Article(id=1240997642382135362, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240997638351409170, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024-0400, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1718380800000, receivedDateStr=2024-06-15, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773806494564, onlineDateStr=2026-03-18, pubDate=1740672000000, pubDateStr=2025-02-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773806494564, onlineIssueDateStr=2026-03-18, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773806494564, creator=13701087609, updateTime=1773806494564, updator=13701087609, issue=Issue{id=1240997638351409170, tenantId=1146029695717560320, journalId=1205117023404326918, year='2025', volume='45', issue='2', pageStart='181', pageEnd='360', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773806493604, creator=13701087609, updateTime=1773810140860, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241012936110560131, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240997638351409170, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241012936110560132, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240997638351409170, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=254, endPage=264, ext={EN=ArticleExt(id=1240997643510403218, articleId=1240997642382135362, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Multi-instrumental approach to investigate the causes of excessive phloem fascicles in commercially available “Ophiopogonis Radix”, columnId=1206272757852074373, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Safety Monitoring, runingTitle=null, highlight=null, articleAbstract=
Objective:

To identify the authenticity of four batches of “Ophiopogonis Radix” in the market by means of multiple means, and to explore the reasons for exceeding the limit of its phloem bundles, so as to provide evidence for its inspection and detection.

Methods:

Based on the relevant provisions of Ophiopogon japonicus (L. f.) Ker-Gawl., a variety in the 2020 edition of Chinese Pharmacopoeia, comparing the common confused products of Ophiopogon japonicus (L. f.) Ker-Gawl., Combined with traditional identification methods (character identification, microscopic identification) and modern analysis techniques (molecular biology ITS 2 sequence), the data of genuine products with mixed products and substandard phloem bundle samples were campared and analyzed.

Results:

The differences between Liriope spicata (Thunb.) Lour. and Ophiopogon japonicus (L. f.) Ker-Gawl. were the surface color, the depth of vertical wrinkles and the thickness of the middle column, and the differences in microscopic cross-sections lay in the number of phloem bundles and whether the inner cortex cells were uniformly thickened. The four batches of “Ophiopogonis Radix” in the market all complied with the relevant regulations under Ophiopogon japonicus (L. f.) Ker-Gawl.. The number of unqualified phloem bundled into microscopic cross-sections. was more than 40%, but the inner cortex cells in the samples showed a comprehensive thickening phenomenon. The results of molecular biology study showed that the four batches of “Ophiopogonis Radix” on the market and the Ophiopogon japonicus (L. f.) Ker-Gawl. were clustered into one, the Liriope spicata (Thunb.) Lour.var. prolifera Y. T. Ma and the Liriope muscari (Decne.) Baily were clustered into one, and showed obvious bar code spacing. The number of phloem bundles was obvious positively correlated with the diameter of wood core by correlation analysis of the number of phloem bundles and the quantitative indexes related to traits.

Conclusion:

These four batches of “Ophiopogonis Radix” on the market are the original of genuine products. Combined with the experimental research, it is speculated that the cause of the unqualified number of phloem bundles may be related to the growth years, and the middle column thickness can roughly predict whether it can meet the requirements of pharmacopoeia. To determine whether Ophiopogonis Radix is an accurate medicinal material based on the original, we should not only make a conclusion based on a certain feature of a certain identification method, but also should combine multiple methods to determine accurately.

, correspAuthors=Lei YANG, Yan GOU, 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=Bi-xing GAO, Qian LI, Jing-liang QI, Cheng-jun HE, Fan-hong LIU, Tian-yuan GAO, Chi GAO, Lei YANG, Yan GOU), CN=ArticleExt(id=1240997647482409245, articleId=1240997642382135362, tenantId=1146029695717560320, journalId=1205117023404326918, language=CN, title=多手段结合探究市售“麦冬”韧皮部束超标的原因*, columnId=1206272758036623764, journalTitle=药物分析杂志, columnName=安全监测, runingTitle=null, highlight=null, articleAbstract=
目的:

多手段结合鉴别4批市售“麦冬”的真伪,探究其韧皮部束超标的原因,为其检验检测提供依据。

方法:

基于2020年版《中华人民共和国药典》(简称《中国药典》)一部麦冬项下的相关规定,对比麦冬常见混淆品山麦冬,结合传统经验鉴别手段(性状、显微鉴别)及现代分析技术手段(分子生物学ITS2序列),针对正品、混淆品及韧皮部束不合格样本进行数据比较分析。

结果:

麦冬与山麦冬性状的差异在于表面颜色、纵皱纹的深浅及中柱粗细,显微横切的差异在于韧皮部束个数及内皮层细胞是否全面增厚。4批市售麦冬性状均符合麦冬项下相关规定,显微横切中韧皮部束的个数不合格批次均达40%以上,但样本中内皮层细胞均呈现全面增厚现象。分子生物学研究发现,4批市售“麦冬”与正品麦冬聚为一支,湖北麦冬、短葶山麦冬各独自聚成一支,并表现出明显的条形码间距,综合判定其应均为麦冬。另外,针对韧皮部束的个数与性状的相关量化指标,进行了相关性分析,发现韧皮部束的个数与木心的直径大小呈显著正相关。

