Article(id=1153433742921163461, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20240923008, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1727020800000, receivedDateStr=2024-09-23, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752929634074, onlineDateStr=2025-07-19, pubDate=1745510400000, pubDateStr=2025-04-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752929634074, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752929634074, creator=13701087609, updateTime=1752929634074, updator=13701087609, issue=Issue{id=1153433737141412332, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='8', pageStart='1', pageEnd='316', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752929632696, creator=13701087609, updateTime=1757293087150, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1171735391666225233, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1171735391666225234, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433737141412332, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=244, endPage=252, ext={EN=ArticleExt(id=1153433743890047713, articleId=1153433742921163461, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Construction of Hengshan Astragalus fingerprint and identification of its origin based on ultra performance liquid chromatography-charged aerosol detector, columnId=1151895321388347923, journalTitle=Journal of Food Safety & Quality, columnName=Food Analysis and Detection, runingTitle=null, highlight=null, articleAbstract=

Objective To construct and compare the fingerprint profiles of Astragalus samples from different origins based on ultra performance liquid chromatography-charged aerosol detector (UPLC-CAD) technology, achieve quality evaluation and origin identification of Astragalus. Methods Using UPLC-CAD technology, this study focused on flavonoid and saponin components to construct the fingerprint of wild-simulated Astragalus membranaceus from the Hengshan Mountains, based on 29 batches of samples collected from multiple geographical origins. Statistical analysis methods such as cluster analysis, principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for analysis and evaluation. Results The fingerprint successfully marked 26 common peaks, and 8 key characteristic peaks were identified, including complanatoside A, ononin, astragaloside I to IV, etc. Cluster analysis clearly divided the samples into two major categories, revealing the influence of regional differences on the quality of Astragalus membranaceus. PCA further extracted 7 principal components. OPLS-DA analysis accurately predicted 8 kinds of key compounds influencing the quality differences, which were closely related to origin, cultivation methods, and growth duration. Conclusion This study successfully constructed a fingerprint capable of distinguishing Astragalus based on its origin, cultivation methods, and growth conditions by integrating UPLC-CAD technology with multivariate chemometric analysis. This method can be used to evaluate the fingerprint similarity of Astragalus membranaceus (Hengshan Mountain wild-simulated), with high specificity and sensitivity, which provids a scientific basis for the quality assessment of wild-simulated Astragalus from the Hengshan Mountains.

, correspAuthors=Juan DU, 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=Ying-Ying ZHAO, Ruo-Yu WANG, Kai-Zhi WU, Yun-Lan LIAN, Zhi-Lai ZHAN, Bao-Peng DING, Li-Ting YANG, Wen-Cui ZHANG, Xiao-Qin ZHAO, Juan DU), CN=ArticleExt(id=1153433783886930020, articleId=1153433742921163461, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=基于超高效液相色谱-电喷雾检测器构建恒山黄芪指纹图谱并鉴别其产地, columnId=1151895321958773274, journalTitle=食品安全质量检测学报, columnName=食品分析与检测, runingTitle=null, highlight=null, articleAbstract=

目的 基于超高效液相色谱-电喷雾检测器(ultra performance liquid chromatography-charged aerosol detector, UPLC-CAD)技术构建不同产地黄芪所含成分的指纹图谱, 并对其进行对比分析, 实现不同产地黄芪的质量评价与产地鉴别。方法 采用UPLC-CAD技术, 聚焦于黄酮类与皂苷类成分, 选取29批多个产地的样本绘制恒山山脉仿野生黄芪指纹图谱。运用聚类分析、主成分分析(principal component analysis, PCA)及正交偏最小二判别分析(orthogonal partial least squares discriminant analysis, OPLS-DA)等统计方法分析评价。结果 指纹图谱成功标记出26个共有峰, 鉴定出包括沙苑子苷A、芒柄花苷、黄芪皂苷Ⅰ至Ⅳ等在内的8个关键特征峰。聚类分析清晰地将样本分为两大类, 揭示了地域差异对黄芪质量的影响。PCA进一步提炼出7个主成分。OPLS-DA分析则精准预测了影响黄芪质量差异的8种关键化合物, 这些差异性化合物与产地、栽培方式及生长年限紧密相关。结论 本研究综合应用UPLC-CAD技术与多元化学计量学分析手段, 成功构建了能够区分不同基原、产地及栽培方式的黄芪指纹图谱, 可对黄芪(恒山山脉仿野生)进行指纹图谱相似度评价, 专属性强, 灵敏度高, 为恒山山脉仿野生黄芪的质量评价提供了科学依据。

, correspAuthors=杜娟, authorNote=null, correspAuthorsNote=
* 杜娟(1984—), 女, 硕士, 工程师, 主要研究方向为中药安全检测。E-mail:
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赵瑛瑛(1985—), 女, 硕士, 高级工程师, 主要研究方向为食品、中药、农产品等相关领域质量安全检测。E-mail:

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赵瑛瑛(1985—), 女, 硕士, 高级工程师, 主要研究方向为食品、中药、农产品等相关领域质量安全检测。E-mail:

