Dereplication by High-Performance Liquid Chromatography (HPLC) with Quadrupole-Time-of-Flight Mass Spectroscopy (qTOF-MS) and Antiviral Activities of Phlorotannins from Ecklonia cava

Mar Drugs. 2019 Mar 4;17(3):149. doi: 10.3390/md17030149.

Abstract

Ecklonia cava is edible seaweed that is found in Asian countries, such as Japan and Korea; and, its major components include fucoidan and phlorotannins. Phlorotannins that are isolated from E. cava are well-known to have an antioxidant effect and strong antiviral activity against porcine epidemic diarrhea virus (PEDV), which has a high mortality rate in piglets. In this study, the bioactive components were determined based on two different approaches: (i) bio-guided isolation using the antiviral activity against the H1N1 viral strain, which is a representative influenza virus that originates from swine and (ii) high-resolution mass spectrometry-based dereplication, including relative mass defects (RMDs) and HPLC-qTOFMS fragmentation analysis. The EC70 fraction showed the strongest antiviral activity and contained thirteen phlorotannins, which were predicted by dereplication. Ten compounds were directly isolated from E. cava extract and then identified. Moreover, the dereplication method allowed for the discovery of two new phlorotannins. The structures of these two isolated compounds were elucidated using NMR techniques and HPLC-qTOFMS fragmentation analysis. In addition, molecular modelling was applied to determine the absolute configurations of the two new compounds. The antiviral activities of seven major phlorotannins in active fraction were evaluated against two influenza A viral strains (H1N1 and H9N2). Six of the compounds showed moderate to strong effects on both of the viruses and phlorofucofuroeckol A (12), which showed an EC50 value of 13.48 ± 1.93 μM, is a potential active antiviral component of E. cava.

Keywords: Ecklonia cava; HPLC-qTOFMS; antiviral; bioactivity-guided isolation; dereplication; phlorotannins; relative mass defect.

MeSH terms

  • Antiviral Agents / chemistry
  • Antiviral Agents / isolation & purification
  • Antiviral Agents / pharmacology*
  • Biological Products / chemistry
  • Biological Products / isolation & purification
  • Biological Products / pharmacology*
  • Chromatography, High Pressure Liquid / methods
  • Influenza A Virus, H1N1 Subtype / drug effects
  • Influenza A Virus, H9N2 Subtype / drug effects
  • Phaeophyta / chemistry*
  • Seaweed / chemistry*
  • Tandem Mass Spectrometry / methods
  • Tannins / chemistry
  • Tannins / isolation & purification
  • Tannins / pharmacology*

Substances

  • Antiviral Agents
  • Biological Products
  • Tannins