## High -performance liquid chromatography with electrospray ionization mass spectrometry (LC/MS) and liquid chromatography with tandem mass spectrometry (LC/MS/MS) were applied to the analysis of the site-specific carbohydrate heterogeneity in erythropoietin (EPO) used as a model of the sialylate
Sheath liquid interface for the coupling of normal-phase liquid chromatography with electrospray mass spectrometry and its application to the analysis of neoflavonoids
✍ Scribed by Laurence Charles; Frédéric Laure; Phila Raharivelomanana; Jean-Pierre Bianchini
- Publisher
- John Wiley and Sons
- Year
- 2005
- Tongue
- English
- Weight
- 163 KB
- Volume
- 40
- Category
- Article
- ISSN
- 1076-5174
- DOI
- 10.1002/jms.777
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✦ Synopsis
Abstract
A novel interface that allows normal‐phase liquid chromatography to be coupled with electrospray ionization (ESI) is reported. A make‐up solution of 60 mM ammonium acetate in methanol, infused at a 5 µl min^−1^ flow‐rate at the tip of the electrospray probe, provides a sheath liquid which is poorly miscible with the chromatographic effluent, but promotes efficient ionization of the targeted analytes. Protonated molecules generated in the ESI source were subjected to tandem mass spectrometric experiments in a triple‐quadrupole mass spectrometer. The main fragmentation reactions were characterized for each analyte and specific mass spectral transitions were used to acquire chromatographic data in the multiple reaction monitoring detection mode. Results obtained during optimization of the sheath liquid composition and flow‐rate suggest that the electrospray process was mainly under the control of the make‐up solution, and that it forms an external charged layer around a neutral chromatographic mobile phase core. This sheath liquid interface was implemented for the analysis of some neoflavonoid compounds and its performance was evaluated. Limits of detection were established for calophillolide, inophyllum B, inophyllum P and inophyllum C at 100, 25, 15 and 100 ng ml^−1^, respectively. Copyright © 2004 John Wiley & Sons, Ltd.
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