Name: hplc coupled with chemical fingerprinting for multi-pattern recognition for identifying the authenticity of clematidis armandii caulis
Uploaded: 2022-11-11T14:40:00
Description: Watch this Scientific Journal Video about HPLC Coupled with Chemical Fingerprinting for Multi-Pattern Recognition for Identifying the Authenticity of Clematidis Armandii Caulis at JoVE.com

这项工作得到了四川省中医药管理局项目（编号：2020JC0088，编号：2021MS203）的支持。

1. 化学指纹检测方法
<ol><li>色谱条件<ol><li>制备乙腈（A）/水（B）流动相。在HPLC软件中设置梯度程序如下：0-20分钟，3%A-10%A;20-25分钟，10%A-13%A;25-65分钟，13%A-25%A;65-75分钟，25%A-40%A;75-76分钟，40%A-3%A;76-85分钟，3%A-3%A。</li>
		<li>将流动相的流速保持在1.0 mL/min。</li>
		<li>在保持在30°C的C18色谱柱（250 mm x 4.6 mm，5 μm）上进行色谱分离。</li>
		<li>将进样体积设置为 10 μL。</li>
		<li>检测波长为...

<ol>
	<li>Chen, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Chinese Medicine Specimen Picture Book. , 116 (1935).</li><li>Xiong, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignans+from+the+stems+of+Clematis+armandii+(&amp;#34;Chuan-Mu-Tong&amp;#34;)+and+their+anti-neuroinflammatory+activities.">Lignans from the stems of Clematis armandii (&amp;#34;Chuan-Mu-Tong&amp;#34;) and their anti-neuroinflammatory activities.</a> Journal of Ethnopharmacology. 153 (3), 737-743 (2014).</li><li>Pan, L. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=a+lignan+from+the+Chinese+medicinal+plant+Clematis+armandii,+inhibits+A431+cell+growth+via+blocking+p70S6/S6+kinase+pathway.">a lignan from the Chinese medicinal plant Clematis armandii, inhibits A431 cell growth via blocking p70S6/S6 kinase pathway.</a> Integrative Cancer Therapies. 16 (3), 351-359 (2017).</li><li>Chinese Pharmacopoeia Commission. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Pharmacopoeia of The People's Republic of China: (Edition 2020). , 38 (2020).</li><li>Wang, D. G., Guo, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Textual+research+on+the+materia+medica+and+authenticity+of+Clematidis+armandii.">Textual research on the materia medica and authenticity of Clematidis armandii.</a> Lishizhen Medicine and Materia Medicine Research. 18 (11), 2696 (2007).</li><li>Fang, Q. M., Zhang, M., Zhong, Y. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+natural+resources+and+exploitation+of+Caulis+Clematidis+Armandii+in+Sichuan.">The natural resources and exploitation of Caulis Clematidis Armandii in Sichuan.</a> West China Journal of Pharmaceutical Sciences. 20 (5), 404-406 (2005).</li><li>Bai, M. N., Dai, Y. X., Wang, Y., Ren, Y. Y., Tan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+on+the+identi&#64257;cation+of+Caulis+Clematidis+Armandii.">Study on the identi&#64257;cation of Caulis Clematidis Armandii.</a> Research and Practice on Chinese Medicines. 31 (3), 13-16 (2017).</li><li>Zhou, P. J., Li, X. F., Fu, D. H., Pu, X. Y., Zhu, G. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacognosy+identification+of+Ethnomedicine+Clematis+ranunculoides+Franchet.">Pharmacognosy identification of Ethnomedicine Clematis ranunculoides Franchet.</a> Journal of Guangzhou University of Traditional Chinese Medicine. 34 (3), 432-436 (2017).</li><li>Guo, J. L., Tang, Y., Pei, J., Wang, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+on+random+amplified+polymorphic+DNA+and+sequence+characterized+amplified+regions+of+Caulis+Clematidis+Armandii.">Study on random amplified polymorphic DNA and sequence characterized amplified regions of Caulis Clematidis Armandii.</a> Journal of Chengdu Medical College. 6 (4), 283-286 (2011).</li><li>Liu, M. Z., Li, M. N., Yao, H., Liu, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+identification+of+Clematidis+Armandii+Caulis+and+its+adulterants+and+closely+related+species+by+ITS2+sequence.">Molecular identification of Clematidis Armandii Caulis and its adulterants and closely related species by ITS2 sequence.</a> Global Traditional Chinese Medicine. 4 (6), 446-450 (2011).</li><li>Liu, J. J., Chen, X., Wei, Z. Q., Li, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+constituents+and+identification+of+the+caules+of+Clematis+armandii+Franch.and+Clematis+montana+Buch.-Ham.">Chemical constituents and identification of the caules of Clematis armandii Franch.and Clematis montana Buch.-Ham.</a> Natural Product Research and Development. 22 (6), 998-1000 (2010).</li><li>Yang, T. R., Xu, Z., Shi, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+four+compounds+from+Clematis+Montana.">Studies on four compounds from Clematis Montana.</a> Journal of Liaoning University of Traditional Chinese Medicine. 10 (4), 142-143 (2008).</li><li>Yan, L. H., Xu, L. Z., Zhou, Z. M., Yang, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+constituents+from+stems+of+Clematis+armandii(I).">Chemical constituents from stems of Clematis armandii(I).