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注: Y1. 甲醇超声提取; Y2. 为80%甲醇超声提取; Y3. 4%氨水的80%甲醇超声提取; Y4. 80%甲醇回流提取。

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注: 。表示数据偏离1倍偏差; 1. 黄芪(恒山山脉仿野生); 2. 黄芪(栽培膜荚黄芪); 3. 黄芪(陕西子洲仿野生); 4. 黄芪(栽培蒙古黄芪)。

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注: S1. 黄芪(恒山山脉仿野生)指纹图谱; S2. 膜荚黄芪; S3. 黄芪(陕西子洲仿野生); S4. 黄芪(内蒙、甘肃栽培)。

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注: C1、C5、C6、C15、C18、C19、C20、C22分别为毛蕊异黄酮葡萄糖苷、沙苑子A、芒丙花素、新芒丙花素、黄芪甲苷Ⅳ、黄芪甲苷Ⅲ、黄芪甲苷Ⅱ、黄芪甲苷Ⅰ。其他组分未采用对照品进行指认。

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注: C7、C18、C1、C22、C23、C2、C4、C20、C11、C24、C8、C15、C5分别组分7、黄芪甲苷Ⅳ、毛蕊异黄酮葡萄糖苷、黄芪甲苷Ⅰ、组分23、组分2、组分4、黄芪甲苷Ⅱ、组分11、组分24、组分8、新芒丙花素、沙苑子A。

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List of sample informations

, figureFileSmall=null, figureFileBig=null, tableContent=
编号 基原 样品名称 生长
年限/年
产地
S1 蒙古黄芪 黄芪(恒山山脉仿野生) 6 山西浑源凌云口
S2 蒙古黄芪 黄芪(恒山山脉仿野生) 8 山西浑源凌云口
S3 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西浑源凌云口
S4 蒙古黄芪 黄芪(恒山山脉仿野生) 12 山西浑源官儿乡
S5 蒙古黄芪 黄芪(恒山山脉仿野生) 12 山西浑源千佛岭乡
S6 蒙古黄芪 黄芪(恒山山脉仿野生) 10 山西浑源青磁窑乡
S7 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西浑源王庄堡乡
S8 蒙古黄芪 黄芪(恒山山脉仿野生) 6 山西浑源大仁庄乡
S9 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西浑源林场乡
S10 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西应县白马石乡
S11 蒙古黄芪 黄芪(恒山山脉仿野生) 9 山西应县白马石乡
S12 蒙古黄芪 黄芪(恒山山脉仿野生) 12 山西应县白马石乡
S13 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西广灵县梁庄镇
S14 蒙古黄芪 黄芪(恒山山脉仿野生) 8 山西广灵县梁庄镇
S15 蒙古黄芪 黄芪(恒山山脉仿野生) 9 山西广灵县梁庄镇
S16 膜荚黄芪 黄芪(内蒙包头栽培) 5 内蒙古包头市
S17 膜荚黄芪 黄芪(内蒙包头栽培) 2 内蒙古包头市
S18 膜荚黄芪 黄芪(内蒙赤峰栽培) 2 内蒙古赤峰市
S19 膜荚黄芪 黄芪(内蒙围场栽培) 1 河北围场
S20 膜荚黄芪 黄芪(河北围场栽培) 1 河北围场
S21 蒙古黄芪 黄芪(陕西子洲仿野生) 7 陕西子洲
S22 蒙古黄芪 黄芪(陕西子洲仿野生) 8 陕西子洲
S23 蒙古黄芪 黄芪(陕西子洲仿野生) 9 陕西子洲
S24 蒙古黄芪 黄芪(内蒙固阳栽培) 2 内蒙固阳
S25 蒙古黄芪 黄芪(内蒙武川栽培) 2 内蒙武川
S26 蒙古黄芪 黄芪(内蒙武川栽培) 2 内蒙武川
S27 蒙古黄芪 黄芪(甘肃陇西栽培) 2 甘肃陇西
S28 蒙古黄芪 黄芪(甘肃定西栽培) 2 甘肃定西
S29 蒙古黄芪 黄芪(甘肃定西栽培) 2 甘肃定西
), ArticleFig(id=1171734020133004156, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=CN, label=表1, caption=

