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Article(id=1218290943371953101, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218290941232861879, articleNumber=1001-2494(2024)15-1429-09, orderNo=null, doi=10.11669/cpj.2024.15.010, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1709740800000, receivedDateStr=2024-03-07, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768392795651, onlineDateStr=2026-01-14, pubDate=1723046400000, pubDateStr=2024-08-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768392795651, onlineIssueDateStr=2026-01-14, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768392795651, creator=13701087609, updateTime=1768392795651, updator=13701087609, issue=Issue{id=1218290941232861879, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='15', pageStart='1361', pageEnd='1452', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768392795141, creator=13701087609, updateTime=1768394622953, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1218298607682376061, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218290941232861879, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1218298607682376062, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1218290941232861879, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1429, endPage=1437, ext={EN=ArticleExt(id=1218290943602639822, articleId=1218290943371953101, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Quality Evaluation of Forsythiae Fructus Based on HPLC Fingerprint and QAMS Multi-Component Content Determination, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To establish the HPLC fingerprint of Forsythiae Fructus and develop a multi-component quantitative analysis method based on quantitative analysis of multi-components by single-marker(QAMS). METHODS The fingerprints of 47 Forsythiae Fructus samples from four provinces, including Henan, Shandong, Shanxi and Shaanxi, were established by high performance liquid chromatography (HPLC), while their similarity was evaluated. Chemical pattern recognition of Forsythiae Fructus was studied by hierarchical cluster analysis (HCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). The content of forsythoside B, forsythoside A, hyperin, arctiin, phillyrin and phillygenin were determined by using QAMS, and the differences of samples from different producing areas were analyzed. RESULTS The fingerprints of 47 Forsythiae Fructus samples were established with similarities ranged from 0.785 to 0.989, and a total of 30 common peaks were identified. According to HCA, the samples were divided into three groups. OPLS-DA analysis can effectively distinguish Forsythiae Fructus from several producing areas (Q2>0.5). There was no significant difference in the content of 6 components in 47 batches of samples by QAMS and external standard method (ESM) (P>0.05), but there was significant difference in the content of active components in samples from different origin (P<0.01). The average contents of forsythoside B, forsythoside A, hyperin, arctiin, phillyrin and phillygenin in samples from different provinces were, Henan(4.50、20.58、0.52、2.39、7.46、1.76 mg·g-1), Shandong(0.90, 9.26、0.36、1.39、3.84、3.87 mg·g-1), Shanxi(0.82、8.74、0.29、1.16、2.64、1.75 mg·g-1), Shaanxi(5.18、55.10、0.88、3.36、13.11、9.25 mg·g-1). CONCLUSION The chemical composition of Forsythiae Fructus samples from different locations was similar, but the origin factors had a significant effect on the content of active ingredients, and the samples from Shaanxi province had the highest content. The HPLC fingerprint combined with the simultaneous determination of multi-component content analysis method established in this study is stable and reliable. The fingerprint combined with the chemical pattern recognition can provide a basis for the identification of origins and quality evaluation of Forsythiae Fructus.

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目的 建立连翘药材的高效液相色谱(HPLC)法指纹图谱及基于“一测多评”(QAMS)技术的多成分定量测定方法。方法 采用HPLC法建立河南、山东、山西、陕西4个省份47批连翘药材的指纹图谱,进行相似度评价,结合聚类分析(HCA)、正交偏最小二乘法-判别分析(OPLS-DA)对连翘进行化学模式识别分析。利用QAMS技术,以连翘酯苷A为内参物,同时对连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷和连翘脂素6种活性成分进行含量测定,分析不同产地样品差异。结果 建立了47批连翘药材的指纹图谱,相似度为0.785~0.989,共标定30个共有峰。聚类分析将47批样品聚为3类,OPLS-DA分析能有效将几个产区连翘药材进行区分(Q2>0.5)。采用QAMS法与外标法(ESM)测定47批样品中6种活性成分的含量结果无明显差异(P>0.05),不同产地样品中活性成分含量存在明显差异(P<0.01)。连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素6种成分在各省样品中的平均含量分别为:河南(4.50、20.58、0.52 、2.39、7.46、1.76 mg·g-1)、山东(0.90、9.26、0.36、1.39、3.84、3.87 mg·g-1)、山西(0.82、8.74、0.29、1.16、2.64、1.75 mg·g-1)、陕西(5.18、55.10、0.88、3.36、13.11、9.25 mg·g-1)。结论 不同来源的连翘药材样品中化学成分近似,但产地因素对有效成分含量有较大的影响,来自陕西的连翘样品中各检测成分含量最高。建立的连翘药材HPLC指纹图谱及多成分含量测定方法稳定、可靠,结合化学识别模式可为连翘药材产地判别及质量评价提供依据。

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邓寒霜,男,硕士,副教授 研究方向:中药质量控制 Tel:(0914)2329816

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邓寒霜,男,硕士,副教授 研究方向:中药质量控制 Tel:(0914)2329816

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Phytochem Anal, 2010, 21(3):253-260., articleTitle=Simultaneous quantification of 14 bioactive constituents in Forsythia suspensa by liquid chromatography-electrospray ionisation-mass spectrometry, refAbstract=null), Reference(id=1218490406774427688, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, doi=null, pmid=null, pmcid=null, year=2014, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[24], rfOrder=23, authorNames=DUAN X Y, journalName=Study on the quality analysis and quality control of Forsythiae Fructus, refType=null, unstructuredReference=DUAN X Y. Study on the quality analysis and quality control of Forsythiae Fructus[D]. Zhengzhou: Henan University of Chinese Medicine, 2014., articleTitle=null, refAbstract=null), Reference(id=1218490406841536553, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, doi=null, pmid=null, pmcid=null, year=2013, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[25], rfOrder=24, authorNames=XU J, journalName=The prelimiary study on quality standard and reasonable harvest period for Forsythia suspensa (Thunb.) Vahl, refType=null, unstructuredReference=XU J. The prelimiary study on quality standard and reasonable harvest period for Forsythia suspensa (Thunb.) Vahl[D]. Taiyuan: Shanxi University, 2013., articleTitle=null, refAbstract=null)], funds=[Fund(id=1218490404916350990, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, awardId=2019ZY-FP-02, language=CN, fundingSource=中央引导地方科技发展专项(2019ZY-FP-02), fundOrder=null, country=null), Fund(id=1218490404975071247, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, awardId=2022JM-487, language=CN, fundingSource=陕西省科技计划项目(2022JM-487), fundOrder=null, country=null), Fund(id=1218490405042180112, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, awardId=20YSZX02, language=CN, fundingSource=商洛学院院士工作站专项基金项目(20YSZX02), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1218490400822711237, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, xref=1, ext=[AuthorCompanyExt(id=1218490400831099846, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, companyId=1218490400822711237, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 College of Biopharmaceutcal and Food Engineering, Shangluo University, Shangluo 726000, China), AuthorCompanyExt(id=1218490400839488455, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, companyId=1218490400822711237, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 商洛学院生物医药与食品工程学院, 陕西 商洛 726000)]), AuthorCompany(id=1218490400894014408, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, xref=2, ext=[AuthorCompanyExt(id=1218490400902403017, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, companyId=1218490400894014408, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 Shaanxi Qinling Industrial Technology Research Institute of Special Biological Resources, Shangluo 726000, China), AuthorCompanyExt(id=1218490400906597322, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, companyId=1218490400894014408, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 陕西秦岭特色生物资源产业技术研究院, 陕西 商洛 726000)]), AuthorCompany(id=1218490400961123276, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, xref=3, ext=[AuthorCompanyExt(id=1218490400969511885, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, companyId=1218490400961123276, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 Shaanxi Xiangju Pharmaceutical Goup Co.,Ltd, Shangluo 726000, China), AuthorCompanyExt(id=1218490400977900494, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, companyId=1218490400961123276, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 陕西香菊药业集团有限公司, 陕西 商洛 726000)])], figs=[ArticleFig(id=1218490403133772791, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Fig.1, caption=HPLC fingerprints for 47 batches of Forsythiae Fructus, figureFileSmall=Wemox7kr0lvq9Zx85HT4wA==, figureFileBig=beDfqWehUy9FBFKPGIBCug==, tableContent=null), ArticleFig(id=1218490403196687352, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=图1, caption=47批连翘药材高效液相色谱(HPLC)指纹图谱, figureFileSmall=Wemox7kr0lvq9Zx85HT4wA==, figureFileBig=beDfqWehUy9FBFKPGIBCug==, tableContent=null), ArticleFig(id=1218490403272184825, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Fig.2, caption=Chromatogram of reference fingerprints of Forsythiae Fructus (A) and mixed reference substances (B)

