Silver Ion-Complexation High-Speed Countercurrent Chromatography Coupled with Prep-HPLC for Separation of Sesquiterpenoids from Germacrene A Fermentation Broth

Huanzhu Zhao,Iftikhar Ali,Li Cui,Xiangyun Song,Qianshan Gao,Andleeb Mehmood,Wenya Ma,Dawei Qin,Manzoor Hussain,Baraa Siyo,Hidayat Hussain,Shahid Aziz,Daijie Wang

doi:10.3390/fermentation7040230

Abstract

A silver ion high-speed counter-current chromatography ([Ag+]-HSCCC) was developed to separate and purify five sesquiterpenoids from germacrene A fermentation broth. The solvent system was consisted of n-hexane-methanol-silver nitrate (3 mol/L) solution (10:9.5:0.5, v/v). By employing this chromatographic protocol, five sesquiterpenoids named β-elemene (1; 54.1 mg), germacrene A (2; 28.5 mg), γ-selinene (3; 4.6 mg), β-selinene (4; 3.4 mg), and α-selinene (5; 1.3 mg) were obtained successfully from 500 mg extracted crude sample with purities of 97.1%, 95.2%, 98.2%, 96.3% and 98.5%, respectively, combined with preparative HPLC. The results reveal that the addition of metal ion in biphasic solvent system significantly improved the HSCCC separation factor of sesquiterpenoids. Meanwhile, our study also provided an alternate approach to separate the compounds with less polarity, also geometrical isomers and various natural product classes.

Highlights

Introduction βElemene (1; β-1-methyl-1-vinyl-2,4-diisopropenyl-cyclohexane), an active noncytotoxic class II antitumor drug derived from Chinese medicinal herb Curcuma phaeocaulis Val., has been reported to possess broad spectrum anti-cancer effects such as lung [1], breast [2], glioma [3], and gastric [4] etc
One of the most important steps in performing an High-speed counter-current chromatography (HSCCC) separation is the selection of an appropriate solvent system, which is not an exception in complexation HSCCC
Due to the small polarity of the target compounds, n-hexane-ethyl acetate-methanol-water was found as the ideal solvent system, which could cover a wide range of polarity by modifying the volume ratio of the four solvents

Summary

Introduction

Elemene (1; β-1-methyl-1-vinyl-2,4-diisopropenyl-cyclohexane), an active noncytotoxic class II antitumor drug derived from Chinese medicinal herb Curcuma phaeocaulis Val., has been reported to possess broad spectrum anti-cancer effects such as lung [1], breast [2], glioma [3], and gastric [4] etc. Pharmacological studies have proved that β-elemene (1). Can play a role through a variety of pharmacological pathways including arresting the cell cycle [5], inhibiting cell proliferation [3], exerting antimetastasis and antiangiogenesis effects [4], and reversing multiple-drug resistance inducing cell apoptosis [6]. Fermentation 2021, 7, 230 by China’s State Food and Drug Administration for the treatment of cancer as a secondclass innovative drug [8] and applied for clinical studies in Food and Drug Administration department at USA. Because of its broad-spectrum, safe, efficient, economical and other outstanding advantages in antitumor activities, this compound has numerous broad clinical applications

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Conclusion