样品信息列表

, figureFileSmall=null, figureFileBig=null, tableContent=
编号 基原 样品名称 生长
年限/年
产地
S1 蒙古黄芪 黄芪(恒山山脉仿野生) 6 山西浑源凌云口
S2 蒙古黄芪 黄芪(恒山山脉仿野生) 8 山西浑源凌云口
S3 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西浑源凌云口
S4 蒙古黄芪 黄芪(恒山山脉仿野生) 12 山西浑源官儿乡
S5 蒙古黄芪 黄芪(恒山山脉仿野生) 12 山西浑源千佛岭乡
S6 蒙古黄芪 黄芪(恒山山脉仿野生) 10 山西浑源青磁窑乡
S7 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西浑源王庄堡乡
S8 蒙古黄芪 黄芪(恒山山脉仿野生) 6 山西浑源大仁庄乡
S9 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西浑源林场乡
S10 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西应县白马石乡
S11 蒙古黄芪 黄芪(恒山山脉仿野生) 9 山西应县白马石乡
S12 蒙古黄芪 黄芪(恒山山脉仿野生) 12 山西应县白马石乡
S13 蒙古黄芪 黄芪(恒山山脉仿野生) 7 山西广灵县梁庄镇
S14 蒙古黄芪 黄芪(恒山山脉仿野生) 8 山西广灵县梁庄镇
S15 蒙古黄芪 黄芪(恒山山脉仿野生) 9 山西广灵县梁庄镇
S16 膜荚黄芪 黄芪(内蒙包头栽培) 5 内蒙古包头市
S17 膜荚黄芪 黄芪(内蒙包头栽培) 2 内蒙古包头市
S18 膜荚黄芪 黄芪(内蒙赤峰栽培) 2 内蒙古赤峰市
S19 膜荚黄芪 黄芪(内蒙围场栽培) 1 河北围场
S20 膜荚黄芪 黄芪(河北围场栽培) 1 河北围场
S21 蒙古黄芪 黄芪(陕西子洲仿野生) 7 陕西子洲
S22 蒙古黄芪 黄芪(陕西子洲仿野生) 8 陕西子洲
S23 蒙古黄芪 黄芪(陕西子洲仿野生) 9 陕西子洲
S24 蒙古黄芪 黄芪(内蒙固阳栽培) 2 内蒙固阳
S25 蒙古黄芪 黄芪(内蒙武川栽培) 2 内蒙武川
S26 蒙古黄芪 黄芪(内蒙武川栽培) 2 内蒙武川
S27 蒙古黄芪 黄芪(甘肃陇西栽培) 2 甘肃陇西
S28 蒙古黄芪 黄芪(甘肃定西栽培) 2 甘肃定西
S29 蒙古黄芪 黄芪(甘肃定西栽培) 2 甘肃定西
), ArticleFig(id=1171734020225278845, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=EN, label=Table 2, caption=

Results of recovery test

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序号 取样量/g 样品中含量/µg 加入量/µg 测得量/µg 回收率/% 平均回收率/% RSDs/%
1 0.5011 200.44 102.50 311.34 108.20 105.56 2.75
2 0.5013 200.52 102.50 309.21 106.04
3 0.5024 200.96 102.50 305.97 102.45
4 0.5023 200.92 205.00 402.21 98.19 98.82 0.59
5 0.5001 200.04 205.00 402.87 98.94
6 0.5003 200.12 205.00 403.76 99.34
7 0.5011 200.44 306.75 508.74 100.51 99.99 0.69
8 0.5012 200.48 306.75 507.99 100.25
9 0.5015 200.60 306.75 504.91 99.20
), ArticleFig(id=1171734020296582014, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=CN, label=表2, caption=

回收率试验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
序号 取样量/g 样品中含量/µg 加入量/µg 测得量/µg 回收率/% 平均回收率/% RSDs/%
1 0.5011 200.44 102.50 311.34 108.20 105.56 2.75
2 0.5013 200.52 102.50 309.21 106.04
3 0.5024 200.96 102.50 305.97 102.45
4 0.5023 200.92 205.00 402.21 98.19 98.82 0.59
5 0.5001 200.04 205.00 402.87 98.94
6 0.5003 200.12 205.00 403.76 99.34
7 0.5011 200.44 306.75 508.74 100.51 99.99 0.69
8 0.5012 200.48 306.75 507.99 100.25
9 0.5015 200.60 306.75 504.91 99.20
), ArticleFig(id=1171734020393051007, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=EN, label=Table 3, caption=

Identification list of standard feature peak

, figureFileSmall=null, figureFileBig=null, tableContent=
峰号 标准品 标准品保留时间/min 样品中相应色谱峰保留时间/min
1 毛蕊异黄酮葡萄糖苷 3.600 3.610
5 沙苑子苷A 10.395 10.389
7 芒柄花苷 12.405 12.407
16 新芒柄花素 32.390 32.392
18 黄芪皂苷Ⅳ 38.065 38.065
19 黄芪皂苷Ⅲ 38.518 38.512
21 黄芪皂苷Ⅱ 40.978 40.974
22 黄芪皂苷Ⅰ 47.592 47.590
), ArticleFig(id=1171734020460159872, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=CN, label=表3, caption=

标准品特征峰指认列表

, figureFileSmall=null, figureFileBig=null, tableContent=
峰号 标准品 标准品保留时间/min 样品中相应色谱峰保留时间/min
1 毛蕊异黄酮葡萄糖苷 3.600 3.610
5 沙苑子苷A 10.395 10.389
7 芒柄花苷 12.405 12.407
16 新芒柄花素 32.390 32.392
18 黄芪皂苷Ⅳ 38.065 38.065
19 黄芪皂苷Ⅲ 38.518 38.512
21 黄芪皂苷Ⅱ 40.978 40.974
22 黄芪皂苷Ⅰ 47.592 47.590
), ArticleFig(id=1171734020531463041, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=EN, label=Table 4, caption=

