六月雪-葎草药对指纹图谱研究及其多指标成分的定量分析

Fingerprints and quantitative analysis of multi-index constituents in Serissa Japonicaand-Humulus Scandens drug pair

目的建立六月雪-葎草(以下简称“雪葎”)药对共煎液HPLC指纹图谱,并进行多成分定量分析,用于评价“雪葎”药对的质量。方法采用Agilent EC-C_(18)色谱柱(150 mm×4.6 mm,4μm)进行检测;以甲醇-0.1%磷酸水溶液为流动相,梯度洗脱;流速为0~30 min:1.0 mL·min^(-1),30~65 min:0.6 mL·min^(-1);检测波长为350,240 nm;柱温为30℃。采用《中药色谱指纹图谱相似度评价系统软件(2012.1版)》对“雪葎”药对共煎液进行相似度评价,并对其中4个指标成分进行定量分析,并将配伍前指标性成分的含量与配伍后相比。结果在特征指纹图谱研究中,共标定12个共有峰,以4号峰为参照峰,10批“雪律”药对共煎液与共有模式对照图谱之间有良好的相似性,相似度均在0.9以上;在建立的色谱条件下,4个成分具有良好的线性关系,平均加样回收率在98.7%~99.6%之间,RSD均小于3%(n=6)。10份“雪葎”药对共煎液中车叶草苷酸、木犀草苷、芦丁和大波斯菊苷含量范围分别为0.2494~0.4028 mg·g^(-1),0.1091~0.2389 mg·g^(-1),0.0459~0.0768 mg·g^(-1),0.0914~0.1737 mg·g^(-1)。雪葎药对配伍后,共煎液中木犀草苷、芦丁和大波斯菊苷含量均降低,降低幅度分别为296.40%,47.86%和25.02%,而车叶草苷酸的含量增加,增长幅度为8.81%,其中大波斯菊苷的含量降低具有统计学意义(P<0.05),木犀草苷和芦丁的含量降低具有显著统计学差异(P<0.01)结论所建立的“雪葎”药对共煎液HPLC指纹图谱和含量测定方法灵敏度高、专属性强,可以用于该药对的质量控制,该方法也为其配伍机制和药理研究奠定了基础。

Objective To establish the HPLC fingerprint of the co-decoction of Serissa japonica and Humulus scandens(hereinafter referred to as“Xuelyu”)and conduct multi-component quantitative analysis to evaluate the quality of“Xuelyu”drug pair.Methods Agilent EC-C_(18)column(4.6 mm×150 mm,4μm)was used for detection.Methanol-0.1%aqueous phosphoric acid solution was used as mobile phase and eluted gradientally;the flow rates were 1.0 mL·min1 for 0-30 min and 0.6 mL·min^(-1)for 30-65 min;the detection wavelengths were 350 nm and 240 nm;the column temperature was 30℃.The"Similarity Evaluation system Software for Traditonal Chinese Medicine Fingerprint(2012.1)Edition"co-decoction solution of“Xuelyu”drug pair was evaluated by similarity evaluation,The four index components were quantitatively analyzed and compared before and after compatibility.Results In the study of characteristic fingerprintgraph,12 common peaks were calibrated,with No.4 peak as the reference peak,10 batches of"Xuelyu"drug pairs had good similarity between the common decoction solution and the common mode control graph,and the similarity was above 0.9;under the established chromatographic conditions,the components had a good linear relationship,and the average dosing recovery rate was between 98.7%and 99.6%.RSD values were all less than 3%(n=6).The contents of plantain glycolate,luteolin,rutin and cosmos in the 10 batches of"Xuelyu"drug pairs in the co-decoction ranges were 0.2494-0.4028 mg·g^(-1),0.1091-0.2389 mg·g^(-1),0.0459-0.0768 mg·g^(-1),0.0914-0.1737 mg·g^(-1),respectively.After the combination of drug pairs,the contents of luteolin,rutin and cosmopolitan in the decoction were all decreased,and the decreasing rates were 296.40%,47.86%and 25.02%,respectively,while the content of plantain glycolate was increased and the increasing rate was 8.81%.The content of cosmopolitan was significantly decreased(P<0.05),and the content of luteolin and rutin was significantly decreased(P<0.01).Conclusion The established HPLC fingerprint and content determination method of co-decoction solution have high sensitivity and strong specificity which can be used for the quality control of the drug pair and also laid the foundation for its compatibility mechanism and pharmacological research.