</a> Chinese Traditional and Herbal Drugs. 38 (3), 340-342 (2007).</li><li>Li, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Traditional+uses,+phytochemistry,+pharmacology,+and+toxicology+of+Akebiae+Caulis+and+its+synonyms:+A+review.">Traditional uses, phytochemistry, pharmacology, and toxicology of Akebiae Caulis and its synonyms: A review.</a> Journal of Ethnopharmacology. 277, 114245 (2021).</li><li>Ye, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diuretic+effect+and+material+basis+of+Clematidis+Armandii+Caulis+in+rats.">Diuretic effect and material basis of Clematidis Armandii Caulis in rats.</a> China Journal of Chinese Materia Medica. 44 (9), 1889-1894 (2019).</li><li>Kuang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Consistency+study+of+Caulis+Clematidis+Armandii+and+its+traditionally+used+products+and+counterfeit+species+based+on+efficacy+and+pharmacognosy.">Consistency study of Caulis Clematidis Armandii and its traditionally used products and counterfeit species based on efficacy and pharmacognosy.</a> Pharmacology and Clinics of Chinese Materia Medica. 36 (3), 115-121 (2020).</li><li>Li, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quality+assessment+of+Asarum+heterotropoides+var.+mandshuricum+based+on+HPLC+multi-component+determination+and+multiple+pattern+recognition+method+of+fingerprint.">Quality assessment of Asarum heterotropoides var. mandshuricum based on HPLC multi-component determination and multiple pattern recognition method of fingerprint.</a> Chinese Traditional and Herbal Drugs. 53 (1), 238-243 (2022).</li><li>Li, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+multi-components+of+Paeoniae+Alba+Radix+based+on+fingerprints+and+chemical+pattern+recognition.">Determination of multi-components of Paeoniae Alba Radix based on fingerprints and chemical pattern recognition.</a> Chinese Traditional and Herbal Drugs. 53 (1), 231-237 (2022).</li><li>Li, H. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+investigation+for+raw+and+processed+products+of+Euodiae+Fructus+based+on+high-performance+liquid+chromatography+fingerprints+and+chemical+pattern+recognition.">Comparative investigation for raw and processed products of Euodiae Fructus based on high-performance liquid chromatography fingerprints and chemical pattern recognition.</a> Chemistry &amp; Biodiversity. 18 (8), 2100281 (2021).</li><li>Dong, L. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+on+suitable+distribution+areas+of+Kawa+Kimichi+produced+in+Sichuan+province-based+on+remote+sensing+and+GIS-A+case+study+of+Clematis+Armandii.">Study on suitable distribution areas of Kawa Kimichi produced in Sichuan province-based on remote sensing and GIS-A case study of Clematis Armandii.</a> Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology. 17 (11), 2398-2404 (2015).</li><li>Tang, S. W., Pei, J., Li, F. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphological+and+histological+identification+of+Chuanmutong.">Morphological and histological identification of Chuanmutong.</a> West China Journal of Pharmaceutical Sciences. 15 (1), 19-21 (2000).</li><li>Li, B., Chen, X., Fang, Q. M., Zhang, B. J., Wei, Z. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quality+Research+of+Chuanmutong.">Quality Research of Chuanmutong.</a> Journal of Sichuan of Traditional Chinese Medicine. 27 (6), 58-60 (2009).</li><li>Wu, W. J., Wan, M. M., Cao, Y. H., Tan, R., Song, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Research+on+the+Quality+Standard+of+Chuanmutong.">Research on the Quality Standard of Chuanmutong.</a> Chinese Archives of Traditional Chinese Medicine. 33 (2), 313-315 (2015).</li><li>Wei, Z. Q., Chen, X., Li, B., Dong, X. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+stigmasterol+in+Clematis+armandii+Franch+and+Clematis+montana+Buch-Ham.+by+HPLC.">Determination of stigmasterol in Clematis armandii Franch and Clematis montana Buch-Ham. by HPLC.</a> Lishizhen Medicine and Materia Medica Research. 20 (3), 574-575 (2009).</li><li>Qing, L. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+oleanolic+acid+in+Caulis+clematidis+armandii+by+HPLC.">Determination of oleanolic acid in Caulis clematidis armandii by HPLC.</a> West China Journal of Pharmaceutical Sciences. 21 (3), 273-274 (2006).</li><li>Yang, S. D., Wang, R., Fu, D. Y., Guo, J. J., Tan, W. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+on+oleanolic+acid+in+caulis+clematidis+armandii+by+microwave+assisted+extraction-HPLC/MS.">Determination on oleanolic acid in caulis clematidis armandii by microwave assisted extraction-HPLC/MS.</a> Anhui Academy of Agricultural Sciences. 38 (13), 6929-6931 (2010).</li></ol>

