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Article(id=1200147840472285993, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1200147837586604797, articleNumber=1001-2494(2024)10-0929-09, orderNo=null, doi=10.11669/cpj.2024.10.010, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1699545600000, receivedDateStr=2023-11-10, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1764067142836, onlineDateStr=2025-11-25, pubDate=1716307200000, pubDateStr=2024-05-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1764067142836, onlineIssueDateStr=2025-11-25, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1764067142836, creator=13701087609, updateTime=1764067142836, updator=13701087609, issue=Issue{id=1200147837586604797, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='10', pageStart='857', pageEnd='950', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1764067142149, creator=13701087609, updateTime=1764067345188, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1200148689244225889, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1200147837586604797, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1200148689244225890, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1200147837586604797, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=929, endPage=937, ext={EN=ArticleExt(id=1200147840778470204, articleId=1200147840472285993, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Study on Quality Evaluation of Arnebiae Radix by HPLC Fingerprint and Components Contents Determination, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To establish the fingerprints of 50 batches of Arnebiae Radix and the methods for determination of the contents of six components. METHODS A high performance liquid chromatographic (HPLC) method was used for the determination of 50 batches of Arnebiae Radix on an Agilent 5 TC-C18(2) column, with acetonitrile-0.05% formic acid aqueous solution as the mobile phase, gradient elution at a flow rate of 1 mL·min-1, detection wavelength of 275 nm, and column temperature of 30 ℃. The contents of six naphthoquinone components, i.e., alkannin, acetylshikonin, deoxyshikonin, β-acetoxyisovalerylshikonin, isobutyryl alkannin, and β,β'-dimethylacrylalkannin were determined. The fingerprints of the herbs were established by HPLC fingerprinting combined with the “Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012A)” to evaluate the similarity and confirm the common peaks. RESULTS The methodological validation results showed that the peak areas of the six components had good linear relationships in the corresponding concentration ranges, and the contents of the six components ranged from 0.006%-0.237%, 0.009%-1.759%, 0.001%-0.129%, 0.002%-0.625%, 0.011%-0.609%, and 0.016%-1.712%, respectively. The HPLC fingerprints of Arnebiae Radix were established, and a total of nine common peaks were calibrated. The similarity fitting was performed by using the TCM fingerprinting software, and it was found that the similarity of the 50 batches of Arnebiae Radix varied greatly. CONCLUSION The HPLC fingerprint of Arnebiae Radix established in this study and the method of simultaneous determination of the contents of six components are simple, stable, accurate, reliable, and reproducible, which can provide a basis for the quality study of Arnebiae Radix.

, correspAuthors=Jian ZHENG, 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=Rui HUANG, Shengyun DAI, Dongxue WU, Jie LIU, Shuangcheng MA, Jian ZHENG), CN=ArticleExt(id=1200147842150007682, articleId=1200147840472285993, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=基于HPLC指纹图谱及多指标成分含量测定的紫草质量评价, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=目的 采用高效液相色谱(HPLC)法建立50批紫草药材指纹图谱及6个成分含量测定方法。方法 采用HPLC法对50批紫草进行测定,采用Agilent 5 TC-C18(2)色谱柱,以乙腈-体积分数0.05%甲酸水溶液为流动相,梯度洗脱,流速1 mL·min-1,检测波长275 nm,柱温30 ℃,并对左旋紫草素、乙酰紫草素、去氧紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰阿卡宁和β,β'-二甲基丙烯酰阿卡宁等6个萘醌类成分进行含量测定。采用HPLC指纹图谱检测并结合“中药色谱指纹图谱相似度评价系统(2012A版)”建立紫草药材的指纹图谱,进行相似度评价和共有峰确认。结果 方法学结果表明,6个成分在相应的浓度范围内与峰面积呈良好的线性关系,6个成分的含量分别为0.006%~0.237%,0.009%~1.759%,0.001%~0.129%,0.002%~0.625%,0.011%~609%,0.016%~1.712%,建立了紫草HPLC指纹图谱,共标定了9个共有峰,采用中药指纹图谱软件进行相似度拟合,发现50批紫草的相似度差异较大。结论 该研究建立的紫草HPLC指纹图谱和同时测定6个成分含量的方法简便、稳定、准确可靠、重复性好,可为紫草药材的质量研究提供参考依据。, correspAuthors=郑健, authorNote=null, correspAuthorsNote=
*郑健,女,博士,研究员 研究方向:民族药质量控制及相关标准研究 Tel:(010)53852080
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黄瑞和戴胜云为共同第一作者

黄瑞,女,硕士研究生 研究方向:民族药质量控制及相关标准研究;

戴胜云,女,博士 研究方向:民族药质量控制及相关标准研究。

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2 China Pharmaceutical University, Nanjing 211198, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1200147843001451439, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, authorId=1200147842724627363, language=CN, stringName=黄瑞, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=1, 2, address=1 中国食品药品检定研究院, 北京 102629
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黄瑞,女,硕士研究生 研究方向:民族药质量控制及相关标准研究;

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黄瑞,女,硕士研究生 研究方向:民族药质量控制及相关标准研究;

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戴胜云,女,博士 研究方向:民族药质量控制及相关标准研究。

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戴胜云,女,博士 研究方向:民族药质量控制及相关标准研究。

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DOI: 10.3389/fphar.2023.1271252., articleTitle=The anti-leukemia activity and mechanisms of shikonin: a mini review, refAbstract=null), Reference(id=1200147851977261220, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, doi=null, pmid=null, pmcid=null, year=2023, volume=105, issue=null, pageStart=105527, pageEnd=null, url=null, language=null, rfNumber=[17], rfOrder=16, authorNames=LI C C, LU S R, XU C Q, journalName=J Funct Foods, refType=null, unstructuredReference=LI C C, LU S R, XU C Q, et al. Shikonin mediated anti-acute lung injury and hemostatic effects via inhibiting Upa[J]. 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Urumqi: Xinjiang Medical University, 2014., articleTitle=null, refAbstract=null), Reference(id=1200147852207947943, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, doi=null, pmid=null, pmcid=null, year=2017, volume=42, issue=13, pageStart=2532, pageEnd=2537, url=null, language=null, rfNumber=[19], rfOrder=18, authorNames=ZAN K, ZHENG H Y, LIU J, journalName=China J Chin Mater Med(中国中药杂志), refType=null, unstructuredReference=ZAN K, ZHENG H Y, LIU J, et al. Simultaneous determination of eight hydroxyl naphthoquinones in different parts of Arnebiae euchroma by HPLC[J]. China J Chin Mater Med(中国中药杂志), 2017, 42(13):2532-2537., articleTitle=Simultaneous determination of eight hydroxyl naphthoquinones in different parts of Arnebiae euchroma by HPLC, refAbstract=null), Reference(id=1200147852581241001, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, doi=null, pmid=null, pmcid=null, year=2016, volume=36, issue=9, pageStart=1526, pageEnd=1535, url=null, language=null, rfNumber=[20], rfOrder=19, authorNames=ZAN K, SU R, TENG A J, journalName=Chin J Pharm Anal(药物分析杂志), refType=null, unstructuredReference=ZAN K, SU R, TENG A J, et al. HPLC specific chromatogram of Arnebia euchroma and determination of six naphthoquinones in Boraginaceous herbs[J]. 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2-Alkannin; 4-Acetylshikonin; 5-Deoxyshikonin; 6-β-Acetoxyisovalerylshikonin; 7-Isobutyryl alkannin; 8-β,β'-Dimethylacrylalkannin; 1,3,9-Unknown.

