Article(id=1240709665894494809, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240709662501294254, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024-0478, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1721664000000, receivedDateStr=2024-07-23, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773737835620, onlineDateStr=2026-03-17, pubDate=1743350400000, pubDateStr=2025-03-31, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773737835620, onlineIssueDateStr=2026-03-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773737835620, creator=13701087609, updateTime=1773737835620, updator=13701087609, issue=Issue{id=1240709662501294254, tenantId=1146029695717560320, journalId=1205117023404326918, year='2025', volume='45', issue='3', pageStart='361', pageEnd='542', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1773737834810, creator=13701087609, updateTime=1773737909503, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1240709975845163177, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240709662501294254, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1240709975845163178, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240709662501294254, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=489, endPage=503, ext={EN=ArticleExt(id=1240709667739988613, articleId=1240709665894494809, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Quality evaluation research of Vernonia amygdalina Del. based on HPLC fingerprint combined with chemical pattern recognition and multi-component determination*, columnId=1239148841803501731, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Quality Control, runingTitle=null, highlight=null, articleAbstract=

Objective: To establish a multi-component determination method and HPLC fingerprint for Vernonia amygdalina Del., combined with chemical pattern recognition to comprehensively evaluate the quality of Vernonia amygdalina Del. from different producing, laying a foundation for the future development of quality control standards for Vernonia amygdalina Del. and other related research on Vernonia amygdalina Del.’s components. Methods: The HPLC using Welch Ultimate® AQ-C18 column (250 mm×4.6 mm,5 µm) and 0.2% phosphoric acid aqueous solution - acetonitrile as moblie phase by gradient elution at the flow rate of 1 mL · min-1, wavelength was set at 258 nm and the column temperature was 35 ℃. Simultaneously establish 24 batches of fingerprint spectra of Vernonia amygdalina Del. from different origins and a multi-component HPLC determination method for 9 components of Vernonia amygdalina Del.. Combine chemical pattern recognition to evaluate the quality of 24 batches of Vernonia amygdalina Del. from different origins. Results: The similarities of the fingerprint spectra of 24 batches of Vernonia amygdalina Del. were above 0.9. The established control fingerprint spectra could stably and effectively distinguish Vernonia amygdalina Del. qualitatively. There were 10 common peaks in the fingerprint spectra, and 9 peaks were identified. The mass fractions of 24 batches of Vernonia amygdalina Del. components that were uracil, chlorogenic acid, luteolin, luteolin-7-O-β-D-glucuronide, isochlorogenic acid B, 3,5-O-dicaffeoyl quinic acid, 1,5-dicaffeoyl quinic acid, apigenin-7-O-β-D-glucopyranoside, 4,5-O-dicaffeoyl quinic acid were 0.007 314–0.084 30 mg · g-1,0.619 6–9.763 mg · g-1,0.303 8–4.031 mg · g-1,0.984 6–8.146 mg · g-1,0.043 29–0.438 7 mg · g-1,0.537 5–11.57 mg · g-1,0.437 6–13.78 mg · g-1,0.032 19–0.720 1 mg · g-1,0.190 0–1.931 mg · g-1. Through cluster analysis, 24 batches of Vernonia amygdalina Del. were divided into 2 categories. Vernonia amygdalina Del.from Guangdong and Hainan were classified as one category, while Vernonia amygdalina Del. from Guangxi were classified as one category alone. The classification results of principal component analysis were consistent with cluster analysis. Further research on the differences in leaf quality among different regions of Vernonia amygdalina Del. was conducted through principal component scores, and it was found that the P5, P24 and P22 regions ranked in the top three with better quality than other regions. Through orthogonal partial least squares discriminant analysis, it was found that the reasons for the differences in leaf quality among different regions of Vernonia amygdalina Del. were ranked in descending order: 1,5-dicaffeoyl quinic acid, chlorogenic acid, and luteolin-7-O-β-D-glucuronide, 3,5-O-dicaffeoyl quinic acid. Conclusion: The fingerprint and multi-component determination methods of Vernonia amygdalina Del. are stable and effective through methodological detection, which can fill the gap in quality control standards for Vernonia amygdalina Del.. Combined with chemical pattern recognition methods, the high-quality production areas of Vernonia amygdalina Del. and the components that should be emphasized in controlling the quality of Vernonia amygdalina Del. from different production areas are pointed out.A comprehensive evaluation of the quality of Vernonia amygdalina Del. has been conducted, providing a solid foundation for future quality control and evaluation of Vernonia amygdalina Del..

, correspAuthors=Hua-liang QIN, Chuan-wu FU, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Jing YANG, Hua-liang QIN, Chuan-wu FU, Shi-hua HU, Qin QIU, Yu-qing LAN, Dong-jie QIN, Min-tao HUANG, Wen-jun ZHONG, Jia XU, Zi-long QIN), CN=ArticleExt(id=1240709669866500892, articleId=1240709665894494809, tenantId=1146029695717560320, journalId=1205117023404326918, language=CN, title=基于高效液相色谱指纹图谱结合化学模式识别及多成分测定的南非叶质量评价研究*, columnId=1239148842025799861, journalTitle=药物分析杂志, columnName=质量分析, runingTitle=null, highlight=null, articleAbstract=

目的:建立南非叶的多成分测定方法及高效液相色谱(HPLC)指纹图谱,并结合化学模式识别的方法,综合评价不同产地的南非叶质量,为今后制定南非叶质量控制标准及其他相关南非叶成分研究奠定基础。方法:使用HPLC法,采用Welch Ultimate® AQ-C18色谱柱(250 mm×4.6 mm,5 µm),以0.2%磷酸水溶液-乙腈为流动相进行梯度洗脱,检测波长258 nm,体积流量1 mL · min-1,柱温35 ℃。同时建立24批不同产地的南非叶指纹图谱及9个成分HPLC测定方法,并结合化学模式识别对各批不同产地的南非叶进行质量评价。结果:24批南非叶的指纹图谱的相似度均在0.9以上,建立的对照指纹图谱可稳定有效地对南非叶进行定性判别,指纹图谱中10个共有峰,指认出9个峰。24批南非叶中尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸的质量分数分别为0.007 314~0.084 30 mg · g-1、0.619 6~9.763 mg · g-1、0.303 8~4.031 mg · g-1、0.984 6~8.146 mg · g-1、0.043 29~0.438 7 mg · g-1、0.537 5~11.57 mg · g-1、0.437 6~13.78 mg · g-1、0.032 19~0.720 1 mg · g-1、0.190 0~1.931 mg · g-1。通过聚类分析,将24批南非叶分为2类,广东和海南产的南非叶为一类,广西产的南非叶独自为一类;主成分分析的分类结果与聚类分析一致,通过主成分得分进一步研究各产地南非叶质量差异,发现P5、P24、P22地区得分位居前三质量优于其他地区。通过正交偏最小二乘-判别分析,发现造成各地南非叶质量差异的影响能力由强到弱排序为1,5-二咖啡酰奎宁酸、绿原酸、木犀草素-7-O-β-D-葡萄糖醛酸苷、3,5-O-二咖啡酰奎宁酸。结论:南非叶指纹图谱及多成分测定方法稳定可靠,可弥补南非叶质量控制标准方面的空白,结合化学模式识别方法,指出了南非叶质量优质的产地及影响不同产地南非叶质量应着重控制的成分,为南非叶质量控制及评价提供了坚实的基础。

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**覃华亮 Tel:18178842081;E-mail:;
符传武 Tel:18007726258;E-mail:
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Guangxi Eco-engineering Vocational and Technical College, Liuzhou 545001, China), AuthorCompanyExt(id=1240722371418575163, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, companyId=1240722371401797945, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4.广西生态工程职业技术学院,柳州 545001)])], figs=[ArticleFig(id=1240722376082641491, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Fig. 1, caption=HPLC fingerprint of 24 batches of V. amygdalina, figureFileSmall=fBFW4EXwuuJx4RzkqeagVA==, figureFileBig=oXObWoGWl5i2netxnDbVGA==, tableContent=null), ArticleFig(id=1240722376162333270, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=图1, caption=24批次南非叶HPLC指纹图谱, figureFileSmall=fBFW4EXwuuJx4RzkqeagVA==, figureFileBig=oXObWoGWl5i2netxnDbVGA==, tableContent=null), ArticleFig(id=1240722376262996574, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Fig. 2, caption=Comparison fingerprint of V. amygdalina, figureFileSmall=gNYHR3J+XLG6smgwf3eyzA==, figureFileBig=reKjmjT5XUpdII40sUCDAw==, tableContent=null), ArticleFig(id=1240722376342688355, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=图2, caption=南非叶对照指纹图谱, figureFileSmall=gNYHR3J+XLG6smgwf3eyzA==, figureFileBig=reKjmjT5XUpdII40sUCDAw==, tableContent=null), ArticleFig(id=1240722376439157352, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Fig. 3, caption=HPLC chromatograms of mixed reference solution (A) and V. amygdalina test solution (B), figureFileSmall=kBzCh8MZSINU5G2ofqeHGg==, figureFileBig=b0gOy3Ux59E4imZlLPiYDQ==, tableContent=null), ArticleFig(id=1240722376539820655, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=图3, caption=混合对照品溶液(A)和南非叶供试品溶液(B)HPLC色谱图

