biopreparatБИОпрепараты. Профилактика, диагностика, лечениеBiological Products. Prevention, Diagnosis, Treatment2221-996X2619-1156Scientific Centre for Expert Evaluation of Medicinal Products10.30895/2221-996X-2023-442biopreparat-442Research ArticleСтатьиОбзор методов определения соединений углеводной природы в биологических лекарственных препаратахQuantification methods for carbohydrate compounds in biologicals: a reviewhttps://orcid.org/0000-0003-0026-7365МинероА. С.MineroA. S.

Минеро Анастасия Сальвадоровна

Петровский б-р, д. 8, стр. 2, Москва, 127051

Anastasia S. Minero

8/2 Petrovsky Blvd, Moscow 127051

minero@expmed.ruhttps://orcid.org/0000-0003-0729-530XРуноваО. Б.RounovaO. B.

Рунова Ольга Борисовна, канд. хим. наук

Петровский б-р, д. 8, стр. 2, Москва, 127051

Olga B. Rounova, Cand. Sci. (Chem.)

8/2 Petrovsky Blvd, Moscow 127051

runova@expmed.ruhttps://orcid.org/0000-0001-5432-1887УстинниковаО. Б.UstinnikovaO. B.

Устинникова Ольга Борисовна, канд. биол. наук

Петровский б-р, д. 8, стр. 2, Москва, 127051

Olga B. Ustinnikova, Cand. Sci. (Biol.)

8/2 Petrovsky Blvd, Moscow 127051

ustinnikova@expmed.ruФедеральное государственное бюджетное учреждение «Научный центр экспертизы средств медицинского применения» Министерства здравоохранения Российской ФедерацииРоссияScientific Centre for Expert Evaluation of Medicinal ProductsRussian Federation202321022023232От традиционных биологических к высокотехнологичным лекарственным препаратам: вопросы разработки и применения194202Copyright © Минеро А.С., Рунова О.Б., Устинникова О.Б., 20232023Минеро А.С., Рунова О.Б., Устинникова О.Б.Minero A.S., Rounova O.B., Ustinnikova O.B.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.biopreparations.ru/jour/article/view/442

Соединения углеводной природы широко используются в качестве наполнителей и стабилизаторов в биологических лекарственных препаратах (БЛП). Наличие данных соединений в составе лекарственного препарата гарантирует стабильность действующего вещества в процессе производства, транспортирования и хранения. При этом нормирование содержания вспомогательных веществ и их количественное определение является фармакопейным требованием к оценке качества БЛП.

Цель работы — выявление перспективных методов для разработки методик количественного определения соединений углеводной природы в биологических лекарственных препаратах.

Проведен анализ нормативных документов зарегистрированных в Российской Федерации БЛП. Показано, что наиболее часто в качестве вспомогательных веществ используются полиолы (сорбитол и маннитол), моносахариды (глюкоза), дисахариды (трегалоза, сахароза, лактоза, мальтоза) как по отдельности, так и в смесях различного состава. На основании данных научной литературы рассмотрены методы количественного определения полиолов, моно- и дисахаридов, применяемые при оценке качества БЛП. Для количественного определения стабилизаторов углеводной природы применяют титриметрические, спектрофотометрические, ферментативные, хроматографические методы. Представлен анализ достоинств и недостатков данных методов. Показаны преимущества метода ионообменной высокоэффективной жидкостной хроматографии (ВЭЖХ) с амперометрическим детектированием и метода гидрофильной ВЭЖХ с рефрактометрическим детектированием и испарительным детектором светорассеяния, обладающих достаточной селективностью и способностью идентификации исходных веществ без дериватизации. Сделан вывод о перспективности разработки методик определения стабилизаторов углеводной природы на основе методов ионообменной и гидрофильной ВЭЖХ.

Carbohydrate compounds are widely used as fillers and stabilisers in biological products. When present, these compounds guarantee that the active pharmaceutical ingredient will remain stable during production, transportation, and storage. At the same time, pharmacopoeias standardise the excipient content and require that excipients should be quantified for assessing the quality of biological products.

The aim of the study was to identify promising methods for the development of quantification procedures for carbohydrate compounds in biological products.