结论:

此4批市售麦冬应为正品基原。结合实验研究推测,韧皮部束的个数不合格原因可能与生长年限有关,中柱粗细可大致推测是否能达到药典规定。判定麦冬是否为基原准确的药材,不能仅凭某一鉴别手段的某一特征下定论,应多手段结合分析,方能准确地进行判定。

, correspAuthors=杨蕾, 苟琰, authorNote=null, correspAuthorsNote=
**杨蕾 Tel:(028)87877181;E-mail:
苟琰 Tel:(028)87877143;E-mail:
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Tel:(028)87877143;E-mail:

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pmid=null, pmcid=null, year=2021, volume=17, issue=6, pageStart=193, pageEnd=null, url=null, language=null, rfNumber=[14], rfOrder=26, authorNames=王丽, 杨乾展, 张加余, journalName=亚太传统医药, refType=null, unstructuredReference=王丽,杨乾展,张加余,等.川、浙麦冬质量差异研究概况 [J].亚太传统医药202117(6):193, articleTitle=川、浙麦冬质量差异研究概况, refAbstract=null), Reference(id=1241033154018988688, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, doi=null, pmid=null, pmcid=null, year=2021, volume=17, issue=6, pageStart=193, pageEnd=null, url=null, language=null, rfNumber=[14], rfOrder=27, authorNames=WANG L, YANG QZ, ZHANG YJ, journalName=Asia Pac Tradit Med, refType=null, unstructuredReference=WANG LYANG QZZHANG YJ,et al.A review on the quality variation of Ophiopogonis Radix cultivated in Sichuan and Zhejiang[J].Asia Pac Tradit Med202117(6):193, articleTitle=A review on the quality variation of Ophiopogonis Radix cultivated in Sichuan and Zhejiang, refAbstract=null)], 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A.麦冬(Ophiopogon japonicus) B.湖北麦冬(Liriope spicata)C.短葶山麦冬(Liriope muscari

, figureFileSmall=3d0wE1v3MveLt+xJdx7gYw==, figureFileBig=rp4yliJDZGax3O7HNwKyDw==, tableContent=null), ArticleFig(id=1241033147215827458, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=EN, label=Fig. 4, caption=Microscopic comparison diagram, figureFileSmall=jEmMeR+mHbAoj/NfqCP8lg==, figureFileBig=qbERUn2nrgNMBbVHxflq/A==, tableContent=null), ArticleFig(id=1241033147287130630, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=CN, label=图4, caption=显微对比图

1.麦冬对口药材(Ophiopogon japonicus) 2.湖北麦冬对口药材(Liriope spicata) 3.短葶山麦冬对口药材(Liriope muscari) 4.韧皮部束不合格样品(failed samples for number of phloem bundles)

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Sample information sheet

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序号(No.)名称(name)编号(serial number)来源(origin)
1麦冬Ophiopogon japonicus(L. f.)Ker-Gawl.MD-1四川省绵阳市三台县(自采)[Santai County, Mianyang City, Sichuan Province(self-collected samples)]
2MD-2四川省绵阳市三台县(自采)[Santai County, Mianyang City, Sichuan Province(self-collected samples)]
3麦冬对照药材Ophiopogon japonicus(L. f.)Ker-Gawl. as controlMD-DZ中国食品药品检定研究院(National Institutes for Food and Drug Control )
4湖北麦冬Liriope spicata(Thunb.)Lour. var. prolifera Y. T. MaHBMD-1湖北省襄阳市九襄镇(自采)[Jiuxiang Town, Xiangyang City, Hubei Province(self-collected samples)]
5HBMD-2湖北省襄阳市九襄镇(自采)[Jiuxiang Town, Xiangyang City, Hubei Province(self-collected samples)]
6HBMD-3湖北省襄阳市宜城市(自采)[Yicheng City, Xiangyang City, Hubei Province(self-collected samples)]
7湖北麦冬对照药材Liriope spicata(Thunb.)Lour. var. prolifera Y. T. Ma as controlHBMD-DZ中国食品药品检定研究院(National Institutes for Food and Drug Control )
8短葶山麦冬Liriope muscari(Decne.)BailyDT四川省药品检验研究院标本室(自采)[Specimen Room,Sichuan Institute for Drug Control(self-collected samples)]
9市场麦冬Ophiopogon japonicus(L. f.)Ker-1企业提供(corporate offerings)
10Gawl. from market2企业提供(corporate offerings)
113企业提供(corporate offerings)
124企业提供(corporate offerings)
), ArticleFig(id=1241033148725776955, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=CN, label=表1, caption=

