UFLC-QTRAP-MS-MS同时测定蚁巢伞属3种鸡[土从]中氨基酸核苷含量及PCA分析

Simultaneous determination of amino acids and nucleosides of three species in Termitomyces by UFLC-QTRAP-MS-MS

建立高分离度快速液相色谱质谱联用(UFLC-QTRAP-MS-MS)同时测定蚁巢伞属3种鸡(土从)中15种氨基酸和12种核苷类成分的方法。采用Waters XBridge Amide色谱柱(2.1 mm×100 mm,3.5μm),流动相为0.2%甲酸水-0.2%甲酸乙腈,梯度洗脱,柱温30℃,流速0.6 mL·min^(-1),采用电喷雾正离子源(ESI+),多反应离子监测(MRM)模式进行检测。采用PCA主成分分析法对不同品种的蚁巢伞属鸡(土从)进行综合评价。结果 15种氨基酸成分和12种核苷成分在一定浓度范围内均呈良好的线性关系,相关系数均大于0.997 3,精密度,重复性和稳定性良好;加样回收率在95.14%~105.0%,RSD均小于5.0%。PCA分析显示在氨基酸中,鸡(土从)综合评价指数分最高;在核苷中,黄鸡(土从)综合评价指数最高。所建立的方法准确、重复性好,可用于蚁巢伞属3种鸡(土从)中15种氨基酸和12种核苷类成分的同时测定,为蚁巢伞属鸡(土从)资源内在质量的综合评价、全面控制以及合理开发应用提供新的方法参考。

A comprehensive analytical method based on UFLC-QTRAP-MS-MS was developed for the simultaneous determination of 15 kinds of amino acids and 12 kinds of nucleosides of three species in Termitomyces. The separation was carried out on a Waters XBridge Amide column （2. 1 mm × 100 mm,3.5 μm） with gradient elution of mobile phase of 0. 2% formic acid in water-0. 2% formic acid in acetonitrile at a flow rate of 0. 6 mL · min^- 1, and column temperature was maintained at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring （MRM） mode. The principal component analysis（PCA） was made to standardized treatment for the comprehensive evaluation of different species in Termitomyces. The 15 kinds of amino acids and 12 kinds of nucleosides multiple constituents showed good linearity （ r 〉 0. 997 3 ） in the range of the tested concentration. The average recoveries ranged from 95. 14% to 105.0% ,and the relative standard deviations were less than 5.0%. The comprehensive evaluation index ob- tained with PCA showed that the Termitomyces albuminosus was significantly higher than others in amino acids and in nucleosides, of which the T. aurantiacus was the best. The developed method with good repeatability and accuracy was suitable for the simultaneous determination of multiple functional substances ,which provided a new basis for the comprehensive assessment and overall control of the quality of Termitomyces fungi.