The authors analysed regulatory documents for biological products approved in the Russian Federation. The most widely used excipients, both individually and in combinations, are polyols (sorbitol and mannitol), monosaccarides (glucose), and disaccharides (trehalose, sucrose, lactose, and maltose). Using literature data, the authors reviewed the methods used for quantifying polyols, monosaccharides, and disaccharides to assess the quality of biological products. Quantitative determination of carbohydrate stabilisers employs titrimetric, spectrophotometric, enzymatic, and chromatographic methods. This review presents an analysis of the advantages and disadvantages of these methods. It highlights the advantages of ionic HPLC with amperometric detection and hydrophilic HPLC with refractometric and evaporative light scattering detection, which are sufficiently selective and can identify substances without prior derivatisation. In conclusion, ionic and hydrophilic HPLC methods are a promising base for the development of quantification procedures for carbohydrate stabilisers.

биологические лекарственные препаратывспомогательные веществасорбитолманнитолглюкозатрегалозалактозасахарозамальтозаВЭЖХbiological productsexcipientssorbitolmannitolglucosetrehaloselactosesucrosemaltoseHPLCРабота выполнена в рамках государственного задания ФГБУ «НЦЭСМП» Минздрава России № 056-00052-23-00 на проведение прикладных научных исследований (номер государственного учета НИР 121022000147-4).The study reported in this publication was carried out as part of publicly funded research project No. 056-00052-23-00 and was supported by the Scientific Centre for Expert Evaluation of Medicinal Products (R&D public accounting 121022000147-4).ReferencesTonnis WF, Mensink MA, De Jager A, Van der Voort Maarschalk K, Frijlink HW, Hinrichs WL. Size and molecular flexibility of sugars determine the dosage stability of freeze-dried proteins. Mol Pharm. 2015;12(3):684–94. https://doi.org/10.1021/mp500423zTonnis WF, Mensink MA, De Jager A, Van der Voort Maarschalk K, Frijlink HW, Hinrichs WL. Size and molecular flexibility of sugars determine the dosage stability of freeze-dried proteins. Mol Pharm. 2015;12(3):684–94. https://doi.org/10.1021/mp500423zБлынская ЕВ, Тишков СВ, Алексеев КВ, Минаев СВ. Создание лиофилизата ГК-2 для приготовления раствора для инъекций с применением полиолов. Разработка и регистрация лекарственных средств. 2018;2:26–31.Blynskaya EV, Tishkov SV, Alekseyev KV, Minaev SV. Creation of liophilisate of GK-2 for preparation of solution for injections with use of polyols. Drug Development & Registration. 2018;(2):26–31 (In Russ.).Chang L, Shepherd D, Sun J, Ouellette D, Grant KL, Tang XC, Pikal MJ. Mechanism of protein stabilization by sugars during freeze-drying and storage: native structure preservation, specific interaction, and/or immobilization in a glassy matrix? J Pharm Sci. 2005;94(7):1427–44. https://doi.org/10.1002/jps.20364Chang L, Shepherd D, Sun J, Ouellette D, Grant KL, Tang XC, Pikal MJ. Mechanism of protein stabilization by sugars during freeze-drying and storage: native structure preservation, specific interaction, and/or immobilization in a glassy matrix? J Pharm Sci. 2005;94(7):1427–44. https://doi.org/10.1002/jps.20364Manas MG, Campos J, Sharninghausen LS, Lin E, Crabtree RH. Selective catalytic oxidation of sugar alcohols to lactic acid. Green Chemistry. 2015;17(1):594–600. https://doi.org/10.1039/C4GC01694GManas MG, Campos J, Sharninghausen LS, Lin E, Crabtree RH. Selective catalytic oxidation of sugar alcohols to lactic acid. Green Chemistry. 