样品来源信息表

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序号(No.)名称(name)编号(serial number)来源(origin)
1麦冬Ophiopogon japonicus(L. f.)Ker-Gawl.MD-1四川省绵阳市三台县(自采)[Santai County, Mianyang City, Sichuan Province(self-collected samples)]
2MD-2四川省绵阳市三台县(自采)[Santai County, Mianyang City, Sichuan Province(self-collected samples)]
3麦冬对照药材Ophiopogon japonicus(L. f.)Ker-Gawl. as controlMD-DZ中国食品药品检定研究院(National Institutes for Food and Drug Control )
4湖北麦冬Liriope spicata(Thunb.)Lour. var. prolifera Y. T. MaHBMD-1湖北省襄阳市九襄镇(自采)[Jiuxiang Town, Xiangyang City, Hubei Province(self-collected samples)]
5HBMD-2湖北省襄阳市九襄镇(自采)[Jiuxiang Town, Xiangyang City, Hubei Province(self-collected samples)]
6HBMD-3湖北省襄阳市宜城市(自采)[Yicheng City, Xiangyang City, Hubei Province(self-collected samples)]
7湖北麦冬对照药材Liriope spicata(Thunb.)Lour. var. prolifera Y. T. Ma as controlHBMD-DZ中国食品药品检定研究院(National Institutes for Food and Drug Control )
8短葶山麦冬Liriope muscari(Decne.)BailyDT四川省药品检验研究院标本室(自采)[Specimen Room,Sichuan Institute for Drug Control(self-collected samples)]
9市场麦冬Ophiopogon japonicus(L. f.)Ker-1企业提供(corporate offerings)
10Gawl. from market2企业提供(corporate offerings)
113企业提供(corporate offerings)
124企业提供(corporate offerings)
), ArticleFig(id=1241033148809663037, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=EN, label=Tab. 2, caption=

GenBank login number information of ITS2 sequence for different origins

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拉丁名(latin name)GenBank 登录号(GenBank registry number)
Ophiopogon japonicus(L. f.)Ker-Gawl.EU930852.1、KF671258.1、MG595277.1、EU930854.1、EU930852.1、MG595276.1、KF671259.1、JF327842.1
Liriope spicata(Thunb.)Lour. var. prolifera Y. T. MaKJ571534.1、EU930857.1、KJ571536.1、KJ571535.1、KF878941.1
Liriope muscari(Decne.)BailyMW714922.1、MF096464.1、MF096463.1、MF096462.1、JF327836.1、JF327833.1、EU930855.1、EU930856.1、
), ArticleFig(id=1241033148906132032, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=CN, label=表2, caption=

不同基原ITS 2序列的GenBank登录号信息

, figureFileSmall=null, figureFileBig=null, tableContent=
拉丁名(latin name)GenBank 登录号(GenBank registry number)
Ophiopogon japonicus(L. f.)Ker-Gawl.EU930852.1、KF671258.1、MG595277.1、EU930854.1、EU930852.1、MG595276.1、KF671259.1、JF327842.1
Liriope spicata(Thunb.)Lour. var. prolifera Y. T. MaKJ571534.1、EU930857.1、KJ571536.1、KJ571535.1、KF878941.1
Liriope muscari(Decne.)BailyMW714922.1、MF096464.1、MF096463.1、MF096462.1、JF327836.1、JF327833.1、EU930855.1、EU930856.1、
), ArticleFig(id=1241033149036155460, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=EN, label=Tab. 3, caption=

Analysis of intraspecific and interspecific genetic distances

, figureFileSmall=null, figureFileBig=null, tableContent=
物种
(species)
距离(distance)
麦冬(Ophiopogon japonicas湖北麦冬(Liriope spicata短葶山麦冬(Liriope muscari
麦冬(Ophiopogon japonicas0~0.0046(0.0008)0.0676~0.0946(0.0824)0.0730~0.0888(0.0795)
湖北麦冬(Liriope spicata0.0676~0.0946(0.0824)0~0.0179(0.0100)0.0180~0.0320(0.0253)
短葶山麦冬(Liriope muscari0.0730~0.0888(0.0795)0.0180~0.0320(0.0253)0~0.0137(0.0041)
), ArticleFig(id=1241033149124235846, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=CN, label=表3, caption=

种内种间遗传距离分析

, figureFileSmall=null, figureFileBig=null, tableContent=
物种
(species)
距离(distance)
麦冬(Ophiopogon japonicas湖北麦冬(Liriope spicata短葶山麦冬(Liriope muscari
麦冬(Ophiopogon japonicas0~0.0046(0.0008)0.0676~0.0946(0.0824)0.0730~0.0888(0.0795)
湖北麦冬(Liriope spicata0.0676~0.0946(0.0824)0~0.0179(0.0100)0.0180~0.0320(0.0253)
短葶山麦冬(Liriope muscari0.0730~0.0888(0.0795)0.0180~0.0320(0.0253)0~0.0137(0.0041)
), ArticleFig(id=1241033149237482059, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=EN, label=Tab. 4, caption=

