysorbates in high protein concentration formulations by solid phase extraction and cobalt-thiocyanate derivatization // Anal. Chim. Acta. 2014. V. 806. P. 144–151.J. Kim, J. Qiu. Quantitation of low concentrations of polysorbates in high protein concentration formulations by solid phase extraction and cobalt-thiocyanate derivatization // Anal. Chim. Acta. 2014. V. 806. P. 144–151.J.A. Van der Hoeve. Analysis of textile auxiliary products // Rec. Trav. Chim. Pays-Bas. 1948. V. 67(8), P. 649–664.J.A. Van der Hoeve. Analysis of textile auxiliary products // Rec. Trav. Chim. Pays-Bas. 1948. V. 67(8), P. 649–664.Analytical Methods Committee. Examination of detergent preparations. Analyst. 1951. V. 76. P. 279–286.Analytical Methods Committee. Examination of detergent preparations. Analyst. 1951. V. 76. P. 279–286.U. Zoller, R. Romano. Determination of nonionic detergents in municipal wastewater // Environ. Int. 1983. V. 9(1). P. 55–61.U. Zoller, R. Romano. Determination of nonionic detergents in municipal wastewater // Environ. Int. 1983. V. 9(1). P. 55–61.E.G. Brown, T.J. Hayes. The absorptiometric determination of polyethyleneglycol mono-oleate // Analyst. V. 80(955). P. 755–767.E.G. Brown, T.J. Hayes. The absorptiometric determination of polyethyleneglycol mono-oleate // Analyst. V. 80(955). P. 755–767.ОФС.1.1.0012.15. Валидация аналитических методик // Государственная фармакопея РФ. XIII изд. Т. 1. – М., 2015.ОФС.1.1.0012.15. Валидация аналитических методик // Государственная фармакопея РФ. XIII изд. Т. 1. – М., 2015.

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