Comparison of procainamide and 2-aminobenzamide labeling for profiling and identification of glycans by liquid chromatography with fluorescence detection coupled to electrospray ionization–mass spectrometry

• Published in: Analytical biochemistry Vol. 486; pp. 38 - 40
• Main Authors: Kozak, Radoslaw P., Tortosa, Concepcion Badia, Fernandes, Daryl L., Spencer, Daniel I.R.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-10-2015
• Subjects: Chromatography, High Pressure Liquid - methods; ESI–MS; Fluorescent Dyes - chemistry; Fluorescent labeling; Glycan derivatization; HILIC–(U)HPLC; Humans; ortho-Aminobenzoates - chemistry; Polysaccharides - analysis; Polysaccharides - chemistry; Procainamide; Procainamide - chemistry; Spectrometry, Fluorescence; Spectrometry, Mass, Electrospray Ionization - methods; Staining and Labeling; HILIC–(U)HPLC; Glycan derivatization; Fluorescent labeling; Procainamide; ESI–MS
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2015.06.006

One of the most widely used methods for glycan analysis is fluorescent labeling of released glycans followed by hydrophilic interaction chromatography–(ultra-)high-performance liquid chromatography [HILIC–(U)HPLC]. Here, we compare the data obtained by (U)HPLC–fluorescence (FLR) coupled to electrospray ionization–mass spectrometry (ESI–MS) for procainamide and 2-aminobenzamide (2-AB)-labeled N-glycans released from human immunoglobulin G (IgG). Fluorescence profiles from procainamide show comparable chromatographic separation to those obtained for 2-AB but gave higher fluorescence intensity as well as significantly improved ESI efficiency (up to 30 times that of 2-AB). Thus, labeling with procainamide increases the ability to identify minor glycan species that may have significant biological activity.