1.尿嘧啶(uracil) 2.绿原酸(chlorogenic acid) 3.木犀草苷(luteolin) 4.木犀草素-7-O-β-D-葡萄糖醛酸苷(luteolin-7-O-β-D-glucuronide)5.异绿原酸B(isochlorogenic acid B) 6. 3,5-O-二咖啡酰奎宁酸(3,5-O-dicaffeoyl quinic acid) 7. 1,5-二咖啡酰奎宁酸(1,5-dicaffeoyl quinic acid)8.芹菜素-7-O-β-D-吡喃葡萄糖苷(apigenin-7-O-β-D-glucopyranoside) 9. 4,5-O-二咖啡酰奎宁酸(4,5-O-dicaffeoyl quinic acid)

, figureFileSmall=kBzCh8MZSINU5G2ofqeHGg==, figureFileBig=b0gOy3Ux59E4imZlLPiYDQ==, tableContent=null), ArticleFig(id=1240722376627901043, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Fig. 4, caption=Dendrogram of 24 batches of V. amygdalina Del., figureFileSmall=1Qfp61AUmg2FzEndwLMHAw==, figureFileBig=LIouRP0sY8MN+ysUl3x4kA==, tableContent=null), ArticleFig(id=1240722376745341560, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=图4, caption=24批次南非叶聚类树状图, figureFileSmall=1Qfp61AUmg2FzEndwLMHAw==, figureFileBig=LIouRP0sY8MN+ysUl3x4kA==, tableContent=null), ArticleFig(id=1240722376854393469, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Fig. 5, caption=PCA scatter plot, figureFileSmall=xklk+BHTBLFyMCz1HFQ1bg==, figureFileBig=zCIxE8EhQXqiV5CVp4H/kA==, tableContent=null), ArticleFig(id=1240722376938279553, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=图5, caption=PCA散点图, figureFileSmall=xklk+BHTBLFyMCz1HFQ1bg==, figureFileBig=zCIxE8EhQXqiV5CVp4H/kA==, tableContent=null), ArticleFig(id=1240722377101857414, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Fig. 6, caption=VIP plot, figureFileSmall=NiZItxBdzyhG9LrvYjHLTQ==, figureFileBig=nbefK02JyHcYzzMOegzW+w==, tableContent=null), ArticleFig(id=1240722377210909322, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=图6, caption=VIP图, figureFileSmall=NiZItxBdzyhG9LrvYjHLTQ==, figureFileBig=nbefK02JyHcYzzMOegzW+w==, tableContent=null), ArticleFig(id=1240722377324155534, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 1, caption=

Informations of V. amygdalina

, figureFileSmall=null, figureFileBig=null, tableContent=
编号(number)来源(source)采收时间(harvest time)
P1广西玉林博白县
(Bobai County,Yulin,Guangxi)
2023年3月份
(March 2023)
P2广西玉林玉州区
(Yuzhou District,Yulin,Guangxi)
2023年3月份
(March 2023)
P3广西南宁马山县(本地)
[Mashan County,Nanning,Guangxi(locality)]
2023年3月份
(March 2023)
P4广西南宁马山县(引种)
[Mashan County,Nanning,Guangxi(introduction)]
2023年3月份
(March 2023)
P5广东东莞森林公园南非叶种植地
V. amygdalina Plantation of Forest Park,Dongguan City,Guangdong)
2023年3月份
(March 2023)
P6广西崇左
(Chongzhuo,Guangxi)
2023年3月份
(March 2023)
P7广西河池
(Heci,Guangxi)
2023年3月份
(March 2023)
P8广西北海
(Beihai,Guangxi)
2023年3月份
(March 2023)
P9广西柳州柳城县
(Liucheng County,Liuzhou,Guangxi)
2023年3月份
(March 2023)
P10广西柳州百草园
(Herb Garden,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P11广西柳州百草园
(Herb Garden,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P12广西柳州百草园
(Herb Garden,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P13广西柳州柳南区
(Liunan District,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P14广西柳州柳江区
(Liujiang District,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P15广西柳州柳南区太阳村
(Sun Village,Liunan District,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P16广西梧州岑溪市
(Cenxi City,Wuzhou,Guangxi)
2023年3月份
(March 2023)
P17广西梧州蒙山县
(Mengshan County,Wuzhou,Guangxi)
2023年3月份
(March 2023)
P18广西梧州藤县
(Teng County,Wuzhou,Guangxi)
2023年3月份
(March 2023)
P19海南五指山市热带雨林基地
(Tropical Rainforest Base,Wuzhishan City,Hainan)
2023年3月份
(March 2023)
P20海南五指山市
(Wuzhishan City,Hainan)
2023年3月份
(March 2023)
P21海南三亚市
(Sanya City,Hainan)
2023年3月份
(March 2023)
P22广东茂名
(Maoming city,Guangdong)
2023年3月份
(March 2023)
P23广东茂名高山镇
(Gaoshan Town,Maoming,Guangdong)
2023年3月份
(March 2023)
P24广东广州
(Guangzhou City,Guangdong)
2023年3月份
(March 2023)
), ArticleFig(id=1240722377458373267, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表1, caption=

南非叶样品信息

, figureFileSmall=null, figureFileBig=null, tableContent=
编号(number)来源(source)采收时间(harvest time)
P1广西玉林博白县
(Bobai County,Yulin,Guangxi)
2023年3月份
(March 2023)
P2广西玉林玉州区
(Yuzhou District,Yulin,Guangxi)
2023年3月份
(March 2023)
P3广西南宁马山县(本地)
[Mashan County,Nanning,Guangxi(locality)]
2023年3月份
(March 2023)
P4广西南宁马山县(引种)
[Mashan County,Nanning,Guangxi(introduction)]
2023年3月份
(March 2023)
P5广东东莞森林公园南非叶种植地
V. amygdalina Plantation of Forest Park,Dongguan City,Guangdong)
2023年3月份
(March 2023)
P6广西崇左
(Chongzhuo,Guangxi)
2023年3月份
(March 2023)
P7广西河池
(Heci,Guangxi)
2023年3月份
(March 2023)
P8广西北海
(Beihai,Guangxi)
2023年3月份
(March 2023)
P9广西柳州柳城县
(Liucheng County,Liuzhou,Guangxi)
2023年3月份
(March 2023)
P10广西柳州百草园
(Herb Garden,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P11广西柳州百草园
(Herb Garden,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P12广西柳州百草园
(Herb Garden,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P13广西柳州柳南区
(Liunan District,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P14广西柳州柳江区
(Liujiang District,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P15广西柳州柳南区太阳村
(Sun Village,Liunan District,Liuzhou,Guangxi)
2023年6月份
(June 2023)
P16广西梧州岑溪市
(Cenxi City,Wuzhou,Guangxi)
2023年3月份
(March 2023)
P17广西梧州蒙山县
(Mengshan County,Wuzhou,Guangxi)
2023年3月份
(March 2023)
P18广西梧州藤县
(Teng County,Wuzhou,Guangxi)
2023年3月份
(March 2023)
P19海南五指山市热带雨林基地
(Tropical Rainforest Base,Wuzhishan City,Hainan)
2023年3月份
(March 2023)
P20海南五指山市
(Wuzhishan City,Hainan)
2023年3月份
(March 2023)
P21海南三亚市
(Sanya City,Hainan)
2023年3月份
(March 2023)
P22广东茂名
(Maoming city,Guangdong)
2023年3月份
(March 2023)
P23广东茂名高山镇
(Gaoshan Town,Maoming,Guangdong)
2023年3月份
(March 2023)
P24广东广州
(Guangzhou City,Guangdong)
2023年3月份
(March 2023)
), ArticleFig(id=1240722377613562522, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 2, caption=