用于研究的样本采集是构建多模式识别识别中药材真伪的第一步。通过市场调研，我们发现四川雅安、凉山、乐山是川木通野生资源的主要产区。同一属的相关品种也具有相同的地理分布6，20;CC、DC、DE、XS和SMT经常被误用为创木通16，21;因此，本研究在上述产地采集了10批次正宗川木桐和5批次混合样品，并确?...

川木通，铁线莲的干花。或铁线莲蒙大拿州Buch.-Ham.，是诊所1，2，3常用的传统中药。它用于治疗泌尿问题，水肿，舌头和口腔溃疡，乳汁分泌减少，关节僵硬和湿热引起的肌肉疼痛4。川木通一直从野生品种获得，主要分布在西南地区，尤其是四川，那里的品质最好5，6。由于正宗品种具有相似的特征，因此很难区分正宗品种及其密切相关的掺杂物7，8，9，10。2020年版《中国药典》中川木通的质量标准仅规定了性质、显微鉴别、薄层鉴定等，没有进行含量测定，不能有效鉴别掺杂物，因此存在潜在风险。此外，很少有报告比较和鉴定川木通和相关植物。因此，一种确保川木通真伪的质量控制方法值得进一步研究。
川木通的化学成分主要由桫椤类五环三萜类化合物及其糖苷、黄酮类化合物和有机酸11，12，13，14组成。其中齐墩果酸、β-谷甾醇、豆甾醇、麦角甾醇具有不同强度的利尿作用，可能是促进利尿、缓解纹尿的潜在药效学物质15，16。化学指纹图谱是通过高效液相色谱（HPLC）、气相色谱（GC）等方法分离和检测样品中所含的许多化学成分而获得的。采用适当的统计分析方法分析川木通的特性，可以确定中药的整体质量控制和科学鉴定17、18、19。
本研究共收集了10批次川木桐正宗品种和5批次掺杂物。采用HPLC指纹图谱法结合多模式识别技术对它们的质量进行比较和分析，包括聚类分析（CA）、主成分分析（PCA）、正交偏最小二乘判别分析（OPLS-CA）和药效学成分含量测定。该方案建立了高特异性正宗品种鉴定方法，是科学鉴定中药材正宗品种和掺杂物的新策略。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Acetic acid</td>
			<td>Zhiyuan Chemical Reagent Co., Ltd., Tianjin, China</td>
			<td>2017381038</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetonitrile</td>
			<td>Sigma-Aldrich&#160; Trading Co., Ltd., Shanghai, China</td>
			<td>WXBD5243V</td>
			<td></td>
		</tr>
		<tr>
			<td>&#946;-Sitosterol</td>
			<td>Meisai Biological Technology Co., Ltd., Chongqing, China</td>
			<td>20210201</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;C18 column</td>
			<td>Yuexu Material Technology Co., Ltd., Shanghai, China</td>
			<td>Welch Ultimate LP</td>
			<td></td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Guoqiang Chinese Herbal Pieces Co., Ltd., Sichuan, China&#160;</td>
			<td>19020103</td>
			<td>CMT-1</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Hongya Wawushan Pharmaceutical Co., Ltd., Sichuan, China&#160;</td>
			<td>200701</td>
			<td>CMT-2</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Hongpu Pharmaceutical Co., Ltd., Sichuan, China&#160;</td>
			<td>200701</td>
			<td>CMT-3</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Hongpu Pharmaceutical Co., Ltd., Sichuan, China&#160;</td>
			<td>200901</td>
			<td>CMT-4</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Xinrentai Pharmaceutical Co., Ltd., Sichuan, China&#160;</td>
			<td>210701</td>
			<td>CMT-5</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Haobo Pharmaceutical Co., Ltd., Sichuan, China&#160;</td>
			<td>210401</td>
			<td>CMT-6</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Xinrentai Pharmaceutical Co., Ltd., Sichuan, China&#160;</td>
			<td>200901</td>
			<td>CMT-7</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Wusheng Pharmaceutical Group Co., Ltd., Sichuan, China&#160;</td>
			<td>201201</td>
			<td>CMT-8</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Limin Chinese Herbal Pieces Co., Ltd., Sichuan, China&#160;</td>
			<td>201001</td>
			<td>CMT-9</td>
		</tr>
		<tr>
			<td>Chuanmutong</td>
			<td>Yuhetang Pharmaceutical Co., Ltd., Sichuan, China</td>
			<td>210501</td>
			<td>CMT-10</td>
		</tr>
		<tr>
			<td>Clematis argentilucida (Levl. et Vant.) W. T. Wang</td>
			<td>Madzi Bridge, Sanlang Township, Tianquan County, Sichuan, China&#160;</td>
			<td>-</td>
			<td>CC</td>
		</tr>
		<tr>
			<td>Clematis apiifolia var. obtusidentata Rehd. et Wils.</td>
			<td>Heilin Village, Qiliping Township, Hongya County, Sichuan, China&#160;</td>
