Preparative isolation of cordycepin, N(6)-(2-hydroxyethyl)-adenosine and adenosine from Cordyceps militaris by macroporous resin and purification by recycling high-speed counter-current chromatography

Zhong Zhang; Tuernisan Tudi; Yanfang Liu; Shuai Zhou; Na Feng; Yan Yang; Chuanhong Tang; Qingjiu Tang; Jingsong Zhang

doi:10.1016/j.jchromb.2016.08.025

Preparative isolation of cordycepin, N(6)-(2-hydroxyethyl)-adenosine and adenosine from Cordyceps militaris by macroporous resin and purification by recycling high-speed counter-current chromatography

J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Oct 15:1033-1034:218-225. doi: 10.1016/j.jchromb.2016.08.025. Epub 2016 Aug 16.

Authors

Zhong Zhang¹, Tuernisan Tudi², Yanfang Liu¹, Shuai Zhou¹, Na Feng¹, Yan Yang¹, Chuanhong Tang¹, Qingjiu Tang³, Jingsong Zhang⁴

Affiliations

¹ Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture, National Engineering Research Center of Edible Fungi, Shanghai 201403, China.
² Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture, National Engineering Research Center of Edible Fungi, Shanghai 201403, China; College of Pharmacognosy, China Pharmaceutical University, Nanjing 210038, China.
³ Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture, National Engineering Research Center of Edible Fungi, Shanghai 201403, China; College of Pharmacognosy, China Pharmaceutical University, Nanjing 210038, China. Electronic address: tangqingjiu@saas.sh.cn.
⁴ Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences, Key Laboratory of Edible Fungi Resources and Utilization (South), Ministry of Agriculture, National Engineering Research Center of Edible Fungi, Shanghai 201403, China. Electronic address: syja16@saas.sh.cn.

PMID: 27567378
DOI: 10.1016/j.jchromb.2016.08.025

Abstract

In this study, cordycepin, N(6)-(2-hydroxyethyl)-adenosine (HEA) and adenosine from the fruiting bodies of Cordyceps militaris were separated by using macroporous resin NKA-II adsorption. The parameters of static adsorption were tested and the optimized conditions were as follow: the total adsorption time was 12h, 50% ethanol was used for desorption and the desorption time was 9h. The crude sample that was prepared by macroporous resin NKA-II contained 3.4% cordycepin, 3.7% HEA and 4.9% adenosine. Then the crude sample was further purified by recycling high-speed counter-current chromatography (HSCCC) with ethyl acetate, n-butanol, 1.5% aqueous ammonium hydroxide (1:4:5, v/v/v) as the optimized two-phase solvent system. Three nucleosides including 15.6mg of cordycepin, 16.9mg of HEA and 23.2mg of adenosine were obtained from 500mg of crude sample in one-step separation. The purities of three compounds were 98.5, 98.3 and 98.0%, respectively, as determined by high performance liquid chromatography.

Keywords: Adenosine; Cordycepin; Cordyceps militaris; High-speed counter-current chromatography; Macroporous resin; N(6)-(2-hydroxyethyl)-adenosine.

MeSH terms

Adenosine / analogs & derivatives*
Adenosine / chemistry
Adenosine / isolation & purification*
Adsorption
Chromatography, High Pressure Liquid
Cordyceps / chemistry*
Countercurrent Distribution / methods*
Deoxyadenosines / chemistry
Deoxyadenosines / isolation & purification*
Porosity
Resins, Synthetic / chemistry*
Solvents

Substances

Deoxyadenosines
N6-(2-hydroxyethyl)adenosine
Resins, Synthetic
Solvents
cordycepin
Adenosine