Eigenvalue and variance contribution rate of principal component

, figureFileSmall=null, figureFileBig=null, tableContent=
主成分 初始特征值 方差贡献率% 累积贡献率%
1 8.122 31.239 31.239
2 4.014 15.438 46.678
3 2.773 10.661 57.342
4 2.305 8.865 66.207
5 2.004 7.706 73.913
6 1.431 5.505 79.418
7 1.167 4.490 83.908
), ArticleFig(id=1171734020602766210, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=CN, label=表4, caption=

主成分特征值和方差贡献率

, figureFileSmall=null, figureFileBig=null, tableContent=
主成分 初始特征值 方差贡献率% 累积贡献率%
1 8.122 31.239 31.239
2 4.014 15.438 46.678
3 2.773 10.661 57.342
4 2.305 8.865 66.207
5 2.004 7.706 73.913
6 1.431 5.505 79.418
7 1.167 4.490 83.908
), ArticleFig(id=1171734020682457987, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=EN, label=Table 5, caption=

List of principal component matrices

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 1 2 6
毛蕊异黄酮葡萄糖苷 0.832
组分2 0.848
组分4 0.814
组分7 0.698
组分11 0.734
黄芪甲苷Ⅳ 0.728
黄芪甲苷Ⅱ 0.789
黄芪甲苷Ⅰ 0.850
组分23 0.834
组分24 0.666
组分8 0.801
新芒丙花素 0.722
沙苑子A 0.781
), ArticleFig(id=1171734020753761156, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433742921163461, language=CN, label=表5, caption=

主成分矩阵列表

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 1 2 6
毛蕊异黄酮葡萄糖苷 0.832
组分2 0.848
组分4 0.814
组分7 0.698
组分11 0.734
黄芪甲苷Ⅳ 0.728
黄芪甲苷Ⅱ 0.789
黄芪甲苷Ⅰ 0.850
组分23 0.834
组分24 0.666
组分8 0.801
新芒丙花素 0.722
沙苑子A 0.781
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基于超高效液相色谱-电喷雾检测器构建恒山黄芪指纹图谱并鉴别其产地
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赵瑛瑛 1 , 王若昱 2, 3 , 武开智 1 , 连云岚 2 , 詹志来 4 , 丁保朋 5 , 杨丽婷 1 , 张文翠 1 , 赵晓琴 1 , 杜娟 2, *
食品安全质量检测学报 | 食品分析与检测 2025,16(8): 244-252
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食品安全质量检测学报 | 食品分析与检测 2025, 16(8): 244-252
基于超高效液相色谱-电喷雾检测器构建恒山黄芪指纹图谱并鉴别其产地
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赵瑛瑛1 , 王若昱2, 3, 武开智1, 连云岚2, 詹志来4, 丁保朋5, 杨丽婷1, 张文翠1, 赵晓琴1, 杜娟2, *
作者信息
  • 1.大同市综合检验检测中心, 大同 037000
  • 2.山西省检验检测中心, 太原 030031
  • 3.山西中医药大学中药与食品工程学院, 晋中 030619
  • 4.中国中医科学院中药资源中心, 北京 100700
  • 5.山西大同大学煤基生态碳汇技术教育部工程研究中心, 大同 037009
  • 赵瑛瑛(1985—), 女, 硕士, 高级工程师, 主要研究方向为食品、中药、农产品等相关领域质量安全检测。E-mail:

通讯作者:

* 杜娟(1984—), 女, 硕士, 工程师, 主要研究方向为中药安全检测。E-mail:
Construction of Hengshan Astragalus fingerprint and identification of its origin based on ultra performance liquid chromatography-charged aerosol detector
Ying-Ying ZHAO1 , Ruo-Yu WANG2, 3, Kai-Zhi WU1, Yun-Lan LIAN2, Zhi-Lai ZHAN4, Bao-Peng DING5, Li-Ting YANG1, Wen-Cui ZHANG1, Xiao-Qin ZHAO1, Juan DU2, *
Affiliations
  • 1. Datong Comprehensive Inspection and Testing Center, Datong 037000, China
  • 2. Shanxi Inspection and Testing Center, Taiyuan 030031, China
  • 3. Shanxi University of Chinese Medicine College of Traditional Chinese Medicine and Food Engineering, Jinzhong 030619, China
  • 4. Chinese Medicine Resource Center, China Academy of Chinese Medicine Sciences, Beijing 100700, China
  • 5. Engineering Research Center of Coal-based Ecological Carbon Sequestration Technology of the Ministry of Education and Key Laboratory of National Forest, Shanxi Datong University, Datong 037009, China
出版时间: 2025-04-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20240923008
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目的 基于超高效液相色谱-电喷雾检测器(ultra performance liquid chromatography-charged aerosol detector, UPLC-CAD)技术构建不同产地黄芪所含成分的指纹图谱, 并对其进行对比分析, 实现不同产地黄芪的质量评价与产地鉴别。方法 采用UPLC-CAD技术, 聚焦于黄酮类与皂苷类成分, 选取29批多个产地的样本绘制恒山山脉仿野生黄芪指纹图谱。运用聚类分析、主成分分析(principal component analysis, PCA)及正交偏最小二判别分析(orthogonal partial least squares discriminant analysis, OPLS-DA)等统计方法分析评价。结果 指纹图谱成功标记出26个共有峰, 鉴定出包括沙苑子苷A、芒柄花苷、黄芪皂苷Ⅰ至Ⅳ等在内的8个关键特征峰。聚类分析清晰地将样本分为两大类, 揭示了地域差异对黄芪质量的影响。PCA进一步提炼出7个主成分。OPLS-DA分析则精准预测了影响黄芪质量差异的8种关键化合物, 这些差异性化合物与产地、栽培方式及生长年限紧密相关。结论 本研究综合应用UPLC-CAD技术与多元化学计量学分析手段, 成功构建了能够区分不同基原、产地及栽培方式的黄芪指纹图谱, 可对黄芪(恒山山脉仿野生)进行指纹图谱相似度评价, 专属性强, 灵敏度高, 为恒山山脉仿野生黄芪的质量评价提供了科学依据。

