Box-Behnken Design for Extraction Optimization Followed by High Performance Countercurrent Chromatography: Production of a Flavonoid-enriched Fraction from Achyrocline Satureioides

Abstract

The biological properties of Achyrocline satureioides have been mostly ascribed to its major flavonoids quercetin (QCT), luteolin (LUT), and 3-O-methylquercetin (3OMQ). The present study aimed to optimize the extraction by dynamic maceration of the major phenolic compounds in order to obtain in a subsequent step a flavonoid-enriched fraction (FEF) using high performance countercurrent chromatography (HPCCC). A 3-level Box-Behnken design (BBD) was applied to maximize the extraction of the substances, using the plant : solvent ratio (X₁ ), extraction time (X₂ ), and ethanol concentration (X₃ ) as factors. One-step HPCCC semipreparative separation with a solvent system composed of hexane : ethyl acetate : methanol : water (0.9 : 0.9 : 0.8 : 1.0, v/v) was employed to obtain the FEF. The second-order polynomial model was able to fit the experimental data adequately. The linear and quadratic terms of X₃ were the most significant factors that affected all the responses. The positive linear term of X₃ indicated a substantial increase in extraction yield, while the negative quadratic term showed a nonlinear tendency. Linear terms of X₁ suggested a tendency to solvent saturation, except for QCT. The terms of X₂ did not affect the responses substantially. The HPCCC method was found to be efficient and rapid for separating the FEF with 71% (w/w) flavonoid content. Overall, the developed extraction procedure coupled with HPCCC proved to be efficient for obtaining an enriched fraction with a very high content of flavonoids from A. satureioides.

Key words

Achyrocline satureioides - Asteraceae - Box-Behnken design - extraction optimization - flavonoid - high performance countercurrent chromatography

