UPLC-MS/MS法同时测定感冒宁颗粒5种成分含量

Simultaneous determination of five compounds in Ganmaoning granules by UPLC-MS/MS

目的建立同时测定感冒宁颗粒中腺苷、绿原酸、咖啡酸、黄芩苷、汉黄芩素含量的方法。方法采用超高效液相色谱-串联质谱(ultra performance liquid chromatography tandem mass spectrometry, UPLC-MS/MS)联用技术,色谱柱为Agilent ZOBAX SB C18 (2.1 mm×150 mm,5 μm),流动相为甲醇-2 mmol/L乙酸铵水溶液,等度洗脱,流速0.2 ml/min,柱温35 ℃,进样量2 μl;采用电喷雾化离子源,正负离子切换模式,多反应监测模式进行定量分析,离子源温度300 ℃,干燥气温度400 ℃,干燥气流量为20 L/min,雾化气压力55 psi,毛细管电压4 000 V。结果感冒宁颗粒中腺苷、绿原酸、咖啡酸、黄芩苷、汉黄芩素检测质量浓度线性范围分别为0.20~12.80 ng (r=0.999 6)、1.00~64.00 ng (r=0.999 8)、0.40~25.60 ng (r=0.999 6)、4.00~256.00 ng (r=0.999 2)、0.20~12.8 ng (r=0.999 1),线性关系良好;检测限分别为0.02、0.10、0.04、0.40、0.02 ng,定量限分别为0.04、0.20、0.08、0.80、0.04 ng;精密度、重复性、稳定性的RSD均≤3.5%;平均加样回收率在99.30%~100.75%之间,RSD为2.09%~3.17%。结论所建立的方法简单、快速、灵敏、准确,可同时测定感冒宁颗粒中5种成分的含量,可用于感冒宁颗粒的质量控制。

Objective To establish a method for the simultaneous determination of adenosine,chlorogenic acid,caffeic acid,baicalin and wogonin in Ganmaoning granules.Methods The UPLC-MS/MS method was adopted. The separation was performed on ZOBAX SB C18 (2.1 mm×150 mm,5 μm) column with mobile phase consisted of methanol-2 mmol/L ammonium acetate aqueous solution (isocratic elution) at the flow rate of 0.2 ml/min. The column temperature was set at 35 ℃,and the injection volume was 2 μl. The electrospray ionization source (ESI) was used. Multiple reaction monitoring (MRM) mode was adopted,using positive and negative ions scanning. The ion source temperature was 300 ℃,the drying gas temperature was 400 ℃,the drying gas flow was 20 L/min,the atomizing gas pressure was 55 psi,and the capillary voltage was 4000 V.Results The linear ranges of adenosine,chlorogenic acid,caffeic acid,baicalin and wogonin in Ganmaoling granules were 0.20-12.80 ng (r=0.999 6),1.00-64.00 ng (r=0.999 8),0.40-25.60 ng (r=0.999 6),4.00-256.00 ng (r=0.999 2),0.20-12.8 ng (r=0.999 1),which showed the linear relationship was good. The limits of detection were 0.02,0.10,0.04,0.40,0.02 ng,respectively. The limits of quantitation were 0.04,0.20,0.08,0.80,0.04 ng,separately. The RSDs of precision,reproducibility and stability (24 h) tests were no more than 3.5%. The recovery rates were 99.3%-100.75% (RSD=2.09%-3.17%).Conclusions The method established in this experiment is simple,rapid,sensitive and accurate. It can be used to simultaneously determine the contents of five components in Ganmaoling granules,and can be used for quality control of Ganmaoning granules.