Article(id=1239173813817111235, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239173808419033435, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024.02.03, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=1700582400000, revisedDateStr=2023-11-22, acceptedDate=null, acceptedDateStr=null, onlineDate=1773371659950, onlineDateStr=2026-03-13, pubDate=1709136000000, pubDateStr=2024-02-29, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773371659950, onlineIssueDateStr=2026-03-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773371659950, creator=13701087609, updateTime=1773371659950, updator=13701087609, issue=Issue{id=1239173808419033435, tenantId=1146029695717560320, journalId=1205117023404326918, year='2024', volume='44', issue='2', pageStart='185', pageEnd='372', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773371658663, creator=13701087609, updateTime=1773371757717, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1239174223944536397, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239173808419033435, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1239174223944536398, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239173808419033435, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=214, endPage=223, ext={EN=ArticleExt(id=1239173814085546702, articleId=1239173813817111235, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Determination and characterization of Poriae Cutis multi-component content based on wavelength conversion RP-HPLC method*, columnId=1239148838318043851, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Ingredient Analys, runingTitle=null, highlight=null, articleAbstract=
Objective:

To comprehensively evaluate the quality of Poriae Cutis, and to establish a dual wavelength switching HPLC method for comparing the characteristic spectra of Poriae Cutis and studying the content of 11 triterpenoid components, to provide reference for the qualitative and quantitative research of Poriae Cutis.

Methods:

Agilent 5 HC-C18(2) column(250 mm×4.6 mm, 5 μm) was adopted. Acetonitrile solution (contain 3% tetrahydrofuran) (A) and 0.1% formic acid aqueous solution (B) were used as the mobile phase with gradient elution at a flow rate of 1.0 mL·min-1. The column temperature was 30 ℃ and the injection volume was 20 μL. The detection wavelengths were 210 and 243 nm.

Results:

The feature profiles developed were effective in identifying the 18 shared peaks. RSD for precision, repeatability and stability (48 h) tests were all less than 3.72%(n=6). The 11 chemical components to be measured were well separated, with good linearity in the mass range examined (all r ≥ 0.999 6). The average recovery rate was 95.4%-105.5%, and the RSD was 1.0%-3.1%. The RSDs of precision, repeatability, and stability (48 h) tests were all less than or equal to 3.0%(n=6). The results of similarity analysis showed that most of the origins of Poriae Cutis were very similar to each other. The results of content determination showed that among the 11 triterpenoid constituents, poricoic acid A accounted for the highest percentage in all batches of Poriae Cutis. In addition, the content of five components, poricoic acid A, dehydrotrametenolic acid, poricoic acid B, dehydroeburicoic acid and trametenolic acid, fluctuated relatively more, while the other components fluctuated more gently. No significant geographic variation in samples from different origins.

Conclusion:

A method for the determination of Poriae Cutis characteristics and multi-component content was established, which laid the foundation for quality control of Poriae Cutis.

, correspAuthors=Ya-zhong ZHANG, 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=Fan XU, Jie YUAN, Jing-zhe PU, Chong HU, Ya-zhong ZHANG, Shou-jin LIU), CN=ArticleExt(id=1239173815452889900, articleId=1239173813817111235, tenantId=1146029695717560320, journalId=1205117023404326918, language=CN, title=基于波长转换RP-HPLC法的茯苓皮多成分含量测定及特征图谱方法研究*, columnId=1206272756476342615, journalTitle=药物分析杂志, columnName=成分分析, runingTitle=null, highlight=null, articleAbstract=
目的:

本文通过建立茯苓皮的双波长切换HPLC特征图谱方法,同时建立茯苓皮中11个三萜类成分含量的测定方法,旨在为茯苓皮全面的质量评价提供定性和定量方法。

方法:

采用Agilent 5 HC-C18(2)色谱柱(250 mm×4.6 mm,5 μm),流动相为乙腈(含3%四氢呋喃)(A)和0.1%磷酸水溶液(B),梯度洗脱,流速1 mL·min-1,柱温35 ℃,进样体积20 μL,检测波长210、243 nm。

结果:

所建立的特征图谱能有效的识别18个共有峰,精密度、重复性、稳定性(48 h)试验的RSD均<3.7%(n=6);11个待测化学成分能够达到良好分离,在考察质量范围内线性关系良好(r均≥0.999 6),平均回收率95.4%~105.5%,RSD 1.0%~3.1%,精密度、重复性、稳定性(48 h)试验的RSD均≤3.0%(n=6);相似度分析结果显示,大部分产地的茯苓皮药材的相似度很高;含量测定结果表明,11个三萜类成分中茯苓酸A在所有批次茯苓皮中占比最高,另外茯苓酸A、松苓新酸、茯苓酸B、去氢齿孔酸、栓菌酸5个成分的含量波动相对较大,其他成分波动较平缓;不同产地茯苓皮样品无明显地域上的差异。

结论:

