不同加工方法中何首乌核苷类成分的含量测定及主成分分析

目的: 建立QTRAP UPLC-MS/MS同时测定何首乌中10种核苷类成分(核苷及碱基)含量的方法,分析不同加工方法对何首乌核苷类成分含量的影响。方法: 样品用超纯水室温下超声提取,提取液经高速离心处理,取上清液,经Waters Atlantis T₃色谱柱(2.1 mm×150 mm,3 μm),以甲醇-5 mmol·L^-1醋酸铵(含0.1%冰醋酸)为流动相,0.4 mL·min^-1梯度洗脱,质谱采用正离子多反应监测(MRM)模式测定不同加工何首乌药材中10种核苷类成分的含量,用主成分分析对核苷类成分进行综合评价。结果: 10种核苷类成分具有良好的线性关系,相关系数均大于0.99,且线性范围满足实验要求;平均加样回收率为98.80%~102.54%, RSD在1.78%~3.02%之间。何首乌核苷类成分中以尿苷、腺嘌呤、鸟苷、胞苷含量较高;不同加工方法对何首乌核苷类成分含量具有一定影响,何首乌厚片80℃烘干样品中核苷类成分综合质量较好。结论: 本文为揭示加工方法对何首乌化学成分的影响及优选何首乌适宜产地和加工方法提供基础资料。

Objective: To establish a method for simultaneous determination of 10 nucleosides and nucleobases in Polygoni Multiflori Radix, and investigate the influence different processing methods on the content of nucleosides and nucleobases in Polygoni Multiflori Radix. Methods: The samples with ultrapure water were extracted by ultrasonic machine at room temperature. The extract was then centrifuged, and the supernatant was harvested. Under optimized chromatographic conditions, good separation for 10 target compounds were obtained on a Waters Atlantis T₃ column (2.1 mm×150 mm, 3 μm) eluted with the mobile phase of methanol-5 mmol·L^-1 ammonium acetate (0.1% acetic acid) at a flow rate of 0.4 mL·min^-1. The positive ion multiple reaction monitoring (MRM) mode was applied for the analysis of 10 kinds of nucleosides and nucleobases in Polygoni Multiflori Radix. In addition, the principal component analysis(PCA) was carried out have a comprehensive evaluation. Results: Good linearity was observed for all the 10 nucleosides and nucleobases . The linearity of the detected nucleosides and nucleobases was excellent more than 0.99 and the limits of detection were all satisfied. The average recoveries of standard addition for the compounds were between 98.80% and 102.54%, and the relative standard deviations were between 1.78% and 3.02%. The content of uridine, adenine, guanosine and cytidine was higher in Polygoni Multiflori Radix. The different processing methods of Polygoni Multiflori Radix showed certain influence on 13 kinds of nucleosides and nucleobases, and 80℃ thick piece had a better quality. Conclusion: Our study has investigated the influence of processing methods on the determination of nucleosides and nucleobases in Polygoni Multiflori Radix, and provided a scientific basis for suitable processing method of Polygoni Multiflori Radix.