祁白芷UPLC指纹图谱的建立及其抗氧化谱效关系研究

Establishment of UPLC Fingerprint and Effects of Antioxidant Related to Its Spectrum Study of Angelica dahurica cv.Qibaizhi

目的建立祁白芷超高效液相色谱(UPLC)指纹图谱,采用超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)对共有峰进行成分鉴定,测定各批次祁白芷抗氧化活性,并探讨祁白芷的化学成分与抗氧化活性的谱效关系。方法采用《中药色谱指纹图谱相似度评价系统(2012版)》建立28批祁白芷的指纹图谱,确认共有峰并进行相似度评价,并对共有峰进行UPLC-Q-TOF-MS分析。以1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除率为抗氧化指标,测定其抗氧化活性。采用灰色关联度和偏最小二乘回归分析祁白芷共有峰与抗氧化活性的谱效关系。结果建立了28批祁白芷药材的指纹图谱,其相似度为0.910~0.997。聚类分析可将不同加工方式的祁白芷分为熏硫和无硫两大类,当平方欧氏距离为10时,可将无硫祁白芷进一步分为两类,与商品规格等级分类相符。其共标定13个共有峰,利用对照品对比和UPLC-Q-TOF-MS解析指认出共有峰分别为奎宁酸、二氢山芹醇、水合氧化前胡素、白当归素、花椒毒素、佛手柑内酯、异茴芹内酯、氧化前胡素、欧前胡素、珊瑚菜素、8-氧甲基异欧前胡内酯、异欧前胡素、镰叶芹酮。通过清除DPPH自由基实验发现,28批祁白芷均有抗氧化能力,谱效分析结果显示,奎宁酸、佛手柑内酯、氧化前胡素、珊瑚菜素、异欧前胡素、镰叶芹酮等与抗氧化能力呈正相关。结论本研究建立了基于化学成分和抗氧化活性的祁白芷质量评价模式,为阐明祁白芷药材中抗氧化活性成分及质量控制提供参考。

OBJECTIVE To establish UPLC fingerprint of Angelica dahurica cv.Qibaizhi,identify the compounds of common peaks by UPLC-Q-TOF-MS technology,determine antioxidant activity of each batch and study the relationship between effects of antioxidant and spectrum of Angelica dahurica cv.Qibaizhi.METHODS The fingerprint of 28 batches of Angelica dahurica cv.Qibaizhi was established by the Similarity Evaluation System of TCM Chromatographic Fingerprint(2012 edition),and the common peaks were identified and evaluated for similarity,and the common peaks were analyzed by UPLC-Q-TOF-MS.The free radical scavenging rate of 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)was used as antioxidant index to determine its antioxidant activity.Grey correlation degree and partial least squares regression were used to analyze the spectral effect relationship between chemical constituents and antioxidant activities of Angelica dahurica cv.Qibaizhi.RESULTS The fingerprints of 28 batches of Radix Angelica were established,the similarity was 0.910-0.997.By cluster analysis,the different processing methods can be divided into two categories:sulfur fumigation and sulphur-free.When the square Euclidean distance is 10,the sulphur-free Angelica dahurica cv.Qibaizhi can be further divided into two categories,which is consistent with the classification of commodity specifications.A total of 13 common peaks were identified.The common peaks were identified by comparison of standards and UPLC-Q-TOF-MS analysis as quinic acid,dihydrooroselol,oxypeucedanin hydrate,byakangelicin,xanthotoxin,bergapten,isopimpinellin,oxypeucedanin,imperatorin,phellopterin,cnidilin,isoimperatorin and falcarinolone.By scavenging DPPH free radicals,it was found that 28 batches of Angelica dahurica cv.Qibaizhi had antioxidant capacity.The spectral analysis results showed that the antioxidant capacity was positively related to quinic acid,bergapten,oxypeucedanin,phellopterin,isoimperatorin and falcarinolone.CONCLUSION In this study,a quality evaluation model of Angelica dahurica cv.Qibaizhi based on chemical composition and activity is established,which provides a reference for elucidating the antioxidant active ingredients and quality control in Angelica dahurica cv.Qibaizhi.