, figureFileSmall=U1+or+8uQEtkcTVaiwoiag==, figureFileBig=qVCD5bStGJtOCEqReFp8wg==, tableContent=null), ArticleFig(id=1200147846273007652, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=图1, caption=紫草的高效液相色谱图(A)及50批样品的指纹图谱(B)

2-左旋紫草素;4-乙酰紫草素;5-去氧紫草素;6-β-乙酰氧基异戊酰阿卡宁;7-异丁酰阿卡宁;8-β,β'-二甲基丙烯酰阿卡宁;1,3,9-未知。

, figureFileSmall=U1+or+8uQEtkcTVaiwoiag==, figureFileBig=qVCD5bStGJtOCEqReFp8wg==, tableContent=null), ArticleFig(id=1200147846545637419, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Fig.2, caption=Reference fingerprint of Arnebiae Radix

2-Alkannin; 4-Acetylshikonin; 5-Deoxyshikonin; 6-β-Acetoxyisovalerylshikonin; 7-Isobutyryl alkannin; 8-β,β'-Dimethylacrylalkannin; 1,3,9-Unknown.

, figureFileSmall=9gqoa5/pLkACQwf0NKd3+w==, figureFileBig=uETCMt4U0bzhJyUPQMGKJQ==, tableContent=null), ArticleFig(id=1200147846667272239, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=图2, caption=紫草的对照指纹图谱

2-左旋紫草素;4-乙酰紫草素;5-去氧紫草素;6-β-乙酰氧基异戊酰阿卡宁;7-异丁酰阿卡宁;8-β,β'-二甲基丙烯酰阿卡宁;1,3,9-未知。

, figureFileSmall=9gqoa5/pLkACQwf0NKd3+w==, figureFileBig=uETCMt4U0bzhJyUPQMGKJQ==, tableContent=null), ArticleFig(id=1200147846788907056, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Fig.3, caption=Total contents of six components in 48 batches of Arnebiae Radix, figureFileSmall=dXX4iGwrZ3DA0E8rTlTlFw==, figureFileBig=LY5h4iaY6sRYjvGfk8CfOw==, tableContent=null), ArticleFig(id=1200147846918930484, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=图3, caption=50批紫草样品中6种成分的总含量, figureFileSmall=dXX4iGwrZ3DA0E8rTlTlFw==, figureFileBig=LY5h4iaY6sRYjvGfk8CfOw==, tableContent=null), ArticleFig(id=1200147847048953912, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.1, caption=

Sample information of Arnebiae Radix

, figureFileSmall=null, figureFileBig=null, tableContent=
Number Source(in Chinese) Origin Number Source(in Chinese) Origin
ZC59 Kyrgyzstan(吉尔吉斯斯坦) - 2158 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC52C Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 2235 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC60 Pakistan(巴基斯坦) - 2154 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC62 Kazakhstan(哈萨克斯坦) - 2224 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC63 Pakistan(巴基斯坦) - 2014 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC69 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. 1707 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC58 Tajikistan(塔吉克斯坦) - 1874 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC53 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 1790 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC61 Afghanistan(阿富汗) - 1252 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC44 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 2156 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC42 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 2155 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC57 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 1375 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC47 Xinjiang, China(中国新疆) Arnebia guttata Bunge 1869 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC68 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. 1871 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC70 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. 1868 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC72 Sichuan, China(中国四川) Onosma paniculatum Bur. et Franch. 2159 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC74 Xinjiang, China(中国新疆) Arnebia guttata Bunge 2222 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC67 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. J95 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC64 Pakistan(巴基斯坦) - J97 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC73 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J92 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC65 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J94 Anhui, China(中国安徽) Arnebia euchroma(Royle) Johnst.
ZC46 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J89 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC66 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J93 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC71 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. J96 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC56B Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J90 Anhui, China(中国安徽) Arnebia euchroma(Royle) Johnst.
), ArticleFig(id=1200147847241891896, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=表1, caption=

紫草药材样品信息

, figureFileSmall=null, figureFileBig=null, tableContent=
Number Source(in Chinese) Origin Number Source(in Chinese) Origin
ZC59 Kyrgyzstan(吉尔吉斯斯坦) - 2158 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC52C Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 2235 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC60 Pakistan(巴基斯坦) - 2154 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC62 Kazakhstan(哈萨克斯坦) - 2224 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC63 Pakistan(巴基斯坦) - 2014 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC69 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. 1707 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC58 Tajikistan(塔吉克斯坦) - 1874 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC53 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 1790 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC61 Afghanistan(阿富汗) - 1252 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC44 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 2156 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC42 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 2155 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC57 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. 1375 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC47 Xinjiang, China(中国新疆) Arnebia guttata Bunge 1869 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC68 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. 1871 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC70 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. 1868 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC72 Sichuan, China(中国四川) Onosma paniculatum Bur. et Franch. 2159 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC74 Xinjiang, China(中国新疆) Arnebia guttata Bunge 2222 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC67 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. J95 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC64 Pakistan(巴基斯坦) - J97 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC73 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J92 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC65 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J94 Anhui, China(中国安徽) Arnebia euchroma(Royle) Johnst.
ZC46 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J89 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC66 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J93 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC71 Yunnan, China(中国云南) Onosma paniculatum Bur. et Franch. J96 Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst.
ZC56B Xinjiang, China(中国新疆) Arnebia euchroma(Royle) Johnst. J90 Anhui, China(中国安徽) Arnebia euchroma(Royle) Johnst.
), ArticleFig(id=1200147847384498238, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.2, caption=

HPLC elution procedure for the determination of Arnebiae Radix

, figureFileSmall=null, figureFileBig=null, tableContent=
t/min Mobile phase A/% Mobile phase B/%
0 57 43
10 65 35
26 65 35
27 57 43
30 57 43
), ArticleFig(id=1200147847560659007, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=表2, caption=

紫草药材高效液相色谱(HPLC)测定洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
t/min Mobile phase A/% Mobile phase B/%
0 57 43
10 65 35
26 65 35
27 57 43
30 57 43
), ArticleFig(id=1200147847745208389, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.3, caption=

Similarity of 50 batches of Arnebiae Radix samples

, figureFileSmall=null, figureFileBig=null, tableContent=
Lot. Similarity Lot. Similarity Lot. Similarity
R 1 S18 0.841 S35 0.995
S2 0.97 S19 0.95 S36 0.937
S3 0.849 S20 0.729 S37 0.898
S4 0.924 S21 0.955 S38 0.982
S5 0.948 S22 0.885 S39 0.981
S6 0.973 S23 0.979 S40 0.982
S7 0.967 S24 0.944 S41 0.98
S8 0.988 S25 0.928 S42 0.945
S9 0.965 S26 0.963 S43 0.977
S10 0.965 S27 0.895 S44 0.983
S11 0.971 S28 0.984 S45 0.985
S12 0.993 S29 0.924 S46 0.961
S13 0.697 S30 0.928 S47 0.969
S14 0.977 S31 0.954 S48 0.911
S15 0.897 S32 0.902 S49 0.969
S16 0.913 S33 0.966 S50 0.99
S17 0.968 S34 0.97 S51 0.928
), ArticleFig(id=1200147847908786248, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=表3, caption=

