苦参的指纹图谱建立及差异性成分含量测定

Establishment of fingerprint and determination of differential components of Sophora flavescens

目的 建立不同产地苦参药材的指纹图谱,筛选差异性成分,并进行含量测定。方法 采用《中药色谱指纹图谱相似度评价系统(2012版)》建立12批苦参药材的高效液相色谱(HPLC)指纹图谱,指认共有峰并进行相似度评价;采用SIMCA 14.1和SPSS 23.0软件进行化学模式识别分析[聚类分析(CA)、主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)],并以变量重要性投影(VIP)值>1为标准筛选影响苦参药材质量的差异性成分;采用同一HPLC法测定4种差异性成分的含量。结果 12批苦参药材的指纹图谱中共有17个共有峰,相似度均大于0.96;共指认了6个共有峰,分别为氧化苦参碱(峰1)、氧化槐果碱(峰2)、苦参碱(峰10)、三叶豆紫檀苷(峰14)、苦参酮(峰16)、降苦参酮(峰17)。CA、PCA、OPLS-DA结果显示,12批苦参药材可按不同产地聚为3类:S1~S7(陕西商州区)聚为一类,S8~S10(河南伊川县)聚为一类,S11~S12(内蒙古赤峰市)聚为一类。苦参碱、降苦参酮、苦参酮、氧化槐果碱以及峰11、峰9所代表的化学成分的VIP值均大于1。12批苦参药材中苦参碱、降苦参酮、苦参酮、氧化槐果碱的含量分别为2.65~4.93、1.54~3.44、9.63~12.94、5.08~6.10 mg/g。结论 本研究成功建立了苦参药材的HPLC指纹图谱,并结合化学模式识别分析筛选出6种差异性成分,可为该药材的质量控制提供参考。

OBJECTIVE To establish the fingerprint of Sophora flavescens, and to screen differential components and determine their contents. METHODS HPLC fingerprints of 12 batches of S. flavescens were established by using Similarity Evaluation System of Chromatographic Fingerprints of TCM(2012 edition);common peaks were identified and their similarities were evaluated. Chemical pattern recognition analysis [cluster analysis(CA),principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA)] were performed with SIMCA 14.1 and SPSS 23.0 software, and differential components which influenced the quality of S. flavescens were screen with variable importance in the projection(VIP)>1 as standard. Meanwhile, the contents of 4 kinds of differential components were determined by the same HPLC method. RESULTS There were 17 common peaks in the fingerprints of 12 batches of S. flavescens,and their similarities were all higher than 0.96. A total of 6 common peaks were identified, i.e. oxymatrine(peak 1), oxysophocarpine(peak 2), matrine(peak 10), trifolirhizin(peak 14), kurarinone(peak 16) and norkurarinone(peak 17). Results of CA, PCA and OPLS-DA showed that 12 batches of S.flavescens were divided into 3 categories according to producing area, i.e. S1-S7(Shangzhou District of Shaanxi Province) were grouped into one category, S8-S10(Yichuan County of Henan Province) into one category and S11-S12(Chifeng City of Inner Mongolia) into one category. VIPs of matrine, norkurarinone, kurarinone and oxysophocarpine and the chemical components represented by peak 11 and 9 were all greater than 1. The contents of matrine, norkurarinone, kurarinone and oxysophocarpine in 12 batches of S. flavescens were 2.65-4.93, 1.54-3.44, 9.63-12.94 and 5.08-6.10 mg/g, respectively. CONCLUSIONS HPLC fingerprint of S. flavescens is established successfully in the study, and can be used to screen 6 differential components by combining with chemical pattern recognition analysis, which can provide reference for quality control of S. flavescens.