2-Pyridylfuran: A New Fluorescent Tag for the Analysis of Carbohydrates

Abstract

We herein report the use of 1,3-di(2-pyridyl)-1,3-propanedione (DPPD) as a fluorogenic labeling reagent for sugars. Reaction of DPPD with the anomeric carbon affords a fluorescent 2-pyridylfuran (2-PF) moiety that permits the sensitive HPLC-based detection of monosaccharides. 2-PF-labeled monosaccharides can be easily separated and analyzed from mixtures thereof, and the reported protocol compares favorably with established labeling reagents such as 2-aminobenzoic acid (2-AA) and 1-phenyl-3-methyl-5-pyrazolone (PMP), ultimately allowing subfemtomole detection of the galactose-derived product. Furthermore, we demonstrate the application of DPPD in the labeling of monosaccharides in complex biological matrices such as blood and milk samples. We envisage that DPPD will prove to be an excellent choice of labeling reagent in monosaccharide and carbohydrate analysis.

Recent liquid chromatographic approaches and developments for the separation and purification of carbohydrates

“…A fluorescent pyridylfuran moiety was formed upon incubation of saccharide and 1,3-di(2-pyridyl)-1,3-propanedione overnight at 110 °C. D-Galactose pyridylfuran derivatives can be detected at femtomole levels, which are significantly lower than traditionally used fluorescent tags 108 . Such tags also permit faster separations, where monosaccharide derivatives can be resolved within 6 minutes with liquid chromatography.…”

Glycomic and glycoproteomic analysis of glycoproteins—a tutorial

“…In yet another novel strategy, monosaccharides were labeled with the fluorescent tag, 2-pyridylfuran (2-PF), introduced by reaction with 1,3-di(2-pyridyl)-1,3-propanedione (DPPD). HPLC analysis of these derivatives achieved sub-femto mole levels of detection [106]. Another recent study reported a new improved label, RapiFluorMS (RFMS), which enables rapid labeling of N-glycans, released as glycosylamine, at their reducing end immediately after PNGase F treatment.…”

Recent Advances in the Analysis of Complex Glycoproteins

“…90 Some of the most commonly used fluorescence reagents for glycan analysis include 2-aminobenzamide (2-AB), anthranilic acid (2-AA) 2-aminopyridine (2-AP), and 1-phenyl-3-methyl-5-pyrazolone (PMP). 90 2-AA-labeled glycans are most often detected with fluorescence, but some reports analyze 2-AA labeled N -glycans with reverse phase LC–MS.…”

“…90 Some of the most commonly used fluorescence reagents for glycan analysis include 2-aminobenzamide (2-AB), anthranilic acid (2-AA) 2-aminopyridine (2-AP), and 1-phenyl-3-methyl-5-pyrazolone (PMP). 90 2-AA-labeled glycans are most often detected with fluorescence, but some reports analyze 2-AA labeled N -glycans with reverse phase LC–MS. 91 Whereas the previously reported literature indicates negligible desialylation during 2-AB labeling of N-linked glycans, a recent report indicates that there is some loss of sialic acid from highly sialylated glycans during 2-AB derivatization or possibly other reductive amination procedures as well.…”