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

• Published in: Analytical and bioanalytical chemistry Vol. 408; no. 6; pp. 1667 - 1678
• Main Authors: Huong, Nguyen Lan, Hoang, Nguyen Huu, Hong, Sung-Yong, Sohng, Jae Kyung, Yoon, Yeo Joon, Park, Je Won
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.02.2016; Springer; Springer Nature B.V
• Subjects: accuracy; Aminoglycosides; Aminoglycosides - analysis; Aminoglycosides - chemistry; Aminoglycosides - metabolism; Analysis; Analytical Chemistry; Antibiotics; Bacteria; Biochemistry; Biosynthesis; Broths; Cations; Characterization and Evaluation of Materials; Chemical properties; Chemistry; Chemistry and Materials Science; Chromatography; Chromatography, High Pressure Liquid - methods; Congeners; Culture; culture media; Fermentation; Food Science; glycopeptides; Gram-positive bacteria; High performance liquid chromatography; Ionization; Laboratory Medicine; Limit of Detection; Liquid chromatography; Magnetic Resonance Spectroscopy; Mass spectrometry; Micromonospora - metabolism; Micromonospora olivasterospora; Monitoring/Environmental Analysis; NMR; Nuclear magnetic resonance; Research Paper; Scientific imaging; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - methods; Japan; South Korea; Micromonospora olivasterospora; HPLC-ESI–ion trap–MS/MS; Pentafluoropropionic acid; Chiral column; Fortimicin aminoglycosides
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-015-9281-2

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.