Length, maximum diameter, mass, number of phloem pieces and core diameter measurements of Ophiopogon japonicus

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编号
(serial number)
最粗端直径
(diameter of the thickest end)/ cm
长度
(lengths)/ cm
质量
(mass)/ g
韧皮部束个数
(number of phloem pieces)
木心直径
(core diameter)/ μm
1-10.5922.5140.403 315714.31
1-20.3221.7260.204 411475.01
1-30.5122.0000.357 422875.20
1-40.2941.2110.197 715530.02
1-50.2821.3220.101 116479.22
1-60.4121.3010.092 710437.51
1-70.4241.5000.192 819750.02
1-80.4261.7110.172 515525.71
1-90.4521.4140.112 516563.24
1-100.3121.2980.081 19412.55
2-10.4221.6920.316 413594.13
2-20.3921.6120.172 97358.87
2-30.4141.7520.242 512500.98
2-40.3261.7140.150 119761.68
2-50.2141.5220.089 010398.69
2-60.4141.5120.192 920863.26
2-70.4021.4820.133 016637.08
2-80.4561.4120.228 315622.89
2-90.3781.4140.125 817665.93
2-100.3921.3020.112 910451.19
3-10.4121.6820.257 99485.49
3-20.5121.6940.312 221824.52
3-30.5922.2420.228 222769.99
3-40.2942.2240.131 919738.95
3-50.4241.3320.116 89472.56
3-60.4921.7940.238 419885.51
3-70.4961.6020.223 718882.11
3-80.3021.5540.070 511450.56
3-90.4761.6940.220 616759.87
3-100.3121.6120.129 49489.24
4-10.4223.0120.448 621961.25
4-20.3141.7020.112 66389.89
4-30.3921.7840.133 519721.62
4-40.4061.7020.144 515537.34
4-50.3961.9120.183 111489.81
4-60.2961.7960.141 013712.61
4-70.3922.3540.161 114604.53
4-80.4041.2940.144 016655.11
4-90.4561.6540.220 918794.92
4-100.4121.7120.193 717673.41
), ArticleFig(id=1241033149321368142, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240997642382135362, language=CN, label=表4, caption=

麦冬的长度、最粗端直径、质量、韧皮部束个数及木心直径测量结果

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(serial number)
最粗端直径
(diameter of the thickest end)/ cm
长度
(lengths)/ cm
质量
(mass)/ g
韧皮部束个数
(number of phloem pieces)
木心直径
(core diameter)/ μm
1-10.5922.5140.403 315714.31
1-20.3221.7260.204 411475.01
1-30.5122.0000.357 422875.20
1-40.2941.2110.197 715530.02
1-50.2821.3220.101 116479.22
1-60.4121.3010.092 710437.51
1-70.4241.5000.192 819750.02
1-80.4261.7110.172 515525.71
1-90.4521.4140.112 516563.24
1-100.3121.2980.081 19412.55
2-10.4221.6920.316 413594.13
2-20.3921.6120.172 97358.87
2-30.4141.7520.242 512500.98
2-40.3261.7140.150 119761.68
2-50.2141.5220.089 010398.69
2-60.4141.5120.192 920863.26
2-70.4021.4820.133 016637.08
2-80.4561.4120.228 315622.89
2-90.3781.4140.125 817665.93
2-100.3921.3020.112 910451.19
3-10.4121.6820.257 99485.49
3-20.5121.6940.312 221824.52
3-30.5922.2420.228 222769.99
3-40.2942.2240.131 919738.95
3-50.4241.3320.116 89472.56
3-60.4921.7940.238 419885.51
3-70.4961.6020.223 718882.11
3-80.3021.5540.070 511450.56
3-90.4761.6940.220 616759.87
3-100.3121.6120.129 49489.24
4-10.4223.0120.448 621961.25
4-20.3141.7020.112 66389.89
4-30.3921.7840.133 519721.62
4-40.4061.7020.144 515537.34
4-50.3961.9120.183 111489.81
4-60.2961.7960.141 013712.61
4-70.3922.3540.161 114604.53
4-80.4041.2940.144 016655.11
4-90.4561.6540.220 918794.92
4-100.4121.7120.193 717673.41
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多手段结合探究市售“麦冬”韧皮部束超标的原因*
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高必兴 , 李倩 , 齐景梁 , 何成军 , 刘繁红 , 高天元 , 高驰 , 杨蕾 ** , 苟琰 **
药物分析杂志 | 安全监测 2025,45(2): 254-264
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药物分析杂志 | 安全监测 2025, 45(2): 254-264
多手段结合探究市售“麦冬”韧皮部束超标的原因*
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高必兴 , 李倩, 齐景梁, 何成军, 刘繁红, 高天元, 高驰, 杨蕾** , 苟琰**
作者信息
  • 四川省药品检验研究院 国家药品监督管理局中成药质量评价重点实验室工业和信息化部产业技术基础公共服务平台,成都 611731
  • Tel:(028)87877143;E-mail:

