HPLC法同时测定中药丹参中水溶性和脂溶性成分的含量
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国家“十一五”科技支撑计划课题(2006BAI08B03-07).


HPLC simultaneously determines multiple hydrophilic and lipophilic bioactive components in Radix et Rhizoma Salviae Miltiorrhizae
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Supported by National Science and Technology Supporting Program of the Ministry of Science and Technology(2006BAI08B03-07).

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    摘要:

    目的:建立同时测定丹参中2种水溶性成分和2种脂溶性成分的方法。方法:HPLC-DAD法,采用Agilent Zorbax TC C18柱(4.6 mm×250 mm,5 μm),以甲醇2%醋酸为流动相进行梯度洗脱:0~15 min,30% B~40% B; 15~20 min,40% B~60% B; 20~25 min,60% B~90% B; 25~40 min,90% B。检测波长为281 nm,柱温:35℃。结果:迷迭香酸、丹酚酸B、隐丹参酮、丹参酮ⅡA浓度分别在3.76~120.20 μg·ml-1 (r=0.999 9)、34.20~1 094.5 μg·ml-1 (r=0.999 9)、0.64~20.32 μg·ml-1 (r=0.999 9)、1.02~32.72 μg·ml-1 (r=0.999 6)范围内呈良好的线性关系。4种成分精密度试验RSD <1%。48 h内稳定性RSD<1%。加样回收率为99.72%~100.63%。结论:该含量测定方法简便,分离效果好,能同时测定丹参中迷迭香酸、丹酚酸B、隐丹参酮及丹参酮ⅡA四种有效成分的含量,结果准确可靠。

    Abstract:

    Objective:To develop a new method for the simultaneous determination of two hydrophilic components and two lipophilic components of Radix et Rhizoma Salviae Miltiorrhizae.Methods: The HPLC-DAD method was employed using a column of Agilent Zorbax TC C18 (4.6 mm×250 mm, 5 μm) with a mobile phase of methanol -2% acetic acid.The gradient elution program was as follow:0-15 min, 30% B-40% B; 15-20 min, 40% B-60% B; 20-25 min, 60% B-90% B;25-40 min, 90% B.The detection wavelength was set at 281 nm and the temperature was 35℃.Results: The linearity was obtained over 3.76-120.20 μg·ml-1(r=0.999 9) for rosmarinic acid, 34.20-109 4.5 μg·ml-1(r=0.999 9) for salviamolic acid B, 0.64-20.32 μg·ml-1(r=0.999 9) for clyptotanshinon,and 1.02-32.72 μg·ml-1(r=0.999 6) for tanshinone ⅡA.The RSDs of precision and stability of the sample were both less than 1% in 48 hours.The average recovery was between 99.72%-100.63%.Conclusion: The present method is simple and has satisfactory efficacy; it can simultaneously determine multiple hydrophilic and lipophilic bioactive components in Salvia miltiorrhiza from different areas.

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  • 收稿日期:2008-09-06
  • 最后修改日期:2008-12-11
  • 录用日期:2008-12-15
  • 在线发布日期: 2009-02-17
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