Planta Med
DOI: 10.1055/a-1228-8473
Original Papers

High-performance Countercurrent Chromatography to Access Rhodiola rosea Influenza Virus Inhibiting Constituents

Julia Langeder
1  Department of Pharmacognosy, University of Vienna, Vienna, Austria
,
1  Department of Pharmacognosy, University of Vienna, Vienna, Austria
,
Kristin Döring
2  Section of Experimental Virology, Department of Medical Microbiology, Jena University, Jena, Germany
,
Mahtab Jafari
3  Department of Pharmaceutical Sciences, University of California, Irvine, CA, USA
,
Christina Ehrhardt
2  Section of Experimental Virology, Department of Medical Microbiology, Jena University, Jena, Germany
,
Michaela Schmidtke
2  Section of Experimental Virology, Department of Medical Microbiology, Jena University, Jena, Germany
,
1  Department of Pharmacognosy, University of Vienna, Vienna, Austria
› Author Affiliations
Supported by: Wilhelm Doerenkamp-Foundation, Chur, Switzerland Natvantage grant (2018)

Dedicated to Professor Arnold Vlietinck on the occasion of his 80th birthday.

Abstract

In a cytopathic effect inhibition assay, a standardized Rhodiola rosea root and rhizome extract, also known as roseroot extract (SHR-5), exerted distinct anti-influenza A virus activity against HK/68 (H3N2) (IC50 of 2.8 µg/mL) without being cytotoxic. For fast and efficient isolation and identification of the extractʼs bioactive constituents, a high-performance countercurrent chromatographic separation method was developed. It resulted in a three-stage gradient elution program using a mobile phase solvent system composed of ethyl acetate/n-butanol/water (1 : 4 : 5 → 2 : 3 : 5 → 3 : 2 : 5) in the reversed-phase mode. The elaborated high-performance countercurrent chromatographic method allowed for fractionation of the complex roseroot extract in a single chromatographic step in a way that only one additional orthogonal isolation/purification step per fraction yielded 12 isolated constituents. They cover a broad polarity range and belong to different structural classes, namely, the phenylethanoid tyrosol and its glucoside salidroside, the cinnamyl alcohol glycosides rosavin, rosarin, and rosin as well as gallic acid, the cyanogenic glucoside lotaustralin, the monoterpene glucosides rosiridin and kenposide A, and the flavonoids tricin, tricin-5-O-β-D-glucopyranoside, and rhodiosin. The most promising anti-influenza activities were determined for rhodiosin, tricin, and tricin-5-O-β-D-glucopyranoside with IC50 values of 7.9, 13, and 15 µM, respectively. The herein established high-performance countercurrent chromatographic protocol enables fast and scalable access to major as well as minor roseroot constituents. This is of particular relevance for extract standardization, quality control, and further in-depth pharmacological investigations of the metabolites of this popular traditional herbal remedy.

Supporting Information



Publication History

Received: 12 June 2020

Accepted after revision: 22 July 2020

Publication Date:
11 August 2020 (online)

Georg Thieme Verlag KG
Stuttgart · New York