Development of a Method to Extract and Purify Target Compounds from Medicinal Plants in a Single Step: Online Hyphenation of Expanded Bed Adsorption Chromatography and Countercurrent Chromatography

Abstract

Pure compounds extracted and purified from natural sources are crucial to lead discovery and drug screening. This study presents a novel two-dimensional hyphenation of expanded bed adsorption chromatography (EBAC) and high-speed countercurrent chromatography (HSCCC) for extraction and purification of target compounds from medicinal plants in a single step. The EBAC and HSCCC were hyphenated via a six-port injection valve as an interface. Fractionation of ingredients of Salvia miltiorrhiza and Rhizoma coptidis was performed on the hyphenated system to verify its efficacy. An amount each of 52.9 mg of salvianolic acid B and 2.1 mg of rosmarinic acid was obtained from Salvia miltiorrhiza by the two-dimensional system in a single step. The purities of the targets were over 96% of salvianolic acid B and 74% of rosmarinic acid. An amount each of 4.6 mg of coptisine and 4.1 mg of berberine was obtained from Rhizoma coptidis each with 98% and 82% purity, respectively. The processing time was nearly 50% that of the multi-step method. These results indicate that the present method is a rapid and green way to harvest targets from medicinal plants in a single step.

Recovery and purification of chitosanase produced by Bacillus cereus using expanded bed adsorption and central composite design

“…Expanded bed adsorption (EBA) is an integrated process that combines solid–liquid separation, volume reduction by protein adsorption, and recovery of the product in a single unit operation . This technique has been successfully employed for biomolecule purification .…”

High-speed counter-current chromatography coupled online to high performance liquid chromatography–diode array detector–mass spectrometry for purification, analysis and identification of target compounds from natural products

“…Trap columns and injection valves have been successfully used as interface in on-line hyphenated system [21][22][23]. Among them, injection valves interface technique is the simplest to be easily operated and widely used.…”

Single‐step purification of cyclotides using affinity chromatography

“…Analyses of complex mixtures or crude extracts may be challenging because the target peptides or proteins are present at low concentrations. Therefore, multistep extraction and separation processes are often used for purification, which may be time‐consuming, costly and decrease yields . Some of these problems can be solved by using affinity chromatography.…”