13-forsythoside B; 14-forsythoside A; 17-hyperin; 20-arctiin; 22-phillyrin; 29-phillygenin; Other peak components unidentified.

, figureFileSmall=0bAx2yMuAwqMFF3BXFTR/A==, figureFileBig=uDUf25CjVLpgjSqHiFqjyA==, tableContent=null), ArticleFig(id=1218490403330905082, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=图2, caption=连翘药材对照指纹图谱(A)和混合对照品色谱图(B)

13-连翘酯苷B;14-连翘酯苷A;17-金丝桃苷;20-牛蒡子苷;22-连翘苷;29-连翘脂素;其他峰成分未知。

, figureFileSmall=0bAx2yMuAwqMFF3BXFTR/A==, figureFileBig=uDUf25CjVLpgjSqHiFqjyA==, tableContent=null), ArticleFig(id=1218490403398013947, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Fig.3, caption=Cluster analysis dendrogram of 47 batches Forsythiae Fructus

47 batches of samples could be clustered into 3 classes(Ⅰ, Ⅱ, Ⅲ), When the clustering distance was 10.

, figureFileSmall=SBtuqkLzkCNGjS6UDS+e5g==, figureFileBig=jMNy7UAXpRpufpO9Yd7DzQ==, tableContent=null), ArticleFig(id=1218490403460928508, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=图3, caption=47批连翘药材聚类树状图

聚类距离为10时,47批样品可聚为3类(Ⅰ、Ⅱ、Ⅲ)。

, figureFileSmall=SBtuqkLzkCNGjS6UDS+e5g==, figureFileBig=jMNy7UAXpRpufpO9Yd7DzQ==, tableContent=null), ArticleFig(id=1218490403519648765, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Fig.4, caption=Scatter diagram of OPLS-DA of Forsythiae Fructus from different origins, figureFileSmall=W+BYeLOQECEs73L8Yf1HIQ==, figureFileBig=DoK04LZ9ioZQbC+BDf+ZNw==, tableContent=null), ArticleFig(id=1218490403574174718, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=图4, caption=不同产地连翘药材正交偏最小二乘法-判别分析(OPLS-DA)散点图, figureFileSmall=W+BYeLOQECEs73L8Yf1HIQ==, figureFileBig=DoK04LZ9ioZQbC+BDf+ZNw==, tableContent=null), ArticleFig(id=1218490403641283583, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Fig.5, caption=VIP values of 30 common peaks of 47 Forsythiae Fructus samples in OPLS-DA model. n=47,$\bar{x}±s$, figureFileSmall=/gjXAKPFOA/j1R/JYn1OKQ==, figureFileBig=jPrNNchBVyCAIZ1b1eSnMw==, tableContent=null), ArticleFig(id=1218490403704198144, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=图5, caption=47批连翘药材30个共有峰在OPLS-DA模型中的VIP值。n=47,$\bar{x}±s$, figureFileSmall=/gjXAKPFOA/j1R/JYn1OKQ==, figureFileBig=jPrNNchBVyCAIZ1b1eSnMw==, tableContent=null), ArticleFig(id=1218490403762917376, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Fig.6, caption=Contents of 6 active ingredients in Forsythiae Fructus samples from different origins, figureFileSmall=WYiK8WP1ruZi+phN03YA9w==, figureFileBig=7kikoSiWW72c+6o4rIhT+Q==, tableContent=null), ArticleFig(id=1218490403838414849, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=图6, caption=不同产地连翘药材中6种活性成分含量测定结果, figureFileSmall=WYiK8WP1ruZi+phN03YA9w==, figureFileBig=7kikoSiWW72c+6o4rIhT+Q==, tableContent=null), ArticleFig(id=1218490403922300930, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Tab.1, caption=

Sample information of Forsythiae Fructus

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Origin(in Chinese) No. Origin(in Chinese) No. Origin(in Chinese) No. Origin(in Chinese)
S1 Xixia,Henan(河南西峡) S13 Dingtao,Shandong(山东定陶) S25 Pingshun,Shanxi(山西平顺) S37 Zhengan,Shannxi(陕西正安)
S2 Xichuan,Henan(河南淅川) S14 Jiaxiang,Shandong(山东嘉祥) S26 Jiangxian,Shanxi(山西绛县) S38 Huazhou,Shannxi(陕西华州)
S3 Lushi,Henan(河南卢氏) S15 Anze,Shanxi(山西安泽) S27 Yongji,Shanxi(山西永济) S39 Danfeng,Shannxi(陕西丹凤)
S4 Lingbao,Henan(河南灵宝) S16 Changzhi,Shanxi(山西长治) S28 Yongji,Shanxi(山西永济) S40 Huanglong,Shannxi(陕西黄龙)
S5 Fangcheng,Henan(河南方城) S17 Changzhi,Shanxi(山西长治) S29 Anzhe,Shanxi(山西安泽) S41 Zhashui,Shannxi(陕西柞水)
S6 Neixiang,Henan(河南内乡) S18 Anzhe,Shanxi(山西安泽) S30 Anzhe,Shanxi(山西安泽) S42 Luonan,Shannxi(陕西洛南)
S7 Zhenping,Henan(河南镇平) S19 Anzhe,Shanxi(山西安泽) S31 Qinyuan,Shanxi(山西沁源) S43 Shangnan,Shannxi(陕西商南)
S8 Tongbai,Henan(河南桐柏) S20 Qinshui,Shanxi(山西沁水) S32 Qinyuan,Shanxi(山西沁源) S44 Shangnan,Shannxi(陕西商南)
S9 Caoxian,Shandong(山东曹县) S21 Qinshui,Shanxi(山西沁水) S33 Qinyuan,Shanxi(山西沁源) S45 Shanyang,Shannxi(陕西山阳)
S10 Juye,Shandong(山东巨野) S22 Yongji,Shanxi(山西永济) S34 Lingchuan,Shanxi(山西陵川) S46 Yaozhou,Shannxi(陕西耀州)
S11 Yuncheng,Shandong(山东郓城) S23 Qinshui,Shanxi(山西沁水) S35 Lantian,Shannxi(陕西蓝田) S47 Yijun,Shannxi(陕西宜君)
S12 Yanggu,Shandong(山东阳谷) S24 Yangcheng,Shanxi(山西阳城) S36 Shangzhou,Shannxi(陕西商州)
), ArticleFig(id=1218490404014575619, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=表1, caption=

