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Separation of simple sugars and carbohydrates

Simple carbohydrates are frequent ingredients in drinks, food, dietary supplements and pharmaceutical formulations. One approach to their analysis is hydrophilic interaction chromatography combined with evaporative light scattering detection, although this might be challenging due to the possibility of reducing sugars to mutarotate and thereby change their conformation into other anomers with different retention.

This application shows separation of the monosaccharides fructose and glucose, the disaccharides sucrose, turanose, maltose and lactose, plus the trisaccharide raffinose. The separation employed HILIC eluent conditions with pure acetonitrile and water, and a bonded hydrophilic stationary phase that accelerates mutarotation, thus avoiding formation of split peaks for reducing sugars.

Simple carbohydrates by HILIC-ELSD and Luna Omega Sugar

Isocratic separation of seven mono-, di- and trisaccharide on a Luna Omega Sugar column (150×4.6 mm) using an online mixed eluent containing 85% acetonitrile in water, pumped at 1.25 mL/min at 25 °C, where 0.4 mL/min was split to the detector. Instrument setup comprised Shimadzu SCL-10Avp controller, DGU-14A degasser, two LC-10ADvp pumps, JetStream Plus column oven, Spark Holland Triathlon 900 autosampler, and Sedex 85 LT ELSD (3.6 bar air, 40 °C) detector, all controlled by Clarity 8.4 software. Injection of 20 µL standard diluted in eluent to concentrations of 1000 mg/L of each saccharide.

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