50批紫草样品相似度

, figureFileSmall=null, figureFileBig=null, tableContent=
Lot. Similarity Lot. Similarity Lot. Similarity
R 1 S18 0.841 S35 0.995
S2 0.97 S19 0.95 S36 0.937
S3 0.849 S20 0.729 S37 0.898
S4 0.924 S21 0.955 S38 0.982
S5 0.948 S22 0.885 S39 0.981
S6 0.973 S23 0.979 S40 0.982
S7 0.967 S24 0.944 S41 0.98
S8 0.988 S25 0.928 S42 0.945
S9 0.965 S26 0.963 S43 0.977
S10 0.965 S27 0.895 S44 0.983
S11 0.971 S28 0.984 S45 0.985
S12 0.993 S29 0.924 S46 0.961
S13 0.697 S30 0.928 S47 0.969
S14 0.977 S31 0.954 S48 0.911
S15 0.897 S32 0.902 S49 0.969
S16 0.913 S33 0.966 S50 0.99
S17 0.968 S34 0.97 S51 0.928
), ArticleFig(id=1200147848072364106, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.4, caption=

Standard curves and linear ranges of six indicator components from Arnebiae Radix

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Regression equation r Linear range/mg·mL-1
Alkannin Y=20 002x-3.575 1 0.999 7 0.001~0.036
Acetylshikonin Y=23 108x-14.911 0.999 8 0.004~0.143
Deoxyshikonin Y=36 152x-9.563 2 0.999 1 0.001~0.018
β-Acetoxyisovalerylshikonin Y=18 183x-12.489 0.999 8 0.005~0.144
Isobutyryl alkannin Y=21 801x-13.381 0.999 8 0.003~0.100
β,β'-Dimethylacrylalkannin Y=20 928x-23.8 0.999 7 0.005~0.144
), ArticleFig(id=1200147848261107791, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=表4, caption=

紫草样品中6种指标成分的标准曲线及线性范围

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Regression equation r Linear range/mg·mL-1
Alkannin Y=20 002x-3.575 1 0.999 7 0.001~0.036
Acetylshikonin Y=23 108x-14.911 0.999 8 0.004~0.143
Deoxyshikonin Y=36 152x-9.563 2 0.999 1 0.001~0.018
β-Acetoxyisovalerylshikonin Y=18 183x-12.489 0.999 8 0.005~0.144
Isobutyryl alkannin Y=21 801x-13.381 0.999 8 0.003~0.100
β,β'-Dimethylacrylalkannin Y=20 928x-23.8 0.999 7 0.005~0.144
), ArticleFig(id=1200147848407908434, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.5, caption=

Results of spiked sample recovery test for 6 chemical.n=6

, figureFileSmall=null, figureFileBig=null, tableContent=
Component m(Sample)/g m(Content)/mg m(Add)/mg m(Measured)/mg Recovery/% Average recovery/% RSD/%
Alkannin 0.250 1 0.147 0 0.099 3 0.242 9 96.56 99.87 2.51
0.252 5 0.148 4 0.099 3 0.249 6 101.81
0.248 9 0.146 3 0.099 3 0.248 2 102.56
0.250 5 0.147 3 0.099 3 0.244 2 97.58
0.250 0 0.147 0 0.090 3 0.238 8 101.72
0.251 0 0.147 6 0.090 3 0.236 9 98.98
Acetylshikonin 0.250 1 0.503 6 0.438 6 0.951 6 102.16 99.93 2.79
0.252 5 0.508 4 0.438 6 0.950 8 100.87
0.248 9 0.501 2 0.421 0 0.918 8 99.21
0.250 5 0.504 4 0.421 0 0.902 8 94.64
0.250 0 0.503 4 0.456 1 0.963 0 100.77
0.251 0 0.505 4 0.456 1 0.970 2 101.91
Deoxyshikonin 0.250 1 0.036 6 0.066 0.103 100.31 99.96 1.51
0.252 5 0.036 9 0.066 0.104 101.98
0.248 9 0.036 4 0.062 0.098 99.36
0.250 5 0.036 6 0.062 0.097 97.52
0.250 0 0.036 6 0.062 0.098 99.77
0.251 0 0.036 7 0.066 0.103 100.85
β-Acetoxyisovaleryl- 0.250 1 0.102 6 0.088 2 0.193 6 103.20 101.03 2.91
shikonin 0.252 5 0.103 5 0.088 2 0.194 7 103.33
0.248 9 0.102 1 0.079 4 0.184 8 104.20
0.250 5 0.102 7 0.079 4 0.182 2 100.09
0.250 0 0.102 5 0.097 0 0.197 3 97.69
0.251 0 0.102 9 0.097 0 0.197 7 97.69
Isobutyryl alkannin 0.250 1 0.191 4 0.225 4 0.424 0 103.21 102.76 1.79
0.252 5 0.193 2 0.225 4 0.424 0 102.40
0.248 9 0.190 5 0.205 8 0.403 2 103.35
0.250 5 0.191 7 0.205 8 0.396 1 99.30
0.250 0 0.191 3 0.225 4 0.425 3 103.80
0.251 0 0.192 1 0.225 4 0.427 7 104.53
β,β'-Dimethylacryl- 1.282 4 1.282 4 1.236 2 2.544 9 102.13 98.50 2.80
alkannin 1.294 6 1.294 6 1.236 2 2.543 8 101.05
1.276 2 1.276 2 1.174 4 2.441 6 99.24
1.284 4 1.284 4 1.174 4 2.402 2 95.18
1.281 8 1.281 8 1.192 0 2.430 7 96.38
1.287 0 1.287 0 1.192 0 2.443 3 97.00
), ArticleFig(id=1200147848550514773, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=表5, caption=

紫草样品中6种化学成分的加样回收率试验结果.n=6

, figureFileSmall=null, figureFileBig=null, tableContent=
Component m(Sample)/g m(Content)/mg m(Add)/mg m(Measured)/mg Recovery/% Average recovery/% RSD/%
Alkannin 0.250 1 0.147 0 0.099 3 0.242 9 96.56 99.87 2.51
0.252 5 0.148 4 0.099 3 0.249 6 101.81
0.248 9 0.146 3 0.099 3 0.248 2 102.56
0.250 5 0.147 3 0.099 3 0.244 2 97.58
0.250 0 0.147 0 0.090 3 0.238 8 101.72
0.251 0 0.147 6 0.090 3 0.236 9 98.98
Acetylshikonin 0.250 1 0.503 6 0.438 6 0.951 6 102.16 99.93 2.79
0.252 5 0.508 4 0.438 6 0.950 8 100.87
0.248 9 0.501 2 0.421 0 0.918 8 99.21
0.250 5 0.504 4 0.421 0 0.902 8 94.64
0.250 0 0.503 4 0.456 1 0.963 0 100.77
0.251 0 0.505 4 0.456 1 0.970 2 101.91
Deoxyshikonin 0.250 1 0.036 6 0.066 0.103 100.31 99.96 1.51
0.252 5 0.036 9 0.066 0.104 101.98
0.248 9 0.036 4 0.062 0.098 99.36
0.250 5 0.036 6 0.062 0.097 97.52
0.250 0 0.036 6 0.062 0.098 99.77
0.251 0 0.036 7 0.066 0.103 100.85
β-Acetoxyisovaleryl- 0.250 1 0.102 6 0.088 2 0.193 6 103.20 101.03 2.91
shikonin 0.252 5 0.103 5 0.088 2 0.194 7 103.33
0.248 9 0.102 1 0.079 4 0.184 8 104.20
0.250 5 0.102 7 0.079 4 0.182 2 100.09
0.250 0 0.102 5 0.097 0 0.197 3 97.69
0.251 0 0.102 9 0.097 0 0.197 7 97.69
Isobutyryl alkannin 0.250 1 0.191 4 0.225 4 0.424 0 103.21 102.76 1.79
0.252 5 0.193 2 0.225 4 0.424 0 102.40
0.248 9 0.190 5 0.205 8 0.403 2 103.35
0.250 5 0.191 7 0.205 8 0.396 1 99.30
0.250 0 0.191 3 0.225 4 0.425 3 103.80
0.251 0 0.192 1 0.225 4 0.427 7 104.53
β,β'-Dimethylacryl- 1.282 4 1.282 4 1.236 2 2.544 9 102.13 98.50 2.80
alkannin 1.294 6 1.294 6 1.236 2 2.543 8 101.05
1.276 2 1.276 2 1.174 4 2.441 6 99.24
1.284 4 1.284 4 1.174 4 2.402 2 95.18
1.281 8 1.281 8 1.192 0 2.430 7 96.38
1.287 0 1.287 0 1.192 0 2.443 3 97.00
), ArticleFig(id=1200147848726675544, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.6, caption=