			<td>-</td>
			<td>DC</td>
		</tr>
		<tr>
			<td>Clematis peterae Hand.-Mazz.</td>
			<td>Huangmu Village, Huangmu Township, Hanyuan County, Sichuan, China&#160;</td>
			<td>-</td>
			<td>DE</td>
		</tr>
		<tr>
			<td>Clematis gouriana Roxb. Var. finetii Rehd. et Wils</td>
			<td>Mixedang Mountain, Huangwan Township, Emei County, Sichuan, China&#160;</td>
			<td>-</td>
			<td>XS</td>
		</tr>
		<tr>
			<td>Clematis finetiana Levl. et Vaniot.</td>
			<td>Wannian Village, Huangwan Township, Emei County, Sichuan, China&#160;</td>
			<td>-</td>
			<td>SMT</td>
		</tr>
		<tr>
			<td>Electronic balance</td>
			<td>Haozhuang Hengping Scientific Instrument Co., Ltd., Shanghai,&#160; China&#160;</td>
			<td>FA1204</td>
			<td></td>
		</tr>
		<tr>
			<td>Ergosterol</td>
			<td>Meisai Biological Technology Co., Ltd, Chongqing, China</td>
			<td>20210201</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Kelon Chemical Co., Ltd., Chengdu, China</td>
			<td>2021112602</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethyl acetate</td>
			<td>Zhiyuan Chemical Reagent Co., Ltd., Tianjin, China</td>
			<td>2017042043</td>
			<td></td>
		</tr>
		<tr>
			<td>Formic acid</td>
			<td>Kelon Chemical Co., Ltd., Chengdu, China</td>
			<td>2016062901</td>
			<td></td>
		</tr>
		<tr>
			<td>High performance liquid chromatography</td>
			<td>Agilent, USA.</td>
			<td>1260</td>
			<td></td>
		</tr>
		<tr>
			<td>IBM SPSS Statistics version 26.0</td>
			<td>International Business Machines Corporation, USA</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Sigma-Aldrich&#160; Trading Co., Ltd., Shanghai, China</td>
			<td>WXBD6409V</td>
			<td></td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Kelon Chemical Co., Ltd., Chengdu, China</td>
			<td>202010302</td>
			<td></td>
		</tr>
		<tr>
			<td>n-butyl alcohol</td>
			<td>Zhiyuan Chemical Reagent Co., Ltd., Tianjin, China</td>
			<td>2020071047</td>
			<td></td>
		</tr>
		<tr>
			<td>Petroleum ether</td>
			<td>Zhiyuan Chemical Reagent Co., Ltd., Tianjin, China</td>
			<td>2020090125</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphoric acid</td>
			<td>Comeo Chemical Reagent Co., Ltd., Tianjin, China</td>
			<td>20200110</td>
			<td></td>
		</tr>
		<tr>
			<td>SESCF-TCM version 2012</td>
			<td>National Pharmacopoeia Commission, China</td>
			<td>-</td>
			<td>http://114.247.108.158:8888/login</td>
		</tr>
		<tr>
			<td>Stigmasterol</td>
			<td>Meisai Biological Technology Co., Ltd., Chongqing, China</td>
			<td>20210201</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloromethane</td>
			<td>Sinopharm Group Chemical Reagent Co., Ltd., Shanghai, China</td>
			<td>20200214</td>
			<td></td>
		</tr>
		<tr>
			<td>Umetrics SIMCA version 14.1.0.2047</td>
			<td>Umetrics, Sweden</td>
			<td>-</td>
			<td>https://www.sartorius.com/en/products/process-analytical-technology/data-analytics-software/mvda-software/simca/simca-free-trial-download</td>
		</tr>
		<tr>
			<td>Ultrapure water machine</td>
			<td>Youpu Ultrapure Technology Co., Ltd., Sichuan, China</td>
			<td>UPH-II-10T</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic cleaner</td>
			<td>Kunshan Hechuang Ultrasound Instrument Co., Ltd., Jiangsu, China</td>
			<td>KH3200E</td>
			<td></td>
		</tr>
	</tbody>
</table>