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**杨蕾 Tel:(028)87877181;E-mail:
苟琰 Tel:(028)87877143;E-mail:
Multi-instrumental approach to investigate the causes of excessive phloem fascicles in commercially available “Ophiopogonis Radix”
Bi-xing GAO , Qian LI, Jing-liang QI, Cheng-jun HE, Fan-hong LIU, Tian-yuan GAO, Chi GAO, Lei YANG** , Yan GOU**
Affiliations
  • Sichuan Institute for Drug Control, Key Laboratory of Quality Evaluation of Chinese Patent Medicines, MIIT Public Service Platforms for Industrial Technological Base, NMPA, Chengdu 611731, China
出版时间: 2025-02-28 doi: 10.16155/j.0254-1793.2024-0400
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目的:

多手段结合鉴别4批市售“麦冬”的真伪,探究其韧皮部束超标的原因,为其检验检测提供依据。

方法:

基于2020年版《中华人民共和国药典》(简称《中国药典》)一部麦冬项下的相关规定,对比麦冬常见混淆品山麦冬,结合传统经验鉴别手段(性状、显微鉴别)及现代分析技术手段(分子生物学ITS2序列),针对正品、混淆品及韧皮部束不合格样本进行数据比较分析。

结果:

麦冬与山麦冬性状的差异在于表面颜色、纵皱纹的深浅及中柱粗细,显微横切的差异在于韧皮部束个数及内皮层细胞是否全面增厚。4批市售麦冬性状均符合麦冬项下相关规定,显微横切中韧皮部束的个数不合格批次均达40%以上,但样本中内皮层细胞均呈现全面增厚现象。分子生物学研究发现,4批市售“麦冬”与正品麦冬聚为一支,湖北麦冬、短葶山麦冬各独自聚成一支,并表现出明显的条形码间距,综合判定其应均为麦冬。另外,针对韧皮部束的个数与性状的相关量化指标,进行了相关性分析,发现韧皮部束的个数与木心的直径大小呈显著正相关。

结论:

此4批市售麦冬应为正品基原。结合实验研究推测,韧皮部束的个数不合格原因可能与生长年限有关,中柱粗细可大致推测是否能达到药典规定。判定麦冬是否为基原准确的药材,不能仅凭某一鉴别手段的某一特征下定论,应多手段结合分析,方能准确地进行判定。

麦冬  /  山麦冬  /  短葶山麦冬  /  性状  /  显微鉴别  /  分子生物学
Objective:

To identify the authenticity of four batches of “Ophiopogonis Radix” in the market by means of multiple means, and to explore the reasons for exceeding the limit of its phloem bundles, so as to provide evidence for its inspection and detection.

Methods:

Based on the relevant provisions of Ophiopogon japonicus (L. f.) Ker-Gawl., a variety in the 2020 edition of Chinese Pharmacopoeia, comparing the common confused products of Ophiopogon japonicus (L. f.) Ker-Gawl., Combined with traditional identification methods (character identification, microscopic identification) and modern analysis techniques (molecular biology ITS 2 sequence), the data of genuine products with mixed products and substandard phloem bundle samples were campared and analyzed.

Results:

The differences between Liriope spicata (Thunb.) Lour. and Ophiopogon japonicus (L. f.) Ker-Gawl. were the surface color, the depth of vertical wrinkles and the thickness of the middle column, and the differences in microscopic cross-sections lay in the number of phloem bundles and whether the inner cortex cells were uniformly thickened. The four batches of “Ophiopogonis Radix” in the market all complied with the relevant regulations under Ophiopogon japonicus (L. f.) Ker-Gawl.. The number of unqualified phloem bundled into microscopic cross-sections. was more than 40%, but the inner cortex cells in the samples showed a comprehensive thickening phenomenon. The results of molecular biology study showed that the four batches of “Ophiopogonis Radix” on the market and the Ophiopogon japonicus (L. f.) Ker-Gawl. were clustered into one, the Liriope spicata (Thunb.) Lour.var. prolifera Y. T. Ma and the Liriope muscari (Decne.) Baily were clustered into one, and showed obvious bar code spacing. The number of phloem bundles was obvious positively correlated with the diameter of wood core by correlation analysis of the number of phloem bundles and the quantitative indexes related to traits.

