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- Chemie (66) (remove)
Organische Chemie
(2020)
In der Arbeit wurde eine GC-MS-Methode zur Quantifizierung von 21 identifizierten Aromastoffen, die aus getoasteten Eichenholzchips in Wein übergehen, entwickelt. Diese Methode wurde für die ausgewählten Substanzen unter Einbeziehung der Festphasenextraktion mittels des internen Standard 2-Octanol kalibriert. Die Analyte zeigten mit Ausnahme von Syringaaldehyd lineares Verhalten bei Korrelationen von 0,986 bis 0,999. Die Nachweisgrenzen, die über das Signal-zu-Rausch-Verhältnis ermittelt wurden, lagen im Bereich von 0,03 bis 0,47 µg/l. Der Test auf Wiederholbarkeit ergab mit Werten von kleiner 10% für die meisten untersuchten Stoffe in Rot- und Weißwein zufrieden stellende Werte. Mit der entwickelten Methode wurden Barriqueweine des Handels und selbst hergestellte Chipsweine untersucht. Bei der Untersuchung der Chipsweine, die nach einem vollen Versuchsplan hergestellt wurden, zeigten sich signifikante Einflüsse durch die Faktoren Kontaktzeit, Toastung und Größe sowie durch die Wechselwirkung Toastung-Größe auf die Konzentrationen der meisten identifizierten Substanzen. Bei der Betrachtung der Konzentrationen der ausgewählten Substanzen in Barriqueweinen zeigten sich signifikante Unterschiede zu den entsprechenden Konzentrationen in den hergestellten Chipsweinen. Diese signifikanten Unterschiede wurden mittels Hauptkomponentenanalyse und anschließender Klassifizierung über die SIMCA-Methode dazu genutzt, um die önolgischen Verfahren Chipsbehandlung und Barriquefasslagerung zu differenzieren.
Multidimensional separation techniques play an increasingly important role in separation science, especially for the analysis of complex samples such as proteins. The combination of reversed-phase liquid chromatography in the nanoscale and CZE is especially beneficial due to their nearly orthogonal separation mechanism and well-suited geometries/dimensions. Here, a heart-cut nano-LC-CZE-MS setup was developed utilizing for the first time a mechanical 4-port valve as LC-CE interface. A model protein mixture containing four different protein species was first separated by nano LC followed by a heart-cut transfer of individual LC peaks and subsequent CZE-MS analysis. In the CZE dimension, various glycoforms of one protein species were separated. Improved separation capabilities were achieved compared to the 1D methods, which was exemplarily shown for ribonuclease B and its different glycosylated forms. LODs in the lower μg/mL range were determined, which are considerably lower compared to traditional CZE-MS. In addition, this study represents the first application of an LC-CE-MS system for intact protein analysis. The nano-LC-CZE-MS system is expected to be applicable to various other analytical challenges.
Coupling imaged capillary isoelectric focusing with mass spectrometry using a nanoliter valve
(2018)
Organische Chemie
(2021)
Thermal metal-free [2+2] cycloaddition of acetylenedicarboxylates to polysubstituted butadienes
(2018)
Online mass spectrometry of CE (SDS)-separated proteins by two-dimensional capillary electrophoresis
(2019)
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is the fundamental technique for protein separation by size. Applying this technology in capillary format, gaining high separation efficiency in a more automated way, is a key technology for size separation of proteins in the biopharmaceutical industry. However, unequivocal identification by online mass spectrometry (MS) is impossible so far, due to strong interference in the electrospray process by SDS and other components of the SDS-MW separation gel buffer. Here, a heart-cut two-dimensional electrophoretic separation system applying an electrically isolated valve with an internal loop of 20 nL is presented. The peak of interest in the CE (SDS) separation is transferred to the CZE-MS, where electrospray-interfering substances of the SDS-MW gel are separated prior to online electrospray ionization mass spectrometry. An online SDS removal strategy for decomplexing the protein-SDS complex is implemented in the second dimension, consisting of the co-injection of organic solvent and cationic surfactant. This online CE (SDS)-CZE-MS system allows MS characterization of proteoforms separated in generic CE (SDS), gaining additional separation in the CZE and detailed MS information. In general, the system can be applied to all kinds of proteins separated by CE (SDS). Here, we present results of the CE (SDS)-CZE-MS system on the analysis of several biopharmaceutically relevant antibody impurities and fragments. Additionally, the versatile application spectrum of the system is demonstrated by the analysis of extracted proteins from soybean flour. The online hyphenation of CE (SDS) resolving power and MS identification capabilities will be a powerful tool for protein and mAb characterization. Graphical abstract Two-dimensional capillary electrophoresis system hyphenated with mass spectrometry for the characterization of CE (SDS)-separated proteins. As first dimension, a generic and high MS-interfering CE (SDS) separation is performed for size separation. After heart-cut transfer of the unknown CE (SDS) protein peak, via a four-port nanoliter valve to a volatile electrolyte system as second dimension, interference-free mass spectrometric data of separated mAb fragments and soybean proteins are obtained.