一测多评法用于茵栀黄传统汤剂12种成分含量测定的适用性研究

Study on Applicability of QAMS for Content Determination of 12 Components in Yinzhihuang Traditional Decoction

目的建立一测多评法同时测定茵栀黄传统汤剂12种成分含量的方法,验证该方法在其质量控制中应用的可行性与适用性。方法采用Agilent Eclipse plus C18色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,流速1.0 mL/min,柱温30℃,新绿原酸、绿原酸、隐绿原酸、异绿原酸A、异绿原酸B、异绿原酸C检测波长为327 nm,栀子苷检测波长为238 nm,对羟基苯乙酮检测波长为275 nm,黄芩苷、千层纸素A苷、汉黄芩苷、黄芩素检测波长为280 nm,以黄芩苷为内参物,建立黄芩苷与新绿原酸、绿原酸、隐绿原酸、栀子苷、对羟基苯乙酮、异绿原酸B、异绿原酸A、异绿原酸C、千层纸素A苷、汉黄芩苷、黄芩素的相对校正因子,计算12种成分含量。将一测多评法测定结果与外标法测定结果对比,验证一测多评法的准确性。结果 12种成分的平均加样回收率为96.86%~103.69%;黄芩苷与新绿原酸、绿原酸、隐绿原酸、栀子苷、对羟基苯乙酮、异绿原酸B、异绿原酸A、异绿原酸C、千层纸素A苷、汉黄芩苷、黄芩素的相对校正因子平均值分别为0.916、1.107、0.962、2.297、3.714、3.294、0.183、0.795、1.798、1.021、5.679;15批样品中12种成分一测多评法的计算值与外标法实测值无明显差异(P>0.05)。结论本研究建立的一测多评方法简便、稳定,可用于茵栀黄传统汤剂中12种指标成分的同步质量控制。

Objective To establish a method for the content determination of 12 components in Yinzhihuang traditional decoction through quantitative analysis of multi-components with single marker(QAMS);To verify the feasibility and applicability of the method in its quality control. Methods Agilent Eclipse plus C18 column(4.6 mm × 250 mm, 5 μm) was used;acetonitrile-0.1% phosphoric acid aqueous solution was used as mobile phase;gradient elution;flow rate was 1.0 m L/min;column temperature was 30 ℃. Detection wavelength for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C was 327 nm;geniposide detection wavelength was 238 nm;p-hydroxyacetophenone detection wavelength was 275 nm;detection wavelength for baicalin, melaleuca A glycoside, wogonin, baicalein was 280 nm. With baicalin as the internal reference, relative correction factors of baicalin and neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, geniposide, p-hydroxyacetophenone, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, melaleuca A glycoside, wogonin and baicalein was established, and the 12 components were determined. The measurement results of QAMS were compared with the results of the external standard method to verify the accuracy of QAMS. Results The average recovery rates of 12 components were 96.86%–103.69%. The relative positive factors of baicalin and neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, geniposide, p-hydroxyacetophenone, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, melaleuca A glycoside, wogonin and baicalei were 0.916, 1.107, 0.962, 2.297, 3.714, 3.294, 0.183, 0.795, 1.798, 1.021, 5.679, respectively. There was no significant difference between the calculated values of 12 components in 15 batches of samples by the QAMS and the measured values of the external standard method(P>0.05). Conclusion The method established in the study is simple and stable, and can be used for the synchronous quality control of 12 index components in Yinzhihuang traditional decoction.