Regression equations and linear range

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
线性范围
(linear range)/(mg · mL–1
r检测限
(LOD) / (ng · mL-1
定量限
(LOQ) / (ng · mL-1
尿嘧啶
(uracil)
Y=3.986×107X-4.866×1030.000 056 31~0.004 5040.996 65.5718.56
绿原酸
(chlorogenic acid)
Y=8.040×106X-5.717×1040.008 216~0.657 20.999 529.8299.40
木犀草苷
(luteolin)
Y=2.211×107X-6.451×1040.003 171~0.253 70.999 520.8069.34
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
Y=2.144×107X-2.101×1050.004 201~0.336 10.997 845.27150.90
异绿原酸B
(isochlorogenic acid B)
Y=7.997×106X-4.564×1030.000 208 4~0.016 670.996 854.46181.53
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
Y=1.024×107X-7.792×1040.004 634~0.370 70.999 090.39301.30
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
Y=9.253×106X-6.312×1040.004 928~0.394 20.999 2101.09336.98
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
Y=1.719×107X-6.937×1030.000 297 5~0.023 800.999 343.51145.03
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
Y=1.172×107X-2.476×1040.000 898 6~0.071 890.998 2104.89349.65
), ArticleFig(id=1240722377747780255, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表2, caption=

线性回归方程及线性范围

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
线性范围
(linear range)/(mg · mL–1
r检测限
(LOD) / (ng · mL-1
定量限
(LOQ) / (ng · mL-1
尿嘧啶
(uracil)
Y=3.986×107X-4.866×1030.000 056 31~0.004 5040.996 65.5718.56
绿原酸
(chlorogenic acid)
Y=8.040×106X-5.717×1040.008 216~0.657 20.999 529.8299.40
木犀草苷
(luteolin)
Y=2.211×107X-6.451×1040.003 171~0.253 70.999 520.8069.34
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
Y=2.144×107X-2.101×1050.004 201~0.336 10.997 845.27150.90
异绿原酸B
(isochlorogenic acid B)
Y=7.997×106X-4.564×1030.000 208 4~0.016 670.996 854.46181.53
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
Y=1.024×107X-7.792×1040.004 634~0.370 70.999 090.39301.30
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
Y=9.253×106X-6.312×1040.004 928~0.394 20.999 2101.09336.98
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
Y=1.719×107X-6.937×1030.000 297 5~0.023 800.999 343.51145.03
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
Y=1.172×107X-2.476×1040.000 898 6~0.071 890.998 2104.89349.65
), ArticleFig(id=1240722377844249252, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 3, caption=

Content of 9 constituents in V. amygdalina

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(number)
含量
(content)/(mg · g-1
尿嘧啶
(uracil)
绿原酸
(chlorogenic acid)
木犀草苷
(luteolin)
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
异绿原酸B
(isochlorogenic acid B)
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
P10.009 8000.619 60.303 80.984 60.046 540.537 50.437 60.032 190.190 0
P20.042 841.1641.4882.5870.248 51.8361.2230.183 31.062
P30.017 890.746 21.0001.8450.077 210.789 10.695 80.135 20.361 9
P40.017 541.0310.882 51.8500.093 460.852 10.965 40.164 70.406 4
P50.008 3065.7123.4315.6520.438 711.5710.860.469 11.931
P60.084 300.646 10.535 51.5850.0482 50.620 60.969 00.181 20.320 1
P70.015 591.2781.5433.0920.167 62.4651.3850.213 70.708 4
P80.007 9961.6711.1142.2200.211 92.2330.965 40.122 50.839 1
P90.059 830.852 20.565 11.8130.165 61.0430.778 00.058 550.726 8
P100.013 133.8372.4134.9000.225 54.4224.7680.271 71.002
P110.013 453.9302.3554.5700.214 64.3214.6650.262 40.944 6
P120.013 504.0212.3664.5870.222 64.3824.7280.255 10.977 4
P130.042 042.2161.8253.7160.0432 91.8175.7970.316 40.245 6
P140.041 082.2331.8793.9330.0502 21.7595.9250.327 60.217 4
P150.044 342.2041.8383.8690.0476 01.7435.8370.284 60.325 8
P160.008 4611.3472.6327.4320.305 44.1002.9470.622 71.056
P170.008 4871.3552.6597.4170.300 64.1602.9920.633 21.046
P180.008 4121.3452.6757.4150.295 44.1552.9660.630 50.908 8
P190.007 3149.1383.6786.9980.122 65.21510.890.712 10.324 7
P200.007 6399.7633.7007.0940.122 55.33011.190.720 10.338 9
P210.007 4259.2703.7017.0900.115 15.29311.020.716 40.343 2
P220.007 5656.1144.0318.0290.190 85.77513.600.714 70.384 2
P230.007 3835.9933.9737.8810.182 75.68613.390.707 80.462 0
P240.007 6766.2294.0098.1460.188 35.79813.780.704 80.410 6
), ArticleFig(id=1240722377936523948, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表3, caption=

南非叶中9个成分的含量(n=2)

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(number)
含量
(content)/(mg · g-1
尿嘧啶
(uracil)
绿原酸
(chlorogenic acid)
木犀草苷
(luteolin)
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
异绿原酸B
(isochlorogenic acid B)
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
P10.009 8000.619 60.303 80.984 60.046 540.537 50.437 60.032 190.190 0
P20.042 841.1641.4882.5870.248 51.8361.2230.183 31.062
P30.017 890.746 21.0001.8450.077 210.789 10.695 80.135 20.361 9
P40.017 541.0310.882 51.8500.093 460.852 10.965 40.164 70.406 4
P50.008 3065.7123.4315.6520.438 711.5710.860.469 11.931
P60.084 300.646 10.535 51.5850.0482 50.620 60.969 00.181 20.320 1
P70.015 591.2781.5433.0920.167 62.4651.3850.213 70.708 4
P80.007 9961.6711.1142.2200.211 92.2330.965 40.122 50.839 1
P90.059 830.852 20.565 11.8130.165 61.0430.778 00.058 550.726 8
P100.013 133.8372.4134.9000.225 54.4224.7680.271 71.002
P110.013 453.9302.3554.5700.214 64.3214.6650.262 40.944 6
P120.013 504.0212.3664.5870.222 64.3824.7280.255 10.977 4
P130.042 042.2161.8253.7160.0432 91.8175.7970.316 40.245 6
P140.041 082.2331.8793.9330.0502 21.7595.9250.327 60.217 4
P150.044 342.2041.8383.8690.0476 01.7435.8370.284 60.325 8
P160.008 4611.3472.6327.4320.305 44.1002.9470.622 71.056
P170.008 4871.3552.6597.4170.300 64.1602.9920.633 21.046
P180.008 4121.3452.6757.4150.295 44.1552.9660.630 50.908 8
P190.007 3149.1383.6786.9980.122 65.21510.890.712 10.324 7
P200.007 6399.7633.7007.0940.122 55.33011.190.720 10.338 9
P210.007 4259.2703.7017.0900.115 15.29311.020.716 40.343 2
P220.007 5656.1144.0318.0290.190 85.77513.600.714 70.384 2
P230.007 3835.9933.9737.8810.182 75.68613.390.707 80.462 0
P240.007 6766.2294.0098.1460.188 35.79813.780.704 80.410 6
), ArticleFig(id=1240722378032992947, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 4, caption=

Correlation analysis of components

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
尿嘧啶
(uracil)
绿原酸
(chlorogenic acid)
木犀草苷
(luteolin)
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
异绿原酸B
(isochlorogenic acid B)
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
尿嘧啶
(uracil)
1-0.457*-0.583**-0.570**-0.436*-0.560**-0.398-0.506*-0.228
绿原酸
(chlorogenic acid)
10.836**0.671**0.0660.684**0.877**0.724**-0.103
木犀草苷
(luteolin)
10.944**0.3880.828**0.910**0.926**0.123
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
10.443*0.731**0.774**0.966**0.137
异绿原酸B
(isochlorogenic acid B)
10.688**0.1150.3060.923**
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
10.740**0.677**0.552**
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
10.799**-0.106
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
1-0.007
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
1
), ArticleFig(id=1240722378125267640, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表4, caption=