连翘药材采集地点信息

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Origin(in Chinese) No. Origin(in Chinese) No. Origin(in Chinese) No. Origin(in Chinese)
S1 Xixia,Henan(河南西峡) S13 Dingtao,Shandong(山东定陶) S25 Pingshun,Shanxi(山西平顺) S37 Zhengan,Shannxi(陕西正安)
S2 Xichuan,Henan(河南淅川) S14 Jiaxiang,Shandong(山东嘉祥) S26 Jiangxian,Shanxi(山西绛县) S38 Huazhou,Shannxi(陕西华州)
S3 Lushi,Henan(河南卢氏) S15 Anze,Shanxi(山西安泽) S27 Yongji,Shanxi(山西永济) S39 Danfeng,Shannxi(陕西丹凤)
S4 Lingbao,Henan(河南灵宝) S16 Changzhi,Shanxi(山西长治) S28 Yongji,Shanxi(山西永济) S40 Huanglong,Shannxi(陕西黄龙)
S5 Fangcheng,Henan(河南方城) S17 Changzhi,Shanxi(山西长治) S29 Anzhe,Shanxi(山西安泽) S41 Zhashui,Shannxi(陕西柞水)
S6 Neixiang,Henan(河南内乡) S18 Anzhe,Shanxi(山西安泽) S30 Anzhe,Shanxi(山西安泽) S42 Luonan,Shannxi(陕西洛南)
S7 Zhenping,Henan(河南镇平) S19 Anzhe,Shanxi(山西安泽) S31 Qinyuan,Shanxi(山西沁源) S43 Shangnan,Shannxi(陕西商南)
S8 Tongbai,Henan(河南桐柏) S20 Qinshui,Shanxi(山西沁水) S32 Qinyuan,Shanxi(山西沁源) S44 Shangnan,Shannxi(陕西商南)
S9 Caoxian,Shandong(山东曹县) S21 Qinshui,Shanxi(山西沁水) S33 Qinyuan,Shanxi(山西沁源) S45 Shanyang,Shannxi(陕西山阳)
S10 Juye,Shandong(山东巨野) S22 Yongji,Shanxi(山西永济) S34 Lingchuan,Shanxi(山西陵川) S46 Yaozhou,Shannxi(陕西耀州)
S11 Yuncheng,Shandong(山东郓城) S23 Qinshui,Shanxi(山西沁水) S35 Lantian,Shannxi(陕西蓝田) S47 Yijun,Shannxi(陕西宜君)
S12 Yanggu,Shandong(山东阳谷) S24 Yangcheng,Shanxi(山西阳城) S36 Shangzhou,Shannxi(陕西商州)
), ArticleFig(id=1218490404098461700, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Tab.2, caption=

Similarities of 47 batches Forsythiae Fructus

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Similarity No. Similarity No. Similarity No. Similarity
S1 0.969 S13 0.874 S25 0.906 S37 0.982
S2 0.954 S14 0.812 S26 0.934 S38 0.948
S3 0.862 S15 0.937 S27 0.865 S39 0.989
S4 0.867 S16 0.922 S28 0.852 S40 0.854
S5 0.935 S17 0.930 S29 0.789 S41 0.979
S6 0.967 S18 0.915 S30 0.851 S42 0.969
S7 0.872 S19 0.874 S31 0.977 S43 0.920
S8 0.889 S20 0.912 S32 0.917 S44 0.947
S9 0.923 S21 0.924 S33 0.974 S45 0.974
S10 0.936 S22 0.897 S34 0.807 S46 0.918
S11 0.881 S23 0.905 S35 0.933 S47 0.898
S12 0.785 S24 0.961 S36 0.976
), ArticleFig(id=1218490404165570565, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=表2, caption=

47批连翘药材HPLC指纹图谱相似度

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Similarity No. Similarity No. Similarity No. Similarity
S1 0.969 S13 0.874 S25 0.906 S37 0.982
S2 0.954 S14 0.812 S26 0.934 S38 0.948
S3 0.862 S15 0.937 S27 0.865 S39 0.989
S4 0.867 S16 0.922 S28 0.852 S40 0.854
S5 0.935 S17 0.930 S29 0.789 S41 0.979
S6 0.967 S18 0.915 S30 0.851 S42 0.969
S7 0.872 S19 0.874 S31 0.977 S43 0.920
S8 0.889 S20 0.912 S32 0.917 S44 0.947
S9 0.923 S21 0.924 S33 0.974 S45 0.974
S10 0.936 S22 0.897 S34 0.807 S46 0.918
S11 0.881 S23 0.905 S35 0.933 S47 0.898
S12 0.785 S24 0.961 S36 0.976
), ArticleFig(id=1218490404253650950, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Tab.3, caption=

Linear relationship of effective fraction content determination in Forsythiae Fructus

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Regression equation r2 Linear range/mg·mL-1 LOQ/mg·mL-1 LOD/mg·mL-1
Forsythoside B Y=7 077 177.963 1X-20 977.504 5 0.999 5 0.0435~0.261 0 0.027 1 0.008 2
Forsythoside A Y=4 800 254.659 5X-20 643.587 4 0.999 4 0.0652~0.625 4 0.037 8 0.011 5
Hyperin Y=11 115 033.145 0X+2 777.047 4 0.999 4 0.0486~0.291 8 0.017 2 0.005 2
Arctiin Y=6 906 738.002 3X-2 940.477 9 0.999 8 0.0237~0.142 4 0.006 6 0.002 0
Phillyrin Y=5 940 465.525 2X-2 503.057 4 0.999 6 0.0398~0.238 7 0.006 7 0.002 0
Phillygenin Y=8 972 467.100 4X-2 246.935 7 0.999 8 0.0415~0.248 8 0.004 4 0.001 3
), ArticleFig(id=1218490404329148423, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=表3, caption=

连翘药材有效成分含量测定线性关系考察

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Regression equation r2 Linear range/mg·mL-1 LOQ/mg·mL-1 LOD/mg·mL-1
Forsythoside B Y=7 077 177.963 1X-20 977.504 5 0.999 5 0.0435~0.261 0 0.027 1 0.008 2
Forsythoside A Y=4 800 254.659 5X-20 643.587 4 0.999 4 0.0652~0.625 4 0.037 8 0.011 5
Hyperin Y=11 115 033.145 0X+2 777.047 4 0.999 4 0.0486~0.291 8 0.017 2 0.005 2
Arctiin Y=6 906 738.002 3X-2 940.477 9 0.999 8 0.0237~0.142 4 0.006 6 0.002 0
Phillyrin Y=5 940 465.525 2X-2 503.057 4 0.999 6 0.0398~0.238 7 0.006 7 0.002 0
Phillygenin Y=8 972 467.100 4X-2 246.935 7 0.999 8 0.0415~0.248 8 0.004 4 0.001 3
), ArticleFig(id=1218490404400451592, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Tab.4, caption=

Relative correction factor(fi/s) and relative retention value(Ri/s) of the components to be tested in Forsythiae Fructus

, figureFileSmall=null, figureFileBig=null, tableContent=
Component fi/s RSD/% Ri/s RSD/%
Forsythoside B 0.981 0.422 0.95 0.326
Forsythoside A 1 - 1 -
Hyperin 0.972 2.651 1.12 0.415
Arctiin 0.251 1.505 1.26 0.216
Phillyrin 0.344 1.665 1.33 0.337
Phillygenin 0.334 1.809 1.55 0.359
), ArticleFig(id=1218490404467560457, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=表4, caption=

连翘药材中各待测成分的相对校正因子(fi/s)与相对保留值(Ri/s)

