Simultaneous purification of minor components in natural products using twin-column recycling chromatography with a step solvent gradient

doi:10.1016/j.cjche.2024.02.004

Abstract

Abstract: The isolation of minor components from complex natural product matrices presents a significant challenge in the field of purification science due to their low concentrations and the presence of structurally similar compounds. This study introduces an optimized twin-column recycling chromatography method for the efficient and simultaneous purification of these elusive constituents. By introducing water at a small flowing rate between the twin columns, a step solvent gradient is created, by which the leading edge of concentration band would migrate at a slower rate than the trailing edge as it flowing from the upstream to downstream column. Hence, the band broadening is counterbalanced, resulting in an enrichment effect for those minor components in separation process. Herein, two target substances, which showed similar peak position in high performance liquid chromatography (HPLC) and did not exceed 1.8% in crude paclitaxel were selected as target compounds for separation. By using the twin-column recycling chromatography with a step solvent gradient, a successful purification was achieved in getting the two with the purity almost 100%. We suggest this method is suitable for the separation of most components in natural produces, which shows higher precision and recovery rate compared with the common lab-operated separation ways for natural products (thin-layer chromatography and prep-HPLC).

Key words: Solvent gradient, Twin-column recycling chromatography, Purification, Minor component, Natural products

摘要： The isolation of minor components from complex natural product matrices presents a significant challenge in the field of purification science due to their low concentrations and the presence of structurally similar compounds. This study introduces an optimized twin-column recycling chromatography method for the efficient and simultaneous purification of these elusive constituents. By introducing water at a small flowing rate between the twin columns, a step solvent gradient is created, by which the leading edge of concentration band would migrate at a slower rate than the trailing edge as it flowing from the upstream to downstream column. Hence, the band broadening is counterbalanced, resulting in an enrichment effect for those minor components in separation process. Herein, two target substances, which showed similar peak position in high performance liquid chromatography (HPLC) and did not exceed 1.8% in crude paclitaxel were selected as target compounds for separation. By using the twin-column recycling chromatography with a step solvent gradient, a successful purification was achieved in getting the two with the purity almost 100%. We suggest this method is suitable for the separation of most components in natural produces, which shows higher precision and recovery rate compared with the common lab-operated separation ways for natural products (thin-layer chromatography and prep-HPLC).

关键词: Solvent gradient, Twin-column recycling chromatography, Purification, Minor component, Natural products

Guangxia Jin, Yuxue Wu, Feng Wei. Simultaneous purification of minor components in natural products using twin-column recycling chromatography with a step solvent gradient[J]. Chinese Journal of Chemical Engineering, 2024, 69(5): 212-219.

