Rapid separation of polysaccharides using a novel spiral coil column by high-speed countercurrent chromatography

Abstract

The separation of polysaccharides is time consuming. We developed and optimized a type-J counter-current chromatography system with a novel tri-rotor spiral coil column for the rapid separation of polysaccharides. The optimal composition of an aqueous PEG1000/K2 HPO4 /KH2 PO4 system was found to be 14:16:14 w/w/w where the lower phase was the mobile phase. Optimal performance was achieved at a column rotational speed, temperature, and flow rate of 1200 rpm, 45°C, and 3.0 mL/min, respectively. The mobile phase was pumped from the inner terminal in a ''head-to-tail'' elution mode. Polysaccharide LCP-1 (10.7 mg) was successfully obtained in high purity in one step from 50.0 mg of a crude polysaccharide extracted from the lychee fruit (Litchi chinensis) within 100 min. LCP-1 possess a number-average molecular weight and weight-average molecular weight of 1.05 × 10(5) and 1.59 × 10(5) kDa, respectively. The monosaccharide composition consists of the molar ratio of glucose, galactose, and arabinose of 1.3:3.5:1.

Gastrodia elata Blume Polysaccharides: A Review of Their Acquisition, Analysis, Modification, and Pharmacological Activities

“…In addition, high-speed countercurrent chromatography (HSCCC) has been widely applied for the separation of natural products [59]. As a new technique, HSCCC has been applied successfully to separate and purify polysaccharides [60,61,62,63,64]. These methods could be used in the separation of G. elata polysaccharides to obtain higher purity G. elata polysaccharides for research.…”

Spiral counter‐current chromatography: Design, development, application, and challenges

“…This is because the aqueous two‐phase system (ATPS) which was usually used to separate biological macromolecules results in a poor retention of stationary phase in the column due to its high viscosity and low interfacial tension. It is a daunting challenge for conventional HSCCC to separate and purify hydrophilic macromolecules . To help solve this problem, a novel HSCCC instrument named spiral CCC was developed .…”

Bioassay‐guided isolation of an active compound with protein tyrosine phosphatase 1B inhibitory activity from Sargassum fusiforme by high‐speed counter‐current chromatography

“…High‐speed counter‐current chromatography (HSCCC), a continuous liquid–liquid separation method which does not require a solid sorbent, eliminates the adsorptive loss of samples compared with conventional methods . So it is an appropriate method to separate the minor active ingredients from natural products . In the present study, a simple and feasible HSCCC method was established for the separation of bioactive compound from Sargassum fusiforme under the bioassay‐guidance of PTP1B inhibitory activity.…”