, figureFileSmall=null, figureFileBig=null, tableContent=
Component fi/s RSD/% Ri/s RSD/%
Forsythoside B 0.981 0.422 0.95 0.326
Forsythoside A 1 - 1 -
Hyperin 0.972 2.651 1.12 0.415
Arctiin 0.251 1.505 1.26 0.216
Phillyrin 0.344 1.665 1.33 0.337
Phillygenin 0.334 1.809 1.55 0.359
), ArticleFig(id=1218490404530475018, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Tab.5, caption=

Sample recovery rate of the components to be tested in Forsythiae Fructus

, figureFileSmall=null, figureFileBig=null, tableContent=
Compounds m(Content)/mg m(Added)/mg m(Found)/mg Recovery/% Mean recover/% RSD/%
Forsythoside B 6.09 6.52 12.21 96.85 98.99 2.67
6.09 6.52 12.25 97.16
6.09 6.52 12.14 96.31
6.09 6.52 12.61 99.98
6.09 6.52 12.99 103.01
6.09 6.52 12.69 100.64
Forsythoside A 24.98 19.14 42.94 97.32 99.46 1.97
24.98 19.14 43.44 98.46
24.98 19.14 43.33 98.21
24.98 19.14 44.11 99.98
24.98 19.14 44.09 99.93
24.98 19.14 45.39 102.87
Hyperin 0.81 0.73 1.48 96.26 99.37 2.71
0.81 0.73 1.52 98.55
0.81 0.73 1.50 97.20
0.81 0.73 1.55 100.86
0.81 0.73 1.60 103.73
0.81 0.73 1.53 99.59
Arctiin 2.97 2.37 5.27 98.62 99.23 3.39
2.97 2.37 5.23 97.87
2.97 2.37 5.18 96.98
2.97 2.37 5.20 97.41
2.97 2.37 5.66 105.98
2.97 2.37 5.26 98.52
Phillyrin 8.57 7.16 15.36 97.65 99.65 2.36
8.57 7.16 15.42 98.00
8.57 7.16 15.33 97.45
8.57 7.16 15.84 100.68
8.57 7.16 15.84 100.68
8.57 7.16 16.26 103.37
Phillygenin 3.27 3.11 6.15 96.35 98.92 1.91
3.27 3.11 6.29 98.62
3.27 3.11 6.22 97.49
3.27 3.11 6.49 101.73
3.27 3.11 6.34 99.37
3.27 3.11 6.38 99.94
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连翘药材中各待测成分的加样回收率

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Compounds m(Content)/mg m(Added)/mg m(Found)/mg Recovery/% Mean recover/% RSD/%
Forsythoside B 6.09 6.52 12.21 96.85 98.99 2.67
6.09 6.52 12.25 97.16
6.09 6.52 12.14 96.31
6.09 6.52 12.61 99.98
6.09 6.52 12.99 103.01
6.09 6.52 12.69 100.64
Forsythoside A 24.98 19.14 42.94 97.32 99.46 1.97
24.98 19.14 43.44 98.46
24.98 19.14 43.33 98.21
24.98 19.14 44.11 99.98
24.98 19.14 44.09 99.93
24.98 19.14 45.39 102.87
Hyperin 0.81 0.73 1.48 96.26 99.37 2.71
0.81 0.73 1.52 98.55
0.81 0.73 1.50 97.20
0.81 0.73 1.55 100.86
0.81 0.73 1.60 103.73
0.81 0.73 1.53 99.59
Arctiin 2.97 2.37 5.27 98.62 99.23 3.39
2.97 2.37 5.23 97.87
2.97 2.37 5.18 96.98
2.97 2.37 5.20 97.41
2.97 2.37 5.66 105.98
2.97 2.37 5.26 98.52
Phillyrin 8.57 7.16 15.36 97.65 99.65 2.36
8.57 7.16 15.42 98.00
8.57 7.16 15.33 97.45
8.57 7.16 15.84 100.68
8.57 7.16 15.84 100.68
8.57 7.16 16.26 103.37
Phillygenin 3.27 3.11 6.15 96.35 98.92 1.91
3.27 3.11 6.29 98.62
3.27 3.11 6.22 97.49
3.27 3.11 6.49 101.73
3.27 3.11 6.34 99.37
3.27 3.11 6.38 99.94
), ArticleFig(id=1218490404677275660, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=EN, label=Tab.6, caption=

Determination results of 6 components in Forsythiae Fructus. %

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No. Forsythoside
ESM		 Forsythoside B Hyperin Arctiin Phillyrin Phillygenin
ESM QAMS ESM QAMS ESM QAMS ESM QAMS ESM QAMS
S1 2.50 0.61 0.61 0.08 0.08 0.30 0.30 0.86 0.88 0.33 0.33
S2 1.40 0.25 0.25 0.03 0.03 0.19 0.19 0.47 0.48 0.09 0.09
S3 1.88 0.72 0.72 0.07 0.07 0.41 0.41 1.20 1.24 0.22 0.22
S4 1.87 0.68 0.68 0.07 0.07 0.39 0.39 1.14 1.18 0.21 0.21
S5 1.95 0.22 0.21 0.03 0.03 0.14 0.14 0.32 0.33 0.08 0.08
S6 2.66 0.60 0.59 0.09 0.09 0.41 0.41 0.95 0.98 0.26 0.26
S7 2.33 0.23 0.23 0.01 0.01 0.02 0.02 0.25 0.26 0.07 0.07
S8 1.87 0.33 0.32 0.02 0.02 0.04 0.04 0.60 0.62 0.15 0.15
S9 1.17 0.14 0.13 0.04 0.04 0.15 0.15 0.39 0.40 0.26 0.26
S10 1.20 0.06 0.05 0.04 0.04 0.15 0.15 0.25 0.26 0.25 0.25
S11 0.94 0.09 0.08 0.04 0.04 0.13 0.13 0.33 0.34 0.31 0.31
S12 0.62 0.09 0.08 0.04 0.04 0.12 0.12 0.43 0.44 0.45 0.45
S13 0.78 0.10 0.09 0.02 0.02 0.15 0.15 0.41 0.42 0.37 0.37
S14 0.85 0.11 0.10 0.04 0.04 0.14 0.14 0.43 0.44 0.68 0.68
S15 0.88 0.14 0.13 0.02 0.02 0.07 0.07 0.18 0.18 0.07 0.07
S16 0.90 0.03 0.02 0.02 0.02 0.07 0.07 0.12 0.12 0.06 0.06
S17 0.71 0.13 0.12 0.02 0.02 0.06 0.06 0.16 0.16 0.07 0.07
S18 0.55 0.07 0.06 0.01 0.01 0.10 0.10 0.21 0.22 0.08 0.08
S19 0.73 0.18 0.17 0.02 0.02 0.17 0.17 0.53 0.55 0.29 0.29
S20 0.59 0.05 0.04 0.01 0.01 0.07 0.07 0.18 0.18 0.09 0.09
S21 0.64 0.04 0.04 0.00 0.00 0.06 0.05 0.17 0.18 0.08 0.08
S22 0.76 0.11 0.10 0.02 0.02 0.16 0.16 0.41 0.43 0.27 0.27
S23 0.65 0.04 0.04 0.01 0.01 0.07 0.07 0.18 0.19 0.11 0.11
S24 0.96 0.04 0.04 0.03 0.03 0.05 0.05 0.16 0.17 0.15 0.15
S25 0.96 0.09 0.08 0.04 0.04 0.18 0.18 0.31 0.32 0.22 0.22
S26 1.25 0.17 0.17 0.04 0.04 0.27 0.27 0.47 0.48 0.41 0.41
S27 0.59 0.16 0.15 0.02 0.02 0.15 0.15 0.48 0.49 0.21 0.21
S28 0.24 0.02 0.01 0.01 0.01 0.03 0.02 0.07 0.08 0.04 0.04
S29 0.35 0.03 0.03 0.01 0.01 0.10 0.10 0.18 0.18 0.17 0.17
S30 0.45 0.04 0.03 0.03 0.03 0.08 0.08 0.26 0.27 0.16 0.16
S31 2.21 0.11 0.11 0.09 0.09 0.24 0.24 0.38 0.39 0.52 0.52
S32 1.54 0.05 0.05 0.04 0.04 0.11 0.11 0.13 0.13 0.05 0.05
S33 1.99 0.06 0.06 0.10 0.10 0.21 0.21 0.28 0.28 0.30 0.30
S34 0.51 0.20 0.19 0.04 0.04 0.07 0.07 0.26 0.27 0.15 0.15
S35 4.29 0.27 0.26 0.02 0.02 0.22 0.22 0.66 0.68 0.55 0.55
S36 5.76 0.70 0.70 0.11 0.11 0.40 0.41 1.56 1.61 1.62 1.62
S37 7.67 0.59 0.59 0.19 0.19 0.53 0.53 1.73 1.78 0.84 0.84
S38 4.88 0.28 0.28 0.05 0.05 0.29 0.29 0.82 0.84 0.82 0.82
S39 6.50 0.70 0.70 0.09 0.09 0.45 0.46 1.78 1.84 1.30 1.30
S40 5.72 0.24 0.23 0.05 0.04 0.08 0.08 0.35 0.36 0.21 0.21
S41 7.78 0.60 0.59 0.16 0.16 0.43 0.43 1.68 1.73 0.86 0.86
S42 4.37 0.62 0.62 0.10 0.10 0.34 0.34 1.82 1.88 0.94 0.95
S43 3.34 0.24 0.24 0.03 0.03 0.07 0.07 0.57 0.59 0.35 0.35
S44 4.57 0.69 0.69 0.15 0.14 0.45 0.46 2.13 2.20 1.65 1.66
S45 6.75 0.76 0.76 0.12 0.12 0.69 0.69 1.95 2.02 2.22 2.23
S46 3.14 0.61 0.61 0.04 0.04 0.15 0.15 0.83 0.86 0.29 0.29
S47 6.87 0.47 0.47 0.05 0.05 0.24 0.24 0.62 0.64 0.35 0.35
), ArticleFig(id=1218490404756967437, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1218290943371953101, language=CN, label=表6, caption=