恒山黄芪  /  超高效液相色谱法  /  电喷雾检测器  /  指纹图谱  /  化学计量学分析

Objective To construct and compare the fingerprint profiles of Astragalus samples from different origins based on ultra performance liquid chromatography-charged aerosol detector (UPLC-CAD) technology, achieve quality evaluation and origin identification of Astragalus. Methods Using UPLC-CAD technology, this study focused on flavonoid and saponin components to construct the fingerprint of wild-simulated Astragalus membranaceus from the Hengshan Mountains, based on 29 batches of samples collected from multiple geographical origins. Statistical analysis methods such as cluster analysis, principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for analysis and evaluation. Results The fingerprint successfully marked 26 common peaks, and 8 key characteristic peaks were identified, including complanatoside A, ononin, astragaloside I to IV, etc. Cluster analysis clearly divided the samples into two major categories, revealing the influence of regional differences on the quality of Astragalus membranaceus. PCA further extracted 7 principal components. OPLS-DA analysis accurately predicted 8 kinds of key compounds influencing the quality differences, which were closely related to origin, cultivation methods, and growth duration. Conclusion This study successfully constructed a fingerprint capable of distinguishing Astragalus based on its origin, cultivation methods, and growth conditions by integrating UPLC-CAD technology with multivariate chemometric analysis. This method can be used to evaluate the fingerprint similarity of Astragalus membranaceus (Hengshan Mountain wild-simulated), with high specificity and sensitivity, which provids a scientific basis for the quality assessment of wild-simulated Astragalus from the Hengshan Mountains.