2015;17(1):594–600. https://doi.org/10.1039/C4GC01694GTarpan MA, De Cooman H, Sagstuen E, Waroquier M, Callens F. Identification of primary free radicals in trehalose dihdrate single crystals X-irradiated at 10 K. Phys Chem Chem Phys. 2011;13(23):11294–302. https://doi.org/10.1039/c0cp02616fTarpan MA, De Cooman H, Sagstuen E, Waroquier M, Callens F. Identification of primary free radicals in trehalose dihdrate single crystals X-irradiated at 10 K. Phys Chem Chem Phys. 2011;13(23):11294–302. https://doi.org/10.1039/c0cp02616fAmchara FZ, Faiz C, Chaouqi S, Khiraoui A, Benhmimou A, Guedira M, Guedira T. Effect of Stevia rebaudiana, sucrose and aspartame on human health: a comprehensive review. J Med Plants Stud. 2018;102(61):102–8.Amchara FZ, Faiz C, Chaouqi S, Khiraoui A, Benhmimou A, Guedira M, Guedira T. Effect of Stevia rebaudiana, sucrose and aspartame on human health: a comprehensive review. J Med Plants Stud. 2018;102(61):102–8.Hernández-López A, Sánchez Félix DA, Zuñiga Sierra Z, García Bravo I, Dinkova TD, Avila-Alejandre AX. Quantification of reducing sugars based on the qualitative technique of Benedict. ACS Omega. 2020;5(50):32403–10. https://doi.org/10.1021/acsomega.0c04467Hernández-López A, Sánchez Félix DA, Zuñiga Sierra Z, García Bravo I, Dinkova TD, Avila-Alejandre AX. Quantification of reducing sugars based on the qualitative technique of Benedict. ACS Omega. 2020;5(50):32403–10. https://doi.org/10.1021/acsomega.0c04467Pons A, Roca P, Aguiló C, Garcia FJ, Alemany M, Palou A. A method for the simultaneous determinations of total carbohydrate and glycerol in biological samples with the antrone reagent. J Biochem Biophys Methods. 1981;4(3–4):227–31. https://doi.org/10.1016/0165-022x(81)90060-9Pons A, Roca P, Aguiló C, Garcia FJ, Alemany M, Palou A. A method for the simultaneous determinations of total carbohydrate and glycerol in biological samples with the antrone reagent. J Biochem Biophys Methods. 1981;4(3–4):227–31. https://doi.org/10.1016/0165-022x(81)90060-9Haas M, Lamour S, Trapp O. Development of an advanced derivatization protocol for the unambiguous identification of monosaccharides in complex mixtures by gas and liquid chromatography. J Chromatogr A. 2018;1568:160–7. https://doi.org/10.1016/j.chroma.2018.07.015Haas M, Lamour S, Trapp O. Development of an advanced derivatization protocol for the unambiguous identification of monosaccharides in complex mixtures by gas and liquid chromatography. J Chromatogr A. 2018;1568:160–7. https://doi.org/10.1016/j.chroma.2018.07.015Alzweiri M, Al-Marabeh S, Bardaweel SK, Alfar R, Al-Hiari YM. Stability determination for cyclized 2,4-dinitrophenyl hydrazone derivative of glucose. J Anal Sci Technol. 2017;8:9. https://doi.org/10.1186/s40543-017-0117-xAlzweiri M, Al-Marabeh S, Bardaweel SK, Alfar R, Al-Hiari YM. Stability determination for cyclized 2,4-dinitrophenyl hydrazone derivative of glucose. J Anal Sci Technol. 2017;8:9. https://doi.org/10.1186/s40543-017-0117-xGonzalez NM, Fitch A, Al-Bazi J. Development of a RP-HPLC method for determination of glucose in Shewanella oneidensis cultures utilizing 1-phenyl-3-methyl-5-pyrazolone derivatization. PLoS One. 2020;15(3):e0229990. https://doi.org/10.1371/journal.pone.0229990Gonzalez NM, Fitch A, Al-Bazi J. Development of a RP-HPLC method for determination of glucose in Shewanella oneidensis cultures utilizing 1-phenyl-3-methyl-5-pyrazolone derivatization. PLoS One. 2020;15(3):e0229990. https://doi.org/10.1371/journal.pone.0229990Debebe A, Temesgen S, Redi-Abshiro M, Chandravanshi BS, Ele E. Improvement in analytical methods for determination of sugars in fermented alcoholic beverages. J Anal Methods Chem. 2018:4010298. https://doi.org/10.1155/2018/4010298Debebe A, Temesgen S, Redi-Abshiro M, Chandravanshi BS, Ele E. Improvement in analytical methods for determination of sugars in fermented alcoholic beverages. J Anal Methods Chem. 2018:4010298. https://doi.org/10.1155/2018/4010298Schenk J, Nagy G, Pohl NLB, Leghissa A, Smuts J, Schug KA. Identification and deconvolution of carbohydrates with gas chromatography-vacuum ultraviolet spectroscopy. J Chromatogr A. 2017;1513:210–21. https://doi.org/10.1016/j.chroma.2017.07.052Schenk J, Nagy G, Pohl NLB, Leghissa A, Smuts J, Schug KA. Identification and deconvolution of carbohydrates with gas chromatography-vacuum ultraviolet