title: Fast carbohydrate analysis via liquid chromatography coupled with ultra violet and electrospray ionization ion trap detection in 96-well format.
abstract: A fast carbohydrate screening platform processible in 96-well format is described. The method is suitable for the determination of various carbohydrates out of complex mixtures as obtained by acidic hydrolysis of carbohydrates polymers. The chromatographic conditions for an efficient separation (12min) and the derivatization process with 1-phenyl-3-methyl-5-pyrazolone (PMP) were optimized for high resolution separation and simultaneous determination of deoxy-, amino-, anhydro-sugars as well as hexoses, pentoses, dimers, uronic acids and degradation products like furfural and hydroxymethylfurfural (HMF). The potential to quantify with UV- and MS-detector in the same range has been demonstrated for 20 different compounds. Finally, the matrix effects of the hydrolysis were positively evaluated. The micro scale hydrolysis and PMP-derivatization without any extraction or drying steps, both in 96-well format, result in a fast and intuitive sample preparation. In combination with a fast liquid chromatography coupled to UV and electrospray ionization ion trap detection (LC-UV-ESI-MS/MS) for the qualification and quantification of various sugars, dimers and degradation products, this method shows great performance in carbohydrate analysis.
authors.affiliation: Chemistry of Biogenic Resources, Technische Universität München, Germany.
authors.forename: Broder
authors.initials: B
authors.lastname: Rühmann
authors.affiliation: Chemistry of Biogenic Resources, Technische Universität München, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Schmid
authors.affiliation: Chemistry of Biogenic Resources, Technische Universität München, Germany. Electronic address: Sieber@tum.de.
authors.forename: Volker
authors.initials: V
authors.lastname: Sieber
mesh_terms: D002241:Carbohydrates / Q000032:analysis*; D002623:Chemistry Techniques, Analytical / Q000379:methods* / Q000592:standards; D002853:Chromatography, Liquid; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet
chemical_list: D002241:Carbohydrates
keywords: 1-Phenyl-3-methyl-5-pyrazolone-derivatization; 96-Well high throughput format; Carbohydrate polymers; Carbohydrates; LC–UV-ESI-MS/MS; Micro scale hydrolysis