Content determination results of 50 batches of Arnebiae Radix. %

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample No. Alkannin Acetylshikonin Deoxyshikonin β-Acetoxyisovalerylshikonin Isobutyryl alkannin β,β'-Dimethylacrylalkannin
ZC60 0.01 0.01 0 0.04 0.01 0.02
ZC62 0.02 0.05 0.01 0.17 0.09 0.74
ZC63 0.04 0.28 0.02 0.26 0.19 0.74
ZC69 0.01 0.27 0 0.01 0.05 0.98
ZC58 0.01 0.08 0.02 0.17 0.12 1.66
ZC2158 0.04 0.18 0.01 0.25 0.16 0.56
ZC2235 0.03 0.13 0.01 0.15 0.14 0.88
ZC2154 0.09 0.66 0.02 0.22 0.49 0.82
ZC2224 0.06 0.28 0.01 0.08 0.17 0.38
ZC2014 0.05 0.34 0.02 0.29 0.16 0.44
ZC53 0.11 1.76 0.11 0.18 0.28 0.88
ZC61 0.02 0.06 0.01 0.05 0.03 0.14
ZC44 0.14 0.4 0.01 0.39 0.54 1.11
ZC42 0.18 0.25 0.01 0.2 0.35 0.72
ZC57 0.08 0.42 0.03 0.13 0.1 0.37
ZC47 0.24 0.17 0.02 0.06 0.18 0.18
ZC68 0.11 0.68 0.04 0.01 0.16 1.12
ZC56B 0.09 1.71 0.13 0.25 0.36 0.83
ZC1707 0.03 0.13 0.01 0.14 0.1 1.01
ZC1874 0.05 0.19 0.02 0.13 0.16 0.97
ZC1790 0.07 0.51 0.02 0.2 0.24 0.55
J95 0.14 0.5 0.02 0.38 0.43 0.64
J97 0.09 0.9 0.02 0.11 0.61 0.89
J92 0.07 0.47 0.03 0.63 0.5 0.59
J94 0.08 0.22 0.02 0.04 0.08 0.58
J89 0.03 0.16 0.01 0.21 0.12 0.37
J93 0.13 1.6 0.11 0.5 0.43 1.1
J96 0.11 0.78 0.03 0.28 0.29 0.97
ZC70 0.02 0.25 0.01 0.01 0.05 0.93
ZC72 0.02 0.7 0.01 0.08 0.17 0.58
ZC74 0.05 0.52 0.03 0.24 0.53 0.74
ZC67 0.01 0.1 0.01 0 0.02 0.37
ZC64 0.02 0.07 0.01 0.15 0.09 0.21
ZC73 0.1 1.16 0.04 0.11 0.51 1.11
ZC65 0.01 0.04 0 0.05 0.01 0.07
ZC46 0.05 0.39 0.01 0.08 0.13 0.75
ZC66 0.13 1.19 0.04 0.19 0.37 0.94
ZC71 0.02 0.28 0.01 0.02 0.07 0.8
ZC1252 0.06 0.29 0.02 0.14 0.17 0.49
ZC2156 0.06 0.18 0.01 0.22 0.24 0.62
ZC2155 0.05 0.22 0.02 0.05 0.12 0.38
ZC1375 0.03 0.11 0.01 0.16 0.1 0.62
ZC1869 0.08 0.41 0.03 0.22 0.39 0.65
ZC1871 0.1 0.44 0.03 0.25 0.44 0.74
ZC1868 0.07 0.46 0.02 0.24 0.33 0.57
ZC2159 0.04 0.24 0.02 0.34 0.23 1.71
ZC2222 0.04 0.18 0.02 0.17 0.17 0.63
J90 0.01 0.06 0.02 0.32 0.1 1.44
ZC52C 0.06 0.15 0.01 0.06 0.04 0.16
ZC59 0.03 0.12 0.01 0.16 0.12 0.2
), ArticleFig(id=1200147848873476188, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=CN, label=表6, caption=

50批紫草样品的含量测定结果. %

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample No. Alkannin Acetylshikonin Deoxyshikonin β-Acetoxyisovalerylshikonin Isobutyryl alkannin β,β'-Dimethylacrylalkannin
ZC60 0.01 0.01 0 0.04 0.01 0.02
ZC62 0.02 0.05 0.01 0.17 0.09 0.74
ZC63 0.04 0.28 0.02 0.26 0.19 0.74
ZC69 0.01 0.27 0 0.01 0.05 0.98
ZC58 0.01 0.08 0.02 0.17 0.12 1.66
ZC2158 0.04 0.18 0.01 0.25 0.16 0.56
ZC2235 0.03 0.13 0.01 0.15 0.14 0.88
ZC2154 0.09 0.66 0.02 0.22 0.49 0.82
ZC2224 0.06 0.28 0.01 0.08 0.17 0.38
ZC2014 0.05 0.34 0.02 0.29 0.16 0.44
ZC53 0.11 1.76 0.11 0.18 0.28 0.88
ZC61 0.02 0.06 0.01 0.05 0.03 0.14
ZC44 0.14 0.4 0.01 0.39 0.54 1.11
ZC42 0.18 0.25 0.01 0.2 0.35 0.72
ZC57 0.08 0.42 0.03 0.13 0.1 0.37
ZC47 0.24 0.17 0.02 0.06 0.18 0.18
ZC68 0.11 0.68 0.04 0.01 0.16 1.12
ZC56B 0.09 1.71 0.13 0.25 0.36 0.83
ZC1707 0.03 0.13 0.01 0.14 0.1 1.01
ZC1874 0.05 0.19 0.02 0.13 0.16 0.97
ZC1790 0.07 0.51 0.02 0.2 0.24 0.55
J95 0.14 0.5 0.02 0.38 0.43 0.64
J97 0.09 0.9 0.02 0.11 0.61 0.89
J92 0.07 0.47 0.03 0.63 0.5 0.59
J94 0.08 0.22 0.02 0.04 0.08 0.58
J89 0.03 0.16 0.01 0.21 0.12 0.37
J93 0.13 1.6 0.11 0.5 0.43 1.1
J96 0.11 0.78 0.03 0.28 0.29 0.97
ZC70 0.02 0.25 0.01 0.01 0.05 0.93
ZC72 0.02 0.7 0.01 0.08 0.17 0.58
ZC74 0.05 0.52 0.03 0.24 0.53 0.74
ZC67 0.01 0.1 0.01 0 0.02 0.37
ZC64 0.02 0.07 0.01 0.15 0.09 0.21
ZC73 0.1 1.16 0.04 0.11 0.51 1.11
ZC65 0.01 0.04 0 0.05 0.01 0.07
ZC46 0.05 0.39 0.01 0.08 0.13 0.75
ZC66 0.13 1.19 0.04 0.19 0.37 0.94
ZC71 0.02 0.28 0.01 0.02 0.07 0.8
ZC1252 0.06 0.29 0.02 0.14 0.17 0.49
ZC2156 0.06 0.18 0.01 0.22 0.24 0.62
ZC2155 0.05 0.22 0.02 0.05 0.12 0.38
ZC1375 0.03 0.11 0.01 0.16 0.1 0.62
ZC1869 0.08 0.41 0.03 0.22 0.39 0.65
ZC1871 0.1 0.44 0.03 0.25 0.44 0.74
ZC1868 0.07 0.46 0.02 0.24 0.33 0.57
ZC2159 0.04 0.24 0.02 0.34 0.23 1.71
ZC2222 0.04 0.18 0.02 0.17 0.17 0.63
J90 0.01 0.06 0.02 0.32 0.1 1.44
ZC52C 0.06 0.15 0.01 0.06 0.04 0.16
ZC59 0.03 0.12 0.01 0.16 0.12 0.2
), ArticleFig(id=1200147849053831268, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1200147840472285993, language=EN, label=Tab.7, caption=

