期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1455, 期 -, 页码 125-132出版社
ELSEVIER
DOI: 10.1016/j.chroma.2016.05.079
关键词
Rhamnolipids; Biosurfactants; Quantification; HPLC-charged-aerosol detection (CAD); Derivatization; HPLC-UV
资金
- Deutsche Bundesstiftung Umwelt (DBU)
- Cluster of Excellence Tailor-Made Fuels from Biomass - Excellence Initiative of the German federal and state governments
A method using high performance liquid chromatography coupled to charged-aerosol detection (HPLC-CAD) was developed for the quantification of rhamnolipid biosurfactants. Qualitative sample composition was determined by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The relative quantification of different derivatives of rhamnolipids including di-rhamnolipids, monorhamnolipids, and their precursors 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs) differed for two compared LC MS instruments and revealed instrument dependent responses. Our here reported HPLC-CAD method provides uniform response. An inverse gradient was applied for the absolute quantification of rhamnolipid congeners to account for the detector's dependency on the solvent composition. The CAD produces a uniform response not only for the analytes but also for structurally different (nonvolatile) compounds. It was demonstrated that n-dodecyl-beta-D-maltoside or deoxycholic acid can be used as alternative standards. The method of HPLC-ultra violet (UV) detection after a derivatization of rhamnolipids and HAAs to their corresponding phenacyl esters confirmed the obtained results but required additional, laborious sample preparation steps. Sensitivity determined as limit of detection and limit of quantification for four mono-rhamnolipids was in the range of 0.3-1.0 and 1.2-2.0 mu g/mL, respectively, for HPLC-CAD and 0.4 and 1.5 mu g/mL, respectively, for HPLC-UV. Linearity for HPLC-CAD was at least 0.996 (R-2) in the calibrated range of about 1-200 mu g/mL. Hence, the here presented HPLC-CAD method allows absolute quantification of rhamnolipids and derivatives. (C) 2016 Elsevier B.V. All rights reserved.
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