References

[1] T. Rodrigues, D. Reker, P. Schneider, G. Schneider, Counting on natural products for drug design, Nat. Chem. 8(6)(2016)531-541.
[2] J.Y. Hong, Natural product synthesis at the interface of chemistry and biology, Chemistry 20(33)(2014)10204-10212.
[3] A.G. Atanasov, S.B. Zotchev, V.M. Dirsch, I.E. Orhan, M. Banach, J.M. Rollinger, D. Barreca, W. Weckwerth, R. Bauer, E.A. Bayer, M. Majeed, A. Bishayee, V. Bochkov, G.K. Bonn, N. Braidy, F. Bucar, A. Cifuentes, G. D'Onofrio, M. Bodkin, M. Diederich, A.T. Dinkova-Kostova, T. Efferth, K. El Bairi, N. Arkells, T.P. Fan, B.L. Fiebich, M. Freissmuth, M.I. Georgiev, S. Gibbons, K.M. Godfrey, C.W. Gruber, J. Heer, L.A. Huber, E. Ibanez, A. Kijjoa, A.K. Kiss, A.P. Lu, F.A. Macias, M.J.S. Miller, A. Mocan, R. Muller, F. Nicoletti, G. Perry, V. Pittala, L. Rastrelli, M. Ristow, G.L. Russo, A.S. Silva, D. Schuster, H. Sheridan, K. Skalicka-Wozniak, L. Skaltsounis, E. Sobarzo-Sanchez, D.S. Bredt, H. Stuppner, A. Sureda, N.T. Tzvetkov, R.A. Vacca, B.B. Aggarwal, M. Battino, F. Giampieri, M. Wink, J.L. Wolfender, J.B. Xiao, A.W.K. Yeung, G. Lizard, M.A. Popp, M. Heinrich, I. Berindan-Neagoe, M. Stadler, M. Daglia, R. Verpoorte, C.T. Supuran, the International Natural Product Sciences Taskforce, Natural products in drug discovery:Advances and opportunities, Nat. Rev. Drug Discov. 20(2021)200-216.
[4] M. Grigalunas, S. Brakmann, H. Waldmann, Chemical evolution of natural product structure, J. Am. Chem. Soc. 144(8)(2022)3314-3329.
[5] F. Rabel, J. Sherma, Review of the state of the art of preparative thin-layer chromatography, J. Liq. Chromatogr. Relat. Technol. 40(4)(2017)165-176.
[6] T. Maswanna, C. Maneeruttanarungroj, Identification of major carotenoids from green alga Tetraspora sp. CU2551:Partial purification and characterization of lutein, canthaxanthin, neochrome, and β-carotene, World J. Microbiol. Biotechnol. 38(8)(2022)129.
[7] P. Dhama, X.T. Ding, A. Sharma, Exploring phytochemicals of Withania somnifera from different vicinity for functional foods, J. Future Foods 3(3)(2023)278-287.
[8] E. Hanan, N. Hasan, S. Zahiruddin, S. Ahmad, V. Sharma, F.J. Ahmad, Metabolite profiling and Ameliorative effect of quince (Cydonia oblonga) leaves against doxorubicin induced cardiotoxicity in Wistar rats, Food Biosci. 53(2023)102691.
[9] N.S. Surve, A.B. Thomas, R.P. Bhole, C.Y. Patil, Flash chromatography and semi-preparative HPLC:Review on the applications and recent advancements over the last decade, Eurasian J. Chem. 109(1)(2023)20-30.
[10] Z. Latif, S.D. Sarker, Isolation of natural products by preparative high performance liquid chromatography (prep-HPLC), Methods Mol. Biol. 864(2012)255-274.
[11] A. Aqel, N. Alkatheri, A. Ghfar, A.M. Alsubhi, Z.A. ALOthman, A.Y. Badjah-Hadj-Ahmed, Preparation of value-added metal-organic frameworks for high-performance liquid chromatography. Towards green chromatographic columns, J. Chromatogr. A 1638(2021)461857.
[12] H.M. Yuan, L.H. Zhang, W.B. Zhang, Z. Liang, Y.K. Zhang, Columns switch recycling size exclusion chromatography for high resolution protein separation, J. Chromatogr. A 1216(42)(2009)7024-7032.
[13] W.Q. Zhao, G. Yang, F.Y. Zhong, N. Yang, X. Zhao, Y.P. Qi, G.R. Fan, Isolation and purification of diastereoisomeric flavonolignans from silymarin by binary-column recycling preparative high-performance liquid chromatography, J. Sep. Sci. 37(17)(2014)2300-2306.
[14] G. Nagy, T.Y. Peng, D.E.K. Kabotso, M.V. Novotny, N.L.B. Pohl, Protocol for the purification of protected carbohydrates:Toward coupling automated synthesis to alternate-pump recycling high-performance liquid chromatography, Chem. Commun. 52(90)(2016)13253-13256.
[15] C. Shi, Z.Y. Gao, Q.L. Zhang, S.J. Yao, N.K.H. Slater, D.Q. Lin, Model-based process development of continuous chromatography for antibody capture:A case study with twin-column system, J. Chromatogr. A 1619(2020)460936.
[16] J. Heinonen, J. Tamper, M. Laatikainen, T. Sainio, Steady-state recycling chromatography in the purification of weakly acidic lignocellulosic hydrolysates, Sep. Purif. Technol. 210(2019)670-681.
[17] F. Wei, Z.W. Yang, Y.X. Zhao, Q. Wang, A twin-column recycling chromatography with solvent gradient for reinforcing the isolation of minor impurities, AIChE. J. 65(2)(2019)702-711.
[18] X. Zhang, F. Wei, Y.X. Zhao, Q. Wang, Fine optimization of twin-column recycling chromatography with a solvent gradient for the removal of minor impurities, J. Chromatogr. A 1609(2020)460443.
[19] F. Gritti, M. Leal, T. McDonald, M. Gilar, Ideal versus real automated twin column recycling chromatography process, J. Chromatogr. A 1508(2017)81-94.
[20] F. Gritti, M. Basile, S. Cormier, M. Fogwill, M. Gilar, T. McDonald, F. Riley, Q. Yan, Semi-preparative high-resolution recycling liquid chromatography, J. Chromatogr. A 1566(2018)64-78.
[21] S.C. Siu, C. Chia, Y. Mok, P. Pattnaik, Packing of large-scale chromatography columns with irregularly shaped glass based resins using a stop-flow method, Biotechnol. Prog. 30(6)(2014)1319-1325.
[22] C. Heuer, A. Seidel-Morgenstern, P. Hugo, Experimental investigation and modelling of closed-loop recycling in preparative chromatography, Chem. Eng. Sci. 50(7)(1995)1115-1127.
[23] H.P. Li, F.Y. Zhang, Q.Y. Jin, T.J. Zhu, Preparative separation and purification of Cyclosporin D from fungus Hypoxylon Spp. by improved closed-loop recycling counter-current chromatography, J. Chromatogr. A 1649(2021)462221.
[24] F. Wei, Y. Hu, Y.X. Zhao, Y.X. Wu, G.X. Jin, Purification and concentration of minor impurity in the bulk drug by step-gradient twin-column recycling chromatography, Ind. Eng. Chem. Res. 62(15)(2023)6241-6250.