Characteristic value and variance contribution rate of the six components from Arnebiae Radix

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Principal
component		 Characteristic
value		 Variance contribution
rate		 Cumulative variance
contribution rate
1 3.133 52.215 52.215
2 0.996 16.606 68.821
3 0.910 15.17 83.991
4 0.593 9.883 93.874
5 0.309 5.144 99.018
6 0.059 0.982 100
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紫草样品中6种成分的特征值和方差贡献率

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Principal
component		 Characteristic
value		 Variance contribution
rate		 Cumulative variance
contribution rate
1 3.133 52.215 52.215
2 0.996 16.606 68.821
3 0.910 15.17 83.991
4 0.593 9.883 93.874
5 0.309 5.144 99.018
6 0.059 0.982 100
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基于HPLC指纹图谱及多指标成分含量测定的紫草质量评价
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黄瑞 1, 2 , 戴胜云 1 , 吴东雪 1 , 刘杰 1 , 马双成 1 , 郑健 1, *
中国药学杂志 | 论著 2024,59(10): 929-937
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中国药学杂志 | 论著 2024, 59(10): 929-937
基于HPLC指纹图谱及多指标成分含量测定的紫草质量评价
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黄瑞1, 2, 戴胜云1, 吴东雪1, 刘杰1, 马双成1, 郑健1, *
作者信息
  • 1 中国食品药品检定研究院, 北京 102629
  • 2 中国药科大学, 南京 211198

    • 黄瑞,女,硕士研究生 研究方向:民族药质量控制及相关标准研究;

    戴胜云,女,博士 研究方向:民族药质量控制及相关标准研究。

通讯作者:

*郑健,女,博士,研究员 研究方向:民族药质量控制及相关标准研究 Tel:(010)53852080
Study on Quality Evaluation of Arnebiae Radix by HPLC Fingerprint and Components Contents Determination
Rui HUANG1, 2, Shengyun DAI1, Dongxue WU1, Jie LIU1, Shuangcheng MA1, Jian ZHENG1, *
Affiliations
  • 1 National Institutes for Food and Drug Control, Beijing 102629, China
  • 2 China Pharmaceutical University, Nanjing 211198, China
出版时间: 2024-05-22 doi: 10.11669/cpj.2024.10.010
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摘要
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目的 采用高效液相色谱(HPLC)法建立50批紫草药材指纹图谱及6个成分含量测定方法。方法 采用HPLC法对50批紫草进行测定,采用Agilent 5 TC-C18(2)色谱柱,以乙腈-体积分数0.05%甲酸水溶液为流动相,梯度洗脱,流速1 mL·min-1,检测波长275 nm,柱温30 ℃,并对左旋紫草素、乙酰紫草素、去氧紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰阿卡宁和β,β'-二甲基丙烯酰阿卡宁等6个萘醌类成分进行含量测定。采用HPLC指纹图谱检测并结合“中药色谱指纹图谱相似度评价系统(2012A版)”建立紫草药材的指纹图谱,进行相似度评价和共有峰确认。结果 方法学结果表明,6个成分在相应的浓度范围内与峰面积呈良好的线性关系,6个成分的含量分别为0.006%~0.237%,0.009%~1.759%,0.001%~0.129%,0.002%~0.625%,0.011%~609%,0.016%~1.712%,建立了紫草HPLC指纹图谱,共标定了9个共有峰,采用中药指纹图谱软件进行相似度拟合,发现50批紫草的相似度差异较大。结论 该研究建立的紫草HPLC指纹图谱和同时测定6个成分含量的方法简便、稳定、准确可靠、重复性好,可为紫草药材的质量研究提供参考依据。
紫草  /  高效液相色谱法  /  指纹图谱  /  成分含量  /  质量评价

OBJECTIVE To establish the fingerprints of 50 batches of Arnebiae Radix and the methods for determination of the contents of six components. METHODS A high performance liquid chromatographic (HPLC) method was used for the determination of 50 batches of Arnebiae Radix on an Agilent 5 TC-C18(2) column, with acetonitrile-0.05% formic acid aqueous solution as the mobile phase, gradient elution at a flow rate of 1 mL·min-1, detection wavelength of 275 nm, and column temperature of 30 ℃. The contents of six naphthoquinone components, i.e., alkannin, acetylshikonin, deoxyshikonin, β-acetoxyisovalerylshikonin, isobutyryl alkannin, and β,β'-dimethylacrylalkannin were determined. The fingerprints of the herbs were established by HPLC fingerprinting combined with the “Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012A)” to evaluate the similarity and confirm the common peaks. RESULTS The methodological validation results showed that the peak areas of the six components had good linear relationships in the corresponding concentration ranges, and the contents of the six components ranged from 0.006%-0.237%, 0.009%-1.759%, 0.001%-0.129%, 0.002%-0.625%, 0.011%-0.609%, and 0.016%-1.712%, respectively. The HPLC fingerprints of Arnebiae Radix were established, and a total of nine common peaks were calibrated. The similarity fitting was performed by using the TCM fingerprinting software, and it was found that the similarity of the 50 batches of Arnebiae Radix varied greatly. CONCLUSION The HPLC fingerprint of Arnebiae Radix established in this study and the method of simultaneous determination of the contents of six components are simple, stable, accurate, reliable, and reproducible, which can provide a basis for the quality study of Arnebiae Radix.

