高效液相色谱指纹图谱结合化学模式识别评价龙葵质量

Quality Evaluation of Solanum Nigrum Based on HPLC Fingerprint and Chemical Pattern Recognition

目的建立评价龙葵质量的高效液相色谱(HPLC)指纹图谱和化学模式识别方法。方法色谱柱为Waters C18柱(150 mm×4.6 mm,5μm),流动相为乙腈-0.3%磷酸溶液(梯度洗脱),流速为1.0 mL/min,检测波长为205 nm,柱温为26℃,进样量为10μL。建立13批样品的HPLC叠加指纹图谱,采用中药色谱指纹图谱相似度评价系统(2004A版)进行相似度评价,确定并指认共有峰,同时测定指认成分的含量。通过层次聚类分析(HCA)、主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)对不同批次样品进行聚类。结果13批样品共标定了9个共有峰,相似度为0.701~0.984;指认出2个成分,分别为澳洲茄碱(6号峰)和澳洲茄边碱(7号峰),其质量浓度均在3.9~500.0μg/mL范围内与峰面积线性关系良好(R^(2)≥0.9950,n=5)。越南产少花龙葵中澳洲茄碱含量(895.57 mg/kg)远高于国内含量最高的广西南宁产少花龙葵(68.05 mg/kg),但越南产少花龙葵中澳洲茄边碱含量(68.05 mg/kg)仅为广西南宁产少花龙葵(867.42 mg/kg)的7.85%。HCA,PCA,OPLS-DA结果显示,13批龙葵药材按基原被分为2类,1类为龙葵,另1类为少花龙葵。结论该方法操作简单、结果准确,能快速、科学、准确地评价龙葵的质量。

Objective To establish a high-performance liquid chromatography(HPLC)fingerprint and chemical pattern recognition method for evaluating the quality of Solanum nigrum.Methods The chromatographic column was Waters C18 column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.3%phosphoric acid solution(gradient elution),and the flow rate was 1.0 mL/min,the detection wavelength was 205 nm,the column temperature was 26℃,and the injection volume was 10μL.HPLC overlay fingerprints of 13 batches of samples were established,and the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2004A)was used for similarity evaluation to determine and identify common peaks,and the content of identified components was determined.The different batches of samples were clustered by the hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares-discriminant analysis(OPLS-DA).Results Nine common peaks were calibrated for 13 batches of samples,with similarity ranging from 0.701 to 0.984.Two components were identified,namely solanen(peak 6)and solamargine(peak 7),with the linear range of 3.9-500μg/mL(R^(2)≥0.9950,n=5).The content of solanenin in Vietnamese-produced Solanum photeinocarpum(895.57 mg/kg)was significantly higher than that in Guangxi Nanning-produced Solanum photeinocarpum(68.05 mg/kg,it was the highest content in China).However,the content of solamargine in Vietnamese-produced Solanum photeinocarpum(68.05 mg/kg)was only 7.85%of that in Guangxi Nanning-produced Solanum photeinocarpum(867.42 mg/kg).The HCA,PCA,OPLS-DA results showed that the 13 batches of Solanum nigrum samples were divided into two categories based on their origins,one group was Solanum nigrum,and the other group was Solanum photeinocarpum.Conclusion The method is simple,accurate,which can quickly,scientifically,and accurately evaluate the quality of Solanum nigrum.