scholarly journals Highly Sensitive and Robust Capillary Electrophoresis-Electrospray Ionization-Mass Spectrometry: Interfaces, Preconcentration Techniques and Applications

2020 ◽  
Vol 39 (1) ◽  
pp. 45-55
Author(s):  
Huanming Wu ◽  
Keqi Tang
Keyword(s):  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
Electrospray Ionization ◽  
Separation Efficiency ◽  
Detection Sensitivity ◽  
Ionization Mass ◽  
Sequence Coverage ◽  
Promising Alternative ◽  
Esi Ms ◽  
Preconcentration Techniques

AbstractCapillary electrophoresis (CE) coupled with mass spectrometry (MS) through electrospray ionization (ESI) is a promising alternative technique to liquid chromatography-ESI-MS (LC-ESI-MS) due to its high separation efficiency and high detection sensitivity. A sensitive and robust interface is essential in CE-ESI-MS. Continued development of CE-ESI-MS interfaces in the last decade, including junction-at-the-tip interfaces and sheathless interfaces, has improved the sensitivity and robustness of CE-ESI-MS significantly. The limited loading capacity of CE, one of major reasons that limits the utility of CE as a routine separation method, has also been addressed effectively by the use of in-capillary sample preconcentration techniques, such as transient CITP/CZE and dynamic pH junction. CE-ESI-MS could yield higher sensitivity as compared with the conventional LC-ESI-MS, and, therefore, is capable of identifying more proteins and peptides when the sample amount is very limited, such as single cell analysis. To improve the protein sequence coverage, CE-ESI-MS can also be used as a complementary technique to LC-ESI-MS, or combined with reversed phase LC to form a two dimensional separation technique. CE-ESI-MS is also effective in quantifying targeted peptides/proteins in complex bio-matrix.

scholarly journals Analysis of Inositol Phosphate Metabolism by Capillary Electrophoresis Electrospray Ionization Mass Spectrometry (CE-ESI-MS)

2020 ◽  
Author(s):  
Danye Qiu ◽  
Miranda S. Wilson ◽  
Verena B. Eisenbeis ◽  
Robert K. Harmel ◽  
Esther Riemer ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
Electrospray Ionization ◽  
Electrospray Ionization Mass Spectrometry ◽  
Inositol Phosphate ◽  
Ionization Mass Spectrometry ◽  
Ionization Mass ◽  
Phosphate Metabolism ◽  
Esi Ms ◽  
Inositol Phosphate Metabolism

AbstractThe analysis of myo-inositol phosphates (InsPs) and myo-inositol pyrophosphates (PP-InsPs) is a daunting challenge due to the large number of possible isomers, the absence of a chromophore, the high charge density, the low abundance, and the instability of the esters and anhydrides. Given their importance in biology, an analytical approach to follow and understand this complex signaling hub is highly desirable. Here, capillary electrophoresis (CE) coupled to electrospray ionization mass spectrometry (ESI-MS) is implemented to analyze complex mixtures of InsPs and PP-InsPs with high sensitivity. Stable isotope labeled (SIL) internal standards allow for matrix-independent quantitative assignment. The method is validated in wild-type and knockout mammalian cell lines and in model organisms. SIL-CE-ESI-MS enables for the first time the accurate monitoring of InsPs and PP-InsPs arising from compartmentalized cellular synthesis pathways, by feeding cells with either [13C6]-myo-inositol or [13C6]-D-glucose. In doing so, we uncover that there must be unknown inositol synthesis pathways in mammals, highlighting the unique potential of this method to dissect inositol phosphate metabolism and signalling.

scholarly journals Analysis of inositol phosphate metabolism by capillary electrophoresis electrospray ionization mass spectrometry

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Danye Qiu ◽  
Miranda S. Wilson ◽  
Verena B. Eisenbeis ◽  
Robert K. Harmel ◽  
Esther Riemer ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
Electrospray Ionization ◽  
Electrospray Ionization Mass Spectrometry ◽  
Inositol Phosphate ◽  
Ionization Mass Spectrometry ◽  
Ionization Mass ◽  
Phosphate Metabolism ◽  
Esi Ms ◽  
Inositol Phosphate Metabolism

AbstractThe analysis of myo-inositol phosphates (InsPs) and myo-inositol pyrophosphates (PP-InsPs) is a daunting challenge due to the large number of possible isomers, the absence of a chromophore, the high charge density, the low abundance, and the instability of the esters and anhydrides. Given their importance in biology, an analytical approach to follow and understand this complex signaling hub is desirable. Here, capillary electrophoresis (CE) coupled to electrospray ionization mass spectrometry (ESI-MS) is implemented to analyze complex mixtures of InsPs and PP-InsPs with high sensitivity. Stable isotope labeled (SIL) internal standards allow for matrix-independent quantitative assignment. The method is validated in wild-type and knockout mammalian cell lines and in model organisms. SIL-CE-ESI-MS enables the accurate monitoring of InsPs and PP-InsPs arising from compartmentalized cellular synthesis pathways, by feeding cells with either [13C6]-myo-inositol or [13C6]-D-glucose. In doing so, we provide evidence for the existence of unknown inositol synthesis pathways in mammals, highlighting the potential of this method to dissect inositol phosphate metabolism and signalling.

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