五匹风高效液相色谱指纹图谱及多指标成分定量分析

HPLC Fingerprint of Potentilla Kleiniana and Quantitative Analysis of Its Multi-Target Components

目的建立五匹风药材的高效液相色谱(HPLC)指纹图谱及相关指标成分含量测定方法。方法色谱柱为Eclipse XDB-C18柱(150 mm×4.6 mm,5μm),流动相为乙腈-0.5%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,检测波长为327 nm,柱温为30℃,进样量为20μL。采用中药色谱指纹图谱相似度评价系统(2012年A版)建立15批药材样品的HPLC指纹图谱,并进行相似度评价。采用HPLC法测定5个指标成分的含量。结果15批药材样品的HPLC指纹图谱共标定6个共有峰,指认了其中5个,分别为3,3’-二甲氧基鞣花酸-4’-O-β-D-吡喃葡萄糖苷、山柰酚-3,7-二-O-α-L-吡喃鼠李糖苷、3,3’-二甲氧基鞣花酸-4’-O-β-D-吡喃木糖苷、3,3’-二甲氧基鞣花酸、3,3’,4’-三甲氧基鞣花酸,相似度介于0.886~0.996。上述5个成分质量浓度分别在22.37~223.72μg/mL(r=0.9993)、6.81~68.08μg/mL(r=0.9992)、5.23~52.34μg/mL(r=0.9993)、4.05~40.46μg/mL(r=0.9995)、17.17~171.74μg/mL(r=0.9993)范围内与峰面积线性关系良好;精密度、稳定性、重复性试验的RSD均小于3.0%;平均加样回收率分别为102.03%,104.09%,95.14%,97.13%,98.63%,RSD分别为2.77%,2.11%,1.24%,2.64%,2.31%(n=6)。15批药材样品中5个成分的含量分别为0.162%~0.879%、0.011%~0.253%、0.019%~0.047%、0.019%~0.209%、0.037%~0.107%。结论所建立的方法简便、稳定,结果准确、可靠,可为五匹风药材的质量控制提供参考。

Objective To establish a high-performance liquid chromatography(HPLC)fingerprint of Potentilla kleiniana and a method for determining the content of its target components.Methods The chromatographic column was Eclipse XDB-C18 column(150 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.5%phosphoric acid aqueous solution(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 327 nm,the column temperature was 30℃,and the injection volume was 20μL.The HPLC fingerprint of 15 batches of medicinal samples was established by the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2012 A),and the similarity evaluation was conducted.The content of five target components was determined by the HPLC method.Results Six common peaks were marked in the HPLC fingerprint of 15 batches of medicinal samples,of which five were identified as 3,3′-di-O-methylellagic acid-4′-O-β-D-glucopyranoside,kaempferol-3,7-di-O-α-L-rhamnopyranoside,3,3′-di-O-methylellagic acid-4′-O-β-D-xylopyranoside,3,3′-di-O-methylellagic acid and 3,3′,4′-tri-O-methylellagic acid,with a similarity in the range of 0.886-0.996.The linear ranges of the above components were 22.37-223.72μg/mL(r=0.9993),6.81-68.08μg/mL(r=0.9992),5.23-52.34μg/mL(r=0.9993),4.05-40.46μg/mL(r=0.9995),17.17-171.74μg/mL(r=0.9993)respectively.The RSDs of precision,stability and repeatability tests were all lower than 3.0%.The average recovery rates were 102.03%,104.09%,95.14%,97.13%,98.63%,with RSDs of 2.77%,2.11%,1.24%,2.64%,2.31%respectively(n=6).The content of five components in 15 batches of medicinal samples was in the range of 0.162%to 0.879%,0.011%to 0.253%,0.019%to 0.047%,0.019%to 0.209%,0.037%to 0.107%.Conclusion The established method is simple,stable,accurate and reliable,which can provide a reference for the quality control of Potentilla kleiniana.