Arnebiae Radix  /  HPLC  /  fingerprint  /  components content  /  quality evaluate
黄瑞, 戴胜云, 吴东雪, 刘杰, 马双成, 郑健. 基于HPLC指纹图谱及多指标成分含量测定的紫草质量评价. 中国药学杂志, 2024 , 59 (10) : 929 -937 . DOI: 10.11669/cpj.2024.10.010
Rui HUANG, Shengyun DAI, Dongxue WU, Jie LIU, Shuangcheng MA, Jian ZHENG. Study on Quality Evaluation of Arnebiae Radix by HPLC Fingerprint and Components Contents Determination[J]. Chinese Pharmaceutical Journal, 2024 , 59 (10) : 929 -937 . DOI: 10.11669/cpj.2024.10.010
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紫草为紫草科植物新疆紫草[Arnebia euchroma(Royle)Johnst.]或内蒙紫草(Arnebia guttata Bunge)的干燥根[1],是一种常见的传统药材,始载于《神农本草经》[2],在我国有着非常悠久的药用历史,具有清热凉血、活血解毒、透疹消斑之功效。临床主要用于治疗压疮、新生儿红臀、皮肤溃疡、皮肤瘙痒、皮炎等常见皮肤病[3]。现代药理学研究表明,紫草素具有明显的抗炎、抗菌、抗病毒等作用,同时紫草素及其衍生物在抗肿瘤、抗氧化、保护心脑血管系统等方面也具有重要作用[4]。紫草资源丰富,在我国除内蒙古、甘肃等地外,全国各地均有分布,其中以新疆、云南、贵州等省区最为集中。随着社会经济发展和药用植物资源的不断开发利用,目前各地紫草存在着品种混杂、质量参差不齐、稳定性差等问题。1990年版到2005年版《中华人民共和国药典》(以下简称《中国药典》)共收录了紫草科的3种植物,即软紫草属的新疆紫草[Arnebia euchroma (Royle) Johnst.]、内蒙紫草(Arnebia guttata Bunge)和紫草属硬紫草(Lithospermum erythrorhizon Sieb. et Zucc.)的干燥根[5-8],而2010年版到2020年版《中国药典》仅收载了前两者紫草的干燥根[9-11]。除药典品种外,滇紫草(Onosma paniculatum Bur. et Franch.)为《云南省中药饮片标准》[11]所收录,长花滇紫草(Onosma hookeri Clarke var. longiflorum Duthie)为《卫生部药品标准》1995年版[12]收载的藏紫草之一,两者均来源于滇紫草属植物,野生紫草来源广泛,且随着现代农业栽培技术的发展,越来越多的紫草栽培品也在市面上流通,造成了紫草资源混乱的问题。研究表明,紫草中含有多种黄酮类化合物,药理活性显著,具有抗氧化、抗肿瘤、烧伤愈合、抗炎、防护热射病等功效[13],其中以紫草素为代表的羟基萘醌类化合物,被认为是紫草的主要活性物质。Sun等[14]研究发现,紫草素可以抗甲状腺癌活性分子从而发挥抗癌的作用。Park等[15]也指出,紫草素通过释放辅激活剂和调节组蛋白甲基化密码来结合和抑制 PPARγ 的活性,调节脂肪生成过程中组蛋白密码的富集来抑制脂肪生成,治疗肥胖相关疾病。此外,紫草素还有抗白血病[16]、抗急性肺损伤和止血作用[17]。然而,紫草中萘醌类成分含量差异较大,导致药材的质量参差不齐[18]。
本课题组前期对紫草的萘醌类成分进行含量测定[19-20],发现存在较大差异,并且在同一产地也会有较大差异。因此,建立科学合理的紫草药材质量评价方法,对保证紫草药材质量的稳定和提高其临床疗效,显得尤为重要。目前对紫草的质量控制主要是以检测β,β'-二甲基丙烯酰阿卡宁的成分含量为评价指标,但紫草化合物组成复杂,单一指标无法准确地控制其质量。基于此,本研究通过建立紫草药材的HPLC指纹图谱和含量测定方法,评价不同产地、基源的紫草药材质量,为制定紫草药材的质量标准提供科学依据,也为进一步探讨紫草资源的开发利用提供基础数据。
1 仪器与材料
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1.1 仪器
Agilent Technologies 1260 Infinity高效液相色谱仪,液相系统包括四元泵、自动进样器、柱温箱、DAD检测器,色谱柱:Agilent 5 TC-C18(2)(4.6 mm×150 mm,5 μm)(美国Agilent公司);SHHW21.600AⅡ型三用恒温水箱(天津市泰斯特仪器有限公司);MS105DU电子分析天平十万分之一(梅特勒-托利多仪器上海有限公司);Eppendorf Centrifuge 5430R型台式平板式离心机;KQ-500B型超声波清洗器(昆山市超声仪器有限公司)。
1.2 材料
紫草药材为自行采收或委托购买,经中国食品药品检定研究院郑健研究员鉴定为紫草科新疆紫草[Arnebia euchroma(Royle)Johnst.]和内蒙紫草(Arnebia guttata Bung)的干燥根、滇紫草(Onosma paniculatum Bur. et Franch.)的根或根皮,由于部分进口品无法用分子生物学技术判定基原,故不列举其基源植物,具体样品信息见表1。β,β'-二甲基丙烯酰阿卡宁(批号:111689-201805,纯度98.0%);左旋紫草素(批号:110769-200506,纯度98.0%)均由中国药品生物制品检定所提供;去氧紫草素(批号:7028,纯度98.0%)、乙酰紫草素(批号:12527,纯度98.0%)、异丁酰阿卡宁(批号:10513,纯度98.0%)、异丁酰紫草素(批号:10398,纯度98.0%)、β-乙酰氧基异戊酰阿卡宁对照品(批号:11099,纯度98.0%)、乙酰阿卡宁(批号:10476,纯度98.0%)、右旋紫草素(批号:6478,纯度98.0%)等(诗丹德标准技术服务有限公司);新疆紫草、藏紫草(长花滇紫草)(中国食品药品检定研究院)。
乙腈、甲醇、甲酸(≥98%)为色谱纯,石油醚为分析纯,水为超纯水和怡宝(华润怡宝有限公司)。
2 方法与结果
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2.1 色谱条件
色谱柱:Agilent 5 TC-C18(2)(4.6 mm×150 mm,5 μm),流动相:乙腈(A)-0.05%甲酸水(B),梯度洗脱,流速1.0 mL·min-1,柱温:30 ℃,检测波长:275 nm,进样量:20 μL。按照表2进行洗脱。
2.2 供试品溶液的制备
取紫草粉末样品约(过四号筛)0.5 g,精密称定,置于具塞锥形瓶,精密加入石油醚(60~90 ℃)25 mL,称定质量,超声处理(功率:500 W,频率:40 kHz)30 min,放冷,再称定质量,用石油醚(60~90 ℃)补足减失质量,摇匀,滤过。精密移取上清液10 mL至蒸发皿,蒸干,残渣加流动相乙腈-水(70∶30)溶解,转移至10 mL量瓶中,加流动相乙腈-水(70∶30)定容至刻度,摇匀,滤过,取续滤液,即得。
2.3 对照品储备液的制备
