基于指纹图谱结合化学模式识别及多成分定量的千斤拔质量评价研究

Determination of multi-components of Moghania Radix based on fingerprints and chemical pattern recognition

目的 建立千斤拔Moghania Radix HPLC指纹图谱及多成分定量方法,为其质量控制提供参考。方法 采用XBridge■C_(18)色谱柱(250 mm×4.6 mm,5μm)为固定相,以乙腈-0.1%乙酸溶液为流动相进行梯度洗脱,体积流量1.0 mL·min^(-1),检测波长260 nm,柱温30℃;对24批千斤拔(编号为S1~S24,其中S2~S9为采集的新鲜全株植物,实验室低温干燥而得,其余样品为市场购买药材或饮片)构建指纹图谱,同时测定相关成分的含量,并结合聚类分析、主成分分析及正交偏最小二乘法进行全面评价。结果 24批不同来源千斤拔指纹图谱共标定11个共有峰,指认出染料木苷、6″-O-丙二酰基染料木苷、染料木素。相似度计算结果表明,各批次相似度在0.707~0.981;聚类分析将24批千斤拔分为2类,S2~S5、S6、S8、S9号样品(均为新鲜全株植物)聚为一类,其他17份样品聚为一类;经主成分分析,主成分1~4是影响样品质量评价的主要因子,选取前4个主因子对不同产地的千斤拔进行评分,24批样品的综合得分在5.34~-3.61,说明各批次质量差异相对较大;样品中综合得分最高是S4,各饮片样品的综合得分较低。24批样品中染料木苷、6″-O-丙二酰基染料木苷、染料木素质量分数分别为0.10~0.57、0.11~2.85、0.15~0.71 mg·g^(-1),不同批次之间3个成分质量分数差异较大,批次之间3个成分含量总体呈现出二低一高的趋势,建议使用3个成分总和作为质量控制指标。结论 建立的HPLC指纹图谱及多成分含量测定方法稳定、可靠,可为千斤拔的质量评价提供依据。

Objective To establish the HPLC fingerprint and muti-component analysis of Moghaniae Radix and provide reference for quality control.Methods Waters XBridge■ C_(18)column(250 mm × 4.6 mm,5 μ m) were performed on HPLC analysis.The mobile phase was acetonitrile(A)-0.1% acetic acid(B) for gradient elution.The flow rate was 1.0 mL· min^(-1),the wavelength was 260 nm and the column temperature was 30 ℃.The content and fingerprint of relative ingredients of 24 batches(the numbers were S1—S24,in which S2—S9 were fresh whole plants collected and dried at low temperature in the laboratory,and the other samples were medicinal materials or decoction pieces purchased from the market) were measured simultaneously,combined with cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis(OPLS-DA),which were applied in comprehensive analysis.Results The similarity calculation results showed that the similarity of each batch was between 0.707—0.981,and 11 common peaks were calibrated,genistin,6 "-O-malonyl genistein,genistein were identified.Cluster analysis classified the 24batches of medicinal materials into two categories,samples S2,S3,S4,S5,S6,S8,S9(all fresh whole plants) were grouped into one group,and the other 17 samples were grouped into one group.According to the principal component analysis,principal component1—4 was the main factor affecting the quality evaluation of the samples.The first four principal factors were selected to score the samples from different producing areas,and the comprehensive score of 24 batches of samples was between 5.34 and-3.61,indicating that the quality of each batch was relatively different.The highest composite score of the samples was S4,while the composite score of each piece was lower.The quality fractions of genistein,6"-O-malondiacylgenistein and genistein in 24 batches of samples ranged from 0.10—0.57 mg·g-1,0.11—2.85 mg·g^(-1) and 0.15—0.71 mg·g^(-1),respectively.The quality fractions of the three batches varied greatly among different batches.The contents of the three items in batches generally show a trend of two low and one high.It was suggested to use the sum of the three items as the quality control index.Conclusion The established HPLC fingerprint and multi-component content determination method are stable and reliable,which can provide a reference for the quality evaluation Moghaniae Radix.