基于UPLC指纹图谱和化学计量学的不同熏硫程度白芷质量评价

Quality determination of Angelicae dahuricae Radix exposed to sulfur fumigation based on UPLC fingerprint and chemometrics

目的建立白芷UPLC指纹图谱,结合化学计量学方法寻找不同熏硫程度白芷质量差异的标志物。方法该药物的分析采用月旭Xtime UPLC C 18色谱柱(4.6 mm×150 mm,1.8μm);流动相乙腈-0.1%乙酸,梯度洗脱;体积流量0.35 mL/min;柱温35℃;检测波长305 nm。采用中药指纹图谱相似度评价软件进行相似度评价,结合聚类分析、主成分分析、正交偏最小二乘法-判别分析对白芷样品进行全面的质量评价。结果共得到11个共有峰,并通过对照品比对出其中7个(白当归素、氧化前胡素、异欧前胡素、欧前胡素、珊瑚菜素、佛手柑内酯、水合氧化前胡素);33批样品指纹图谱的相似度为0.735~0.997。聚类分析仅将白芷样品聚为3类;主成分分析将样品分为3类,与4种熏硫程度分类一致。正交偏最小二乘法-判别分析筛选出5种差异性成分,经对照品比对,确定了其中2种差异性成分,分别为氧化前胡素、水合氧化前胡素。3种主成分含量测定结果为欧前胡素0.444~3.725 mg/g,珊瑚菜素0.241~0.631 mg/g,异欧前胡素0.179~0.508 mg/g。结论该方法稳定可靠可用于不同熏硫程度白芷的质量控制。

AIM To establish the UPLC fingerprint of Angelicae dahuricae Radix,in combination with chemometrics to search markers for the quality identification of sulfur-fumigated Angelicae dahuricae Radix.METHODS The analysis of this drug was performed on a 35℃thermostatic Welch Xtime UPLC C 18 column(4.6 mm×150 mm,1.8μm),with mobile phase comprising of acetonitrile-0.1%acetic acid flowing at 0.35 mL/min in a gradient elution manner,and the detection wavelength was set at 305 nm.Angelicae dahuricae Radix had its similarity assessed by the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine,and its quality evaluated comprehensively by cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis.RESULTS Among the eleven common peaks totally obtained,seven were determined to be byakangelicin,oxypeucedanin,isoimperatorin,imperatorin,phellopterin,bergapten,and oxypeucedanin hydrate.With fingerprint similarities range of 0.735-0.997,thirty-three batches of Angelicae dahuricae Radix were basically categorized into three groups by cluster analysis;and classified into four groups by principal component analysis complying with the four different degrees of sulfur fumigation.Among the five differential markers screened out by orthogonal partial least squares-discriminant analysis,the two chemicals of oxypeucedanin and oxypeucedanin hydrate were identified by reference substances.In the quantitative determination of the three main components,0.444-3.725 mg/g imperatorin,0.241-0.631 mg/g phellopterin,and 0.179-0.508 mg/g isoimperatorin were obtained.CONCLUSION This stable and reliable method can be used for the quality control of differently sulfur-fumigated Angelicae dahuricae Radix.