不同产地地参核苷类成分的UPLC-MS-MS定性定量分析

Qualitative and Quantitative Analysis of Nucleosides in Dried Rhizome of Lycopus lucidus var.hirtus Regel from Different Habitats by UPLC-MS-MS

该文建立1种地参中胞苷、尿嘧啶、腺嘌呤、尿苷、鸟苷和腺苷6种核苷类成分的超高效液相色谱-串联质谱(ultra performance liquid chromatograph tandem mass spectrometer,UPLC-MS-MS)定性定量分析方法。采用纯水超声提取,以甲酸水溶液(pH4.0)-甲醇为流动相,在Agilent SB-C_(18)色谱柱上梯度洗脱,正离子多反应监测模式测定,并采用差异性分析、聚类分析和相关性分析对测定结果进行比较。结果显示:6种核苷类成分在各自的质量浓度范围内线性关系良好,相关系数为0.9952~0.9999,检出限为0.07 ng/mL~8.34 ng/mL,加标回收率为96.30%~103.71%,相对标准偏差值为0.68%~2.32%。核苷含量结果显示不同产地间存在一定差异性(p<0.05),其中江苏徐州(S7)样品中核苷总量最高;6种核苷类成分中尿苷和鸟苷含量居首,分别占比核苷总量的35.60%和34.60%,是地参中主要的核苷类成分。该方法定性定量准确、分析速度快、灵敏度高,可用于不同产地地参中6种核苷类成分的分析。

This study aimed to develop an ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS-MS)method for simultaneous determination of six nucleosides(cytosine,uracil,adenine,uridine,guanosine,and adenosine)in dried rhizome of Lycopus lucidus var.hirtus Regel from different habitats.To be specific,the samples were extracted by sonication with pure water.UPLC-MS-MS conditions were as follows:formic acid solution(pH4.0)and methanol as mobile phase,Agilent SB-C_(18) column,positive ion mode,multiple reaction monitoring.The determination results were subjected to difference analysis,cluster analysis,and correlation analysis.The result suggested that the 6 nucleosides showed good linear relationship within the respective range of mass concentration,with correlation coefficients of 0.9952-0.9999 and limits of detection of 0.07 ng/mL-8.34 ng/mL.The spiked recoveries fell in the range of 96.30%-103.71%,and the relative standard deviations were 0.68%-2.32%.The nucleoside content was different among habitats(p<0.05)and the highest content was found in the sample from Xuzhou of Jiangsu province(S7).Uridine and guanosine topped the 6 nucleosides in terms of the content in dried rhizome of L.lucidus,which accounted for 35.60%and 34.60%of the total nucleosides,respectively.This method was accurate,fast,and sensitive,which was appli-cable to the analysis of 6 nucleosides in dried rhizome of L.lucidus from different habitats.