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

Refractive 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 mu m 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 withR(2) > 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 whiteAgaricus bisporus, mannitol was the most abundant sugar (7.575 mg/g dw), whereas trehalose forPleurotus 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.

Automated summary

The refractive index detector lacks sensitivity, reproducibility, and is susceptible to interference. This study introduced a new method using HPLC-CAD for identifying and quantifying free sugars in mushrooms. The developed method showed good linearity and reproducibility, successfully detecting fructose, mannitol, and glucose in mushroom samples, with mannitol being the most abundant sugar in white Agaricus bisporus and trehalose in Pleurotus ostreatus.