Nat Prod Sci.  2018 Dec;24(4):288-292. 10.20307/nps.2018.24.4.288.

Efficient Isolation of Dihydrophaseic acid 3′-O-β-D-Glucopyranoside from Nelumbo nucifera Seeds Using High-performance Countercurrent Chromatography and Reverse-phased High-performance Liquid Chromatography

• Rho T
• Yoon KD

• KMID: 2432425
• DOI: http://doi.org/10.20307/nps.2018.24.4.288

Abstract

High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed to isolate dihydrophaseic acid 3"²-O-β-D-glucopyranoside (DHPAG) from the extract of Nelumbo nucifera seeds. Enriched DHPAG sample (2.3 g) was separated by HPCCC using ethyl acetate/n-butanol/water system (6:4:10, v/v/v, normal-phase mode, flow rate: 4.0 mL/min) to give 23.1 mg of DHPAG with purity of 88.7%. Further preparative RP-HPLC experiment gave pure DHPAG (16.3 mg, purity > 98%). The current study demonstrates that utilization of CCC method maximizes the isolation efficiency compared with that of solid-based conventional column chromatography.

Keyword

Nelumbo nucifera; Dihydrophaseic acid 3′-O-β-D-glucopyranoside; High-performance countercurrent chromatography; Isolation efficiency

MeSH Terms

Chromatography
Chromatography, Liquid*
Countercurrent Distribution*
Methods
Nelumbo*

Figure

• 

  Fig. 1. Chemical structure of dihydrophaseic acid 3′-O-β-D-glucopyranoside (DHPAG).

• 

  Fig. 2. HPLC analyses of NNE samples. (A) 25% ethanol extract, (B) ethyl acetate soluble extract, (C) n-butanol soluble extract and (D) enriched DHPAG extract.

• 

  Fig. 3. HPCCC experiment on enriched DHPAG extract (A), HPLC analyses of target HPCCC fraction (B) and purified DHPAG by preparative RP-HPLC (C). HPCCC parameters are described in Experimental section.

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