Direct from solid natural products to pure compounds in a single step: Coupling online extraction with high‐speed counter‐current chromatography

Marinobufagenin is a bufadienolide compound detected mainly in skin and parotoid gland secretions of Rhinella marina (L.) toad. Bufadienolides regulate the Na+/K+‐ATPase pump by inhibiting the cardiotonic steroid dependent‐site and act as cardiac inotropes with vasoconstrictive properties. Marinobufagenin and other bufadienolides, such as telocinobufagin and bufalin, are thought to be found endogenously in mammals in salt‐sensitive hypertensive states such as essential hypertension, congestive heart‐failure, and preeclampsia. The role of marinobufagenin as antimicrobial agent and its cytotoxic potential have also been recognized. The particular interest around marinobufagenin prompts us to consider the Rhinella marina toad venom as a possible source for molecules with pharmacological and/or diagnostic potential. In this article, two different approaches of extraction and purification of marinobufagenin from Rhinella marina (L.) venom are studied: (i) Preparative thin‐layer chromatography combined to mass spectrometry and/or ultraviolet detection and (ii) solid‐phase extraction coupled with fractionation on high‐performance liquid chromatography. Different chromatographic conditions are tested for each approach. The solid‐phase extraction combined with high‐performance liquid chromatography fractionation approach was preferred as it offered a greater yield, was less time‐consuming and allowed us to selectively isolate marinobufagenin. Both protocols aim to provide efficient and convenient methods for toad venom extraction, based on an easily automatable and systematized strategy.

Section: Discussionmentioning

confidence: 99%

Marinobufagenin extraction from Rhinella marina toad glands: Alternative approaches for a systematized strategy

Lenaerts

Wells

Hambye

et al. 2019

“…Natural products have been used in medical purposes for hundreds of years, which are good exogenous candidates (e.g. antioxidant, anti‐inflammatory, antimicrobial, and antitumor drugs) [1]. However, natural products usually contain a large number of compounds, and the traditional bioassay‐guided procedure for purification of bioactive compounds is labor and resource intensive [2].…”

Section: Introductionmentioning

confidence: 99%

“…HPLC–QTOF‐MS/MS has been widely used for identification of bioactive compounds with high separation resolution, unambiguous precursor, and product ions information [1,4]. Nevertheless, produced massive MS data make manual annotation a rather tedious and difficult work [4].…”

Section: Introductionmentioning

confidence: 99%

“…Comparing with repeated silica gel column chromatography, HSCCC eliminates complications, such as irreversible adsorption, peak tailing, deactivation, and contamination from the solid support [10‒13]. Moreover, online extraction (OLE) technology coupling with HSCCC was a rapid and high‐yield alternative technique to simultaneously extract and purify bioactive compounds from natural products [1]. Li et al.…”

Section: Introductionmentioning

confidence: 99%

“…Guo et al. designed OLE−HSCCC system for efficient purification of two phenolic compounds from Mangnolia officinalis and three alkaloids from Piper nigrum [1]. Results demonstrated that OLE−HSCCC can largely improve separation efficiency and sample recovery.…”

Section: Introductionmentioning

confidence: 99%

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Diagnostic ion filtering targeted screening and isolation of anti‐inflammatory iridoid glycosides from Hedyotis diffusa

Shi

Tong

et al. 2021

Self Cite

Efficient and targeted screening and isolation of bioactive compounds from complex natural products is still a challenging work. Herein, diagnostic ion filtering based high‐performance liquid chromatography–quadrupole time‐of‐flight−tandem mass spectrometry was firstly developed to screen six main iridoid glycosides from Hedyotis diffusa. Then, online extraction‒high‐speed counter current chromatography was proposed for targeted enrichment and preparative isolation using ethyl acetate/n‐butanol/water (4.5:0.5:5, v/v/v) as solvent system. After that, Sephadex LH‐20 column chromatography using methanol as solvent system was selected for further purification of six iridoid glycosides with purities over 98%. They were finally identified as monotropein, desacetylasperuloside acid, asperuloside, 6‐O‐(Z)‐p‐coumaroyl scandoside methyl ester, 6‐O‐(Z)‐feruloyl scandoside methyl ester, and 6‐O‐(E)‐p‐coumaroyl scandoside methyl ester. And their anti‐inflammatory activities were evaluated and confirmed by lipopolysaccharide activated RAW 264.7 macrophages. Obviously, the results provide a scientific basis for the potential applications of H. diffusa, and the developed methodology is efficient and reliable for targeted screening and isolation of bioactive compounds from natural products.

Effective on‐line high‐speed shear dispersing emulsifier technique coupled with high‐performance countercurrent chromatography method for simultaneous extraction and isolation of carotenoids from Lycium barbarum L. fruits

Gong

Huang

Liu

et al. 2020

An efficient combination strategy based on high‐speed shear dispersing emulsifier technique and high‐performance countercurrent chromatography was developed for on‐line extraction and isolation of carotenoids from the fruits of Lycium barbarum. In this work, the high‐speed shear dispersing emulsifier technique has been employed to extract crude extracts using the upper phase of high‐performance countercurrent chromatography solvent system composed of n‐hexane−dichloromethane−acetonitrile (10:4:6.5, v/v) as the extraction solvent. At the separation stage, the high‐performance counter‐current chromatography process adopts elution–extrusion mode and the upper phase of the solvent system as stationary phase (reverse‐phase mode). As a result, three compounds including zeaxanthin, zeaxanthin monopalmitate, and zeaxanthin dipalmitate with purities of 89, 90, and 93% were successfully obtained in one extraction‐separation operation within 120 min. The targeted compounds were analyzed and identified by high‐performance liquid chromatography, mass spectrometry, and NMR spectroscopy. The results indicated that the present on‐line combination method could serve as a simple, rapid, and effective way to achieve weak polar and unstable compounds from natural products.