Hengshan Astragalus  /  ultra performance liquid chromatography  /  charged aerosol detector  /  fingerprint  /  stoichiometric analysis
赵瑛瑛, 王若昱, 武开智, 连云岚, 詹志来, 丁保朋, 杨丽婷, 张文翠, 赵晓琴, 杜娟. 基于超高效液相色谱-电喷雾检测器构建恒山黄芪指纹图谱并鉴别其产地. 食品安全质量检测学报, 2025 , 16 (8) : 244 -252 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20240923008
Ying-Ying ZHAO, Ruo-Yu WANG, Kai-Zhi WU, Yun-Lan LIAN, Zhi-Lai ZHAN, Bao-Peng DING, Li-Ting YANG, Wen-Cui ZHANG, Xiao-Qin ZHAO, Juan DU. Construction of Hengshan Astragalus fingerprint and identification of its origin based on ultra performance liquid chromatography-charged aerosol detector[J]. Journal of Food Safety & Quality, 2025 , 16 (8) : 244 -252 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20240923008
《中国药典》2020年版一部中, 黄芪来源为豆科植物蒙古黄芪或膜荚黄芪的干燥根。具有补气升阳, 固表止汗, 利水消肿, 生津养血, 行滞通痹, 托毒排脓, 敛疮生肌的作用[1]。现代药理学研究表明, 黄芪具有抗衰老[2-4]、抗肿瘤[5-7]、免疫调节[8-9]、降糖调脂[10-12]、抗纤维化[13-16]等多种药理作用。黄芪不仅有着悠久的药用历史, 目前在人们的日常生活中也应用广泛。2023年国家卫生健康委员会、国家市场监督管理总局将其纳入《按照传统既是食品又是中药材的物质目录》[17]。自宋代以来医家普遍认同山西、内蒙所产绵黄芪品质最佳[18], 而久负盛名的黄芪(恒山山脉仿野生)曾作为进献朝廷的直贡药材“贡品”, 来源于蒙古黄芪[19]。2014年4月16日, 国家质量监督检验检疫总局批准“恒山黄芪”实施地理标志产品保护[20], 因此有必要对恒山黄芪质量特性进行研究。
目前黄芪的有效成分研究主要集中于以毛蕊异黄酮葡萄糖苷为代表的黄酮类成分[21-22]和以黄芪甲苷等为代表的皂苷类成分[23-24], 但黄酮类成分多具有紫外吸收而皂苷类成分为末端吸收, 无法做到同时对两种成分的同时评价与研究。随着现代检测技术的发展, 电喷雾检测器(charged aerosol detector, CAD)[25]作为一种高灵敏度的通用性检测器, 具备同时检测具有强紫外吸收特性的黄酮类成分和不具备紫外吸收特性的皂苷类成分的技术优势[26-27]。目前多采用高效液相色谱法、液相色谱-质谱法等方法, 分别建立黄芪指纹图谱, 但皂苷类成分很难达到理想的响应和检测[28-30]。鉴于此, 本研究通过超高效液相色谱-CAD (ultra performance liquid chromatography- CAD, UPLC-CAD)技术同时对黄芪中两大类有效成分进行分析, 建立(恒山山脉仿野生)的指纹图谱, 通过现代分析和检测技术, 对黄芪中的化学信息以图形(图像)方式进行表征, 从中提取有用的信息, 并结合化学计量学手段评估分析黄芪(恒山山脉仿野生)与其他产地以及不同栽培方式黄芪的质量差异, 以期为恒山黄芪的质量评价提供理论依据。
Vanquish系列超高效液相色谱仪、Thermo Hypersil GOLDTMVANQUISH色谱柱(2.1 mm×100 mm, 1.9 µm)(美国赛默飞世尔科技公司); LC-350A超声清洗机(山东济宁鲁超声设备有限公司); HH-6单列六孔电热恒温水浴锅(金坛区白塔新宝仪器厂); MS204S电子分析天平(精度0.1 mg)、MS105DU电子分析天平(精度0.01 mg)(瑞士Mettler Toledo公司)。
黄芪皂苷Ⅰ(批号: 230225, 纯度≥98%)、黄芪皂苷Ⅱ(批号: 230206, 纯度≥98%)、黄芪皂苷Ⅲ(批号: 230112, 纯度≥98%)、黄芪皂苷Ⅳ(批号: 202219, 纯度≥98%)、沙苑子苷(批号: 221117, 纯度≥98%)、芒柄花苷(批号: 220914, 纯度≥98%)、新芒柄花素(批号: 221014, 纯度≥98%)、毛蕊异黄酮葡萄糖苷(批号: 202308, 纯度≥96.9%)(伟业计量-国家标准物质研究中心); 乙腈、甲醇(色谱纯, 德国默克集团公司); 甲醇、氨水(分析纯, 国药集团化学试剂有限公司); 纯化水(超纯水, 默克Milli-Q® IQ 7000水纯化系统)。
研究用黄芪样品均从产地收集, 其中黄芪(恒山山脉仿野生)15批, 黄芪(陕西子洲仿野生)3批, 膜荚黄芪5批, 黄芪(内蒙、甘肃栽培的蒙古黄芪)6批。均经山西省检验检测中心连云岚主任药师鉴定为豆科植物蒙古黄芪或膜荚黄芪的干燥根。样品信息见表1
分别精密称取黄芪皂苷Ⅰ、Ⅱ、Ⅲ、Ⅳ, 沙苑子苷A, 芒柄花苷, 新芒柄花素及毛蕊异黄酮葡萄糖苷标准品适量, 加80%甲醇分别制成20 μg/mL的标准品溶液, 即得。
提取溶剂研究: 取本品粉末(过四号筛)约1 g, 精密称定, 置具塞锥形瓶中, 分别精密加入①甲醇; ②80%甲醇溶液; ③含4%氨水的80%甲醇溶液[31]50 mL, 密塞, 称定重量, 加热回流[32]1 h, 放冷, 再称定重量, 用80%甲醇溶液补足减失的重量, 摇匀, 滤过, 取续滤液, 即得。
