Development and validation of an HPLC-DAD-MS method for determination of four nucleoside compounds in the New Zealand native mushroom Hericium sp.

To identify and quantify the content of nucleoside compounds in the New Zealand native edible mushroom Hericium sp., a high-performance liquid chromatography coupled with a triple quadrupole detector mass method was developed and validated. Four nucleoside substitutes, namely cytidine, uridine, adenosine, and guanosine, were identified. Optimization was conducted to study the effect of extraction method type, solvent pH, and extraction time. The optimal conditions were obtained using ultrasonic treatment in water at pH 3.8 for 30 min. For chromatographic separation, a C18 column was applied using 0.1% formic acid (pH 3.4) as the mobile phase with detection at 260 nm. The total concentration of the four nucleoside compounds was high, at 10.7 mg/g dry weight, indicating a potential benefit for human health. The excellent validation results based on selectivity, linearity, precision, accuracy and robustness revealed the reliability of the newly developed analytical method, which could be applied routinely in research laboratories.