摘要目的 建立应用液相色谱-质谱联用技术(liquid chromatography-mass spectrometry,HPLC-MS/MS)全面监测和控制银黄胶囊中金银花提取物、黄芩提取物的投料情况,并建立可同时检测银黄胶囊中绿原酸和黄芩苷含量的方法 .方法采用Inertsil ODS-3色谱柱(2.1 mm×150 mm,5μm),柱温40℃,流动相为甲醇-10 mmol/L醋酸铵溶液,梯度洗脱;采用HPLC法分离绿原酸和黄芩苷,采用电喷雾离子源负离子模式检测,选择多反应离子监测方式进行定性监测原料药的投料情况及有效成分的定量分析.结果 通过分析供试品与混合对照提取物、对照品的一级质谱图、二级质谱图有相同的特征峰,可判断该中成药是按处方工艺投料的;通过测得的HPLC-MS/MS色谱峰面积,计算出6批次供试品定量测定结果均符合标准规定.方法学验证结果表明,绿原酸和黄芩苷分别在0.60~4.80μg/ml(r=0.9989)、2.87~14.40μg/ml(r=0.9986)浓度内线性关系良好;精密度试验的RSD分别为0.69%、1.24%;平均回收率分别为95%~102%、95%~103%.结论 该法快速简便、准确可靠、灵敏度高,适用于全面监测和控制银黄胶囊中金银花提取物、黄芩提取物的投料情况及质量.

abstractsObjective To develop a HPLC-MS/MS method for comprehensive monitoring and control of the raw material feeding (honeysuchle flowers extract and baikal skullcap root extract) , and simultaneous determination of chlorogenic acid and baicalin in Yinhuang capsules. Methods The separation was performed on an Inertsil ODS-3 (2.1 mm × 150 mm, 5 μm) analytical column with the mobile phase consisting of methanol- 10 mmol/L ammonium acetate solution by gradient elution program, and the column temperature was 40 ℃. Active ingredients were separated by HPLC, and the Electrospray Ionization Mass (API) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results The samples and the mixed Extract have the same characteristic peak in MS and MS/MS. According to the prescription feeding process, the proprietary Chinese medicine wasdetermined. The results showed that the palladium residue of 6 batches samples were up to the standard by HPLC/MS/MS chromatographic peak areas. The calibration cruve of chlorogenic acid and baicalin were linear: 0.60-4.80 μg/ml (r = 0.9989),2.87-14.40 μg/ml (r = 0.9986), with the relative standard deviation of repeatability by 0.69% and 0.69% respectively, and the mean recovery rate were 95%-102%, 95%-103%, respectively. Conclusions The method was proven to be simple, accurate, reliable, high sensitive and can be used for determination and control of the raw material feeding (honeysuchle flowers extract and baikal skullcap root extract) and quality in Yinhuang capsules.