Get access

Analysis of nucleosides and monophosphate nucleotides from mushrooms with reversed-phase HPLC



The retention characteristics of five stationary phases were tested by using a selection of 5′-mononucleotides and nucleosides with the aim to develop a simple, rapid and sensitive reversed-phase liquid chromatography method without ion-pair reagent usage. The method was optimized by changes in temperature, pH and ionic strength on a column showing a superior performance. The mobile phase consisted of a mixture of 0.05 M phosphate buffer and methanol, delivered at a flow rate of 0.4 mL/min and based on a gradient program. UV detection was used at a 254 nm wavelength. The method was validated for a quantitative analysis of 5′-mononucleotides and nucleosides in wild edible mushrooms. For all nucleosides and nucleotides, the LOD and LOQ were less than 0.02 and 0.07 μg/mL, respectively. Validation parameters yielded recovery rates between 68.6 and 98.2%, with a precision expressed as a relative standard deviation of 7.6–15.3%. The content of 5′-mononucleotides and nucleosides was determined for 10 samples of wild edible mushrooms found in Croatia and, accordingly, the equivalent umami concentrations were calculated.

Get access to the full text of this article