分别取左旋紫草素、乙酰紫草素、去氧紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰阿卡宁和β,β'-二甲基丙烯酰阿卡宁约10 mg,精密称定,置于10 mL量瓶中,用甲醇溶解并定容至刻度,得质量浓度分别为0.903、0.895、0.899、0.900、1.000、0.901 mg·mL-1的储备液。
2.4 方法学考察
2.4.1 精密度考察
按“2.2”项下方法制备供试品溶液(J94),按“2.1”项下方法连续进样6针,记录色谱图,以乙酰紫草素为参照峰,计算共有峰相对保留时间及主要色谱峰(占总峰面积的3%以上)的相对峰面积,以及相应相对标准偏差(RSD)值。计算得到各共有峰相对保留时间、相对峰面积的RSD分别为0.02%~0.12%、0.05%~0.89%,说明仪器精密度良好,符合指纹图谱分析要求。
2.4.2 稳定性考察
按“2.2”项下方法制备供试品溶液(J94),按“2.1”项下检测条件,分别在0、5、6、10、22、24 h进样分析,记录色谱图,以乙酰紫草素为参照峰,计算共有峰相对保留时间及主要色谱峰(占总峰面积的3%以上)的相对峰面积,以及相应RSD值。计算得到各共有峰相对保留时间、相对峰面积的RSD分别为0.03%~0.26%、0.15%~0.86%,说明紫草24 h内的稳定性较好。
2.4.3 重复性考察
取紫草药材粉末样品(批号:J94),精密称定,按“2.2”项下方法制备供试品溶液,平行操作6份,按“2.1”项下色谱条件,进样分析,以乙酰紫草素为参照峰,计算共有峰相对保留时间及主要色谱峰(占总峰面积的3%以上)的相对峰面积,以及相应的RSD值。计算得到各共有峰相对保留时间、相对峰面积的RSD分别为0.05%~0.42%、0.38%~1.54%,表明该方法重复性良好,符合指纹图谱分析要求。
以上方法学考察结果显示,建立的紫草指纹图谱分析方法精密度、重复性、稳定性均较好,能够满足其指纹图谱的测定要求。
2.5 指纹图谱的建立及相似度评价
取15 批紫草样品,分别按“2.2”项下的方法制备供试品溶液并按“2.1”项下色谱检测方法检测,记录指纹图谱。紫草混合对照品和样品色谱图见图1。初步确定了9个共有色谱峰作为紫草的特征指纹峰,保留时间分别为4.017,6.019,8.237,10.067,13.859,14.598,16.677,19.469,21.34 min,这些特征峰可作为控制紫草药材的指标之一。通过与对照品比对,确定2号峰为左旋紫草素,4号峰为乙酰紫草素,5号峰为去氧紫草素,6号峰为β-乙酰氧基异戊酰阿卡宁,7号峰为异丁酰阿卡宁,8号峰为β,β'-二甲基丙烯酰阿卡宁。其中4号峰的峰面积适中,峰形较好,可作为参照峰。另9号峰峰形明显,响应较大,参考文献可知该成分可能为2-甲基丁基酰紫草素[20]。取50批紫草样品,按上述“2.2”项下方法制备供试品溶液,按“2.1”项下色谱条件,采集指纹图谱。将其中15批样品色谱图导入由国家药典委员会主持开发的“中药色谱指纹图谱相似度评价软件2012版”中进行数据处理分析,生成紫草的对照指纹图谱,结果见图2。并以此为对照,计算各批次样品与对照指纹图谱的相似度,结果见表3,结果在0.697~1.000之间,说明不同批次间样品的质量差异较大。
2.6 紫草6种萘醌类成分的含量测定
2.6.1 线性关系考察
分别精密吸取“2.3”项下对照品储备液适量,用体积分数50%甲醇配制成浓度分别为0.036、0.143、0.018、0.144、0.100、0.144 mg·mL-1的混标溶液,采用逐级稀释法制得6个成分系列浓度的标准工作液,即:左旋紫草素:0.036、0.018、0.009、0.005、0.002、0.001 mg·mL-1;乙酰紫草素:0.143、0.072、0.036、0.018、0.009、0.004 mg·mL-1;去氧紫草素:0.018、0.009、0.005、0.002、0.001、0.001 mg·mL-1;β-乙酰氧基异戊酰阿卡宁:0.144、0.072、0.036、0.018、0.009、0.005 mg·mL-1;异丁酰阿卡宁:0.100、0.050、0.025、0.013、0.006、0.003 mg·mL-1;β,β'-二甲基丙烯酰阿卡宁:0.144、0.072、0.036、0.018、0.009、0.005 mg·mL-1。按“2.1”项下色谱条件,分别进样分析。以各成分的峰面积Y(A)对浓度x(mg·mL-1)进行线性回归,得各成分的回归方程表4。
2.6.2 精密度考察
分别取“2.3”项下对照品溶液(C3)和“2.2”项下供试品溶液(批号:ZC59),按“2.1”项下色谱条件分别连续进样6次,记录峰面积并计算RSD。结果显示对照品溶液左旋紫草素、乙酰紫草素、去氧紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰阿卡宁和β,β'-二甲基丙烯酰阿卡宁的峰面积RSD值分别为0.19%、0.11%、0.16%、0.15%、0.09%、0.13%,均小于2%,供试品溶液6种成分峰面积的RSD值分别为0.23%、0.14%、1.02%、0.08%、0.12%、0.40%,均小于2%,说明仪器精密度良好。
2.6.3 重复性考察
取紫草药材粉末样品(批号:J94),按“2.2”项下方法制备供试品溶液,平行操作6份,按“2.1”项下色谱条件进样测定,记录峰面积,并计算含量。结果显示左旋紫草素、乙酰紫草素、去氧紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰阿卡宁和β,β'-二甲基丙烯酰阿卡宁含量的RSD值分别为1.22%、1.12%、1.48%、2.96%、1.02%、0.96%(n=6),表明方法的重复性良好。
2.6.4 稳定性考察
取紫草药材粉末样品(批号:J94),按“2.2”项下方法制备供试品溶液,分别于制备后0,2,6,12,24,48 h,按“2.1”项下色谱条件分别注入色谱仪,记录6种成分峰面积,计算RSD。结果显示左旋紫草素、乙酰紫草素、去氧紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰阿卡宁和β,β'-二甲基丙烯酰阿卡宁峰面积的RSD分别为1.32%,0.50%,1.94%,0.76%,1.80%,2.16%,表明仪器稳定性良好。
2.6.5 耐用性考察
本实验对不同品牌色谱柱和仪器进行了考察,将Waters e2695 高效液相色谱仪、Agilent Technologies 1260 Infinity高效液相色谱仪和Waters Xselect HSS T3(4.6 mm×250 mm,5 μm)、Agilent 5 TC-C18(2)(4.6 mm×150 mm,5 μm)色谱柱进行比较,发现该实验最终选择的仪器与色谱柱有更好的灵敏度与分离效果。
同时,本实验也对色谱条件中流速、柱温等因素进行考察,分别在流速0.9、1.0、1.1 mL·min-1和柱温25、28、30、32、35 ℃下进行实验,结果显示,流速、柱温的微小变动不会对6种萘醌类成分的分离度、峰形及含量测定结果产生显著影响。说明流速在0.9~1.1 mL·min-1、柱温在25~35 ℃条件下,方法的耐用性较好。
2.6.6 加样回收率试验
