High-Performance Liquid Chromatography Determination of Free Sugars and Mannitol in Mushrooms Using Corona Charged Aerosol Detection

Abstract

AbstractRefractive index detector is usually used in the analysis of sugars in mushrooms, which is characterized by poor sensitivity, reproducibility, and susceptibility to interference from co-eluting sample components. In the current study, identification and determination of free sugars in mushroom samples by high-performance liquid chromatography coupled to corona charged aerosol detector (HPLC-CAD) were presented for the first time. The best chromatographic separation was performed on a Shodex Asahipak NH2P-50 4E 5 μm and mobile phase composed of 75% acetonitrile and 25% water with flow rate was 1 mL/min. The developed method offers good linearity in concentration range 0.001–0.01 or 0.01–0.2 mg/mL for tested compounds with R2 > 0.99. Limit of detection (LOD) for analytes was in the range of 7.1–120.2 ng on column. HPLC-CAD method showed very good reproducibility (RSD < 5.1%). Fructose, mannitol, and glucose were detected in all examined mushroom samples. For white Agaricus bisporus, mannitol was the most abundant sugar (7.575 mg/g dw), whereas trehalose for Pleurotus ostreatus (3.426 mg/g dw). The developed method was successfully applied for quantification of free sugars and mannitol in mushrooms. The optimized method proved to be sensitive, reproducible, and accurate.