• References

• 1 Simões CMO, Schenkel EP, Bauer L, Langeloh A. Pharmacological investigations on Achyrocline satureioides (Lam.) D.C. Compositae . J Ethnopharmacol 1988; 22: 281-293
• 2 De Souza KCB, Bassani VL, Schapoval EES. Influence of excipients and technological process on anti-inflammatory activity of QCT and Achyrocline satureioides (Lam.) D.C. extracts by oral route. Phytomedicine 2007; 14: 102-108
• 3 Oliveira SGD, Moura FRR, Demarco FF, Nascente OS, Del Pino FAB, Lund RG. An ethnomedicinal survey on phytotherapy with professionals and patients from basic care units in the brazilian unified health system. J Ethnopharmacol 2012; 140: 428-437
• 4 Retta D, Dellacassa E, Villamil J, Suárez AS, Bandoni AL. Marcela, a promising medicinal and aromatic plant from Latin America: a review. Ind Crops Prod 2012; 38: 27-38
• 5 Kadarian C, Broussalis AM, Miño J, Lopez P, Gorzalczany S, Ferraro G, Acevedo C. Hepatoprotective activity of Achyrocline satureioides (Lam.) D.C. Pharmacol Res 2002; 45: 57-61
• 6 Sabini MC, Escobar FM, Tonn CE, Zanon SM, Contigiani MS, Sabini LI. Evaluation of antiviral activity of aqueous extracts from Achyrocline satureioides against western equine encephalitis virus. Nat Prod Res 2012; 26: 405-415
• 7 Balestrin LA, Bidone J, Bortolin RC, Moresco KS, Moreira JC, Teixeira HF. Protective effect of a hydrogel containing Achyrocline satureioides extract-loaded nanoemulsion against UV-induced skin damage. J Photochem Photobiol B 2016; 163: 269-276
• 8 Souza PO, Bianchi SE, Figueiró F, Heimfarth L, Moresco KS, Gonçalves RM, Hoppe JB, Klein CP, Salbego CG, Gelain DP, Bassani VL, Zanotto Filho A, Moreira JCF. Anticancer activity of flavonoids isolated from Achyrocline satureioides in gliomas cell lines. Toxicol In Vitro 2018; 4: 23-33
• 9 Bianchi SE, Kaiser S, Pittol V, Doneda E, De Souza KCB, Bassani VL. Semipreparative isolation and purification of phenolic compounds from Achyrocline satureioides (Lam) D.C. by high-performance counter-current chromatography. Phytochem Anal 2019; 30: 182-192
• 10 Goltz C, Ávila S, Barbieri JB, Igarashi-Mafra L, Mafra MR. Ultrasound-assisted extraction of phenolic compounds from macela (Achyrolcine satureioides) extracts. Ind Crops Prod 2018; 115: 227-234
• 11 Montgomery DC. Design and Analysis of Experiments, 8th edition. New Jersey: John Wiley & Sons, Inc.; 2013: 478-553
• 12 Kaiser S, Verza SG, Moraes RC, Pittol V, Peñaloza EMC, Pavei C, Ortega GG. Extraction optimization of polyphenols, oxindole alkaloids and quinovic acid glycosides from catʼs claw bark by Box–Behnken design. Ind Crops Prod 2013; 48: 153-161
• 13 Wang X, Wu Y, Chen G, Yue W, Liang Q, Wu Q. Optimisation of ultrasound assisted extraction of phenolic compounds from Sparganii rhizoma with response surface methodology. Ultrason Sonochem 2013; 20: 846-854
• 14 Jang S, Lee AY, Lee AR, Choi G, Kim HK. Optimization of ultrasound-assisted extraction of glycyrrhizic acid from licorice using response surface methodology. Integr Med Res 2017; 6: 388-394
• 15 Ferreira SLC, Bruns RE, Ferreira HS, Matos GD, David JM, Brandão GC, da Silva EGP, Portugal LA, dos Reis PS, Souza AS, dos Santos WNL. Box-Behnken design: an alternative for the optimization of analytical methods. Anal Chim Acta 2007; 597: 179-186
• 16 Senna EL, Petrovick PR, Ortega GG, Bassani VL. Preparation and characterization of spray-dried powders from Achyrocline satureioides (Lam.) DC extracts. Phytother Res 1997; 11: 123-127
• 17 Bidone J, Argenta DF, Kratz J, Pettenuzzo LF, Horn AP, Koester LS, Bassani VL, Simões CMO, Teixeira HF. Antiherpes activity and skin/mucosa distribution of flavonoids from Achyrocline satureioides extract incorporated into tropical nanoemulsions. BioMed Res Int 2015; 2015: 1-7
• 18 Carini JP, Leitão GG, Schneider PH, Santos CC, Costa FN, Holzschuh MH, Klamt F, Bassani VL. Isolation of ACB from Achyrocline satureioides by high-speed countercurrent chromatography. Curr Pharm Biotechnol 2015; 16: 66-71
• 19 Duo-Long D, Yuan-Yuan Z, Xiao-Fen C, Xin-Yi H, Shi-Lan F. Advances in application of high-speed countercurrent chromatography in separation and purification of flavonoids. Chin J Anal Chem 2011; 39: 269-275
• 20 Friesen JB, McAlpine JB, Chen SN, Pauli GF. Countercurrent separation of natural products: an update. J Nat Prod 2015; 78: 1765-1796
• 21 Bojczuk M, Zyzelewicz D, Hodurek P. Centrifugal partition chromatography – a review of recent applications and some classic references. J Sep Sci 2017; 40: 1597-1609
• 22 Ito Y. Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography. J Chromatogr A 2005; 1065: 145-168
• 23 Conway WD. Counter-current chromatography: simple process and confusing terminology. J Chromatogr A 2011; 1218: 6015-6023
• 24 Berthod A, Ruiz-Angel MJ, Carda-Broch S. Elution-extrusion countercurrent chromatography. Use of the liquid nature of the stationary phase to extend the hydrophobicity window. Anal Chem 2003; 75: 5886-5894
• 25 Marston A, Hostettmann K. Developments in the application of countercurrent chromatography to plant analysis. J Chromatogr A 2006; 1112: 181-194
• 26 Ćujić N, Šavikin K, Janković T, Pljevljakusić D, Zdunić G, Ibrić S. Optimization of polyphenols extraction from dried chokeberry using maceration as traditional technique. Food Chem 2016; 194: 135-142
• 27 Jovanović AA, Dordević VB, Zdunić GM, Pljevljakušić DS, Savikin KP, Godevac DM, Bugarski BM. Optimization of the extraction process of polyphenols from Thymus serpyllum L. herb using maceration, heat- and ultrasound-assisted techniques. Sep Purif Technol 2017; 179: 369-380
• 28 Chizzola R, Michitsch H, Franz C. Antioxidative properties of Thymus vulgaris leaves: comparison of different extracts and essential oil chemotypes. J Agric Food Chem 2008; 56: 6897-6904
• 29 Sutherland IA. Recent progress on the industrial scale-up of counter-current chromatography. J Chromatogr A 2007; 1151: 6-13
• 30 Costa FN, Leitão GG. Strategies of solvent system selection for the isolation of flavonoids by countercurrent chromatography. J Sep Sci 2010; 33: 336-347
• 31 Bidone J, Bica VC, Petrovick PR, Simoes CM, Koester LS, Bassani VL, Teixeira HF. Simultaneous quantification of flavonoids from Achyrocline satureioides by a polar-reversed phase LC method-application to skin permeation/retention studies. Pharmazie 2014; 69: 5-9