各成分相关性分析

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
尿嘧啶
(uracil)
绿原酸
(chlorogenic acid)
木犀草苷
(luteolin)
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
异绿原酸B
(isochlorogenic acid B)
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
尿嘧啶
(uracil)
1-0.457*-0.583**-0.570**-0.436*-0.560**-0.398-0.506*-0.228
绿原酸
(chlorogenic acid)
10.836**0.671**0.0660.684**0.877**0.724**-0.103
木犀草苷
(luteolin)
10.944**0.3880.828**0.910**0.926**0.123
木犀草素-7-O-β-D-葡萄糖醛酸苷
(luteolin-7-O-β-D-glucuronide)
10.443*0.731**0.774**0.966**0.137
异绿原酸B
(isochlorogenic acid B)
10.688**0.1150.3060.923**
3,5-O-二咖啡酰奎宁酸
(3,5-O-dicaffeoyl quinic acid)
10.740**0.677**0.552**
1,5-二咖啡酰奎宁酸
(1,5-dicaffeoyl quinic acid)
10.799**-0.106
芹菜素-7-O-β-D-吡喃葡萄糖苷
(apigenin-7-O-β-D-glucopyranoside)
1-0.007
4,5-O-二咖啡酰奎宁酸
(4,5-O-dicaffeoyl quinic acid)
1
), ArticleFig(id=1240722378251096764, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 5, caption=

The PCA of eigenvalues and contribution rates

, figureFileSmall=null, figureFileBig=null, tableContent=
主成分
(principal component)
特征值
(eigenvalue)
贡献率
(contribution rate)/%
累积贡献率
(cumulative contribution rate)/%
PC15.63462.60362.603
PC22.09823.31085.912
PC30.6146.82292.735
PC40.4635.15097.884
), ArticleFig(id=1240722378322399935, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表5, caption=

主成分分析特征值及贡献率

, figureFileSmall=null, figureFileBig=null, tableContent=
主成分
(principal component)
特征值
(eigenvalue)
贡献率
(contribution rate)/%
累积贡献率
(cumulative contribution rate)/%
PC15.63462.60362.603
PC22.09823.31085.912
PC30.6146.82292.735
PC40.4635.15097.884
), ArticleFig(id=1240722378406286019, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 6, caption=

The PCA of factor load matrix

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
主成分(principal component)
PC1(principal component 1)PC2(principal component 2)
尿嘧啶(uracil)-0.661-0.170
绿原酸(chlorogenic acid)0.817-0.386
木犀草苷(luteolin)0.981-0.141
木犀草素-7-O-β-D-葡萄糖醛酸苷(luteolin-7-O-β-D-glucuronide)0.931-0.075
异绿原酸B(isochlorogenic acid B)0.5110.845
3,5-O-二咖啡酰奎宁酸(3,5-O-dicaffeoyl quinic acid)0.8990.300
1,5-二咖啡酰奎宁酸(1,5-dicaffeoyl quinic acid)0.866-0.380
芹菜素-7-O-β---吡喃葡萄糖苷(apigenin-7-O-β-D-glucopyranoside)0.904-0.224
4,5-O-二咖啡酰奎宁酸(4,5-O-dicaffeoyl quinic acid)0.2560.946
), ArticleFig(id=1240722378490172104, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表6, caption=

主成分分析因子载荷矩阵

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
主成分(principal component)
PC1(principal component 1)PC2(principal component 2)
尿嘧啶(uracil)-0.661-0.170
绿原酸(chlorogenic acid)0.817-0.386
木犀草苷(luteolin)0.981-0.141
木犀草素-7-O-β-D-葡萄糖醛酸苷(luteolin-7-O-β-D-glucuronide)0.931-0.075
异绿原酸B(isochlorogenic acid B)0.5110.845
3,5-O-二咖啡酰奎宁酸(3,5-O-dicaffeoyl quinic acid)0.8990.300
1,5-二咖啡酰奎宁酸(1,5-dicaffeoyl quinic acid)0.866-0.380
芹菜素-7-O-β---吡喃葡萄糖苷(apigenin-7-O-β-D-glucopyranoside)0.904-0.224
4,5-O-二咖啡酰奎宁酸(4,5-O-dicaffeoyl quinic acid)0.2560.946
), ArticleFig(id=1240722378582446795, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=EN, label=Tab. 7, caption=

principal component values and comprehensive principal component values

, figureFileSmall=null, figureFileBig=null, tableContent=
编号(number) Z1 Z2 Z排名(ranking)
P1-3.26-0.62-2.1923
P2-1.841.51-0.8016
P3-2.65-0.37-1.7521
P4-2.53-0.24-1.6420
P53.613.633.111
P6-3.78-0.98-2.5924
P7-1.460.65-0.7714
P8-1.681.23-0.7715
P9-3.160.57-1.8422
P100.311.050.4411
P110.160.900.3113
P120.211.000.3612
P13-1.50-1.44-1.2719
P14-1.40-1.46-1.2217
P15-1.53-1.29-1.2618
P161.091.631.078
P171.121.581.079
P181.071.340.9810
P192.65-1.751.257
P202.80-1.811.335
P212.71-1.791.286
P223.04-1.151.643
P232.95-1.051.604
P243.07-1.141.662
), ArticleFig(id=1240722378666332879, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240709665894494809, language=CN, label=表7, caption=

主成分得分及综合主成分得分

, figureFileSmall=null, figureFileBig=null, tableContent=
编号(number) Z1 Z2 Z排名(ranking)
P1-3.26-0.62-2.1923
P2-1.841.51-0.8016
P3-2.65-0.37-1.7521
P4-2.53-0.24-1.6420
P53.613.633.111
P6-3.78-0.98-2.5924
P7-1.460.65-0.7714
P8-1.681.23-0.7715
P9-3.160.57-1.8422
P100.311.050.4411
P110.160.900.3113
P120.211.000.3612
P13-1.50-1.44-1.2719
P14-1.40-1.46-1.2217
P15-1.53-1.29-1.2618
P161.091.631.078
P171.121.581.079
P181.071.340.9810
P192.65-1.751.257
P202.80-1.811.335
P212.71-1.791.286
P223.04-1.151.643
P232.95-1.051.604
P243.07-1.141.662
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基于高效液相色谱指纹图谱结合化学模式识别及多成分测定的南非叶质量评价研究*
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杨婧 1 , 覃华亮 1, ** , 符传武 1, ** , 胡士华 2 , 丘琴 3 , 蓝榆清 1 , 覃冬杰 1 , 黄敏桃 4 , 钟文俊 1 , 徐嘉 1 , 覃子龙 1
药物分析杂志 | 质量分析 2025,45(3): 489-503
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药物分析杂志 | 质量分析 2025, 45(3): 489-503
基于高效液相色谱指纹图谱结合化学模式识别及多成分测定的南非叶质量评价研究*
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杨婧1 , 覃华亮1, ** , 符传武1, ** , 胡士华2, 丘琴3, 蓝榆清1, 覃冬杰1, 黄敏桃4, 钟文俊1, 徐嘉1, 覃子龙1
作者信息
  • 1.柳州市质量检验检测研究中心,柳州 545001
  • 2.柳州铁道职业技术学院,柳州 545001
  • 3.广西中医药大学,南宁 530001
  • 4.广西生态工程职业技术学院,柳州 545001
  • Tel:13557329896;E-mail:

通讯作者:

