不同采收期太子参中13种核苷类成分的含量测定研究

 目的 建立超高效液相色谱-三重四级杆线性离子肼复合质谱(QTRAP UPLC-MS/MS)同时测定太子参中13种核苷类成分含量的方法,分析不同生长期太子参中核苷类成分积累的动态变化。方法 采用Waters Atlantis T3色谱柱(2.1 mm×150 mm,3 μm),以5 mmol·mL^-1醋酸铵(B相)-甲醇(A相)为流动相,0.4 mL·min^-1梯度洗脱:0~4.5 min,3%~4%A;4.5~8 min,4%~18% A;8~10 min,18% A;10~10.1 min,18%~3% A;10.1~13 min,3% A。正离子多反应监测(MRM)模式测定。结果 13种核苷类成分的浓度与峰面积的线性关系良好(r>0.991 0);加样回收率为95.52%~105.07%。太子参核苷类成分中,以胞苷、尿苷、肌苷、鸟苷、胸苷含量较高;不同生长期太子参核苷类成分含量有所差异,6月中旬含量相对较高。结论 该方法简便、灵敏,结果准确、可靠,为探索太子参药材的品质形成机制及确定药材适宜采收期提供依据。

Quantitative Determination of Thirteen Nucleosides and Nucleobases in Pseudostellariae Radix in Different Harvest Periods by QTRAP LC-MS/MS

OBJECTIVE To establish a QTRAP UPLC-MS/MS method for simultaneous determination of thirteen nucleosides and nucleobases in Pseudostellariae Radix, and analyze their dynamic changes at different periods. METHODS The separation was carried out on a Waters Atlantis T3 column (2.1 mm×150 mm,3 μm)eluted by a mobile phase of 5 mmol·mL^-1 of ammonium acetate (B)-methanol(A) at a flow rate of 0.4 mL·min^-1. The elution program is as follows. 0-4.5 min,3%-4%A;4.5-8 min,4%-18% A;8-10 min,18%A;10-10.1 min,18%-3% A;10.1-13 min,3%A. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. RESULTS The calibration curves of the 13 nucleosides and nucleobases showed good linearity (r>0.991 0) in the ranges of the tested concentrations, and the average recoveries were between 96.46%-105.07%. The contents of cytidine, uridine, inosine, guanosine, and thymidine in Pseudostellariae Radix harvested at different periods showed some differences, and the contents were higher in mid-June. CONCLUSION The method is simple, sensitive, accurate and credible, and provides the basis for exploring the quality forming mechanism of Pseudostellariae Radix medicinal materials and the suitable harvest time.