取紫草粉末样品(批号:J94)约0.25 g,精密称定,平行操作6份,分别置于25 mL量瓶中,精密加入0.903 mg·mL-1的左旋紫草素对照品适量、0.895 mg·mL-1的乙酰紫草素适量、0.090 mg·mL-1的去氧紫草素适量、0.900 mg·mL-1的β-乙酰氧基异戊酰阿卡宁适量、1.000 mg·mL-1的异丁酰阿卡宁适量,以及0.901 mg·mL-1的β,β'-二甲基丙烯酰阿卡宁适量,超声处理(功率:500 W,频率:40 kHz)30 min,放冷,石油醚定容至刻度,精密移取上清液10 mL至蒸发皿,蒸干,残渣加流动相乙腈-水(70∶30)溶解,转移至10 mL量瓶中,加流动相乙腈-水(70∶30)定容至刻度,摇匀,滤过,取续滤液,即得。按“2.1”项下色谱条件进样测定,记录峰面积,并计算加样回收率。结果发现,6个指标性成分的平均回收率分别为99.87%、99.93%、99.96%、101.03%、102.76%、98.50%,平均回收率RSD范围为1.51%~2.91%,见表5,表明该方法的回收率较好,均符合样品含量测定的要求。
2.7 样品的含量测定
分别取50批市售紫草药材粉末样品0.5 g,精密称定,按“2.2”项下方法制备供试品溶液。按“2.1”项下色谱条件进样测定,分别记录各成分峰面积,并计算其含量。结果见表6。结果发现,50批紫草中左旋紫草素的含量范围为0.006%~0.237%、乙酰紫草素0.009%~1.759%、去氧紫草素0.001%~0.129%,β-乙酰氧基异戊酰阿卡宁0.002%~0.625%、异丁酰阿卡宁0.011%~0.609%,β,β'-二甲基丙烯酰阿卡宁0.016%~1.712%,说明不同批次样品间含量差异极大,亦说明在市场上药材质量存在参差不齐的现象根据相似度评价可知该结论与指纹图谱结果一致。
2.8 含量测定结果分析
50批紫草样品存在含量差异,6个成分总含量的结果见图3。不同批次的紫草6个成分含量差异较大,其平均含量从大到小的排列顺序依次β,β'-二甲基丙烯酰阿卡宁>乙酰紫草素>异丁酰阿卡宁>β-乙酰氧基异戊酰阿卡宁>左旋紫草素>去氧紫草素,含量平均值依次为0.70%,0.41%,0.22%,0.18%,0.06%,0.02%。见表6和图3。
2.9 主成分分析
主成分分析(principal component analysis,PCA)是一种无监督的多元处理技术,可对数据处理中变异数据进行总结,它将会产生一个视觉散点图,可定性分析多元数据中的异同。为了更加清晰地区分50批紫草样品,将6种化合物的数据进行了主成分分析。将50批样品6种化合物的数据导入统计软件SPSS21.0中,进行主成分分析,计算相关系数特征值和方差贡献率,结果见表7。
以上6种成分按得分排序依次是乙酰紫草素、异丁酰阿卡宁、去氧紫草素、左旋紫草素、β-乙酰氧基异戊酰阿卡宁、和β,β'-二甲基丙烯酰阿卡宁,主成分分析中提取1~3个主成分即可代表紫草6个成分84%的信息量,说明前3个主成分即乙酰紫草素、异丁酰阿卡宁、去氧紫草素作为评价紫草的质量的指标性成分。
3 讨论
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本实验参考2020年版《中国药典》一部含量测定项下色谱条件选择和溶液制备方法,在其基础上进行梯度优化,建立了6个成分的HPLC分析方法。测定的6个萘醌类成分极性较小,试验前期对提取方式(回流提取法、超声提取法)、提取溶剂(乙醇、甲醇、石油醚)、复溶的溶剂体积分数70%乙腈、100%乙腈进行了考察,根据指纹图谱中色谱峰的分离度等信息,最终确定上述提取方法。另外课题组前期筛选了检测波长,结果显示当波长为275 nm时,均能检测到上述6种成分,且各峰的分离度良好,因此确定检测波长为275 nm[20]。
本实验以2020年版《中国药典》紫草项下检测方法“新疆紫草药材中β,β'-二甲基丙烯酰阿卡宁含量在0.3%以上”为评价标准,筛选符合条件的15批紫草药材,进行指纹图谱研究,确定共有峰,生成标准指纹图谱。并以此为标准进行市售药材的质量评价,发现50批样品的相似度结果在0.746~0.999内,说明紫草药材的质量差异较大,在成分含量上有一定的区别,但未呈现一定的规律性,这与含量测定的结果相一致。可以看出同来源于新疆维吾尔自治区的50批紫草成分含量差异较大,《中国药典》规定,紫草药材中指标性成分β,β'-二甲基丙烯酰阿卡宁不得少于0.30%,ZC52C、ZC59、ZC60均低于上述要求,在含量方面不符合规定,未达到药用要求。
通过主成分分析以及成分的方差贡献率,可以确定乙酰紫草素、异丁酰阿卡宁、去氧紫草素为评价紫草的质量的指标性成分。紫草为多来源多产地药材,主要的产区为我国新疆维吾尔自治区与内蒙古自治区,其道地产区面积辽阔,野生品与栽培品混杂,生长年限不确定,故紫草药材的质量不稳定、差异性较大。近年来随着栽培品的大力度种植,不同地区、不同生长年限的栽培品在市场上流通,加大了紫草药材质量控制的难度,后续需继续对不同来源、不同产区、不同生长年限的紫草野生品与栽培品进行质量控制。
4 结论
收起
实验旨在建立紫草的HPLC指纹图谱评价方法,以Agilent 5 TC-C18为色谱柱,乙腈-体积分数0.05%甲酸水溶液为流动相,30 min内梯度洗脱,发现该方法色谱峰分离度良好,精密度、重复性、稳定性均符合要求。通过化学模式分析技术,获得3个与原始变量关系最为密切的目标物,可推断是区分不同批次样品的关键成分,可加强对3个关键成分(乙酰紫草素、异丁酰阿卡宁、去氧紫草素)的考察,从而有效地保障紫草质量的一致性。综上,本实验所建立的多指标成分HPLC含量测定和指纹图谱分析方法,均可用于紫草药材的质量评价,可为制定科学合理、稳定可控的紫草质量标准提供基础数据。后续研究应对紫草的来源、产地、生长年限进行合理筛选找出符合《中国药典》规定的药材,以解决紫草资源匮乏的问题。
基金
收起
  • 紫草类民族药材的品质评价示范研究项目资助(2021X4)
  • 国家药品监督管理局药品注册司专项“特色民族药检验方法示范性研究”资助
  • 新疆维吾尔自治区药品监督管理局智力援疆创新拓展人才计划——“新疆特色民族药研究(以新疆紫草为例)团队”项目资助
文献
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2024年第59卷第10期
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文章信息
doi: 10.11669/cpj.2024.10.010
  • 接收时间：2023-11-10
  • 首发时间：2025-11-25
  • 出版时间：2024-05-22
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  • 收稿日期：2023-11-10
基金
紫草类民族药材的品质评价示范研究项目资助(2021X4)
国家药品监督管理局药品注册司专项“特色民族药检验方法示范性研究”资助
新疆维吾尔自治区药品监督管理局智力援疆创新拓展人才计划——“新疆特色民族药研究(以新疆紫草为例)团队”项目资助
作者信息
    1 中国食品药品检定研究院, 北京 102629
    2 中国药科大学, 南京 211198

通讯作者:

*郑健,女,博士,研究员 研究方向:民族药质量控制及相关标准研究 Tel:(010)53852080
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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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