hplc coupled with chemical fingerprinting for multi-pattern recognition for identifying the authenticity of clematidis armandii caulis

以 铁线莲 为例，构建了中药材及其相关掺杂物的鉴别方法。采用高效液相色谱（HPLC）指纹图谱结合化学计量学方法，对10批次真品川木通品种和5批次相关掺杂物进行分析比较，包括聚类分析（CA）、主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）。此外，测定了β-谷甾醇的含量。建立川木通对照化学指纹图谱，鉴定出12个共同峰。10批正品川木通品种指纹与对照指纹的相似度为0.910-0.989，而5批次掺杂物的相似度仅为0.133-0.720。基于色谱图中的常见峰，将15批次样品通过PCA分为3个含量等级，并按CA归集为4类，明确区分正品川木通和川木通掺杂物。此外，通过OPLS-DA发现了7种能够有效识别川木桐正宗和川木掺杂物的差异成分。10批次纯川木通品种的β-谷甾醇含量为97.53-161.56 μg/g，而5批次掺杂物的β-谷甾醇含量差异较大，其中 铁线莲 的β-谷甾醇含量为马兹。和 铁线莲 gouriana Roxb。Var. finetii Rehd.等威尔斯。明显低于川木通正宗品种。本研究建立的HPLC指标组分含量和化学指纹多模式识别方法为有效鉴定正宗中药材及相关掺杂物提供了新的策略。

川木通色谱指纹图谱及相似度分析（SA） 精密度、重复性和稳定性相对保留时间的RSD值分别低于0.46%、1.65%和0.53%;相对峰面积的RSD值分别低于4.23%、3.56%和3.96%。 如图1A，B所示，10份真品川木通样品的HPLC指纹图中有12个不同的共同峰（从峰1到峰12）。由于10号峰面积比较大，分辨率较好;由于它是每个样品中存在的组分，因此将其用作参考峰来研究指?...

