High performance liquid chromatography-charged aerosol detection applying an inverse gradient for quantification of rhamnolipid biosurfactants
| Autor: | Matthias Baune, Beate Behrens, Janek Jungkeit, Lars M. Blank, Heiko Hayen, Till Tiso |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Analyte Analytical chemistry Phenacyl Tandem mass spectrometry 01 natural sciences Biochemistry High-performance liquid chromatography Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Limit of Detection Tandem Mass Spectrometry Sample preparation Derivatization Chromatography High Pressure Liquid Detection limit Aerosols Chromatography 010401 analytical chemistry Organic Chemistry Solid Phase Extraction Rhamnolipid General Medicine Reference Standards 0104 chemical sciences 030104 developmental biology chemistry Spectrophotometry Ultraviolet Glycolipids |
| Zdroj: | Journal of chromatography. A. 1455 |
| ISSN: | 1873-3778 |
| Popis: | 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, mono-rhamnolipids, 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-β- 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 μg/mL, respectively, for HPLC-CAD and 0.4 and 1.5 μg/mL, respectively, for HPLC-UV. Linearity for HPLC-CAD was at least 0.996 (R2) in the calibrated range of about 1⿿200 μg/mL. Hence, the here presented HPLC-CAD method allows absolute quantification of rhamnolipids and derivatives. |
| Databáze: | OpenAIRE |
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