Characterization of fortimicin aminoglycoside profiles produced from Micromonospora olivasterospora DSM 43868 by high-performance liquid chromatography-electrospray ionization-ion trap-mass spectrometry

Nguyen Lan Huong, Nguyen Huu Hoang, Sung Yong Hong, Jae Kyung Sohng, Yeo Joon Yoon, Je Won Park

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3 Citations (Scopus)


In this study, an efficient high-performance liquid chromatography (HPLC)-electrospray ionization (ESI)-ion trap-tandem mass spectrometry (MS/MS) was developed for the identification of the biosynthetic congeners involved in the aminocyclitol aminoglycosidic fortimicin pathway from Micromonospora olivasterospora fermentation. The usage of both acid extraction (pH ∼2.5) followed by an cationic-exchanging SPE cleanup and pentafluoropropionic acid mediated ion-pairing chromatography with ESI-ion trap-MS/MS detection was determined to be sufficiently practical to profile the fortimicin (FOR) congeners produced in a culture broth. The limit of the quantification for the fortimicin A (FOR-A) standard spiked in the culture broth was ∼1.6 ng mL-1. The average recovery rate was 93.6 %, and the intra- and inter-day precisions were <5 % with accuracy in the range from 87.1 to 94.2 %. Moreover, the epimeric mixtures including FOR-KH, FOR-KR, and FOR-B were separately resolved through a macrocyclic glycopeptide (teicoplanin)-bonded chiral column. As a result, ten natural FOR pseudodisaccharide analogs were identified and semi-quantified in descending order as follows: FOR-A, FOR-B, DCM, FOR-KH plus FOR-KR, FOR-KK1, FOR-AP, FOR-KL1, FOR-AO, and FOR-FU-10. This is the first report on both the simultaneous characterization of diverse structurally closely related FORs derived from bacterial fermentation using HPLC-ESI-ion trap-MS/MS analysis and the chromatographic separation of the three FOR epimers.

Original languageEnglish
Pages (from-to)1667-1678
Number of pages12
JournalAnalytical and Bioanalytical Chemistry
Issue number6
Publication statusPublished - 2016 Feb 1



  • Chiral column
  • Fortimicin aminoglycosides
  • HPLC-ESI-ion trap-MS/MS
  • Micromonospora olivasterospora
  • Pentafluoropropionic acid

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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