Watch this Scientific Journal Video about HPLC Coupled with Chemical Fingerprinting for Multi-Pattern Recognition for Identifying the Authenticity of Clematidis Armandii Caulis at JoVE.com

HPLC Coupled with Chemical Fingerprinting for Multi-Pattern Recognition for Identifying the Authenticity of Clematidis Armandii Caulis

在这里，我们提出了一种建立高效液相色谱（HPLC）的协议，结合化学指纹多模式识别，为有效鉴定铁 线莲 及其掺杂物的真正品种提供了一种新策略。

高效液相色谱结合化学指纹图谱进行多模式识别，鉴定金线莲真伪

A method for identifying Chinese medicinal materials and their related adulterants was constructed by taking Clematidis Armandii Caulis (Chuanmutong, a ...

该协议可以为有效鉴定铁线莲及其掺杂物的真伪材料提供新的策略。该方法可用于药材质量研究，为研究其他中药材质量标准提供参考。要开始样品制备，请将原材料通过具有适当内径的尼龙网，将其研磨至均匀的粒径。将准确称取2克磨碎的原料转移到带塞的50毫升锥形烧瓶中，然后向烧瓶中加入50毫升甲醇。将塞子放在烧瓶上，以600瓦和40千赫兹的超声波加热30分钟。在孵育结束时，通过滤纸过滤药提取物溶液。过滤后，将4毫升含有药用提取物的甲醇溶液转移到10毫升容量瓶中。然后加入六毫升水并混合溶液。让溶液沉淀 10 分钟。用于样品的HPLC分析。首先制备流动相，并按照手稿中所述设置HPLC梯度程序。通过0.45微米微孔滤膜过滤样品并进行HPLC分析。对于每个样品，每天执行六次分析。为了评估样品溶液的稳定性，请在制备后分析在室温下储存0、2、4、6、8、12和24小时的相同样品溶液。对同一样品进行六次重复，并在HPLC中检测其指纹。将相关数据导入名为《中药色谱指纹相似度评价系统》或SESCF TCM 2012版的软件。将10批次正宗川木通样品和5批次掺杂物的保留时间和峰面积导入SESCF中药进行分析。在主菜单中，点击设置参考光谱，进入参数设置窗口，将川木桐正宗物种的参考色谱指纹图谱设置为参考。接下来，将控制频谱生成方法设置为时间窗口宽度为 0.5 的中位数方法。然后，单击主菜单中的多点校准，选择峰，然后选择峰匹配选项作为标记峰。最后，点击计算相似度，基于川木通参比色谱图谱对样品进行相似性分析。进行系统聚类分析，打开相关统计分析软件，点击文件，将10批次真实样品和5批次掺杂物数据导入软件。在菜单中的分类选项卡下，单击分析和系统聚类。选择10批次真实样品和5批次掺杂物的共同峰面积作为变量。将聚类数设置为 4。接下来，单击“方法”以选择“聚类方法”作为组间连接。选择测量间隔作为 Pearson 相关性，然后使用确定选项卡绘制聚类分析地图。对于主成分分析或 PCA，请打开数据分析软件。单击菜单上的文件，然后创建一个新的常规项目。从HPLC系统导入Excel或类似电子表格中12个常见峰的峰面积。打 结束通话 按钮完成数据导入。要使用 PCA 设置模型类型，请单击“新建”创建新模型，然后使用自动调整和添加选项卡拟合数据。转到分数以获取 PCA 分数图。使用正交偏最小二乘判别分析或 OPLSDA。生成 PCA 分数图，如前所述。使用 OPLSDA 设置模型类型，方法是单击“新建”和“新建”作为模型一 在进行缩放之前，为所有标准杆选择标准杆，然后点击分数自动拟合以生成 OPLSDA 分数图。要估计变量重要性和预测，请转到数据分析软件菜单并转到分析，然后将每个突变下的数字设置为 200。从OPLSDA分数中获取R平方和Q平方值，然后单击VIP和VIP预测以获取VIP地图。10份真品川木通样品的HPLC指纹图谱显示12个不同的共同峰，其中峰相对较大且分离度较好。使用峰数10作为参考峰来估计其余峰的相对保留时间。真实样本的相似度非常接近1，这意味着高度相似，一致性良好。但5批掺杂物与对照川木通的相似度较低。观察到五种掺杂物的HPLC指纹图谱与真实样品不同，主要在保留时间为28至55分钟的区域。除掺杂物CC外，PCA与掺杂物品种之间有明显的相似性，与掺杂物的差异性也相似，分类距离为20时，聚类分析也得到相似结果。OPLSDA生成的评分矩阵表明，真实样品的采样点与掺杂物之间没有交集。同样，掺假CC比其他掺杂物更像正宗品种。根据VIP图，数值大于1的峰被确定为导致真实样品和掺杂物差异的主要标记成分。在川木桐中发现的活性成分β-谷甾醇的含量在所有5批掺杂物中均得到检测，但含量差异较大。该协议建立了多种分析方法，从多个角度反映有关药材的所有信息，并全方位估计川木通药材的真实性。

