Characterization and identification of pradimicin analogs from Actinomadura hibisca using liquid chromatography-tandem mass spectrometry

Je Won Park, Sung Ryeol Park, Ah Reum Han, Yeon Hee Ban, Young Ji Yoo, Eunji Kim, Beom Seok Kim, Jae Kyung Sohng, Yeo Joon Yoon

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Microbial cultures produce complex and potentially interesting mixtures of biosynthetic intermediates and derivatives of metabolites. These mixtures' reliable identification is important and so too is the development of techniques for their analysis. Here, a simple and highly selective method of detecting the biosynthetic congeners involved in the pentangular polyphenol pradimicin (PR) pathway from Actinomadura hibisca fermentation was developed. Solid-phase extraction (SPE) cleanup using an OASIS HLB cartridge was a simple and reliable tool for the extraction of PRs from a fermentation broth. The separation of each natural PR analog - eluted with a gradient system of aqueous acetonitrile through a reversed-phase C18 column containing ammonium acetate and acetic acid as additives - allowed their simultaneous profiling. The combined use of SPE cleanup and chromatographic separation, coupled with electrospray ionization-tandem mass spectrometry (ESI-MS/MS) detection was demonstrated to be sufficiently accurate and reliable to analyze the natural PR analogs produced from A. hibisca. Ten natural PRs were identified: four alanine-containing (PRA, PRC, PRL, and PRB), two glycine-substituted (PRD and PRE), and four serine-substituted (PRFA-1, PRFA-2, PRFL, and PRFB). This report demonstrates the first use of both SPE cleanup and HPLC-ESI-MS/MS to profile a wide range of structurally closely related PRs in a bacterial fermentation broth.

Original languageEnglish
Pages (from-to)2284-2291
Number of pages8
JournalJournal of Chromatography A
Volume1218
Issue number16
DOIs
Publication statusPublished - 2011 Apr 22

Keywords

  • Actinomadura hibisca
  • HPLC-ESI-MS/MS
  • Metabolite profiling
  • Pradimicins

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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