title: Characterization of fortimicin aminoglycoside profiles produced from Micromonospora olivasterospora DSM 43868 by high-performance liquid chromatography-electrospray ionization-ion trap-mass spectrometry.
abstract: 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.
authors.affiliation: Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University, Chungnam, 336-708, Republic of Korea.
authors.forename: Nguyen Lan
authors.initials: NL
authors.lastname: Huong
authors.affiliation: Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University, Chungnam, 336-708, Republic of Korea.
authors.forename: Nguyen Huu
authors.initials: NH
authors.lastname: Hoang
authors.affiliation: School of Biosystem and Biomedical Science, Korea University, Seoul, 136-713, Republic of Korea.
authors.forename: Sung-Yong
authors.initials: SY
authors.lastname: Hong
authors.affiliation: Department of Biotechnology Convergent Pharmaceutical Engineering, SunMoon University, Chungnam, 336-708, Republic of Korea.
authors.forename: Jae Kyung
authors.initials: JK
authors.lastname: Sohng
authors.affiliation: Department of Chemistry and Nano Sciences, Ewha Womans University, Seoul, 136-750, Republic of Korea.
authors.forename: Yeo Joon
authors.initials: YJ
authors.lastname: Yoon
authors.affiliation: School of Biosystem and Biomedical Science, Korea University, Seoul, 136-713, Republic of Korea. jewonpark@korea.ac.kr.
authors.forename: Je Won
authors.initials: JW
authors.lastname: Park
mesh_terms: D000617:Aminoglycosides / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005285:Fermentation; D057230:Limit of Detection; D009682:Magnetic Resonance Spectroscopy; D008847:Micromonospora / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000617:Aminoglycosides; C023385:fortimicin B; C023521:fortimicins; C023384:fortimicin A
keywords: Chiral column; Fortimicin aminoglycosides; HPLC-ESI–ion trap–MS/MS; Micromonospora olivasterospora; Pentafluoropropionic acid