HPLC法测定丹参及其炮制品中5种成分的含量

Contents of five components in Salviae Miltiorrhizae Radix et Rhizoma and its processed products determined by HPLC method

目的建立丹参及酒丹参中二氢丹参酮Ⅰ、四氢丹参酮Ⅰ、隐丹参酮、丹参酮ⅡA、丹参新酮5种丹参酮类成分含量的测定方法。方法采用高效液相色谱法(HPLC)测定丹参和酒丹参中二氢丹参酮Ⅰ、四氢丹参酮Ⅰ、隐丹参酮、丹参酮ⅡA、丹参新酮含量。色谱柱为Phenomenex Luna-C 18(250 mm×4.6 mm,5μm),柱温为30℃,流动相为乙腈-0.01%磷酸溶液,梯度洗脱,流速为1.0 ml/min,检测波长为270 nm。丹参和酒丹参供试品采用甲醇超声提取。结果二氢丹参酮Ⅰ、四氢丹参酮Ⅰ、隐丹参酮、丹参酮ⅡA、丹参新酮分别在0.6625~66.25μg/ml,0.5236~52.36μg/ml,3.1208~312.08μg/ml,1.9663~196.63μg/ml,0.1252~12.52μg/ml范围内质量浓度与峰面积呈良好线性关系,回归方程分别为Y=63.57X-15.52,Y=61.84 X+11.35,Y=285.83X+119.38,Y=163.36X-181.42,Y=18.53X+2.54。精密度、稳定性、重复性、加样回收率均符合分析要求。3个批次的丹参中二氢丹参酮Ⅰ、四氢丹参酮Ⅰ、隐丹参酮、丹参酮ⅡA、丹参新酮的平均含量均高于酒丹参中含量。结论建立的HPLC方法快速准确、操作简便、专属性好、灵敏度高,可为丹参及其酒炙品的质量评价提供依据。

Objective To establish a method for determining the contents of dihydrotanshinoneⅠ,tetrahydro tanshinoneⅠ,cryptotanshinone,tanshinoneⅡA and miltirone in Salviae Miltiorrhizae Radix et Rhizoma and its wine-processed product.Methods The contents of dihydrotanshinoneⅠ,tetrahydro tanshinoneⅠ,cryptotanshinone,tanshinoneⅡA,and miltirone in Salviae Miltiorrhizae Radix et Rhizoma and its wine-processed product were determined by high-performance liquid chromatography(HPLC)on Phenomenex Luna-C 18 column(250 mm×4.6 mm,5μm)with acetonitrile-0.1%phosphoric acid solution as the mobile phase at a flow rate of 1.0 ml/min.The column temperature was set at 30℃,and the detection wavelength was set at 270 nm.The test samples of Salviae Miltiorrhizae Radix et Rhizoma and its wine-processed product were extracted with methanol using ultrasonic extraction method.Results DihydrotanshinoneⅠ,tetrahydro tanshinoneⅠ,cryptotanshinone,tanshinoneⅡA and miltirone showed a good linear relationship with peak area in the concentration ranges of 0.6625-66.25μg/ml,0.5236-52.36μg/ml,3.1208-312.08μg/ml,1.9663-196.63μg/ml,0.1252-12.52μg/ml,respectively,and the regression equations were Y=63.57X-15.52,Y=61.84X+11.35,Y=285.83X+119.38,Y=163.36X-181.42,Y=18.53X+2.54,respectively.The precision,the stability,the repeatability,and the sample recovery rate all met the analysis requirements.The average contents of dihydrotanshinoneⅠ,tetrahydro tanshinoneⅠ,cryptotanshinone,tanshinoneⅡA,and miltirone in three batches of Salviae Miltiorrhizae Radix et Rhizoma were lower than those in its wine-processed product,respectively.Conclusion The established HPLC method is fast,accurate,simple,specific and sensitive,and can provide a basis for the quality evaluation of Salviae Miltiorrhizae Radix et Rhizoma and its wine-processed product.