**覃华亮 Tel:18178842081;E-mail:;
符传武 Tel:18007726258;E-mail:
Quality evaluation research of Vernonia amygdalina Del. based on HPLC fingerprint combined with chemical pattern recognition and multi-component determination*
Jing YANG1 , Hua-liang QIN1, ** , Chuan-wu FU1, ** , Shi-hua HU2, Qin QIU3, Yu-qing LAN1, Dong-jie QIN1, Min-tao HUANG4, Wen-jun ZHONG1, Jia XU1, Zi-long QIN1
Affiliations
  • 1. Liuzhou Quality Inspection and Testing Research Center, Liuzhou 545001, China
  • 2. Liuzhou Railway Vocational Technical College, Liuzhou 545001, China
  • 3. Guangxi University of Chinese Medicine, Nanning 530001, China
  • 4. Guangxi Eco-engineering Vocational and Technical College, Liuzhou 545001, China
出版时间: 2025-03-31 doi: 10.16155/j.0254-1793.2024-0478
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目的:建立南非叶的多成分测定方法及高效液相色谱(HPLC)指纹图谱,并结合化学模式识别的方法,综合评价不同产地的南非叶质量,为今后制定南非叶质量控制标准及其他相关南非叶成分研究奠定基础。方法:使用HPLC法,采用Welch Ultimate® AQ-C18色谱柱(250 mm×4.6 mm,5 µm),以0.2%磷酸水溶液-乙腈为流动相进行梯度洗脱,检测波长258 nm,体积流量1 mL · min-1,柱温35 ℃。同时建立24批不同产地的南非叶指纹图谱及9个成分HPLC测定方法,并结合化学模式识别对各批不同产地的南非叶进行质量评价。结果:24批南非叶的指纹图谱的相似度均在0.9以上,建立的对照指纹图谱可稳定有效地对南非叶进行定性判别,指纹图谱中10个共有峰,指认出9个峰。24批南非叶中尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸的质量分数分别为0.007 314~0.084 30 mg · g-1、0.619 6~9.763 mg · g-1、0.303 8~4.031 mg · g-1、0.984 6~8.146 mg · g-1、0.043 29~0.438 7 mg · g-1、0.537 5~11.57 mg · g-1、0.437 6~13.78 mg · g-1、0.032 19~0.720 1 mg · g-1、0.190 0~1.931 mg · g-1。通过聚类分析,将24批南非叶分为2类,广东和海南产的南非叶为一类,广西产的南非叶独自为一类;主成分分析的分类结果与聚类分析一致,通过主成分得分进一步研究各产地南非叶质量差异,发现P5、P24、P22地区得分位居前三质量优于其他地区。通过正交偏最小二乘-判别分析,发现造成各地南非叶质量差异的影响能力由强到弱排序为1,5-二咖啡酰奎宁酸、绿原酸、木犀草素-7-O-β-D-葡萄糖醛酸苷、3,5-O-二咖啡酰奎宁酸。结论:南非叶指纹图谱及多成分测定方法稳定可靠,可弥补南非叶质量控制标准方面的空白,结合化学模式识别方法,指出了南非叶质量优质的产地及影响不同产地南非叶质量应着重控制的成分,为南非叶质量控制及评价提供了坚实的基础。

南非叶  /  指纹图谱  /  多成分  /  化学模式识别  /  质量评价

Objective: To establish a multi-component determination method and HPLC fingerprint for Vernonia amygdalina Del., combined with chemical pattern recognition to comprehensively evaluate the quality of Vernonia amygdalina Del. from different producing, laying a foundation for the future development of quality control standards for Vernonia amygdalina Del. and other related research on Vernonia amygdalina Del.’s components. Methods: The HPLC using Welch Ultimate® AQ-C18 column (250 mm×4.6 mm,5 µm) and 0.2% phosphoric acid aqueous solution - acetonitrile as moblie phase by gradient elution at the flow rate of 1 mL · min-1, wavelength was set at 258 nm and the column temperature was 35 ℃. Simultaneously establish 24 batches of fingerprint spectra of Vernonia amygdalina Del. from different origins and a multi-component HPLC determination method for 9 components of Vernonia amygdalina Del.. Combine chemical pattern recognition to evaluate the quality of 24 batches of Vernonia amygdalina Del. from different origins. Results: The similarities of the fingerprint spectra of 24 batches of Vernonia amygdalina Del. were above 0.9. The established control fingerprint spectra could stably and effectively distinguish Vernonia amygdalina Del. qualitatively. There were 10 common peaks in the fingerprint spectra, and 9 peaks were identified. The mass fractions of 24 batches of Vernonia amygdalina Del. components that were uracil, chlorogenic acid, luteolin, luteolin-7-O-β-D-glucuronide, isochlorogenic acid B, 3,5-O-dicaffeoyl quinic acid, 1,5-dicaffeoyl quinic acid, apigenin-7-O-β-D-glucopyranoside, 4,5-O-dicaffeoyl quinic acid were 0.007 314–0.084 30 mg · g-1,0.619 6–9.763 mg · g-1,0.303 8–4.031 mg · g-1,0.984 6–8.146 mg · g-1,0.043 29–0.438 7 mg · g-1,0.537 5–11.57 mg · g-1,0.437 6–13.78 mg · g-1,0.032 19–0.720 1 mg · g-1,0.190 0–1.931 mg · g-1. Through cluster analysis, 24 batches of Vernonia amygdalina Del. were divided into 2 categories. Vernonia amygdalina Del.from Guangdong and Hainan were classified as one category, while Vernonia amygdalina Del. from Guangxi were classified as one category alone. The classification results of principal component analysis were consistent with cluster analysis. Further research on the differences in leaf quality among different regions of Vernonia amygdalina Del. was conducted through principal component scores, and it was found that the P5, P24 and P22 regions ranked in the top three with better quality than other regions. Through orthogonal partial least squares discriminant analysis, it was found that the reasons for the differences in leaf quality among different regions of Vernonia amygdalina Del. were ranked in descending order: 1,5-dicaffeoyl quinic acid, chlorogenic acid, and luteolin-7-O-β-D-glucuronide, 3,5-O-dicaffeoyl quinic acid. Conclusion: The fingerprint and multi-component determination methods of Vernonia amygdalina Del. are stable and effective through methodological detection, which can fill the gap in quality control standards for Vernonia amygdalina Del.. Combined with chemical pattern recognition methods, the high-quality production areas of Vernonia amygdalina Del. and the components that should be emphasized in controlling the quality of Vernonia amygdalina Del. from different production areas are pointed out.A comprehensive evaluation of the quality of Vernonia amygdalina Del. has been conducted, providing a solid foundation for future quality control and evaluation of Vernonia amygdalina Del..

