Abstract: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-C18column，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 0.995 2-0.999 9 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 applicable to the analysis of 6 nucleosides in dried rhizome of L.lucidus from different habitats.