Conclusion:

These four batches of “Ophiopogonis Radix” on the market are the original of genuine products. Combined with the experimental research, it is speculated that the cause of the unqualified number of phloem bundles may be related to the growth years, and the middle column thickness can roughly predict whether it can meet the requirements of pharmacopoeia. To determine whether Ophiopogonis Radix is an accurate medicinal material based on the original, we should not only make a conclusion based on a certain feature of a certain identification method, but also should combine multiple methods to determine accurately.

Ophiopogon japonicus (L. f.) Ker-Gawl.  /  Liriope spicata (Thunb.) Lour.  /  Liriope muscari (Decne.) Baily  /  character  /  microscopic identification  /  molecular biology
高必兴, 李倩, 齐景梁, 何成军, 刘繁红, 高天元, 高驰, 杨蕾, 苟琰. 多手段结合探究市售“麦冬”韧皮部束超标的原因*. 药物分析杂志, 2025 , 45 (2) : 254 -264 . DOI: 10.16155/j.0254-1793.2024-0400
Bi-xing GAO, Qian LI, Jing-liang QI, Cheng-jun HE, Fan-hong LIU, Tian-yuan GAO, Chi GAO, Lei YANG, Yan GOU. Multi-instrumental approach to investigate the causes of excessive phloem fascicles in commercially available “Ophiopogonis Radix”[J]. Chinese Journal of Pharmaceutical Analysis, 2025 , 45 (2) : 254 -264 . DOI: 10.16155/j.0254-1793.2024-0400
麦冬来源于百合科植物麦冬Ophiopogon japonicus(L.f.)Ker-Gawl.的干燥块根,可养阴生津、润肺清心,用于肺燥干咳、阴虚痨嗽、喉痹咽痛、津伤口渴、内热消渴、心烦失眠、肠燥便秘[1],收载于2020年版《中华人民共和国药典》(简称《中国药典》)一部,主要培育方式为栽培,按产地分为川麦冬及浙麦冬。川麦冬主要分布于我国四川省绵阳市涪江流域,1年生,以三台县为其最大产区;浙麦冬主要分布于浙江省慈溪、台州、宁波等地,2~3年生,但市场占有量少,价格高[2-3]。除麦冬外,2020年版《中国药典》一部还收载了山麦冬,为来源于百合科植物湖北麦冬Liriope spicata (Thunb.) Lour. var. prolifera Y. T. Ma或短葶山麦冬Liriope muscari (Decne.) Baily的干燥块根[1],以基原湖北麦冬市场占比大,主产于湖北省襄阳市等地。山麦冬价格远低于麦冬,市场上常有山麦冬混入麦冬使用[4]。显微鉴别中的韧皮部束数量一直为区分麦冬(标准规定为16~22个)与山麦冬(标准规定为7~15个)的主要鉴别特征,前期检验发现,有4批麦冬样品因韧皮部束个数未符合标准规定,本研究基于传统鉴别手段(性状、显微鉴别)及现代分析技术手段(分子生物学ITS 2序列[5-7]),分析韧皮部束个数不合规的原因,判断是否为山麦冬的混入或其他因素而导致,同时为其检验检测提供依据。
SZX 16体式显微镜、BX 63光学显微镜(奥林巴斯公司),C 1000型聚合酶链式反应(PCR)仪、PowerPac Basic型电泳仪、Gel Doc XR+型全自动凝胶成像系统(Bio-Rad公司);XP 205型十万分之一电子分析天平(Mettler Toledo公司);MM 400型球磨仪(Retsch公司);MiniSpin型高速离心机(Eppendorf公司);Milli-Q Advantage A10型纯水仪(Millipore公司)。
