The applicability of high‐speed counter‐current chromatography for preparative separation of biosynthesis products: Glycosylation products as example

Abstract

Biosynthesis is a promising way to manufacture desired products, however, the purification of its final products is a tough work due to the huge amount of reaction matrix. Liquid stationary phase of high-speed counter-current chromatography could easily avoid the commonly disadvantages that occurred in traditional column chromatography in the field of biosynthesized products purification. This characteristic makes high-speed counter-current chromatography particularly applicable for final products separation in biosynthesis. In this study, the glycosylation products of Silybin B by one-pot glycosylation were successfully purified by high-speed counter-current chromatography to show the applicability of high-speed counter-current chromatography for preparative separation of biosynthesis products. An optimized n-hexane/ethyl acetate/methanol/water (2:5:2:3, v/v/v/v) system was applied in this study. As a result, four Silybin B glycosylation products, including 7 mg of Silybin B-5-O-β-D-glucoside (SG-1), 12 mg of Silybin B-3-O-β-D-glucoside (SG-2), 10 mg of Silybin B-7-O-β-D-glucoside (SG-3), and 24 mg of Silybin B-20-O-β-D-glucoside (SG-4), were simultaneously separated from 200 mg of glycosylation crude products, with the purity of 89.3, 95.2, 96.4, and 97.5%, respectively. Their structures were identified by spectroscopic analysis.

Impact of Preparative Isolation of C-Glycosylflavones Derived from Dianthus superbus on In Vitro Glucose Metabolism

“…Thus, traditional approaches are ineffective for isolating high-purity compounds. Recent studies have highlighted high-speed counter-current chromatography as an innovative method for differentiating standard compounds from natural products [ 14 , 15 , 16 ]. Nevertheless, this approach continues to be hindered by insufficient separation resolution, the requirement for estimating the partition coefficient, and its capability to isolate only moderately polar compounds.…”

Efficient Synthesis and In Vitro Hypoglycemic Activity of Rare Apigenin Glycosylation Derivatives

“…Compared with synthetic glycosylation, biotransformation catalyzed by glycosyltransferases or produced by microorganisms avoids repetitive, multistep, complex, and dangerous steps. The products of biotransformation are also more easily isolated and purified than products of chemical synthesis [ 8 , 9 ]. The 7-hydroxycoumarin derivatives esculetin, skimmin, and herniarin produced by Bacillus licheniformis DSM 13 and UDP-glycosyltransferase YjiC were found to have greater antibacterial and anticancer activity than 7-hydroxycoumarin [ 10 ].…”

An efficient strategy for large‐scale preparation of low polarity gingerols directly from ginger crude extract by high‐speed countercurrent chromatography with different rotation mode

“…HSCCC is a unique support-free liquid-liquid partition chromatography, which shows high recovery and efficiency as well as elimination of irreversible adsorption of the sample on the solid stationary phase occurred in the conventional chromatographic column and low risk of sample denaturation [15,16]. It is also very efficient for the separation of target components from crude extracts without any pretreatment and high sample loading is possible [17][18][19]. It is obtaining increasing popularity as a means for the separation of pharmaceutical substances, especially for the preparative separation of bioactive components from crude extracts of natural products.…”