连翘药材中6种成分含量测定结果。%

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Forsythoside
ESM		 Forsythoside B Hyperin Arctiin Phillyrin Phillygenin
ESM QAMS ESM QAMS ESM QAMS ESM QAMS ESM QAMS
S1 2.50 0.61 0.61 0.08 0.08 0.30 0.30 0.86 0.88 0.33 0.33
S2 1.40 0.25 0.25 0.03 0.03 0.19 0.19 0.47 0.48 0.09 0.09
S3 1.88 0.72 0.72 0.07 0.07 0.41 0.41 1.20 1.24 0.22 0.22
S4 1.87 0.68 0.68 0.07 0.07 0.39 0.39 1.14 1.18 0.21 0.21
S5 1.95 0.22 0.21 0.03 0.03 0.14 0.14 0.32 0.33 0.08 0.08
S6 2.66 0.60 0.59 0.09 0.09 0.41 0.41 0.95 0.98 0.26 0.26
S7 2.33 0.23 0.23 0.01 0.01 0.02 0.02 0.25 0.26 0.07 0.07
S8 1.87 0.33 0.32 0.02 0.02 0.04 0.04 0.60 0.62 0.15 0.15
S9 1.17 0.14 0.13 0.04 0.04 0.15 0.15 0.39 0.40 0.26 0.26
S10 1.20 0.06 0.05 0.04 0.04 0.15 0.15 0.25 0.26 0.25 0.25
S11 0.94 0.09 0.08 0.04 0.04 0.13 0.13 0.33 0.34 0.31 0.31
S12 0.62 0.09 0.08 0.04 0.04 0.12 0.12 0.43 0.44 0.45 0.45
S13 0.78 0.10 0.09 0.02 0.02 0.15 0.15 0.41 0.42 0.37 0.37
S14 0.85 0.11 0.10 0.04 0.04 0.14 0.14 0.43 0.44 0.68 0.68
S15 0.88 0.14 0.13 0.02 0.02 0.07 0.07 0.18 0.18 0.07 0.07
S16 0.90 0.03 0.02 0.02 0.02 0.07 0.07 0.12 0.12 0.06 0.06
S17 0.71 0.13 0.12 0.02 0.02 0.06 0.06 0.16 0.16 0.07 0.07
S18 0.55 0.07 0.06 0.01 0.01 0.10 0.10 0.21 0.22 0.08 0.08
S19 0.73 0.18 0.17 0.02 0.02 0.17 0.17 0.53 0.55 0.29 0.29
S20 0.59 0.05 0.04 0.01 0.01 0.07 0.07 0.18 0.18 0.09 0.09
S21 0.64 0.04 0.04 0.00 0.00 0.06 0.05 0.17 0.18 0.08 0.08
S22 0.76 0.11 0.10 0.02 0.02 0.16 0.16 0.41 0.43 0.27 0.27
S23 0.65 0.04 0.04 0.01 0.01 0.07 0.07 0.18 0.19 0.11 0.11
S24 0.96 0.04 0.04 0.03 0.03 0.05 0.05 0.16 0.17 0.15 0.15
S25 0.96 0.09 0.08 0.04 0.04 0.18 0.18 0.31 0.32 0.22 0.22
S26 1.25 0.17 0.17 0.04 0.04 0.27 0.27 0.47 0.48 0.41 0.41
S27 0.59 0.16 0.15 0.02 0.02 0.15 0.15 0.48 0.49 0.21 0.21
S28 0.24 0.02 0.01 0.01 0.01 0.03 0.02 0.07 0.08 0.04 0.04
S29 0.35 0.03 0.03 0.01 0.01 0.10 0.10 0.18 0.18 0.17 0.17
S30 0.45 0.04 0.03 0.03 0.03 0.08 0.08 0.26 0.27 0.16 0.16
S31 2.21 0.11 0.11 0.09 0.09 0.24 0.24 0.38 0.39 0.52 0.52
S32 1.54 0.05 0.05 0.04 0.04 0.11 0.11 0.13 0.13 0.05 0.05
S33 1.99 0.06 0.06 0.10 0.10 0.21 0.21 0.28 0.28 0.30 0.30
S34 0.51 0.20 0.19 0.04 0.04 0.07 0.07 0.26 0.27 0.15 0.15
S35 4.29 0.27 0.26 0.02 0.02 0.22 0.22 0.66 0.68 0.55 0.55
S36 5.76 0.70 0.70 0.11 0.11 0.40 0.41 1.56 1.61 1.62 1.62
S37 7.67 0.59 0.59 0.19 0.19 0.53 0.53 1.73 1.78 0.84 0.84
S38 4.88 0.28 0.28 0.05 0.05 0.29 0.29 0.82 0.84 0.82 0.82
S39 6.50 0.70 0.70 0.09 0.09 0.45 0.46 1.78 1.84 1.30 1.30
S40 5.72 0.24 0.23 0.05 0.04 0.08 0.08 0.35 0.36 0.21 0.21
S41 7.78 0.60 0.59 0.16 0.16 0.43 0.43 1.68 1.73 0.86 0.86
S42 4.37 0.62 0.62 0.10 0.10 0.34 0.34 1.82 1.88 0.94 0.95
S43 3.34 0.24 0.24 0.03 0.03 0.07 0.07 0.57 0.59 0.35 0.35
S44 4.57 0.69 0.69 0.15 0.14 0.45 0.46 2.13 2.20 1.65 1.66
S45 6.75 0.76 0.76 0.12 0.12 0.69 0.69 1.95 2.02 2.22 2.23
S46 3.14 0.61 0.61 0.04 0.04 0.15 0.15 0.83 0.86 0.29 0.29
S47 6.87 0.47 0.47 0.05 0.05 0.24 0.24 0.62 0.64 0.35 0.35
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基于HPLC指纹图谱和一测多评多成分定量分析的连翘质量评价研究
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邓寒霜 1, 2 , 李筱玲 1, 2 , 王欣 1 , 张浪 1 , 孙金娟 3
中国药学杂志 | 论著 2024,59(15): 1429-1437
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中国药学杂志 | 论著 2024, 59(15): 1429-1437
基于HPLC指纹图谱和一测多评多成分定量分析的连翘质量评价研究
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邓寒霜1, 2, 李筱玲1, 2, 王欣1, 张浪1, 孙金娟3
作者信息
  • 1 商洛学院生物医药与食品工程学院, 陕西 商洛 726000
  • 2 陕西秦岭特色生物资源产业技术研究院, 陕西 商洛 726000
  • 3 陕西香菊药业集团有限公司, 陕西 商洛 726000