Vernonia amygdalina Del.  /  fingerprint  /  multiple component  /  chemical pattern recognition  /  quality
杨婧, 覃华亮, 符传武, 胡士华, 丘琴, 蓝榆清, 覃冬杰, 黄敏桃, 钟文俊, 徐嘉, 覃子龙. 基于高效液相色谱指纹图谱结合化学模式识别及多成分测定的南非叶质量评价研究*. 药物分析杂志, 2025 , 45 (3) : 489 -503 . DOI: 10.16155/j.0254-1793.2024-0478
Jing YANG, Hua-liang QIN, Chuan-wu FU, Shi-hua HU, Qin QIU, Yu-qing LAN, Dong-jie QIN, Min-tao HUANG, Wen-jun ZHONG, Jia XU, Zi-long QIN. Quality evaluation research of Vernonia amygdalina Del. based on HPLC fingerprint combined with chemical pattern recognition and multi-component determination*[J]. Chinese Journal of Pharmaceutical Analysis, 2025 , 45 (3) : 489 -503 . DOI: 10.16155/j.0254-1793.2024-0478
原产于热带地区的南非叶是菊科斑鸠菊属多年旱生型灌木、小乔木扁桃斑鸠菊(Vernonia amygdalina Del.)的叶子,又名将军叶、苦茶、桃叶斑鸠菊、苦胆叶、杏叶斑鸠菊叶、抗癌草等[1-2]。它原来分布于热带非洲西部的加纳、喀麦隆、尼日利亚至东部的坦桑尼亚和埃塞俄比亚等地[2]。现广泛分布于马来西亚、吉隆坡等东南亚国家,由这些国家引入我国,在我国南方地区大量种植,大多分布于广东、广西、海南等地[3]。王子恒等[3]曾以中医中药理论对南非叶进行性味归经中药药性探讨,认为南非叶苦,甘、寒,归脾、肝、胃、大肠经,有清热燥湿、泻火解毒、杀虫截疟的功效。
从南非叶分离出的化合物中研究较多的是甾体皂苷类化合物及倍半萜类化合物,其他主要活性物质还有黄酮、酚、单宁、香豆素、木脂素、生物碱等类成分。王菁[4]通过使用不同溶剂对南非叶95%乙醇提取物进行萃取,再运用不同色谱柱对萃取液进行分离提纯,分离鉴定出木犀草素-7-O-β-D-葡萄糖苷、咖啡酸、芹菜素-7-O-β-葡萄糖苷等黄酮类化合物和其他甾体皂苷类化合物。江燕[5]对南非叶的85%乙醇水和水提取部位进行了膜分离技术和柱色谱技术的分离提取,并通过多项波谱检测技术对分离得的化合物进行结构鉴定,分离鉴定出木犀草苷、尿嘧啶及甾醇类、倍半萜类、脂肪酸类等8个化合物,王永霞等[6]通过对南非叶的同属植物驱虫斑鸠菊的研究,成功分离提纯3,4-O-双咖啡酰奎宁酸等6个咖啡酰奎宁酸类化合物。同时袁珂等[7]也从南非叶的同属植物少花斑鸠菊中发现1,5-二咖啡酰奎宁酸、绿原酸等7个苯丙素类化合物。
南非叶的抗癌作用较强,尤其对鼻咽癌等的作用,因此也有抗癌草药的称呼。随着人们对南非叶的研究,近年来发现其还有新的药效用途。Djeujo等[8]通过对南非叶叶子与根浸出物的化合物进行研究,发现其中木犀草素具有较高的抗自由基活性。Gyebi等[9]采用基于结构的虚拟筛选方法,筛选了大量来自南非叶的化合物,发现南非叶中木犀草素和斑鸠菊内酯(vernolide)等6个化合物与冠状病毒的3-糜蛋白酶样蛋白酶活性位点的平衡构象表现出最高的结合趋势。这些化合物可以稳定柔性氧阴离子环的构象,从而干扰酶蛋白水解过程中四面体氧阴离子中间体的形成,以达到治疗的作用。
2020年版《中华人民共和国药典》提出中药药效的整体作用性,中药材及中药饮片通常以中医药理论为基础归纳总结药效药性,中药相关成分发挥作用的特点即为一个整体的作用,不能单一就某个成分进行检测作为衡量中药质量的标准,因而对中药材及中药饮片进行指纹图谱及多成分含量测定研究是非常重要的,通过指纹图谱及多成分测定多方面综合地评价中药材及饮片的质量,以提高对中药材及饮片有效性的检测标准。同时化学模式识别结合指纹图谱全面分析中药材质量在时下运用得也非常广泛。目前南非叶的质量控制方面研究还处于空白,本课题组致力于研究南非叶的特征指纹及多成分测定,为综合地、全面地、有效地评价南非叶的质量奠定基础。同时用南非叶指纹图谱及多成分指标结合化学模式识别的方法进行全面分析,对可能造成南非叶质量差异的成分或指纹图谱峰进行探究,为今后南非叶的质量标准的建立提供实验数据的支撑。
LC-20A高效液相色谱仪(岛津公司);XS205DU十万分之一电子分析天平(梅特勒-托利多公司);AE200S万分之一电子分析天平(梅特勒-托利多公司);SK8200HP超声波清洗器(上海科导超声仪器有限公司);GWB-2E超纯水仪(北京普析通用仪器有限责任公司)。
对照品尿嘧啶(批号100469-201302,含量99.6%)、绿原酸(批号110753-202119,含量96.3%)、木犀草苷(批号111720-200905,含量96.5%)、木犀草素-7-O-β-D-葡萄糖醛酸苷(批号111968-201602,含量97.7%)、3,5-O-二咖啡酰奎宁酸(批号111782-201706,含量97.3%)、4,5-O-二咖啡酰奎宁酸(批号111894-202205,含量94.9%)均购自中国食品药品检定研究院,异绿原酸B(批号2203553,含量97.5%)购自上海安谱实验科技股份有限公司,1,5-二咖啡酰奎宁酸(批号M06GB147633,含量99.2%)、芹菜素-7-O-β-D-吡喃葡萄糖苷(批号O20GB164741,含量99.8%)购自上海源叶生物科技有限公司。
色谱纯甲醇购自上海安谱实验科技股份有限公司,色谱纯乙腈购自恩科化学有限公司,水为自制超纯水,除此以外其他试剂均为分析纯。
南非叶植物分别收集于广西、广东、海南三省,共24批次,见表1。所有样品经柳州市质量检验检测研究中心药品科科长副主任药师覃华亮鉴定均为菊科斑鸠菊属植物扁桃斑鸠菊(Vernonia amygdalina Del.)的叶子。样品经过高速粉碎机打粉后过3号筛,在干燥条件下贮存备用。
分别精密称取尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸的对照品适量,再用70%甲醇水分别溶解稀释上述对照品,配成质量浓度分别为0.090 09、5.258、3.382、2.689、1.112、1.236、3.942、1.058和0.958 5 mg · mL-1的单一对照品储备液。再精密量取各单一对照品储备液适量,置于同一20 mL棕色量瓶中,用70%甲醇水稀释定容至刻度并摇匀,即得尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸质量浓度分别为0.004 504、0.657 2、0.253 7、0.336 1、0.016 67、0.370 7、0.394 2、0.023 80和0.071 89 mg · mL-1的混合对照品溶液。
取处理好的南非叶粉末约0.5 g,精密称定,置20 mL的棕色量瓶中,加入70%甲醇水15 mL,超声(频率53 kHz,功率100 W)60 min,放置至室温,用70%甲醇水稀释至刻度,摇匀,取适量上清液,用0.45 μm微孔滤膜过滤,取续滤液,即得。
采用月旭(Welch)Ultimate® AQ-C18(4.6 mm×250 mm,5 µm)色谱柱,以0.2%磷酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0~2 min,1% B;2~5 min,1%B→4%B;5~27 min,4%B→12% B;27~33 min,12%B→15%B;33~85 min,15%B→17%B;85~95 min,17%B→19%B),体积流量1 mL · min-1,进样量10 μL,柱温35 ℃,检测器为PDA,检测波长258 nm。
取编号为P10的南非叶样品的粉末约0.5 g,精密称定,按“2.2”项下方法制备供试品溶液,按“2.3”项下的色谱条件,连续进样6次,记录色谱图,以3号峰(木犀草苷)为参照峰计算各个共有峰的相对保留时间和相对峰面积,计算得出各峰的相对保留时间RSD值均<0.08%,各峰的相对峰面积RSD值均<2.1%,这表明了此方法的精确性较高。
取编号为P10的南非叶样品的粉末约0.5 g,精密称定,按“2.2”项下方法制备供试品溶液,按“2.3”项下的色谱条件,分别在制样后的0、2、4、6、8、12 h测定,记录色谱图,以3号峰(木犀草苷)为参照峰计算各个共有峰的相对保留时间和相对峰面积,计算得出各峰的相对保留时间RSD均<0.07%,各峰的相对峰面积RSD均<1.7%,这表明了此样品制样后在室温下放置12 h内稳定性良好。
取编号为P10的南非叶样品的粉末约0.5 g,共6份,精密称定,分别按“2.2”项下方法制备供试品溶液,按“2.3”项下的色谱条件进行测定,记录色谱图,以3号峰(木犀草苷)为参照峰计算各个共有峰的相对保留时间和相对峰面积,计算得出各峰的相对保留时间RSD均<0.14%,各峰的相对峰面积RSD值均<2.0%,这表明了此方法重复性良好。