水合氯醛、丙三醇(成都市科隆化学品有限公司),植物基因组DNA提取试剂盒、2×Taq Master Mix缓冲液、Gene Red核酸染料、Taq酶PCR扩增试剂盒、琼脂糖、ITS2引物(ITS2F: 5’-ATGCGATACTTGGTGTGAAT-3’;ITS3R: 5’-GACGCTTCTCCAGACTACAAT-3’)均由生工生物工程(上海)股份有限公司购买及合成。
收集对口药材8批,其中麦冬3批,山麦冬(湖北麦冬)4批,山麦冬(短葶山麦冬)1批,经四川省药品检验研究院齐景梁副主任中药师鉴定为基原准确的药材。同时于GenBank数据库中下载21条ITS 2序列,包括麦冬8条,山麦冬(湖北麦冬)5条,山麦冬(短葶山麦冬)8条。样品来源及其ITS 2序列信息分别见表12
取实验样品4批,观察其性状,照2020年版《中国药典》一部麦冬项下性状的规定并对比市场常见混淆品湖北麦冬及短葶山麦冬进行观察。采用四分法分取样品并各批次随机挑取10根,使用单反相机进行拍照,同时使用游标卡尺及十万分之一天平记录每批次10根麦冬的长度、最粗端直径及质量,后将每根分为两部分,一部分照2020年版《中国药典》四部2001显微鉴定法进行显微横切研究,使用显微成像系统并结合偏光拍摄,记录韧皮部束个数,并使用显微软件测量工具(已通过成都计量院计量合格)测量木心直径,同时对比湖北麦冬及短葶山麦冬的显微横切。相应示意图见图1图2
正品麦冬较湖北麦冬表面颜色深,纵皱纹明显,同时中柱粗,较短葶山麦冬饱满,纵皱纹以短葶山麦冬更为明显。依据上述结果,结合2020年版《中国药典》一部麦冬性状项下的相关规定,因其与正品麦冬更为接近,故应为麦冬药材[8]。三者主要差异点表面颜色、纵皱纹及中柱的形态描述如图3所示。
3种麦冬的横切主要鉴别点在韧皮部束个数及内皮层细胞是否增厚,韧皮部束个数麦冬与短葶山麦冬相似,但都多于湖北麦冬;内皮层细胞除麦冬为全面增厚外,湖北麦冬及短葶山麦冬均三面增厚或侧壁部分增厚。市售4批次韧皮部束个数(规定应为16~22个)不合格的数量分别为6根、6根、4根、5根,但无论韧皮部束个数合格与否,其内皮层细胞均全面增厚。通过对口药材显微主要鉴别点无法准确地确定其是否为麦冬,故结合分子生物学进行进一步验证,以便针对显微鉴别特征给予正确的依据[9-10]。3种麦冬的横切区别及市售4批次挑选“麦冬”的性状、显微镜下明场及偏光的木心特征分别如图4~图8所示。
挑选市场麦冬1号样品全部根数(包含合格与不合格)、4批市场麦冬韧皮部全部不合格根数、麦冬对口药材2批、山麦冬(湖北麦冬)对口药材3批、山麦冬(短葶山麦冬)对口药材1批、麦冬对照药材、湖北麦冬对照药材,照2020年版《中国药典》四部9107中药材DNA条形码分子鉴定法指导原则进行研究,使用核糖体DNA第二内部转录间隔区(ITS 2)为主体条形码序列鉴定中药材的方法体系进行评判,并从GenBank上下载相关序列21条[(麦冬8条,山麦冬(湖北麦冬)5条,山麦冬(短葶山麦冬)8条]。具体研究方法及结果如下。
用75%乙醇擦拭样品表面,晾干,液氮冷冻后用球磨仪研磨成粉末,称取粉末约20 mg,照植物基因组DNA提取试剂盒说明书(生工生物工程(上海)股份有限公司)进行操作,提取DNA,-20 ℃保存备用。
选取通用引物ITS2F(5’-ATGCGATACTTGGTGTGAAT-3’)和ITS3R(5’-GACGCTTCTCCAGACTACAAT-3’)对样品DNA进行扩增。反应体系为50 μL,包含2×Taq Master Mix缓冲液25 μL、上下游引物各2 μL、DNA模板5 μL,加灭菌超纯水16 μL;阴性对照为无模板DNA的反应体系;以10 000 r·min-1离心5 s,置于PCR仪中。反应程序:95 ℃预变性4 min,94 ℃变性30 s,56 ℃退火30 s,72 ℃延伸45 s,35个循环;72 ℃延伸5 min。PCR产物用1.5%琼脂糖凝胶电泳检测以确定成功率及目的条带片段大小,扩增产物委托生工生物工程(上海)股份有限公司进行双向测序[5]
应用CodonCode Aligner V6.0对测序所获得的数据进行校对拼接,去除引物及低质量区,基于隐马尔科夫模型(HMMer)的注释方法去除5.8 S和28 S区段,即可获得ITS 2序列信息。
进入美国国家生物技术信息中心(NCBI,https://blast.ncbi.nlm.nih.gov/Blast.Cgi)和中药材DNA条形码鉴定系统(http://barcode.ndctcm.org/china/),应用相似性搜索法对实验样品的不同单倍型序列进行BLAST鉴定分析。
应用MEGA 6.0软件进行序列比对,对序列的变异位点进行分析,不同物种的单倍型为A1、B1,同一物种存在变异位点的单倍型为A1、A2;基于K2P模型计算种内、种间遗传距离,利用邻接法(Neighbor-joining,NJ)构建系统聚类树,同时以Bootstrap自展支持率(1 000次)重复检验各分支的支持率[5]
基于ITS 2序列的NJ系统聚类树,观察各基原准确的三类麦冬、市售麦冬韧皮部束数合格批次及全部不合格批次间的聚类情况,Bootstrap 1 000次重复,支上数值仅显示自展支持率≥50%。可以看出,市售麦冬韧皮部束数合格批次及全部不合格批次均能与基原准确的麦冬(自采麦冬、麦冬对照药材及网上下载麦冬的数据)以100%的自展支持率聚为1支,表现出了很好的单系性。同时,自采及下载的湖北麦冬、自采及下载的短葶山麦冬以大于50%的自展支持率各自聚为1支,表现出了良好的单系性。说明市售麦冬韧皮部束数不合格批次应为麦冬。如图9所示。