    • 邓寒霜,男,硕士,副教授 研究方向:中药质量控制 Tel:(0914)2329816

Quality Evaluation of Forsythiae Fructus Based on HPLC Fingerprint and QAMS Multi-Component Content Determination
Hanshuang DENG1, 2, Xiaoling LI1, 2, Xin WANG1, Lang ZHANG1, Jinjuan SUN3
Affiliations
  • 1 College of Biopharmaceutcal and Food Engineering, Shangluo University, Shangluo 726000, China
  • 2 Shaanxi Qinling Industrial Technology Research Institute of Special Biological Resources, Shangluo 726000, China
  • 3 Shaanxi Xiangju Pharmaceutical Goup Co.,Ltd, Shangluo 726000, China
出版时间: 2024-08-08 doi: 10.11669/cpj.2024.15.010
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摘要
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目的 建立连翘药材的高效液相色谱(HPLC)法指纹图谱及基于“一测多评”(QAMS)技术的多成分定量测定方法。方法 采用HPLC法建立河南、山东、山西、陕西4个省份47批连翘药材的指纹图谱,进行相似度评价,结合聚类分析(HCA)、正交偏最小二乘法-判别分析(OPLS-DA)对连翘进行化学模式识别分析。利用QAMS技术,以连翘酯苷A为内参物,同时对连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷和连翘脂素6种活性成分进行含量测定,分析不同产地样品差异。结果 建立了47批连翘药材的指纹图谱,相似度为0.785~0.989,共标定30个共有峰。聚类分析将47批样品聚为3类,OPLS-DA分析能有效将几个产区连翘药材进行区分(Q2>0.5)。采用QAMS法与外标法(ESM)测定47批样品中6种活性成分的含量结果无明显差异(P>0.05),不同产地样品中活性成分含量存在明显差异(P<0.01)。连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素6种成分在各省样品中的平均含量分别为:河南(4.50、20.58、0.52 、2.39、7.46、1.76 mg·g-1)、山东(0.90、9.26、0.36、1.39、3.84、3.87 mg·g-1)、山西(0.82、8.74、0.29、1.16、2.64、1.75 mg·g-1)、陕西(5.18、55.10、0.88、3.36、13.11、9.25 mg·g-1)。结论 不同来源的连翘药材样品中化学成分近似,但产地因素对有效成分含量有较大的影响,来自陕西的连翘样品中各检测成分含量最高。建立的连翘药材HPLC指纹图谱及多成分含量测定方法稳定、可靠,结合化学识别模式可为连翘药材产地判别及质量评价提供依据。

连翘  /  指纹图谱  /  高效液相色谱法  /  化学模式识别  /  一测多评  /  质量评价

OBJECTIVE To establish the HPLC fingerprint of Forsythiae Fructus and develop a multi-component quantitative analysis method based on quantitative analysis of multi-components by single-marker(QAMS). METHODS The fingerprints of 47 Forsythiae Fructus samples from four provinces, including Henan, Shandong, Shanxi and Shaanxi, were established by high performance liquid chromatography (HPLC), while their similarity was evaluated. Chemical pattern recognition of Forsythiae Fructus was studied by hierarchical cluster analysis (HCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). The content of forsythoside B, forsythoside A, hyperin, arctiin, phillyrin and phillygenin were determined by using QAMS, and the differences of samples from different producing areas were analyzed. RESULTS The fingerprints of 47 Forsythiae Fructus samples were established with similarities ranged from 0.785 to 0.989, and a total of 30 common peaks were identified. According to HCA, the samples were divided into three groups. OPLS-DA analysis can effectively distinguish Forsythiae Fructus from several producing areas (Q2>0.5). There was no significant difference in the content of 6 components in 47 batches of samples by QAMS and external standard method (ESM) (P>0.05), but there was significant difference in the content of active components in samples from different origin (P<0.01). The average contents of forsythoside B, forsythoside A, hyperin, arctiin, phillyrin and phillygenin in samples from different provinces were, Henan(4.50、20.58、0.52、2.39、7.46、1.76 mg·g-1), Shandong(0.90, 9.26、0.36、1.39、3.84、3.87 mg·g-1), Shanxi(0.82、8.74、0.29、1.16、2.64、1.75 mg·g-1), Shaanxi(5.18、55.10、0.88、3.36、13.11、9.25 mg·g-1). CONCLUSION The chemical composition of Forsythiae Fructus samples from different locations was similar, but the origin factors had a significant effect on the content of active ingredients, and the samples from Shaanxi province had the highest content. The HPLC fingerprint combined with the simultaneous determination of multi-component content analysis method established in this study is stable and reliable. The fingerprint combined with the chemical pattern recognition can provide a basis for the identification of origins and quality evaluation of Forsythiae Fructus.

