Ultra-high-performance liquid chromatography fingerprinting method for chemical screening of metabolites in cultivation broth
Jazyk angličtina Země Nizozemsko Médium print-electronic
Typ dokumentu hodnotící studie, časopisecké články, práce podpořená grantem
PubMed
20832070
DOI
10.1016/j.chroma.2010.08.031
PII: S0021-9673(10)01102-7
Knihovny.cz E-zdroje
- MeSH
- Actinobacteria chemie metabolismus MeSH
- antibakteriální látky analýza metabolismus MeSH
- extrakce na pevné fázi MeSH
- fermentace MeSH
- kultivační média chemie metabolismus MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibakteriální látky MeSH
- kultivační média MeSH
A fingerprinting method for chemical screening of microbial metabolites, potential antibiotics, in spent cultivation broths is described. The method is based on high-throughput ultra-high-performance liquid chromatography (UHPLC) separation with UV detection (photodiode array detector). Thirteen antibiotic standards and four cultivation broths were used for the method development. The comparison of ten liquid-liquid and solid phase extraction protocols for sample clean-up and pre-concentration revealed that Oasis HLB C18 sorbent gives the best recoveries. The Acquity BEH C18 chromatographic column was chosen for the samples separation with respect to its universality, selectivity, efficiency and robustness. The method is presented by two 3D fingerprints for every sample that was obtained under different, acidic and alkaline, UHPLC conditions. The acidic mobile phase consisted of 0.5% phosphoric acid with methanol and the alkaline mobile phase of 1mM ammonium formate, pH 9 with acetonitrile. Each pair of 3D fingerprints includes the following physico-chemical information: polarity (retention time), presence and characterization of chromophores (UV spectra), compound concentration (detector response), and acid-base properties (influence of different pH of the aqueous parts of mobile phases on retention times). The sample extraction and method validation were assessed with relative standard deviation (RSD) of 0.5, 5.0 and 20.0% for retention times, peak areas and minor compound peak areas, respectively.
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