不同“发汗”条件加工玄参中核苷类成分的含量测定及灰色关联度分析

Content determination and grey relational analysis of nucleosides and nucleobases in different “sweating” processed products of Scrophulariae Radix

目的:建立QTRAP-UPLC-MS/MS同时测定玄参中10种核苷类成分含量的方法,分析不同"发汗"加工条件对玄参核苷类成分含量的影响。方法:采用Atlantis T_3色谱柱(2.1 mm×150 mm,3μm),流动相为甲醇-5 mmol·L^(-1)醋酸铵(含0.1%冰醋酸),流速0.4 mL·min^(-1)梯度洗脱,用正离子多反应监测(MRM)模式测定样品中核苷类成分含量;用灰色关联度分析对核苷类成分进行综合评价。结果:玄参中10种核苷类成分的浓度与峰面积的线性关系良好(r≥0.997 3);平均加样回收率(n=6)为98.7%~101.0%,RSD为0.96%~2.44%。玄参核苷类成分中以鸟苷、尿苷、腺苷、尿嘧啶含量较高;不同"发汗"加工条件对玄参核苷类成分含量具有一定影响,以玄参完整主根、烘箱干燥(35℃)、发汗3 d的"发汗"加工样品中核苷相对关联度质量较优。结论:本法简便、快速、灵敏、准确,为揭示"发汗"加工条件对玄参化学成分的影响及建立规范、标准的玄参"发汗"初加工方法提供基础资料。

Objective： To establish a QTRAP-UPLC-MS/MS method for content determination of 10 nucleosides and nucleobases in Scrophulariae Radix, and investigate the influence of nucleosides and nucleobases of the different ＂sweating＂ processing methods of Scrophulariae Radix. Methods： The analysis was carried out on an Waters Atlantis T3 column（2.1 mm×150 mm,3μm ）with a mobile phase of methanol-5 mmol · L^-1 ammonium acetate （0.1% acetic acid ）at the flow rate of 0.4 mL· min^-1. The target compounds were analyzed by the positive ion multiple reaction monitoring （ MRM ） mode. In addition, grey relational analysis was carrid out for comprehensive evaluation. Results： All the 10 kinds of nucleosides and nucleobases showed good linearity （ r≥ 0.997 3 ）in the range of the tested concentration. The average recovery （ n=6 ）was 98.7%-101.0%, and RSD was 0.96%-2.44%. The contents of guanosine, uridine, adenosine and uracil in Scrophulariae Radix were higher. There are differences between nucleosides and nucleobases in Scrophulariae Radix with different ＂sweating＂ processing methods, and grey incidence degree method analysis was used for comprehensive evaluation. Relative relation grade of the processing method of 35 （2 oven drying and sweating for 3 days was better. Conclusion： The established method is convenient, rapid, sensitive and reliable. The study has investigated the influence of ＂sweating＂ processing methods by the determination of nucleosides and nucleobases in Scrophulariae Radix, and provided a scientific basis for Scrophulariae Radix normative and standard ＂sweating＂ processing method.