使用采集来的24批次不同产地的南非叶粉末,分别按“2.2”项下方法制样,随后按“2.3”项下的色谱条件进样测定。记录色谱图,得到24批次南非叶样品的色谱图,将24批南非叶样品的色谱图导入由国家药典委员会发布的“中药色谱指纹图谱相似度评价系统(2012.130723版)”软件中,设置编号P10样品的色谱图为参考图谱,其色谱图在软件中为编号S10。设定以中位数法为对照图谱的生成方式,时间宽度为0.1 min,采用多点校正的方式对24批次色谱图中的各个峰进行Mark峰自动匹配如图1。选取峰形较好且峰面积较大的峰作为共有峰,共选取了10共有峰。从而确定南非叶指纹图谱共有峰,并生成南非叶对照指纹图谱如图2,这可为今后未知植物判定是否为南非叶提供对照指纹图谱。
在“2.5.1”项的操作基础上,对24批次南非叶样品的色谱峰与生成的对照指纹图谱峰的进行相似度计算,24批次样品色谱峰与对照指纹图谱之间的相似度依次为0.937、0.947、0.931、0.961、0.930、0.902、0.958、0.945、0.933、0.993、0.991、0.991、0.983、0.983、0.984、0.943、0.942、0.942、0.984、0.980、0.984、0.988、0.988和0.989。这24批次不同产地南非叶指纹图谱相似度都达到0.9以上,证明这几个产地的南非叶指纹图谱相似性很高。此指纹图谱可成为今后的南非叶质量标准中指纹图谱部分检验检测标准中的对照图谱,为今后南非叶植物的定性提供一定的支持。
通过查阅南非叶及其同属植物相关成分分离的文献,大致了解南非叶植物中可能含有成分。通过对照品的保留时间及光谱图与南非叶指纹图谱各峰的保留时间及光谱图进行对比确定,从而指认南非叶各峰。取“2.1”项下各对照品储备液适量,分别用70%甲醇水稀释20倍,制成混合对照溶液,再取“2.2”项下供试品溶液,按“2.3”项下色谱条件进样分析,得到色谱图,如图3。从而确定南非叶指纹图谱中的9个峰对应的成品分别是尿嘧啶(峰1)、绿原酸(峰2)、木犀草苷(峰3)、木犀草素-7-O-β-D-葡萄糖醛酸苷(峰4)、异绿原酸B(峰5)、3,5-O-二咖啡酰奎宁酸(峰6)、1,5-二咖啡酰奎宁酸(峰7)、芹菜素-7-O-β-D-吡喃葡萄糖苷(峰8)、4,5-O-二咖啡酰奎宁酸(峰10)。南非叶指纹图谱各峰对应的成分确认,有助于后续的多成分测定研究。
分别精密吸取“2.1”项下尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸9个对照品储备液1、2.5、1.5、2.5、0.3、6、2、0.45、1.5 mL置于同一20 mL棕色量瓶中,用70%甲醇水定容至刻度线,摇匀,得到混合对照溶液;再分别精密吸取混合对照溶液10、5、2.5、1.25和0.25 mL置于5个20 mL棕色量瓶中,加70%甲醇水稀释定容至刻度线,摇匀,即得到6个不同质量浓度的混合系列对照溶液。在“2.3”项的色谱条件下测定,以质量浓度(X)为横坐标,峰面积(Y)为纵坐标,并将2个变量进行线性回归。结果表明,9个成分在相应的范围内线性关系良好。分别以信噪比为3 ∶ 1和10 ∶ 1计算检出下限和定量下限,结果见表2
取编号为P10的南非叶样品的粉末,按“2.2”项下方法制备供试品溶液并按“2.3”项下色谱条件,连续进样6次,记录色谱图,计算尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸的峰面积RSD分别为2.0%、0.75%、0.17%、0.18%、0.65%、0.34%、0.41%、0.65%和0.82%。表明该方法有较好的精密度。
取编号为P10的南非叶样品的粉末,按“2.2”项下方法制备供试品溶液,分别于0、2、4、6、8、12 h按“2.3”项下色谱条件测定,记录色谱图,计算尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸的峰面积RSD分别为1.4%、1.7%、0.27%、0.12%、0.55%、0.22%、0.34%、1.0%和1.6%。表明供试品溶液的上述成分在12 h内具有较好的稳定性。
取编号为P10的南非叶样品的粉末,按“2.2”项下的方法制备南非叶供试品溶液6份,并在“2.3”项色谱条件下测定,记录色谱图及峰面积,计算尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸9个成分平均含量分别为0.013 13、3.837、2.413、4.900、0.225 5、4.422、4.768、0.271 7、1.002 mg · mL-1,RSD分别为1.0%、0.45%、0.65%、0.17%、1.4%、0.27%、0.26%、1.3%和0.57%。表明该方法的重复性良好。
取已知含量的南非叶样品粉末(样品编号为P10)9份,每份0.25 g,精密称取,平均分为3组,分别置于20 mL棕色量瓶中,加入“2.1”项下一定量的尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸单一对照品储备液,使得第1组样品中加入上诉对照品的量与已知含量样品中上述成分的量大约为0.5 ∶ 1的比例,第2组比例大约为1 ∶ 1,第3组比例大约为1.5 ∶ 1,制成低中高3个水平的加标样品。按“2.2”项下方法制备供试溶液,并在“2.3”项色谱条件下进样测定,记录各成分峰面积并结合各成分的加入量计算加样回收率,结果尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸回收率分别为92.6%、91.1%、91.5%、105.8%、93.5%、98.6%、91.1%、104.7%、97.0%,RSD分别为1.9%、0.75%、1.1%、1.1%、1.5%、2.0%、0.85%、0.85%、1.5%。南非叶中9个成分的回收率符合2020年版《中华人民共和国药典》对中药分析方法验证指导原则中回收率限度的要求,表明本方法回收率良好。
分别精密称取24批次不同产地的南非叶样品0.5 g,按“2.2”项下方法制备供试品溶液,再按照“2.3”项下色谱条件进样,记录峰面积,并按标准曲线法计算样品中各个成分的含量,同时用2020年版《中华人民共和国药典》四部通则0832水分测定法,取“2.2”项下不同产地的南非叶样品2.5 g,平铺于干燥至恒重的扁形称量瓶中,不盖瓶盖在105 ℃干燥5 h,将瓶盖盖好,移至干燥器中,放冷30 min,精密称定,再在上述温度干燥1 h,放冷,称量,至连续2次称量差异不超过5 mg为止,并计算24批次样品的水分,样品中各成分的含量以干燥品计,结果见表3
将24批次不同产地南非叶样品测得含量数据,导入IBM SPSS Statistics 23统计分析软件中,进行聚类分析,以平方欧式距离作为样品聚类距离对这24批次南非叶样品分类,绘出树状图,见图4。分析数据发现24批次不同产地的样品可以分为2大类,P1~P4、P6~P18这17个产地的南非叶样品可以分为第1类;P5、P19~P24这7个产地的南非叶样品可以分为第2类。通过产地分析,广西地区的南非叶整体质量较为相似,而广东和海南地区的南非叶整体质量较为相似。
表3的数据导入IBM SPSS Statistics 23统计分析软件中,进行皮尔森(Pearson)相关性分析,皮尔森相关性系数是反映2个随机变量线性相关程度的统计量[10-12],皮尔森相关性分析可以帮助研究人员确定变量之间的因果关系。尽管相关性不能证明因果关系,但它可以提供关于变量之间关系的线索。当皮尔森相关性系数为0时,2个变量之间没有线性关系;当皮尔森相关性系数为0~1时,2个变量之间为正的线性关系,即其中1个变量增长另一个变量也随之增长,反之,亦然;当皮尔森相关性系数为-1~0时,2个变量关系为负的线性关系,即其中1个变量增长另一个变量随之在降低,反之,亦然;当皮尔森相关性系数绝对值越大,其2个变量的线性相关程度就越强[11]。当相关系数>0.8即认为线性相关程度极高。将数据处理后,结果见表4。数据中标记“**”的数据,表示此数据经过皮尔森相关性显著检验,P<0.01,2个变量相关性极为显著。通过表4可知在南非叶9个成分中,绿原酸与木犀草苷、1,5-二咖啡酰奎宁酸呈极显著正相关;木犀草苷与木犀草素-7-O-β-D-葡萄糖醛酸苷、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸呈极显著正相关;木犀草素-7-O-β-D-葡萄糖醛酸苷与芹菜素-7-O-β-D-吡喃葡萄糖苷呈极显著正相关;异绿原酸B与4,5-O-二咖啡酰奎宁酸呈极显著正相关。其中木犀草素-7-O-β-D-葡萄糖醛酸苷与芹菜素-7-O-β-D-吡喃葡萄糖苷这2个成分的皮尔森相关性系数为0.966,是所有变量中相关系数最大的。通过查阅文献了解到,Shui等[13]曾通过Illumina测序技术等手段对南非叶进行基因分析研究,发现南非叶中芹菜素与木犀草素会在特定的时期进行优先积累但其作用未明,某些特定的基因参与表达了这2个成分的形成。由此推测南非叶中的各成分之间可能由于生长环境或生长时限的不同及基因表达的因素存在相互影响的情况,从而影响南非叶的质量。其次绿原酸与木犀草苷、1,5-二咖啡酰奎宁酸呈极显著相关可能与其南非叶的药理作用抗菌及抗病毒作用相关,有文献指出发现南非叶中木犀草素等10个化合物与3种相关冠状病毒株有很强的相互作用。
