Validation of an Aqueous Normal Phase Chromatography Method for the Analysis of Ergothioneine in Commercial Mushrooms

Abstract

An analytical HPLC method for the identification and quantification of ergothioneine (EGT) has been developed using a silica hydride-based column. A simplified sample preparation method was developed. Several types of commercially available mushrooms were analyzed for quantification of ergothioneine using a Cogent Diamond Hydride column (100 x 4.6 mm) at a flow rate of 1 mL/min, with a deionized water:acetonitrile:0.1% formic acid mobile phase using an aqueous normal phase (ANP) gradient. EGT was detected at 254 nm and was identified by matching retention times of the verified standard. The standard ergothioneine content ranged from 0.01 mg to 0.1 mg/mL with excellent linearity correlation (R2 = 0.99999). EGT recoveries were within 96.5–100.3%. The developed HPLC method was simple, accurate, reproducible, sensitive and rugged.