腹水草HPLC指纹图谱研究及香草酸等6种成分的含量测定

目的：建立腹水草茎叶的HPLC指纹图谱和同时测定咖啡酸、香草酸、阿魏酸、原儿茶酸、木犀草素和金合欢素含量的方法，为腹水草药材的质量控制提供可靠参考。方法：采用Zobax C₁₈色谱柱（250 mm×4.6 mm，5μm），以乙腈-0.1%磷酸溶液为流动相，进行梯度洗脱，流速1.0 mL·min^-1，检测波长230 nm，柱温30℃，建立腹水草茎叶指纹图谱，使用中国药典委员会“中药色谱指纹图谱相似度评价系统2004A版”确定共有峰，计算相似度；并对咖啡酸、香草酸、阿魏酸、原儿茶酸、木犀草素和金合欢素的含量测定方法进行方法学验证。结果：10批不同产地腹水草茎叶指纹图谱中有27个共有峰，与对照品比较，指认其中咖啡酸、香草酸、阿魏酸、原儿茶酸、木犀草素和金合欢素6个成分，10批样品的相似度均在0.900以上；上述6个成分含量测定方法经验证，线性范围分别为3.0~60.0、10.0~200.0、5.0~100.0、10.0~200.0、10.0~200.0和5.0~100.0 μg·mL^-1，平均加样回收率分别为102.0%、102.8%、101.2%、97.5%、98.0%和101.1%。10批腹水草样品中咖啡酸、香草酸、阿魏酸、原儿茶酸、木犀草素和金合欢素含量分别在22.1~38.4、39.9~58.8、23.8~59.1、43.6~68.0、53.8~89.6、36.6~62.7 μg·g^-1范围内。结论：本文所建立的腹水草茎叶HPLC指纹图谱及主要成分含量测定方法，可用于腹水草药材质量控制等研究。

Objective: To establish an HPLC fingerprint,determine 6 components in the stems and leaves of Veronicastrum axillare Yamazaki including caffeic acid,vanillic acid,ferulic acid,protocatechuic acid,luteolin and acacetin and provide a reference for quality control. Methods: Zobax C₁₈ column (250 mm×4.6 mm,5μm) was used,and the mobile phase consisted of acetonitrile-0.1% phosphoric acid solution with gradient elution at the flow rate of 1.0 mL·min^-1,the detection wavelength was set at 230 nm and the column temperature was 30℃. The HPLC fingerprint was evaluated by chromatographic fingerprint evaluation system software (2004A Edition) by determining the common peaks and calculating the similarity. Besides,the analyical method of caffeic acid,vanillic acid,ferulic acid,protocatechuic acid,luteolin and acacetin was validated. Results: The 27 common peaks of fingerprint from 10 batches were found,and caffeic acid,vanillic acid,ferulic acid,protocatechuic acid,luteolin and acacetin were identified by using standard chemicals. The similar degree of 10 batches samples were more than 0.900. In quality determination,the good linear was observed in the ranges of 3.0-60.0 μg·mL^-1,10.0-200.0 μg·mL^-1,5.0-100.0 μg·mL^-1,10.0-200.0 μg·mL^-1,10.0-200.0 and 5.0-100.0 μg·mL^-1,respectively,and the average recoveries were 102.0%,102.8%,101.2%,97.5%,98.0% and 101.1%,respectively. The content ranges of caffeic acid,vanillic acid,ferulic acid,protocatechuic acid,luteolin and acacetin were 22.1-38.4 μg·g^-1,39.9-58.8 μg·g^-1,23.8-59.1 μg·g^-1,43.6-68.0 μg·g^-1,53.8-89.6 μg·g^-1 and 36.6-62.7 μg·g^-1,respectively. Conclusion: The HPLC fingerprint of Veronicastrum axillare Yamazaki and determination method of caffeic acid,vanillic acid,ferulic acid,protocatechuic acid,luteolin and acacetin are simple,accurate and reproduceable,which can provide a reference for the quality control for Veronicastrum axillare Yamazaki.

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