不同产地川乌炮制前后差异性成分研究

Different components between Aconiti Radix and its processed product from different regions

关键词:  川乌, 高效液相色谱(HPLC), 指纹图谱, 化学计量学方法, 含量测定, 质量控制

Key words:  Aconiti Radix, high performance liquid chromatography(HPLC), fingerprints, chemometric method, content determination, quality control

摘要:

为建立川乌及制川乌的高效液相色谱(HPLC)指纹图谱,并结合化学计量学方法和主成分含量测定和比较3个不同产地川乌炮制前后的差异成分。该文采用HPLC法,利用中药色谱指纹图谱相似度评价系统(2012版)计算相似度,并结合聚类分析(HCA)、主成分分析(PCA)、偏最小二乘判别分析(PLS-DA)、正交偏最小二乘判别分析(OPLS-DA)及主成分含量测定结果进行综合分析。结果表明:(1)建立了3个产地15批川乌炮制前后的HPLC指纹图谱,分别标定了25个和14个共有峰,并通过对照品指认出6个共有峰,分别为苯甲酰新乌头原碱(峰16)、苯甲酰乌头原碱(峰17)、苯甲酰次乌头原碱(峰18)、新乌头碱(峰21)、次乌头碱(峰22)、乌头碱(峰23)。(2)化学计量学方法分析显示,川乌炮制前后的30批样品被明显分为2类,15批川乌被分为3类,15批制川乌被分为3类,新乌头碱、次乌头碱等6个成分可能是不同产地川乌炮制前后的潜在差异成分。(3)炮制前后主成分含量测定结果均有不同程度的变化,四川江油所产川乌炮制前后部分成分含量明显高于另外两个产地所产川乌。该研究建立的指纹图谱精密度、重复性、稳定性均较好,结合化学计量学分析方法和主成分含量测定分析,可为不同产地川乌炮制前后质量控制提供参考。

Abstract:

To evaluate the differences of Aconiti Radix and Aconiti Radix Preparata from three different regions by chemometrics and principal component content determinations. The high performance liquid chromatography(HPLC)method was established on a Waters SunFire C₁₈(4.6 mm×150 mm, 5 μm), and acetonitrile-glacial acetic acid(0.2%)solution(triethylamine adjusted pH to 6.20)was adopted as the mobile phase for gradient elution, with the flow rate of 1.0 mL·min^-1 and the detection wavelength of 235 nm. The fingerprints were analyzed by the software of “Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012)”. The discriminant analysis of hierarchical cluster analysis(HCA), principal component analysis(PCA), orthogonal partial least squares-discriminant analysis(OPLS-DA)and partial least squares-discriminant analysis(PLS-DA)were employed, combined with chemometrics and the determination results of principal components. The results were as follows:(1)HPLC fingerprints of 15 batches of Aconiti Radix and its processed products from three areas were established, 25 and 14 common fingerprint peaks were calibrated in 15 batches of raw and processed, six peaks were identified by the mixed reference product. They were benzoylmesaconitine(peak 16), benzoylaconitine(peak 17), benzoylhypaconitine(peak 18), mesaconitine(peak 21), hypaconitine(peak 22), aconitine(peak 23).(2)Chemometrics showed that 30 batches of samples were clustered into two categories, 15 batches of Aconiti Radix were classified into three classes, 15 batches of Aconiti Radix Preparata were classified into three classes. Six components containing mesaconitine and hypaconitine may be the potential different components of Aconiti Radix from different regions before and after processing.(3)The content determination results of principal components changed in varying degrees before and after processing, and the determination results of some principal components in Jiangyou of Sichuan producing area before and after processing were significantly higher than those in the other two producing areas. The fingerprint method which established in the study is accurate, repeatable, and reliable. Furthermore, combined with chemometrics and principal component content determination, it could be used for the quality control of Aconiti Radix and its processed product from different regions.