基于指纹图谱和化学模式识别分析的蒙药森登-4质量评价研究

Quality evaluations of Mongolian medicine Sendeng-4 based upon fingerprint and chemical pattern recognition

目的:建立森登-4高效液相色谱指纹图谱,结合化学模式识别分析比较不同批次样品的质量差异,为其质量评价提供依据。方法:采用Agilent Eclipse XDB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,流动相为甲醇(A)-0.2%磷酸水(B),梯度洗脱,流速1 mL·min^(–1),柱温35℃,进样量10μL,检测波长254 nm,采用《中药色谱指纹图谱相似度评价系统(2012版)》对17批森登-4进行相似度评价;采用微生信平台进行聚类分析(CA),采用SIMCA 14.1软件进行主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA),并以变量重要性投影(VIP)值大于1.0为标准,筛选差异性成分,采用Graph Pad Prism 8.0软件对差异成分进行方差分析。结果:17批森登-4指纹图谱共有24个共有峰,相似度均≥0.992,指认了9个共有峰,分别为没食子酸(2号峰)、京尼平苷酸(4号峰)、二氢杨梅素(11号峰)、栀子苷(12号峰)、柯里拉京(13号峰)、诃子酸(18号峰)、鞣花酸(20号峰)、杨梅素(21号峰)、西红花苷Ⅰ(22号峰);CA和PCA将17批森登-4聚为2大类:S1、S3~S14、S16聚为一类,S2、S15、S17聚为一类,OPLS-DA模式下筛选出10个潜在差异标志物,且均具有显著性差异(P<0.01)。结论:建立了森登-4指纹图谱,结合化学模式识别可用于森登-4整体质量评价,柯里拉京等10个成分可能是影响森登-4质量的潜在差异标志物。

OBJECTIVE The fingerprint of Senden-4 was established by HPLC,and the quality difference of different batches of samples was compared by chemical pattern recognition analysis to provide basis for quality evaluation.METHODS The chromatography column was Agilent Eclipse XDB C_(18) (4.6 mm×250 mm,5μm),with methanol (A)-0.2%phosphoric acid (B) solution as a mobile phase for gradient elution.Flow rate was 1 mL·min^(–1),column temperature 35℃,injection volume10μL and detection wavelength 254 nm.The Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine (Edition 2012) was employed for evaluating the similarity of 17 batches of Sendeng-4.Bioinformatic platform was utilized for cluster analysis (CA) and SIMCA 14.1 software for principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA).Variable importance in projection (VIP) value>1.0 was utilized as an index for screening the differential components and Graph Pad Prism 8.0 software for examining variance of different components.RESULTS There were a total of 24 common peaks in fingerprints of 17 batches of Sendeng-4 and the similarities surpassed0.992;nine common peaks were identified,including gallic acid (peak 2),geniposide acid (peak 4),dihydromyricetin (peak11),geniposide (peak 12),corilatin (peak 13),chebulic acid (peak 18),ellagic acid (peak 20),myricetin (peak 21) and crocin I(peak 22).According to the results of CA/PCA,17 batches of samples could be clustered into two categories of S1,S3–S14,S16 and S2,S15,S17.Under the OPLS-DA model,10 differential biomarkers were screened based upon VIP value>1 and the differences were significant (P<0.01).CONCLUSION In conjunctions with chemical pattern recognition techniques,HPLC fingerprinting of Sendeng-4 may be employed for quality evaluations of Sendeng-4.Ten components such as Corilagin are differential markers affecting the quality of Sendeng-4.