目的: 建立快速、准确、可同时测定矮地茶中10个成分(3个儿茶素类、4个黄酮、2个酚酸和1个异香豆素类)的HPLC-ESI-MS/MS分析方法,并用于不同于产地矮地茶药材的分析。方法: 采用Diamonsil-C18(150 mm×4.6 mm,5 μm,Dikma)色谱柱,以甲醇-0.02%甲酸水为流动相,梯度洗脱,流速0.8 mL·min-1;采用ESI负离子化监测,源喷射电压(IS)-4 500 V,离子源温度650 ℃,雾化气(Gas1)413 kPa,加热气(Gas2)448 kPa,帘气172 kPa。由MRM-IDA-EPI模式对矮地茶中没食子酸、表儿茶素、岩白菜素、原儿茶酸、表没食子儿茶素没食子酸酯、表儿茶素没食子酸酯、杨梅苷、芦丁、杨梅素、槲皮素10个成分的结构进行确证,采用MRM扫描模式建立测定其含量的方法,用所建立的方法对25批不同产地的矮地茶药材进行定量测定,将不同组分的含量,应用SPSS软件经过标准化转换后做散点图进行分析。结果: 10个化学成分的峰面积与质量浓度在测定范围内均具有良好的线性关系,检测限、定量限、精密度和稳定性均符合要求,加样回收率范围为97.7%~103.6%,RSD为0.43%~3.3%。样品测定结果表明,不同产地矮地茶有效成分的含量差异较大,矮地茶的道地药材(来自于湖南、湖北和广东)与对照药材趋于一致。结论: 本法简便、快速,灵敏度高,专属性好,可用于矮地茶中10个成分的含量测定,可为矮地茶药材质量控制提供强有力的工具。
Objective: To develop a sensitive and selective high performance liquid chromatography-tandem mass spectrometric method (HPLC-ESI-MS/MS) in order to qualitatively identify and quantitatively determine the ten components including three catechins, four flavonoids, two phenolic acids and one isocoumarin in Ardisiae Japonicae Herba from different habitats. Methods: Chromatographic separation was performed on a C18 column with linear gradient elution of methanol and 0.2‰ formic acid at a flow rate of 0.8 mL·min-1. The identification and quantification of those ten analytes were achieved on a hybrid quadrupole linear ion trap mass spectrometer. The detection was accomplished in the negative mode using multiple-reaction monitoring. The operating conditions were as follows: the ion spray voltage was -4 500 V; the turbo spray temperature was 650 ℃; nitrogen was used as the nebulizer gas (413 kPa) and heater gas (448 kPa); the curtain gas was kept at 172 kPa and interface heater was on. The results of different components were standardized by SPSS software, and analyzed using scatter diagram. Results: The linear relationships, linearity range, precision, stability, LOD and LOQ of the method were good for the 10 components. The average recoveries were found between 97.7%-103.6% and the precision in terms of RSDs were in the range of 0.43%-3.3%. The proposed method was successfully applied for the qualitative and quantitative analysis of ten chemical compositions in twenty-five batches of Ardisiae Japonicae Herba samples from different habitats which had great variation on the contents. The result of scatter diagram analysis demonstrated that the samples from authentic source (Hunan, Hubei and Guangdong) had closer relationship to the reference sample than samples from other sources. Conclusion: The method was simple, rapid, sensitive and specific. Simultaneous quantification of bioactive components by HPLC-ESI-MS/MS coupled with chemometric techniques would be a well-acceptable strategy to comprehensively control the quality of Ardisiae Japonicae Herba.
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