Simultaneous Determination of the Absolute Configuration of Twelve Monosaccharide Enantiomers from Natural Products in a Single Injection by a UPLC-UV/MS Method

Abstract

In natural product chemistry, it is often crucial to determine sugar composition as well as the absolute configuration of each monosaccharide in glycosides. An ultra-performance liquid chromatography method using both photodiode array (PDA) and mass spectrometry detectors (UPLC-UV/MS) was developed for qualitative analysis of the absolute configuration of monosaccharide enantiomers. Within a single injection, 16 monosaccharide derivatives including 6 pairs of aldose enantiomers (D/L-glucose, D/L-galactose, D/L-allose, D/L-arabinose, D/L-xylose, and D/L-fucose) and 4 other monosaccharides (L-rhamnose, 2-deoxy-D-glucose, 6-deoxy-D-glucose, and 2-deoxy-D-glalactose) were identified in less than 25 minutes. It was found that the structures of derivatives of sugar enantiomers correlated with retention time. Among derivatives of sugar enantiomers in the current study, the stereoisomer of R-configuration at C-3' retained longer than the corresponding S-configuration isomer. The UPLC-MS method can increase sensitivity of detection of the saccharides by more than 10 times compared to previously reported methods. The one-pot reaction of monosaccharide with L-cysteine methyl ester and phenyl isothiocyanate is easily reproduced, clean, and relatively simple in a screw-capped reaction vial. The developed method was successfully applied for the analysis of different types of glycosides, viz., glycosides of triterpenes, steroids, and flavonoids, that commonly exist in nature. The absolute configurations of composed monosaccharides are clearly characterized from tested samples.

NMR characterization of novel pyranoanthocyanins derived from the pulp of Panax quinquefolius L. (North American ginseng)

“…Other HMBC peaks allow for the assignment of the carbons within the aglycone, which have been well described in the literature. [12,18] Using an established HPLC method, [14,15] the sugars were both found to be the D-enantiomer (UPLC profile of the standard sugars and the sugars from 1 can be found in the Supporting information).…”

“…[12] Sugar enantiomer determination To determine if the attached sugars were D or L, a chromatographic methodology was used. [14,15] The sugars were hydrolyzed from the anthocyanins by heating 4 mg anthocyanin in 1.0 mL of 0.1M HCl for 12 h at 70°C. The sugars were isolated from the anthocyanin aglycones using a C18 cartridge.…”

Monitoring vaccine protein glycosylation: analytics and recent developments

“…A combination of techniques is required. [22] HPAEC-PAD Relatively newly applied to oligosaccharides [24] CE-LIF Amination [32] CE-LIF: Capillary electrophoresis with laser-induced fluorescence; CID: Collision-induced dissociation; HPAEC-PAD: High-pH anion exchange chromatography with amperometric detection.…”

Simultaneous Enantioseparation of Monosaccharides Derivatized with l-Tryptophan by Reversed Phase HPLC

“…Wang et al 25 reported a simultaneous enantioseparation of monosaccharides derivatized with L-cysteine methyl ester and phenyl isothiocyanate by ultraperformance liquid chromatography (UPLC) with mass spectrometry, where enantiomeric pairs of three hexoses (Glc, Gal and allose), three pentoses and four deoxy-D-hexoses were simultaneously separated.…”