提取方法研究: 取本品粉末(过四号筛)约1 g, 精密称定, 置具塞锥形瓶中, 精密加入80%甲醇溶液50 mL, 密塞, 称定重量, 加热回流1 h或超声提取(250 W, 33 kHz) 1 h, 放冷, 再称定重量, 用80%甲醇溶液补足减失的重量, 摇匀, 滤过, 取续滤液, 即得。
色谱柱: Thermo Hypersil GOLDTMVANQUISH (2.1 mm× 100 mm, 1.9 µm); 以0.2%甲酸水溶液为流动相A, 以乙腈为流动相B, 进行梯度洗脱: 0~15 min, 15%~20% B; 15~25 min, 20%~25% B; 25~30 min, 25% B; 30~40 min, 25%~37% B; 40~50 min, 37% B; 50~55 min, 37%~46% B; 流速: 0.3 mL/min; 柱温: 30 ℃; 电喷雾式检测器检测, 雾化温度为35 ℃, 数据采集频率为10 Hz, 过滤常数(F)为1.0 (S)。理论板数按黄芪甲苷IV峰计, 均不得低于10000。进样量为2 μL。
运用“中药色谱指纹图谱相似度评价系统”2.0版本, 以15批次黄芪(恒山山脉仿野生)为原始图谱进行多点校正, 自动化匹配等构建黄芪(恒山山脉仿野生)指纹图谱。
本研究所呈现的实验数据, 均为3次测量后, 得出的平均值±标准偏差。在开展数据的统计和分析工作时, 使用的软件是SPSS 21.0, 在实施试验数据的绘图作业时, 采用的软件是OriginPro 2021、SIMCA 14.1。
通过对比甲醇、80%甲醇及含4%氨水的80%甲醇3种溶剂的提取效果, 结果表明, 80%甲醇作为提取溶剂时, 其色谱图中不仅色谱峰的数目多, 且峰面积较大。进一步对比加热回流与超声两种提取方式结果表明, 两者色谱图中色谱峰的数量较为一致, 但加热回流提取的色谱图中峰面积相对较大。结果见图1, 最终选取以80%甲醇为溶剂, 加热回流1 h, 作为本方法的供试品溶液制备方法。
选取黄芪(编号: S1)作为方法学研究用样品。采用1.3.2项下方法制备供试品溶液, 按1.3.3项下色谱条件连续进样6次, 以黄芪甲苷I色谱峰做为参照峰(S), 计算各共有峰的相对保留时间和相对峰面积, 相对标准偏差(relative standard deviations, RSDs)为0.03%~0.06%、0.9%~1.7%, 表明仪器精密度良好。
取精密度试验下供试品溶液, 在室温下放置0、4、8、12、16、24 h进行测定, 以22号峰黄芪甲苷I色谱峰为参照峰(S), 计算各共有峰的相对保留时间和相对峰面积, RSDs分别为0.04%~0.87%、0.26%~2.87%, 说明该供试品溶液在24 h内稳定。
取同一批样品, 根据1.3.2项下方法平行制备6份供试品溶液, 以22号峰黄芪甲苷I的相对保留时间和相对峰面积做参照, 计算其RSD值, 得到各共有峰的相对保留时间的RSDs为0.02%~0.5%, 相对峰面积的RSDs为1.7%~2.1%, 表明该方法的重复性良好。
通过22号峰黄芪甲苷I对方法的线性关系进行考察。精密量取黄芪皂苷Ⅰ对照品溶液适量, 用甲醇逐级稀释, 得到系列对照品溶液(质量浓度分别为4.10、10.25、13.67、20.5、30.75 μg/mL), 进样, 记录峰面积。以质量浓度为横坐标(X, μg/mL), 峰面积为纵坐标(Y), 进行线性回归, 结果质量浓度在4.10~30.75 μg/mL, 线性范围内, Y=-0.000002X2+ 0.0076X+0.0041 (r2=0.9999), 线性关系良好。
取S1号黄芪样品约0.5 g, 分别精密加入低、中、高质量浓度的黄芪皂苷Ⅰ对照品溶液(2.05、4.10、6.14 μg/mL) 50 mL, 按1.3.2项下方法平行制备9份供试品溶液, 进样测定, 结果平均回收率为99.82%~105.56%, 见表2。精密度与准确度满足实验要求。
选择黄芪(恒山山脉仿野生)15批次样品, 按既定方法制备和测定供试品, 并记录色谱峰, 将原始数据导入“中药色谱指纹图谱相似度评价系统”2.0版本, 以其中稳定出现, 分离度好, 保留时间一致的26个共有峰进行标记, 生成黄芪(恒山山脉仿野生)指纹图谱。选择黄芪的质量标志物22号峰黄芪皂苷I作为参照峰(见图2)。
按既定方法测定标准品和样品溶液, 并记录色谱图, 通过与标准品的比对, 对26个共有峰中8个指纹峰进行了指认, 其中, 1号峰为毛蕊异黄酮葡萄糖苷, 5号峰为沙苑子苷A, 7号峰为芒柄花苷, 16号峰为新芒柄花素, 18号峰为黄芪皂苷Ⅳ, 19号峰为黄芪皂苷Ⅲ, 21号峰为黄芪皂苷Ⅱ, 22号峰为黄芪皂苷Ⅰ, 结果见表3
采用本研究建立的方法将其余产地的14批样品的指纹图谱与对照指纹图谱进行自动匹配, 其相似度为0.930~0.995(见图3)。黄芪(恒山山脉仿野生)的相似度主要为0.978~0.995, 膜荚黄芪的相似度主要为0.948~0.958, 黄芪(陕西子洲仿野生)的相似度主要为0.930~0.941, 该方法可以较好地区分不同基源的蒙古黄芪与膜荚黄芪, 也可以较好地区分不同产地蒙古黄芪的黄芪(恒山山脉仿野生)与黄芪(陕西子洲仿野生); 栽培的黄芪主要为内蒙、甘肃产的蒙古黄芪整体相似度跨度较大, 主要为0.957~0.958与黄芪(恒山山脉仿野生)质量相似度也可以得到较好地区分。不同基原、产地的与栽培黄芪的典型图谱详见图4
使用OriginPro 2021软件, 以29批黄芪样品指纹图谱的共有峰的峰面积为变量进行热图分析, 得到指纹图谱热图聚类分析图(见图5), 当类间距为25时, 样品被分为两类, S2~S13、S15、S22、S23聚为一类; S1、S14、S16~S21、S24~S29聚为一类。结果表明, 黄芪(恒山山脉仿野生)与其他产地黄芪(不论野生还是栽培)所含有的化学成分存在一定差异, 可通过指纹图谱各色谱峰的峰面积对不同基源的黄芪进行初步分类。