spectroscopy. J Chromatogr A. 2017;1513:210–21. https://doi.org/10.1016/j.chroma.2017.07.052Ruiz-Matutea AI, Hernández-Hernández O, Rodríguez-Sánchez S, Sanz ML, Martínez-Castro I. Derivatization of carbohydrates for GC and GC–MS analyses. J Chromatogr B Analyt Technol Biomed Life Sci. 2011;879(17–18):1226–40. https://doi.org/10.1016/j.jchromb.2010.11.013Ruiz-Matutea AI, Hernández-Hernández O, Rodríguez-Sánchez S, Sanz ML, Martínez-Castro I. Derivatization of carbohydrates for GC and GC–MS analyses. J Chromatogr B Analyt Technol Biomed Life Sci. 2011;879(17–18):1226–40. https://doi.org/10.1016/j.jchromb.2010.11.013Mechri B, Tekaya M, Cheheb H, Hammami M. Determination of mannitol sorbitol and myo-inositol in olive tree roots and rhizospheric soil by gas chromatography and effect of severe drought conditions on their profiles. J Chromatogr Sci. 2015;53(10):1631–8. https://doi.org/10.1093/chromsci/bmv066Mechri B, Tekaya M, Cheheb H, Hammami M. Determination of mannitol sorbitol and myo-inositol in olive tree roots and rhizospheric soil by gas chromatography and effect of severe drought conditions on their profiles. J Chromatogr Sci. 2015;53(10):1631–8. https://doi.org/10.1093/chromsci/bmv066Akyüza E, Başkan KS, Tütem E, Apak R. High performance liquid chromatographic method with post-column detection for quantification of reducing sugars in foods. J Chromatogr A. 2021;1660:462664. https://doi.org/10.1016/j.chroma.2021.462664Akyüza E, Başkan KS, Tütem E, Apak R. High performance liquid chromatographic method with post-column detection for quantification of reducing sugars in foods. J Chromatogr A. 2021;1660:462664. https://doi.org/10.1016/j.chroma.2021.462664Hadjikinova R, Petkova N, Hadjikinov D, Denev P, Hrusavov D. Development and validation of HPLC-RID method for determination of sugars and polyols. J Pharm Sci Research. 2017;9(8):1263–9.Hadjikinova R, Petkova N, Hadjikinov D, Denev P, Hrusavov D. Development and validation of HPLC-RID method for determination of sugars and polyols. J Pharm Sci Research. 2017;9(8):1263–9.Liu W, Xiao X, Song Z, Cheng X, Wei F, Liu Y, Zhang Y. Research on determination of mannitol in Bailing capsules by HPLC-RID instead of titration. Chin J Pharm Anal. 2014;34(1):159–62.Liu W, Xiao X, Song Z, Cheng X, Wei F, Liu Y, Zhang Y. Research on determination of mannitol in Bailing capsules by HPLC-RID instead of titration. Chin J Pharm Anal. 2014;34(1):159–62.Forget R, Spagnoli S. Excipient quantitation and drug distribution during formulation optimization. J Pharm Biomed Anal. 2006;41(3):1051–5. https://doi.org/10.1016/j.jpba.2006.01.039Forget R, Spagnoli S. Excipient quantitation and drug distribution during formulation optimization. J Pharm Biomed Anal. 2006;41(3):1051–5. https://doi.org/10.1016/j.jpba.2006.01.039Simonzadeh N, Ronsen B. An isocratic HPLC method for the determination of sorbitol and glycerol in pharmaceutical formulations. J Chromatogr Sci. 2012;50(7):644–7. https://doi.org/10.1093/chromsci/bms044Simonzadeh N, Ronsen B. An isocratic HPLC method for the determination of sorbitol and glycerol in pharmaceutical formulations. J Chromatogr Sci. 2012;50(7):644–7. https://doi.org/10.1093/chromsci/bms044Fang T, Cai Y, Yang Q, Ogutu CO, Liao L, Han Y Analysis of sorbitol content variation in wild and cultivated apples. J Sci Food Agric. 2020;100(1):139–44. https://doi.org/10.1002/jsfa.10005Fang T, Cai Y, Yang Q, Ogutu CO, Liao L, Han Y Analysis of sorbitol content variation in wild and cultivated apples. J Sci Food Agric. 2020;100(1):139–44. https://doi.org/10.1002/jsfa.10005Sławińska A, Jabłońska-Ryś E, Stachniuk A. High-performance liquid chromatography determination of free sugars and mannitol in mushrooms using corona charged aerosol detection. Food Anal Meth. 2021;14:209–16. https://doi.org/10.1007/s12161-020-01863-8Sławińska A, Jabłońska-Ryś E, Stachniuk A. High-performance liquid chromatography determination of free sugars and mannitol in mushrooms using corona charged aerosol detection. Food Anal Meth. 2021;14:209–16. https://doi.org/10.1007/s12161-020-01863-8Pitsch J, Weghuber J. Hydrophilic interaction chromatography coupled with charged aerosol detection for simultaneous quantitation of carbohydrates, polyols and ions in food and beverages. Molecules. 