Determination of sibiromycin and its natural derivatives using new analytical and structural approaches.
A new separation and quantification method using ultra high-performance liquid chromatography (UHPLC) with UV detection was developed for the detection of sibiromycin in fermentation broth of Streptosporangium sibiricum. The solid phase extraction method based on cation-exchange was employed to pre-concentrate and purify fermentation broth containing sibiromycin prior to UHPLC analysis. The whole assay was validated and showed a linear range of detector response for the quantification of sibiromycin in a concentration from 3.9 to 250.0 μg mL⁻¹, with correlation coefficient of 0.999 and recoveries ranging from 71.66±3.55% to 74.76±5.18%. Method limit of quantification of the assay was determined as 0.18 μg mL⁻¹ and was verified with resulting RSD of 9.6% and accuracy of 97.6%. The developed assay was used to determine the sibiromycin production in 12 different fermentation broths. Moreover, several natural sibiromycin analogues/derivatives were described with pilot characterization using off-line mass spectrometry: the previously described dihydro-sibiromycin (DH-sibiromycin) and tentative bis-glycosyl forms of sibiromycin and its dihydro-analogue.
Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic.
Miroslav
M
Sulc
Iva
I
Fadrhoncová
Markéta
M
Jelínková
Miriam
M
Chudomelová
Jürgen
J
Felsberg
Jana
J
Olšovská
D000192:Actinomycetales / Q000378:metabolism; D000617:Aminoglycosides / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017077:Culture Media, Conditioned / Q000737:chemistry; D004355:Drug Stability; D005285:Fermentation; D016014:Linear Models; D013058:Mass Spectrometry / Q000379:methods*; D015394:Molecular Structure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction
D000617:Aminoglycosides; D017077:Culture Media, Conditioned; C100298:sibiromycin