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[摘要]
目的 建立了HPLC-ESI-MS/MS法同时测定八珍益母丸中盐酸水苏碱、盐酸益母草碱、芍药苷、阿魏酸、甘草苷、党参炔苷、毛蕊花糖苷、白术内酯I和猪苓酸C 9种有效成分的分析方法。方法 HPLC法采用色谱柱Waters Atlantis T3柱(150 mm×4.6 mm,3.0μm),流动相为甲醇-0.1%甲酸水溶液,梯度洗脱,体积流量0.5 mL/min,进样量10 μL;质谱采用电喷雾离子源、负离子模式(ESI−),通过多反应监测(MRM)同时对八珍益母丸中的9种有效成分进行定量分析。结果 八珍益母丸中9种有效成分盐酸水苏碱、盐酸益母草碱、芍药苷、阿魏酸、甘草苷、党参炔苷、毛蕊花糖苷、白术内酯I和猪苓酸C的线性范围分别为0.04~40.00 μg/mL(r=0.999 2)、0.04~40.00 μg/mL(r=0.999 3)、1.0~100.0 μg/mL(r=0.999 1)、0.2~20.0 μg/mL(r=0.999 6)、0.2~20.0 μg/mL(r=0.997 5)、0.05~5.00 μg/mL(r=0.999 4)、0.1~10.0 μg/mL(r=0.999 4)、0.1~10.0 μg/mL(r=0.999 2)、0.1~10.0 μg/mL(r=0.999 2),平均加样回收率分别为99.7%、98.1%、98.5%、101.5%、99.5%、98.4%、99.1%、101.2%、100.1%,RSD值分别为0.52%、0.64%、1.08%、1.12%、0.39%、0.74%、0.91%、0.54%、0.47%。9批八珍益母丸样品中9种有效成分的质量分数分别为0.423~0.752、0.505~0.722、0.613~1.300、0.102~0.184、0.195~0.255、0.021~0.035、0.034~0.072、0.039~0.063、0.051~0.095 mg/g。结论 所建立的分析方法简单、灵敏度高、专属性好,可应用于不同厂家、不同生产批次的八珍益母丸中有效成分的测定及质量控制。
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[Abstract]
Objective To develop and validate a high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of nine major bioactive components (stachydrine hydrochloride, leonurine hydrochloride, paeoniflorin, ferulic acid, liquiritin, lobetyolin, verbascoside, atracylenolide I, and polyporenic acid C) in Bazhen Yimu Pills (BYP). Methods The chromatographic separation was performed on an Waters Atlantis T3 column (150 mm×4.6 mm, 3.0 μm) with a gradient elution of methanol and 0.1% formic acid in water at a flow rate of 0.5 mL/min, and the injection volume was 10 μL. The nine major bioactive components were detected using an electrospray ionization source in negative ionization mode (ESI−) and quantified by multiple reaction monitor (MRM) scanning at the same time. Results The linear ranges of stachydrine hydrochloride, leonurine hydrochloride, paeoniflorin, ferulic acid, liquiritin, lobetyolin, verbascoside, atracylenolide I, and polyporenic acid C were 0.04-40.00 μg/mL (r=0.999 2), 0.04-40.00 μg/mL (r=0.999 3), 1.0-100.0 μg/mL (r=0.999 1), 0.2-20.0 μg/mL (r=0.999 6), 0.2-20.0 μg/mL (r=0.997 5), 0.05-5.00 μg/mL (r=0.999 4), 0.1-10.0 μg/mL (r=0.999 4), 0.1-10.0 μg/mL (r=0.999 2), 0.1-10.0 μg/mL (r=0.999 2), and the average recoveries were 99.7% (RSD=0.52%), 98.1% (RSD=0.64%), 98.5% (RSD=1.08%), 101.5% (RSD=1.12%), 99.5% (RSD=0.39%), 98.4% (RSD=0.74%), 99.1% (RSD=0.91%), 101.2% (RSD=0.54%), and 100.1% (RSD=0.47%), respectively. The content of nine batches of the nine major bioactive components were 0.423-0.752, 0.505-0.722, 0.613-1.300, 0.102-0.184, 0.195-0.255, 0.021-0.035, 0.034-0.072, 0.039-0.063, and 0.051-0.095 mg/g, respectively. Conclusion The developed method is simple, specific, and sensitive, and it can be applied for the determination of nine major bioactive components and the quality control of BYP collected from different production batches.
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