A Practical Method for Determination of Nine Nucleosides in Tricholoma matsutake by UPLC/MS and Quantitative Analysis of Multicomponents Using Single Marker Method.

Li Yong,Zhi-Xiang Yang,Ying Xue,An-Qin Leng,Marcia F Mesko

doi:10.1155/2021/9571329

Abstract

Nucleosides can be used as quality evaluation indicators of Tricholoma matsutake. In this work, a new ultra-performance liquid chromatography-tandem mass spectrometry (UPLC/MS) strategy for quantitative analysis of multiple components using a single marker (QAMS) was proposed to determine nine nucleosides (adenosine, cytidine, guanosine, inosine, uridine, 2′-deoxyadenosine, 2′-deoxycytidine, 2′-deoxyguanosine, and 2′-deoxyuridine) in T. matsutake. Guanosine was set as the internal reference substance, whose content in T. matsutake was determined using the conventional external standard method. Relative correction factors (RCFs) between guanosine and eight other nucleosides were measured. The concentrations of the eight components were calculated with the obtained RCFs by QAMS. An ultrasonic extraction method is used for sample preparation. This method was validated to be sensitive, precise, and accurate with the LODs of 0.31–1.9 ng, the overall intraday and interday variations less than 4.08%, and the overall recovery over 89.0%. The correlation coefficients (r2) of the calibration curves were higher than 0.9918. The values of vector angle analysis were above 0.99845, which indicates no significant differences between the conventional external standard method and the present QAMS method. As far as we know, this is also the first report of UPLC/MS analysis of nucleoside compounds by QAMS, providing an efficient and feasible quality assessment method for other natural products containing nucleosides.

Highlights

Tricholoma matsutake is a wild edible fungus endemic to East Asia. e Hengduan Mountain Region of Southwest China, especially Sichuan Province and Yunnan Province, is the world’s foremost production center of T. matsutake
Guanosine was set as the internal reference substance, whose concentration in T. matsutake was determined using an external standard. e concentration’s relative correction factors (RCFs) between guanosine and eight other nucleosides were determined using external standards. is method was validated to be sensitive, precise, and accurate with the LODs of 0.31–1.9 ng, the overall intraday and interday variations less than 4.08%, and the overall recovery over 89.0%. e correlation coefficients (r2) of the calibration curves were higher than 0.9918. en, the concentrations of the components were calculated with the obtained RCFs by QAMS
For confidence in the boiling water method, we carried out a comparative experiment by six replicate. e results of the six samples are given in Table 2. rough one-way ANOVA, the results showed no significant difference between the two extraction methods (P-value 0.99). e subsequent method validation results further verified that both methods can be used to extract nucleosides from T. matsutake

Summary

Introduction

Tricholoma matsutake is a wild edible fungus endemic to East Asia. e Hengduan Mountain Region of Southwest China, especially Sichuan Province and Yunnan Province, is the world’s foremost production center of T. matsutake. Nucleic acid constituents that can regulate various physiological processes in vivo through the purine/pyrimidine receptors are considered suitable markers for the quality evaluation of T. matsutake [7, 8]. Sichuan Food and Drug Administration commissioned researchers from several units, including our group, to jointly draft local standards for the safety of T. matsutake and its products, which were implemented on July 20, 2018. On this basis, our team conducted an in-depth study on the active ingredients in T. matsutake and proposed for the first time that nucleic acid compounds be used as the quality control markers for commercial T. matsutake products. The external standard method, as a classical quantitative test method, requires the purchase of all reference substances and preparation of each corresponding solution as well as other operations, which may be considered wasteful of time

Methods

Results

Conclusion