麦冬ITS 2序列长度均为219 bp,湖北麦冬ITS 2序列长度均为226 bp,短葶山麦冬ITS 2序列长度均为226 bp。所有样品鸟嘌呤和胞嘧啶(GC)的含量为73.0%~78.0%,平均GC含量为74.9%。3种麦冬间存在25个变异位点,种内麦冬存在2个变异位点,形成3个单倍型;湖北麦冬存在3个变异位点,形成4个单倍型;短葶山麦冬存在1变异位点,形成2个单倍型。如图10所示。
3种麦冬的种内和种间遗传距离的结果(表3)显示,种内最大遗传距离远小于种间最小遗传距离,具有明显的条形码间距(barcoding gap),说明K2P距离可很好地区分开此3种麦冬。
针对4批次10根麦冬(共计40根麦冬)的长度、最粗端直径及质量进行测量,同时对40根麦冬的长度、最粗端直径、质量、木心直径与韧皮部个数使用SPSS进行相关性分析发现:韧皮部束的个数与长度(r=0.321,P=0.043)、最粗端直径(r=0.467,P=0.002)、质量(r=0.431,P=0.006)、木心直径(r=0.907,P=0.000 2)均具有显著性差异,存在一定的正相关,且与木心直径最为相关。见表4图11
本研究基于正品及混淆品,使用传统经验鉴别手段(性状及显微鉴别)寻找差异,并结合现代分子生物学技术,探究4批市售“麦冬”的真伪及部分“麦冬”韧皮部束不合格的原因,旨在为麦冬的检验检测提供实验依据。前期调研发现,市场上常见的混淆品有湖北麦冬及短葶山麦冬,但以湖北麦冬最为常见。4批麦冬性状中,表面颜色黄白色,不规则纵纹较深且木心较粗者,应为麦冬;但显微横切中韧皮部束批次1~4不合格率分别为60%、60%、40%、50%。所有不合格麦冬中柱内皮层细胞均全面增厚。同时,通过分子生物学研究发现,4批市售麦冬均应为麦冬。前期针对4批市售麦冬中所含山麦冬特有性成分(山麦冬皂苷B及短葶山麦冬皂苷C)进行了检测,结果均未检出。结合性状、显微、分子生物学研究及特有性成分的检测,判定4批市售“麦冬”应为正品基原。提示检验者在判断麦冬是否为正品时,不能拘泥于某一检项中某一部分的结果而草草下定结论,应结合多种鉴别手段共同判定。
当传统鉴别方法面临挑战时,可以依据2020年版《中国药典》四部通则[4]收载的中药材DNA条形码分子鉴定法指导原则,采用ITS 2序列作为鉴定手段。这种方法不受样本个体形态特征和完整性的限制,易于实现方法的统一和标准化,是传统鉴定方法的有力补充[8]。结合正伪品及下载序列,使用聚类分析和K2P遗传距离分析,可作为鉴定的有效补充。
本研究同时对正品麦冬韧皮部束不合格的原因进行了初步的探索及思考。研究发现,韧皮部束的个数与木心直径最为相关,相关系数达0.907,说明麦冬韧皮部束个数不合格与木心的直径存在着一定的相关性。经验鉴别中麦冬的木心直径比湖北麦冬的大,韧皮部束个数麦冬的比湖北麦冬的多(麦冬规定为16~22个,湖北麦冬规定为7~15个)。这些数据不仅验证了实验的准确性,也反映了传统经验在鉴别麦冬与湖北麦冬方面的可靠性。
前期研究发现,浙麦冬韧皮部束个数及木心直径往往超过川麦冬,甚至会超过标准的限度。川麦冬通常生长1年,浙麦冬则生长2年以上,因此,推测正品麦冬韧皮部束不合格的原因可能与其生长年限有关。但是,这不能作为判定浙麦冬的依据,而应结合多指标综合判定[14]。中药的种植过程复杂多变,具体原因仍需继续深入调研探究。
  • *四川省药品监督管理局2025年药品科技计划项目(2025001)
  • 国家药品监督管理局药品监管科学体系建设重点项目“新技术新方法在中药质量控制中的应用”(RS2024Z006)
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2025年第45卷第2期
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doi: 10.16155/j.0254-1793.2024-0400
  • 接收时间:2024-06-15
  • 首发时间:2026-03-18
  • 出版时间:2025-02-28
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*四川省药品监督管理局2025年药品科技计划项目(2025001)
国家药品监督管理局药品监管科学体系建设重点项目“新技术新方法在中药质量控制中的应用”(RS2024Z006)
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    四川省药品检验研究院 国家药品监督管理局中成药质量评价重点实验室工业和信息化部产业技术基础公共服务平台,成都 611731

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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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