Forsythiae Fructus  /  fingerprint  /  HPLC  /  chemical pattern recognition  /  QAMS  /  quality evaluation
邓寒霜, 李筱玲, 王欣, 张浪, 孙金娟. 基于HPLC指纹图谱和一测多评多成分定量分析的连翘质量评价研究. 中国药学杂志, 2024 , 59 (15) : 1429 -1437 . DOI: 10.11669/cpj.2024.15.010
Hanshuang DENG, Xiaoling LI, Xin WANG, Lang ZHANG, Jinjuan SUN. Quality Evaluation of Forsythiae Fructus Based on HPLC Fingerprint and QAMS Multi-Component Content Determination[J]. Chinese Pharmaceutical Journal, 2024 , 59 (15) : 1429 -1437 . DOI: 10.11669/cpj.2024.15.010
正文
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连翘为木犀科植物连翘[Forsythia suspensa (Thunb.) Vahl]的干燥果实[1],味苦、性微寒,具有清热解毒,消痈散结,疏散风热等功效,常用于治疗风热感冒、温病初起、温热入营、高热烦渴、热淋尿闭、痈疽、瘰疬、乳痈、丹毒和肿毒等疾病[2]。秋季连翘果实初熟尚带绿色时采收,蒸熟,晒干为青翘;果实熟透时采收,晒干为老翘。连翘主产于我国河南、山西、陕西等省[3-5]。现代药学研究发现连翘中化学成分较多,主要为苯乙醇苷类(连翘酯苷A、B、C、D、E等20余种)、木脂素类(连翘苷、连翘脂素、牛蒡子苷等40余种)和挥发油类(相对含量较多的有α-蒎烯、β-蒎烯、水芹烯、对聚伞花烯、柠檬烯、孟二烯、伞花烃等),具有抗炎、抑菌、抗病毒、抗氧化、保肝及抗肿瘤等活性[6-11]。基于高效液相色谱(HPLC)的中药指纹图谱能够较为全面地反映中药的化学特征,能很好体现中药的均一性、稳定性,有利于从整体上控制中药质量,确保中药产品质量稳定。“一测多评”(quantitative analysis of multi-components by single-marker, QAMS)技术是由Wang等[12]提出,它利用了样品中多种化学成分与内参物间的相对校正因子,实现了只用一种对照品完成多种化学成分的定量分析,可节省检测成本,提高工作效率。目前QAMS技术已被《中国药典》收录用于丹参、淫羊藿、黄连、生姜等多个品种的含量测定,有文献报道该技术在中药材及中成药的质量控制中都得到了很好的应用[13~19]。为比较不同产地连翘品质差异,本研究从陕西、山西、河南、山东等地收集47份样品,建立了连翘药材指纹图谱和基于QAMS技术,可同时测定连翘酯苷A、连翘酯苷B、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素6种成分的含量,为综合评价连翘药材质量提供了数据支持。
1 仪器与药品
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1.1 仪器
岛津LC-20A高效液相色谱仪(包括在线脱气机、四元泵、柱温箱、PDA检测器等)、色谱柱WondSil C18 Superd(4.6 mm×250 mm,5 μm)(日本岛津公司);LA230S型电子分析天平(赛多利斯天平公司);SB25-12DTDN型超声波提取仪(宁波新芝生物科技股份有限公司)等。
1.2 试剂
连翘苷(批号200406,中国食品药品检定研究院);金丝桃苷(批号171224,北京世纪奥科生物技术有限公司);连翘酯苷A(批号210507)、连翘酯苷B(批号210204)、连翘脂素(批号210807)、牛蒡子苷(批号210505)(北京中科质检公司),以上对照品纯度均≥98%;甲醇为色谱纯;甲醇、磷酸为分析纯;纯净水(杭州娃哈哈公司)。
1.3 药材
47批药材样品收集自山西、河南、陕西、山东等地,经商洛学院李筱玲副教授/执业药师鉴定为木犀科连翘属植物连翘[Forsythia suspensa (Thunb.) Vahl]的干燥果实。样品信息见表1。
2 方法与结果
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2.1 指纹图谱的建立
2.1.1 供试品溶液的制备
取连翘干燥药材样品适量,除去杂质后,粉碎,过3号标准药典筛。精确称量连翘药材粉末1.0 g,置具塞平底烧瓶内,精密加入体积分数70%甲醇25 mL,塞密,称定重量,超声(功率250 W,频率40 kHz)处理30 min,取出,放冷,再次称定重量,以70%甲醇补足减失重量,摇匀,快速滤纸过滤,取滤液,再以0.45 μm微孔滤膜过滤,即得。
2.1.2 对照品溶液的制备
精密称取各对照品适量,制成浓度为连翘酯苷A 0.625 4 mg·mL-1、连翘酯苷B 0.261 0 mg·mL-1、连翘苷0.238 7 mg·mL-1、连翘脂素0.248 8 mg·mL-1、金丝桃苷0.291 8 mg·mL-1、牛蒡子苷0.142 4 mg·mL-1的混合对照品溶液,0.45 μm微孔滤膜过滤,即得。
2.1.3 色谱条件
WondSil C18 Superd色谱柱(4.6 mm×250 mm,5μm)。流动相为甲醇(A)-0.4%磷酸(B),梯度洗脱程序0~30 min,20%~35%A;30~50 min,35%~60%A;50~55 min,60%A;55~57 min,60%~20%A;57~60 min,20%A。流速1.0 mL·min-1,柱温35 ℃,进样量10 μL,检测波长230 nm。
2.1.4 精密度试验
取S1号样品适量,按照“2.1.1”项下方法制备供试品溶液,按照“2.1.3”项下方法连续进样6次,以连翘酯苷A为对照,计算其他共有峰的相对保留值与相对峰面积。结果显示,各共有峰的相对保留值与相对峰面积的相对标准偏差(RSD)值均小于2.0%,表明仪器精密度良好。
2.1.5 稳定性试验
取S1号样品适量,按照“2.1.1”项下方法制备供试品溶液,分别放置0、4、10、24、48、72 h后照“2.1.3”项下方法进样,以连翘酯苷A为对照,计算其他共有峰的相对保留值与相对峰面积。结果显示,各共有峰的相对保留值与相对峰面积的RSD值均小于3.0%,表明供试品溶液在72 h内具有良好的稳定性。
2.1.6 重复性试验
取S1号样品适量,按照“2.1.1”项下方法,同步制备6份供试品溶液,按照“2.1.3”项下方法进样,以连翘酯苷A为对照,计算其他共有峰的相对保留值与相对峰面积。结果显示,各共有峰的相对保留值与相对峰面积的RSD值均小于3.0%,表明该实验方法具有良好的重复性。
2.1.7 指纹图谱的建立
取各批样品适量,按照“2.1.1”项下方法,分别制备供试品溶液,按照“2.1.3”项下方法进样分析,记录色谱图。将所得47批样品色谱数据导入“中药色谱指纹图谱相似度评价系统”(版本号2012.130723),选择共有峰峰面积在全部样品中处于中间水平的S1为参照图谱,中位数为基准,时间窗口0.5,得到47批连翘样品指纹图谱及对照指纹图谱(图1~2)。经匹配后确认共有峰30个,其中13号峰为连翘酯苷B、14号峰为连翘酯苷A、17号峰为金丝桃苷、20号峰牛蒡子苷、22号峰为连翘苷、29号峰为连翘脂素。
2.1.8 相似度分析
将47批连翘药材样品HPLC色谱数据导入中药指纹图谱相似度评价系统,计算相似度。所有样品相似度在0.785~0.989之间,表明产地因素对连翘药材质量有一定的影响,原因应与产地气候环境及产地加工技术相关。相似度≥0.9的样品占总数的63.8%,表明多数连翘样品中的化学成分近似,连翘药材整体质量较稳定。各样品与对照指纹图谱相似度计算结果见表2。
2.2 化学模式识别分析
2.2.1 聚类分析(HCA)
以47批连翘药材色谱指纹图中30个共有峰峰面积为变量,利用SPSS 23.0软件,以平方欧氏距离为区间,采用组间联接法进行聚类分析,结果见图3。
当类间距离小于10时,47批连翘药材可聚为3类。Ⅰ类共有34个样品,全部来自陕西省外产区;Ⅱ类6个样品、Ⅲ类7个样品全部来源于陕西省,其中Ⅲ类样品全部来自连翘药材的传统道地产区陕西省商洛市,表明产于该地区的连翘药材在品质上有一定的特点。
2.2.2 正交偏最小二乘法-判别分析(OPLS-DA)
为进一步判断产地因素对连翘药材质量的影响,寻找造成陕西道地产区连翘药材独特品质差异的标志物,以47批连翘药材指纹图谱30个共有峰的峰面积为变量,应用SIMCA 14.1软件进行OPLS-DA分析(图4)。由图4可知,陕西省样品与其他样品得到了较好的分离;得OPLS-DA模型的 R Y 2=0.714,Q2=0.585,均大于0.5,表明模型稳定可靠,可用于陕西连翘传统道地产区与其他产地间的判别。
