目的: 建立一测多评法同时测定丁公藤及其炮制品中7个活性成分含量,并分析炮制对其影响。方法: 采用超高效液相色谱法(UPLC),以绿原酸为参照物,建立新绿原酸、东莨菪苷、东莨菪内酯、异绿原酸B、异绿原酸A和异绿原酸C的相对校正因子。分别采用外标一点法和一测多评法测定上述7个活性成分含量,比较计算值与实测值的差异,以验证一测多评法的准确性和和可行性。结果: 新绿原酸、东莨菪苷、绿原酸、东莨菪内酯、异绿原酸B、异绿原酸A、异绿原酸C分别在0.028~1.420、0.019~0.956、0.027~1.324、0.014~0.720、0.017~0.824、0.010~0.500和0.013~0.672 μg·μL-1范围内呈现良好线性关系,平均加样回收率(n=6)分别为98.9%、99.0%、100.6%、101.2%、100.8%、101.7%和100.5%,RSD分别为1.1%、1.7%、1.5%、1.6%、0.55%、1.6%和1.5%。采用校正因子计算的7个活性成分含量值与外标法实测值之间无显著差异。不同丁公藤饮片中新绿原5个有机酸(新绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C)含量变化趋势基本一致,Y5(甘草汁-盐水煮品)含量最高,Y10(甘草汁-盐水晒品Ⅱ)含量最低,东莨菪苷含量以Y5含量最高(3.53 mg·g-1),Y10最低(0.31 mg·g-1)。东莨菪内酯含量以Y7(甘草汁晒品Ⅰ)含量最高(1.48 mg·g-1),Y3(甘草汁煮品Ⅱ)含量最低(0.30 mg·g-1)。结论: 以绿原酸为参照物建立的一测多评法可用于丁公藤饮片质量评价,加热和加辅料炮制均对丁公藤的化学成分产生了一定影响。
Objective: To establish a method and validate its feasibility for quality evaluation of Erycibes Caulis and its processed products, and analyze the effect on the contents of the seven active ingredient before and after processing. Methods: The contents of components were determined by ultra performance liquid chromatography (UPLC). Chlorogenic acid was chosen as the internal reference substances, the relative correction factors (RCFs) of neochlorogenic acid scopolin, scopoletin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C and to chlorogenic acid were established. The contents of these seven active ingredients were determined by external standard method and quantitative analysis of multi-components by single-marker (QAMS) method. The method was evaluated by comparison of the quantitative results between external standard method and quantitative analysis of multi-components by single-marker (QAMS). Results: The peaks of neochlorogenic acid, scopolin, chlorogenic acid, scopoletin, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C in the sample showed good linear relationship between 0.028-1.420 μg·μL-1, 0.019-0.956 μg·μL-1, 0.027-1.324 μg·μL-1, 0.014-0.720 μg·μL-1, 0.017-0.824 μg·μL-1, 0.010-0.500 μg·μL-1 and 0.013-0.672 μg·μL-1, respectively. The average recoveries of them(n=6)were as follows 98.9%, 99.0%, 100.6%, 101.2%, 100.8%, 101.7% and 100.5%, with the RSDs of 1.1%, 1.7%, 1.5%, 1.6%, 0.55%, 1.6% and 1.5%, respectively. It was showed that no significant difference was found in the quantitative results of seven ingredients by external standard method and QAMS method. The content of 5 kinds of organic acids (neochlorogenic acid, chlorogenic acid, isochlorogenic acid B, isochlorogenic acid A and isochlorogenic acid C) in Erycibes Caulis with different processing technology showed the same change trend. The sample of Y5 (boiled products with licorice sauce and brine) had the highest contents, and the sample of Y10 (dried products Ⅱ soaked in licorice sauce and brine) had the lowest. The content of scopoline was the highest in Y5(3.53 mg·g-1), while the lowest was Y10(0.31 mg·g-1). The content of scopoletin in Y7(dried products Ⅰ with licorice sauce) was the highest(1.48 mg·g-1) and that in Y3(boiled products Ⅱ with licorice sauce) was the lowest(0.30 mg·g-1). Conclusion: The RCFs established in the QAMS methods with chlorogenic acid as the internal reference substances is accurate and feasible. It can be used to control the quality of Erycibes Caulis. Both heating and excipients preparation have a certain effect on the active ingredients in Erycibes Caulis.
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