Resolving Isomeric Structures of Native Glycans by Nanoflow Porous Graphitized Carbon Chromatography–Mass Spectrometry

Glycosylation is one of the most common and complex post-translational modifications of proteins. However, there are other carbohydrates such as free oligosaccharides and glycosphingolipids-glycans that are associated with important biological and clinical roles. To analyze these molecules using liquid chromatography coupled with mass spectrometry (LC-MS), the permethylation approach was utilized. Although permethylation is a commonly utilized glycan derivatization technique, separation of permethylated glycans released from glycosphingolipid (GSL) by LC-MS has never been previously demonstrated. Here, a nanoflow porous graphitized carbon (PGC) column coupled with a high-resolution mass spectrometer was used to achieve isomeric separation of these permethylated glycans. We demonstrate the separation of free reducing end and reduced end O-glycans, free oligosaccharides derived from human milk, and GSL glycans derived from the MDA-MB-231BR cancer cell line using PGC-LC-MS.

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Introducing porous graphitized carbon liquid chromatography with evaporative light scattering and mass spectrometry detection into cell wall oligosaccharide analysis

Journal of Chromatography A ◽

10.1016/j.chroma.2009.12.005 ◽

2010 ◽

Vol 1217 (5) ◽

pp. 689-695 ◽

Cited By ~ 33

Author(s):

Y. Westphal ◽

H.A. Schols ◽

A.G.J. Voragen ◽

H. Gruppen

Keyword(s):

Mass Spectrometry ◽

Cell Wall ◽

Liquid Chromatography ◽

Light Scattering ◽

Mass Spectrometry Detection ◽

Porous Graphitized Carbon ◽

Graphitized Carbon ◽

Evaporative Light Scattering

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Multiresidue Analysis of Pesticides in Vegetables and Fruits by Gas Chromatography/Mass Spectrometry after Gel Permeation Chromatography and Graphitized Carbon Column Cleanup

Journal of AOAC International ◽

10.1093/jaoac/87.4.1003 ◽

2004 ◽

Vol 87 (4) ◽

pp. 1003-1015 ◽

Cited By ~ 28

Author(s):

Eiji Ueno ◽

Harumi Oshima ◽

Isao Saito ◽

Hiroshi Matsumoto ◽

Yoshihiro Yoshimura ◽

...

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Sequential Analysis ◽

Gel Permeation Chromatography ◽

Gas Chromatography Mass Spectrometry ◽

Graphitized Carbon ◽

Chromatography Mass Spectrometry ◽

Relative Standard ◽

Fruit Samples ◽

Gel Permeation

Abstract A multiresidue method for pesticides that enables quantitative, sequential analysis of a large number of vegetable and fruit samples by gas chromatography/mass spectrometry has been developed. First, 89 important target compounds were selected for monitoring, and then the appropriate internal standards for these pesticides, 14 stable isotopically labeled pesticides (surrogates), were used. The sample was extracted with acetonitrile, and the extract was cleaned up by a salting-out step followed by redissolution in ethyl acetate. Coextractives were removed automatically by gel permeation chromatography with a graphitized carbon column, and then by use of a tandem silica-gel/PSA cartridge column. Recoveries of 82 of the 89 pesticides from fortified spinach, tomato, apple, and strawberry were within a range from 70 to 120%, and the relative standard deviation values of 80 of the 89 pesticides were <5%. The method was applied to 188 commercial vegetable and fruit samples to demonstrate its use in routine analysis.

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