表3的不同产地南非叶的各成分数据导入软件IBM SPSS Statistics 23统计分析软件中,在建立分析模型之前,需要对主成分数据结构进行检验,这里分别用到了KMO检验及Bartlett’s检验。通过软件计算得出KMO度量为0.588,此值大于0.5认为南非叶样品每个数据满足主成分分析要求;同时Bartlett’s检验P<0.001即认为南非叶样品数据可以进行主成分提取[14-17]。通过数据结构检验,将数据进行主成分分析,得到各个主成分特征值及各贡献率值见表5。对特征值>1的主成分进行提取,同时提取的主成分应可以累计诠释60%~70%以上的数据,是比较多学者所认同的[14-17]。通过结果分析发现2个主成分PC1和PC2可以满足要求,同时从累计贡献率来看这2个主成分可以累计诠释85.912%的原始数据。故选用这2个主成分来评价南非叶整体的综合质量是非常稳妥的。
通过表6的因子载荷矩阵表可以观察出,主成分1主要诠释尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷这7个成分的数据会比较好,主成分2诠释异绿原酸B、4,5-O-二咖啡酰奎宁酸这2个成分的数据会比较好。通过以PC1和PC2建立的坐标系如图5。此图为以主成分PC1和PC2表达下的各产地南非叶的散点图,由这些散点将各产地分为2类,P1~P4、P6~P18这17个产地的南非叶样品可以分为第1类;P5、P19~P24这7个产地的南非叶样品可以分为第2类。此结果与聚类分析一致。
根据1933年霍特林提出的主成分分析模型及一系列推导[14-15],最终主成分与原始数据矩阵标准为原数据矩阵主成分分析后生成的相关矩阵R的第i个主成分特征值对应的单位特征向量,Xc为原始数据矩阵进行标准化得到标准化矩阵中第c个成分的数据,而wci为原始数据矩阵中化后的矩阵关系为Zi = aciXci = 1,2,…,P,其中i为提取的主成分中第i个主成分,c为原始数据的成分数,acic个成分第i个主成分特征值对应的特征向量即表6中的载荷值,λi为第i个主成分的特征值。而综合主成分得分公式为 ,其中μi为第i个主成分的贡献率[14],即本实验的主成分得分Z = 0.626 03×Z1+0.233 1×Z2,由此可以得出各个产地的南非叶主成分综合得分即表7。根据各个产地样品的主成分得分情况看,P5、P24、P22分别位居前三,质量相对其他地方的较优。结合产地分析,P5样品来自广东东莞森林公园南非叶种植基地,P24样品来自广东广州,P22样品来自广东茂名。由于广东地区较早引进南非叶进行大规模种植培养,并且形成了南非叶茶叶的产业链。广东地区的样品质量好于其他地区,可能与广东的地理位置、广东地区南非叶培育基地对南非叶的选种育种规范种植方式及广东地区气候日照等条件有关。由于此次采集广西地区的南非叶样品的种植规模较小,没有专人培育,多是家庭单颗植物种植,选种及种植方式可能有所欠缺,所以其主成分得分排名较低。近年来广西也在逐步大力引进中药材南非叶种植项目,龙合镇惠布村就是其中1个种植南非叶致富的案例。
将数据导入软件SIMCA 14.1中,并将24批样品中的9个成分尿嘧啶、绿原酸、木犀草苷、木犀草素-7-O-β-D-葡萄糖醛酸苷、异绿原酸B、3,5-O-二咖啡酰奎宁酸、1,5-二咖啡酰奎宁酸、芹菜素-7-O-β-D-吡喃葡萄糖苷、4,5-O-二咖啡酰奎宁酸含量分别赋值于D1~D9,做正交偏最小二乘-判别分析,得模型参数可知数据矩阵的解释率参数R2X = 0.967,模型分区参数R2Y = 1,模型预测参数Q2 = 0.573,均大于0.5,表明建立的数学模型稳定且预测能力较强[18]。通过软件做出变量重要性投影值(VIP)图,见图6,从中可以观察出造成不同产地南非叶质量差异的成分影响能力的排序。取VIP>1的成分观察,成分影响能力由强到弱的顺序为D7(1,5-二咖啡酰奎宁酸)、D2(绿原酸)、D4(木犀草素-7-O-β-D-葡萄糖醛酸苷)、D6(3,5-O-二咖啡酰奎宁酸)。在今后的南非叶质量控制中应该多注意对这些成分的控制。
南非叶作为南非国家的民间草药,常用来治疗发烧、肠胃等疾病,近代药理研究发现其对鼻咽癌、乳腺癌等癌症细胞有DNA抑制作用,同时具有抗虫、抗痢疾、降血糖、降血压等功效[2]。王菁[4]通过分离提取南非叶中的化合物并进行药理实验,发现芹菜素-7-O-β-葡萄糖苷对A549细胞(人肺腺癌细胞系)有很强的抑制作用,其半抑制浓度(IC50)为12.68 μmol · L-1。江燕[5]通过多项分离技术及波谱检测技术提取分离检测出8个化合物,其中尿嘧啶作为首次在扁桃斑鸠菊植物中发现的化合物,并且其是抗癌活性物5-氟尿嘧啶的前体物,在南非叶抗癌药理作用起到一定作用。刘忠祥[19]通过实验分离提取南非叶中豆甾醇型甾体皂苷有明显的黄嘌呤氧化酶抑制活性及NF-κB抑制活性,南非叶有潜在的抗痛风活性。近年来,也有学者在南非叶的化合物中筛选抗新冠肺炎的物质并还发现南非当地医生利用南非叶治疗疟疾。南非叶药理作用多样,但仍有进一步研究的空间,我国正在大力引进这些疗效好的外来药,以中医药理论解释其药效。
本实验组探究了南非叶的指纹图谱及多成分测定,通过对Symmetry C18、Ultimate AQ-C18等多个色谱柱的使用,回流提取、超声提取、冷浸提取、索氏提取等多种提取方式尝试,以水、乙酸乙酯、石油醚、正丁醇、无水乙醇、无水甲醇、30%甲醇、50%甲醇、70%甲醇等多种溶剂作为提取溶液及不同的流动相不同梯度的流动相比例为色谱条件的预实验对比探究,找出了南非叶指纹图谱制样及色谱条件的最优解,成功提取出了不同产地的南非叶10个共有峰,并且确定了9个峰的成分,为之后的南非叶质量控制奠定了坚实的基础。
通过对南非叶指纹图谱中的9个已知成分进行各项化学模式识别的分析,探究了这24批南非叶样品中9个化学成分之间的关系,化学成分与产地之间的关系。首先使用了聚类分析方法将24批次样品进行了分类,发现广东与海南地区的南非叶质量较为相近,广西的样品则独立为一类。再通过相关性分析及主成分分析,探究出各成分之间相关性关系和对24批南非叶样品进行主成分评分,其中较为有趣的是木犀草素-7-O-β-D-葡萄糖醛酸苷与芹菜素-7-O-β-D-吡喃葡萄糖苷这2个成分的皮尔森相关性系数最大且是正相关。同时有研究指出这2个成分会在南非叶特定时期积累,则本实验组探究的多成分测定方法会为今后的南非叶黄酮类物质合成机理的相关研究奠定基础。同时有研究指出南非叶中木犀草苷与冠状病毒的3-糜蛋白酶样蛋白酶活性位点的平衡构象表现出最高的结合趋势,以达到干扰冠状病毒蛋白酶的作用,以达到治疗新型冠状病毒感染的作用。从南非叶新的药效方面来讲,木犀草苷应该作为南非叶质量评价的重要指标。通过正交偏最小二乘-判别分析,观察到对这24批南非叶的质量差异的影响能力从强到弱排序为1,5-二咖啡酰奎宁酸、绿原酸、木犀草素-7-O-β-D-葡萄糖醛酸苷、3,5-O-二咖啡酰奎宁酸。因此若是从减少检验成本方面来说,可以考虑对这些成分进行重点的含量控制,制定南非叶质量控制标准。
  • *柳州市科技计划项目(2024SZ0505G001自)
  • 广西高校中青年教师科研基础能力提升项目(2023KY1257)
  • 国家自然科学基金资助项目(81460601)
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doi: 10.16155/j.0254-1793.2024-0478
  • 接收时间:2024-07-23
  • 首发时间:2026-03-17
  • 出版时间:2025-03-31
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  • 收稿日期:2024-07-23
基金
*柳州市科技计划项目(2024SZ0505G001自)
广西高校中青年教师科研基础能力提升项目(2023KY1257)
国家自然科学基金资助项目(81460601)
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    1.柳州市质量检验检测研究中心,柳州 545001
    2.柳州铁道职业技术学院,柳州 545001
    3.广西中医药大学,南宁 530001
    4.广西生态工程职业技术学院,柳州 545001

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**覃华亮 Tel:18178842081;E-mail:;
符传武 Tel:18007726258;E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

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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