摘要
目的 建立达原饮的超高效液相色谱(UPLC)指纹图谱,结合化学模式识别评价其质量.方法 色谱柱为Acquity UPLC BEH C18柱(50 mm×2.1mm,1.7μm),流动相为乙腈-0.1％磷酸溶液(梯度洗脱),流速为0.5mL/min,柱温为30 ℃,检测波长为190～400 nm,图谱提取波长为240 nm,进样量为2μL.建立18批达原饮样品的UPLC指纹图谱,应用中药色谱指纹图谱相似度评价系统(2012版)进行相似度评价,并对共有峰进行归属与指认.采用层次聚类分析(HCA)法、主成分分析(PCA)法及正交偏最小二乘判别分析(OPLS-DA)法对指纹图谱进行评价.结果 18批达原饮样品的指纹图谱相似度较好(0.988～0.999),标定了 30个共有峰,指认了 8个色谱峰,即峰7(芒果苷)、峰9(芍药苷)、峰11(甘草苷)、峰17(黄芩苷)、峰24(黄芩素)、峰27(甘草酸铵)、峰29(和厚朴酚)、峰30(厚朴酚).经HCA和PCA可将18批达原饮样品分为3类;通过变量投影重要性(VIP)值筛选出4个差异性成分,即峰17、峰22、峰23(归属于黄芩)和峰3(归属于知母).结论 所建立的达原饮UPLC指纹图谱及其化学模式识别评价方法稳定可行,可用于达原饮的质量控制与评价,方中黄芩和知母饮片的质量优劣对达原饮指纹图谱影响较大.
Abstract
Objective To establish the ultra-high performance liquid chromatography(UPLC)fingerprint of Dayuanyin,and to evaluate its quality by combining chemical pattern recognition.Methods The chromatographic column was an Acquity UPLC BEH C18 column(50 mm × 2.1 mm,1.7 μm),the mobile phase was acetonitrile-0.1％phosphoric acid solution(gradient elution),the flow rate was 0.5 mL/min,the column temperature was 30 ℃,the detection wavelength was 190-400 nm,the detection wavelength was 240 nm,and the injection volume was 2 µL.The UPLC fingerprints of eighteen batches of Dayuanyin samples were established,and the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2012)was used for similarity evaluation.The common peaks were assigned and identified.The fingerprints were evaluated by the hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares-discriminant analysis(OPLS-DA).Results The fingerprints of eighteen batches of samples showed good similarity(0.988-0.999),with thirty common peaks calibrated and assigned,and eight chromatographic peaks identified,namely peak 7(mangiferin),peak 9(paeoniflorin),peak 11(liquiritin),peak 17(baicalin),peak 24(baicalein),peak 27(monoammonium glycyrrhizinate),peak 29(honokiol),and peak 30(magnolol).The eighteen batches of samples could be divided into three categories by the HCA and PCA methods,and four differential components,namely peak 17,peak 22,peak 23(from Scutellariae Radix),and peak 3(from Anemarrhenae Rhizoma),were judged by the variable importance in projection(VIP)value.Conclusion The established UPLC fingerprint and chemical pattern recognition evaluation method for Dayuanyin are stable and feasible,which can be used for quality control and evaluation of Dayuanyin.The quality of Scutellariae Radix and Anemarrhenae Rhizoma decoction pieces in the formula has a significant impact on the fingerprint of Dayuanyin.
维普
万方数据