Research

JoVE Journal

Chemistry

HPLC Coupled with Chemical Fingerprinting for Multi-Pattern Recognition for Identifying the Authenticity of Clematidis Armandii Caulis

Preparation of Highly Porous Coordination Polymer Coatings on Macroporous Polymer Monoliths for Enhanced Enrichment of Phosphopeptides

对磷酸增强富集制备高度多孔配位聚合物涂层的孔聚合物石柱的

We describe a protocol for the preparation of hybrid materials based on highly porous coordination polymer coatings on the internal surface of macroporous ...

科研

化学

Facile Synthesis of Worm-like Micelles by Visible Light Mediated Dispersion Polymerization Using Photoredox Catalyst

使用Photoredox催化剂蠕虫状胶束的简便合成可见光介导的分散聚合

Presented herein is a protocol for the facile synthesis of worm-like micelles by visible light mediated dispersion polymerization. This approach begins ...

Determining the Chemical Composition of Corrosion Inhibitor/Metal Interfaces with XPS: Minimizing Post Immersion Oxidation

确定缓蚀剂/金属界面的化学成分与XPS：最小化后浸入氧化

An approach for acquiring more reliable X-ray photoelectron spectroscopy data from corrosion inhibitor/metal interfaces is described. More specifically, ...

A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography (HPLC)

A Straightforward Method for Glucosinolate Extraction and Analysis with High-pressure Liquid Chromatography HPLC

简捷方法对硫甙的提取与分析用高压液相色谱法（HPLC）

Glucosinolates are a well-studied and highly diverse class of natural plant compounds. They play important roles in plant resistance, rapeseed oil ...

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

Chemical Precipitation Method for the Synthesis of Nb2O5 Modified Bulk Nickel Catalysts with High Specific Surface Area

化学沉淀法合成铌2O5改性高比表面积镍催化剂

We demonstrate a method for the synthesis of NixNb1-xO catalysts with sponge-like and fold-like nanostructures. By varying the Nb:Ni ratio, a series of ...

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery

用于靶向质粒DNA输送的美聚氨酸功能化生物相容性块共聚物

Reversible addition-fragmentation chain transfer (RAFT) polymerization integrates the advantages of radical polymerization and living polymerization. This ...

Extraction of Lignin with High &#946;-O-4 Content by Mild Ethanol Extraction and Its Effect on the Depolymerization Yield

轻度乙醇萃取高β-o-4 的木质素及其对解聚产率的影响

Lignin valorization strategies are a key factor for achieving more economically competitive biorefineries based on lignocellulosic biomass. Most of the ...

A Two-Step Pyrolysis-Gas Chromatography Method with Mass Spectrometric Detection for Identification of Tattoo Ink Ingredients and Counterfeit Products

一种用于检测纹身油墨成分和假冒产品的两步热解-气相色谱法

Tattoo inks are complex mixtures of ingredients. Each of them possesses different chemical properties which have to be addressed upon chemical analysis. ...

Tuning the Acidity of Pt/ CNTs Catalysts for Hydrodeoxygenation of Diphenyl Ether

调整二苯醚氢氧化的Pt/CNT催化剂的酸度

We herein present a method for the synthesis of HNbWO6, HNbMoO6, HTaWO6 solid acid nanosheet modified Pt/CNTs. By varying the weight of various solid acid ...

Integrated Cell Manipulation Platform Coupled with the Single-probe for Mass Spectrometry Analysis of Drugs and Metabolites in Single Suspension Cells

集成细胞操作平台与单悬架细胞中药物和代谢物质谱分析的单探针相结合

Single cell mass spectrometry (SCMS) enables sensitive detection and accurate analysis of broad ranges of cellular species on the individual-cell level. ...