以29批黄芪样品的指纹图谱共有峰的峰面积为变量, 运用SPSS 21.0软件对其进行主成分分析(principal component analysis, PCA), 得到凯撒-米尔-奥尔金检验值(Kaiser-Meyer-Olkin, KMO)>0.5, 表明变量之间是存在相关性的, 此分析模型可靠, 可以用于区分不同基源的黄芪品种。根据特征值和累计贡献率以及初始特征值>1为标准, 得到7个主成分, 累计贡献率达到83.908%, 表明这7个主成分可以代表变量包含的大部分信息, 结果见表4。依据绝对值分析, 第1主成分的贡献度来自毛蕊异黄酮葡萄糖苷、组分2、组分4、组分7、组分11、黄芪甲苷Ⅳ、黄芪甲苷Ⅱ、黄芪甲苷Ⅰ、组分23、组分24、组分8; 第2主成分的贡献度来自8号峰、新芒丙花素; 第6主成分的贡献度来自沙苑子A, 结果见表5。以贡献度较大的成分的峰面积作为变量, 运用SIMCA14.1软件分析, 得到PCA得分图(见图6), 图中显示, 29批黄芪分为两组, S1、S16~S18、S19~S21、S24~S29为一组, S2~S15、S22、S23为一组。
为深入分析不同产地不同基源样品间的差异性成分, 建立正交偏最小二乘判别分析(orthogonal partial least squares discriminant analysis, OPLS-DA)分析模型(见图7), t1轴将样品分为2部分。自变量拟合数R2X为0.66, 因变量拟合数R2Y为0.85, 模型预测指数Q2为0.768, 认为R2Q2超过0.5表示模型拟合结果可接受, 能用于不同品种、不同产地、不同栽培方式的黄芪区分。经过200次置换检验, 比较R2Q2值的位置, 表明该模型不存在过度拟合, 可对不同基源、不同栽培方式的黄芪进行有效判别(见图8)。进一步以变量重要性投影(variable importance projection, VIP)值> 1为筛选标准(见图9), 得到7号峰、黄芪皂苷Ⅳ、黄芪皂苷Ⅰ、毛蕊异黄酮葡萄糖苷、2号峰、4号峰、黄芪皂苷Ⅱ、23号峰等8个差异性化合物。
本研究采用CAD检测器, 共检测到26个出峰稳定且可重现的指纹峰, 建立了恒山黄芪指纹图谱, 与传统的紫外检测器相比, CAD检测器[33]在黄芪皂苷类成分的检测中展现出了独特的优势。鉴定出包括沙苑子苷A、芒柄花苷、黄芪皂苷Ⅰ至Ⅳ等在内的8个关键特征峰。在建立指纹图谱时, 采用了“中药色谱指纹图谱相似度评价系统”2.0版本, 并对29批次黄芪的相识度进行自动匹配, 其相似度为0.930~0.995。其中黄芪(恒山山脉仿野生)的相似度主要为0.978~0.995, 膜荚黄芪的相似度主要为0.948~0.958, 黄芪(陕西子洲仿野生)的相似度主要为0.930~0.941; 栽培的黄芪主要为内蒙、甘肃产的蒙古黄芪整体相似度跨度较大, 主要为0.957~0.958, 与黄芪(恒山山脉仿野生)质量相似度也可以得到较好地区分。聚类分析清晰地将样本分为两大类, 黄芪(恒山山脉仿野生)与其他产地黄芪(不论野生还是栽培)所含有的化学成分存在一定差异, 通过PCA、OPLS-DA分析则精准预测了影响黄芪质量差异的8种关键化合物, 而恒山黄芪在这8个成分量值方面表现为质量分析与评价评判优异, 揭示了地域差异对黄芪质量的影响, 这些差异与产地、栽培方式及生长年限紧密相关。
本研究综合应用UPLC-CAD技术与多元化学计量学分析手段, 通过26个指纹峰成功构建了能够区分不同基原、产地及栽培方式的黄芪指纹图谱, 可对黄芪(恒山山脉仿野生)进行指纹图谱相似度评价, 与其他检测方法[34-35]相比该方法专属性强, 灵敏度高, 可为恒山山脉仿野生黄芪的质量分析与研究提供参考依据和方法, 为黄芪野生、仿野生资源的保护和利用提供技术支撑。
  • 大同市重点研发计划项目(2022022)
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2025年第16卷第8期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20240923008
  • 接收时间:2024-09-23
  • 首发时间:2025-07-19
  • 出版时间:2025-04-25
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  • 收稿日期:2024-09-23
基金
大同市重点研发计划项目(2022022)
作者信息
    1.大同市综合检验检测中心, 大同 037000
    2.山西省检验检测中心, 太原 030031
    3.山西中医药大学中药与食品工程学院, 晋中 030619
    4.中国中医科学院中药资源中心, 北京 100700
    5.山西大同大学煤基生态碳汇技术教育部工程研究中心, 大同 037009

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* 杜娟(1984—), 女, 硕士, 工程师, 主要研究方向为中药安全检测。E-mail:
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2种不同金属材料的力学参数

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

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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