结合变量重要性投影值(variable importancd for ghte projection, VIP),以VIP>1筛选出差异成分(图5)。标记出差异性成分11种,按贡献由大到小分别是,峰30、峰13(连翘酯苷B)、峰23、峰27、峰29(连翘脂素)、峰19、峰28、峰18、峰14(连翘酯苷A)、峰1、峰2。这些成分是造成陕西连翘与其他连翘品质差异的主要标志性成分,生产中可加强上述成分的含量控制,作为区别连翘产区的依据之一。
2.3 连翘药材QAMS含量测定
2.3.1 色谱条件及供试品、对照品溶液制备
同“2.1.1”“2.1.2”“2.1.3”项下方法。
2.3.2 线性关系考察
分别精密吸取“2.1.2”项下混合对照品溶液适量,用甲醇稀释成不同浓度,进样分析,记录色谱图。以对照品溶液浓度为横坐标(X)、峰面积为纵坐标(Y)绘制校准曲线,以信噪比10:1计算定量限(LOQ),信噪比3:1计算检测限(LOD),结果见表3。
2.3.3 精密度考察
精密吸取“2.1.2”项下混合对照品溶液10 μL,按“2.1.3”项下色谱条件连续进样6次,记录各对照品色谱峰面积,得连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素峰面积RSD值分别为0.271%、0.203%、0.215%、0.312%、0.223%、0.278%,表明所用仪器精密度良好。
2.3.4 稳定性试验
取“2.1.2”项所得混合对照品溶液适量,分别放置0、4、10、24、48、72 h后按照“2.1.3”项下方法进样,记录各对照品色谱峰面积,得连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素峰面积RSD值分别为0.309%、0.219%、0.387%、0.420%、0.432%、0.324%,表明所得混合对照溶液在72 h内稳定。
2.3.5 重复性试验
精密称取S1号连翘药材粉末适量,按照“2.1.1”项下方法,平行制备6份供试品溶液,按照“2.1.3”项下方法进样,记录各待测组分色谱峰面积,得连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素峰面积RSD值分别为1.191%、1.267%、1.604%、1.517%、1.078%、1.121%,表明所用方法具有良好的重复性。
2.3.6 相对校正因子与相对保留值的计算
取“2.1.2”项下所得混合对照品溶液,按“2.1.3”项下方法,连续进样12次,记录各对照品色谱峰面积,选择连翘酯苷A为内参物,按公式1计算各对照品的相对校正因子(fi/s);按公式2计算各对照品的相对保留值(Ri/s),结果见表4。
fi/s=fi/fs=Asρi/Aiρs
Ri/s=tRi/tRs
式中,fi为待测成分斜率,fs为内参物斜率,As为内参物峰面积,ρs为内参物质量浓度,Ai为待测成分峰面积,ρi为待测成分质量浓度;tRs为内参物保留时间,tRi为待测成分保留时间。
2.3.7 加样回收试验
取已用外标法(external standard method,ESM)测知含量的S01号样品粉末适量,精确称量6份,每份1.0 g,精密加入与样品含量相当的连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素对照品,按“2.1.1”项下方法制备供试品溶液,按“2.1.3”项中方法进样分析,计算各组分加样回收率,结果见表5。
2.3.8 样品测定
取各批次样品适量,按照“2.1.1”项下方法制备供试品溶液,按“2.1.3”项下方法进样分析,记录每个样品色谱图中各待测组分的色谱峰面积。每个样品平行处理3份,取平均值。采用QAMS法按照公式3计算含量并与ESM法比较,结果见表6。
M(%)=fi/sAiρs/As×25/1 000×100%
式中,M(%)为待测组分百分含量,As为连翘酯苷A对照品峰面积,ρs为连翘酯苷A对照品质量浓度,Ai为待测成分峰面积,fi/s为待测组分相对校正因子。
利用SPSS 23.0软件对除内参物(连翘酯苷A)外的5个成分的2组检测数据进行F检验,结果显示两种方法没有显著差异(P>0.05),连翘酯苷B、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素的P值分别为0.910、0.937、0.990、0.865、0.995,表明可用QAMS替代ESM对连翘药材进行含量测定。
连翘酯苷B、连翘酯苷A、金丝桃苷、牛蒡子苷、连翘苷、连翘脂素6种成分在各省样品中的平均含量为:河南(4.50、20.58、0.52、2.39、7.46、1.76 mg·g-1)、山东(0.90、9.26、0.36、1.39、3.84、3.87 mg·g-1)、山西(0.82、8.74、0.29、1.16、2.64、1.75 mg·g-1)、陕西(5.18、55.10、0.88、3.36、13.11、9.25 mg·g-1)。利用SPSS 23.0软件对来自不同产地的47批样品中6个成分的总量及单个成分含量进行了Pearson相关性分析,结果不同产地样品间存在极显著差异(P<0.01)。4个省份样品中6种有效成分含量比较见图6。来自陕西的连翘样品中有效成分含量最高,其次为河南样品。
3 讨论
收起
连翘药材主要含有苯乙醇苷类、木脂素类、挥发油等成分,2020年版《中国药典》一部规定,连翘药材需进行挥发油、连翘苷、连翘酯苷A的含量测定,其中后两种成分测定采用的是HPLC法,但这两种成分所用的提取方法与色谱条件均不相同。为确保目标成分的提取率与分离效果,课题组对连翘药材提取方法与色谱分离条件进行了考察。主要比较了不同提取方式(超声、回流)、不同提取溶剂(水、甲醇、70%甲醇、50%甲醇、乙醇、70%乙醇、50%乙醇)、不同料液比(1:5、1:10、1:25、1:50、1:100)。发现以70%甲醇为溶剂,按料液比1:25,采用超声处理制备的供试品溶液所含色谱峰较多,且经济、省时。对流动相组成(甲醇:0.4%磷酸、乙腈:0.4%磷酸)、检测波长(190、230、277、330 nm)进行考察,发现以甲醇:0.4%磷酸溶液为流动相系统,进行梯度洗脱得到的色谱图中,连翘药材各成分得到了良好的分离;以230 nm作检测波长时,连翘药材色谱图基线平稳,各目标成分色谱峰分离度、峰面积均较为理想,能较为全面地体现出连翘药材中所包含的化学信息。连翘药材按采收期不同可分为青翘与老翘,两种规格的连翘药材品质上有一定的差异性。有学者认为老翘中某些活性成分如梾木苷甲基醚的含量高于青翘[20],特别是老翘中比例更高的有益糖类,能促进肠道有益菌群生长,具有更好的解毒保肝作用[21]。也有文献报道青翘中苯乙醇苷类成分含量更高,青翘品质更好[22-25],在生产实践中一般青翘价格更高。本研究所用样品全部为青翘,在后续的工作中,课题组将针对连翘采收期与其内在化学成分的变化关系进行研究,以期建立起连翘药材采收时间与内在化学信息间的动态指纹图谱,为更客观、准确地评价连翘药材质量奠定基础。
基金
收起
  • 中央引导地方科技发展专项(2019ZY-FP-02)
  • 陕西省科技计划项目(2022JM-487)
  • 商洛学院院士工作站专项基金项目(20YSZX02)
文献
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2024年第59卷第15期
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doi: 10.11669/cpj.2024.15.010
  • 接收时间：2024-03-07
  • 首发时间：2026-01-14
  • 出版时间：2024-08-08
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  • 收稿日期：2024-03-07
基金
中央引导地方科技发展专项(2019ZY-FP-02)
陕西省科技计划项目(2022JM-487)
商洛学院院士工作站专项基金项目(20YSZX02)
作者信息
    1 商洛学院生物医药与食品工程学院, 陕西 商洛 726000
    2 陕西秦岭特色生物资源产业技术研究院, 陕西 商洛 726000
    3 陕西香菊药业集团有限公司, 陕西 商洛 726000
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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