2019;24(23):4333. https://doi.org/10.3390/molecules24234333Pitsch J, Weghuber J. Hydrophilic interaction chromatography coupled with charged aerosol detection for simultaneous quantitation of carbohydrates, polyols and ions in food and beverages. Molecules. 2019;24(23):4333. https://doi.org/10.3390/molecules24234333Cintrón JM, Risley DS. Hydrophilic interaction chromatography with aerosol-based detectors (ELSD, CAD, NQAD) for polar compounds lacking a UV chromophore in an intravenous formulation. J Pharm Biomed Anal. 2013;78–79:14–8. https://doi.org/10.1016/j.jpba.2013.01.022Cintrón JM, Risley DS. Hydrophilic interaction chromatography with aerosol-based detectors (ELSD, CAD, NQAD) for polar compounds lacking a UV chromophore in an intravenous formulation. J Pharm Biomed Anal. 2013;78–79:14–8. https://doi.org/10.1016/j.jpba.2013.01.022Koh DW, Park JW, Lim JH, Yea MJ, Bang DY. A rapid method for simultaneous quantification of 13 sugars and sugar alcohols in food products by UPLC-ELSD. Food Chem. 2018;240:694–700. https://doi.org/10.1016/j.foodchem.2017.07.142Koh DW, Park JW, Lim JH, Yea MJ, Bang DY. A rapid method for simultaneous quantification of 13 sugars and sugar alcohols in food products by UPLC-ELSD. Food Chem. 2018;240:694–700. https://doi.org/10.1016/j.foodchem.2017.07.142Risley DS, Yang WQ, Peterson JA. Analysis of mannitol in pharmaceutical formulations using hydrophilic interaction liquid chromatography with evaporative light-scattering detection. J Sep Sci. 2006;29(2):256–64. https://doi.org/10.1002/jssc.200500253Risley DS, Yang WQ, Peterson JA. Analysis of mannitol in pharmaceutical formulations using hydrophilic interaction liquid chromatography with evaporative light-scattering detection. J Sep Sci. 2006;29(2):256–64. https://doi.org/10.1002/jssc.200500253Pissmennõi D, Kiritsenko V, Marhivka J, Kütt ML, Vilu R. Development and optimisation of HILIC-LC-MS method for determination of carbohydrates in fermentation samples. Molecules. 2021;26(12):3669. https://doi.org/10.3390/molecules26123669Pissmennõi D, Kiritsenko V, Marhivka J, Kütt ML, Vilu R. Development and optimisation of HILIC-LC-MS method for determination of carbohydrates in fermentation samples. Molecules. 2021;26(12):3669. https://doi.org/10.3390/molecules26123669Bennett R, Olesik SV. Gradient separation of oligosaccharides and suppressing anomeric mutarotation with enhance-fluidity liquid hydrophilic interaction chromatography. Anal Chim Acta. 2017;960:151–9. https://doi.org/10.1016/j.aca.2017.01.006Bennett R, Olesik SV. Gradient separation of oligosaccharides and suppressing anomeric mutarotation with enhance-fluidity liquid hydrophilic interaction chromatography. Anal Chim Acta. 2017;960:151–9. https://doi.org/10.1016/j.aca.2017.01.006Chen H, Chen W, Hong B, Zhang Y, Hong Z, Yi R. Determination of trehalose by ion chromatography and its application to a pharmacokinetic study in rats after intramuscular injection. Biomed Chromatogr. 2018;32(12):e4355. https://doi.org/10.1002/bmc.4355Chen H, Chen W, Hong B, Zhang Y, Hong Z, Yi R. Determination of trehalose by ion chromatography and its application to a pharmacokinetic study in rats after intramuscular injection. Biomed Chromatogr. 2018;32(12):e4355. https://doi.org/10.1002/bmc.4355Pico J, Martínez MM, Martín MT, Gómez M. Quantification of sugars in wheat flours with an HPAEC-PAD method. Food Chem. 2015;173:674–81. https://doi.org/10.1016/j.foodchem.2014.10.103Pico J, Martínez MM, Martín MT, Gómez M. Quantification of sugars in wheat flours with an HPAEC-PAD method. Food Chem. 2015;173:674–81. https://doi.org/10.1016/j.foodchem.2014.10.103

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