建立了茯苓皮特征图谱和多成分含量测定方法,为茯苓皮的质量控制奠定了基础。

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Quality evaluation of Poria based on specific chromatogram and quantitative analysis of multicomponents[J]. China J Chin Mater Med201944(7): 1371, articleTitle=Quality evaluation of Poria based on specific chromatogram and quantitative analysis of multicomponents, refAbstract=null)], funds=[Fund(id=1239173823619199129, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, awardId=2019YFC1711500, language=CN, fundingSource=*国家重点研发计划项目-中药材净切制关键技术与智能设备研究及应用(2019YFC1711500), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1239173815679382337, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, xref=1., ext=[AuthorCompanyExt(id=1239173815683576642, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173815679382337, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.School of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230012, China), AuthorCompanyExt(id=1239173815691965251, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173815679382337, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.安徽中医药大学药学院,合肥 230012)]), AuthorCompany(id=1239173815763268427, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, xref=2., ext=[AuthorCompanyExt(id=1239173815771657037, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173815763268427, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.Anhui Institute for Food and Drug Control, Hefei 230051, China), AuthorCompanyExt(id=1239173815775851344, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173815763268427, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.安徽省食品药品检验研究院,合肥 230051)]), AuthorCompany(id=1239173815947817811, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, xref=3., ext=[AuthorCompanyExt(id=1239173815964595030, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173815947817811, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.State Drug Administration Key Laboratory of Quality Research and Evaluation of Chinese Medicine, Hefei 230051, China), AuthorCompanyExt(id=1239173815972983639, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173815947817811, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3.国家药品监督管理局中药质量研究与评价重点实验室,合肥 230051)]), AuthorCompany(id=1239173817331938143, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, xref=4., ext=[AuthorCompanyExt(id=1239173817336132450, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173817331938143, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4.Anhui Province Key Laboratory of Traditional Chinese Medicine Decoction Pieces of New Manufacturing Technology, Hefei 230012, China), AuthorCompanyExt(id=1239173817344521057, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, companyId=1239173817331938143, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=4.中药饮片制造新技术安徽省重点实验室,合肥 230012)])], figs=[ArticleFig(id=1239173820397973569, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Fig.1, caption=HPLC specific chromatogram of Poriae Cutis, figureFileSmall=aYpkcsLq87QEIVznumgFXA==, figureFileBig=AdEve09QBXJS8PTIkbK37A==, tableContent=null), ArticleFig(id=1239173821882757191, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=图1, caption=茯苓皮药材HPLC特征图谱

3.茯苓酸B(poricoic acid B) 4.去氢土莫酸(dehydrotumulosic acid) 6.茯苓酸A(poricoic acid A) 7.多孔菌酸C(polyporenic acid C) 8. 3-表去氢土莫酸(3-epidehydrotumulosic acid) 9.3-O-乙酰-16α-羟基松苓新酸(3-O-acetyl-16 alpha-hydroxydehydrotrametenolic acid) 11.去氢茯苓酸(dehydropachymic acid) 12.茯苓酸(pachymic acid) 15.松苓新酸(dehydrotrametenolic acid) 16.栓菌酸(trametenolic acid) 17.去氢齿孔酸(dehydroeburicoic acid)

, figureFileSmall=aYpkcsLq87QEIVznumgFXA==, figureFileBig=AdEve09QBXJS8PTIkbK37A==, tableContent=null), ArticleFig(id=1239173822054723669, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Fig.2, caption=HPLC overlay chart of Poriae Cutis, figureFileSmall=KSZQ6h8fegFuHraT7Nxoyw==, figureFileBig=x3tzxtsM3vf1A2THDMtPGA==, tableContent=null), ArticleFig(id=1239173822121832537, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=图2, caption=茯苓皮药材HPLC叠加图, figureFileSmall=KSZQ6h8fegFuHraT7Nxoyw==, figureFileBig=x3tzxtsM3vf1A2THDMtPGA==, tableContent=null), ArticleFig(id=1239173822188941404, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Fig.3, caption=HPLC chromatograms of Poriae Cutis(A) and mixed reference substances(B), figureFileSmall=ny7VUQNDhUAviB9vGAbRig==, figureFileBig=VTMQudDX9Z+JfFYlbtf08g==, tableContent=null), ArticleFig(id=1239173822277021793, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=图3, caption=茯苓皮样品(A)与混合对照品(B)HPLC图, figureFileSmall=ny7VUQNDhUAviB9vGAbRig==, figureFileBig=VTMQudDX9Z+JfFYlbtf08g==, tableContent=null), ArticleFig(id=1239173822402850918, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Fig.4, caption=Cluster analysis of the content of 11 components in 21 batches of Poriae Cutis, figureFileSmall=hgF7j6qmtNojolmaL8dwhQ==, figureFileBig=R22hI6tjcVHVkDDhLqy7CQ==, tableContent=null), ArticleFig(id=1239173822503514223, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=图4, caption=21批茯苓皮样品中11个成分含量的聚类分析, figureFileSmall=hgF7j6qmtNojolmaL8dwhQ==, figureFileBig=R22hI6tjcVHVkDDhLqy7CQ==, tableContent=null), ArticleFig(id=1239173822633537650, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Fig.5, caption=Principal component analysis of 21 batches of Poriae Cutis samples, figureFileSmall=ZJgCrC9ROR5BFkiQ0QiWVQ==, figureFileBig=RlQvYRsO057MU46OTr39VA==, tableContent=null), ArticleFig(id=1239173822738395256, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=图5, caption=21批茯苓皮样品主成分分析图, figureFileSmall=ZJgCrC9ROR5BFkiQ0QiWVQ==, figureFileBig=RlQvYRsO057MU46OTr39VA==, tableContent=null), ArticleFig(id=1239173822834864253, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Tab.1, caption=

Regression equation and linear range of 11 components

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
线性范围
(linear range)/μg
r
茯苓酸B(poricoic acid B)Y=1 681.9X+15.10.184 7~5.541 01.000
去氢土莫酸(dehydrotumulosic acid)Y=1 716.3X-14.80.077 9~2.337 00.999 7
茯苓酸A(poricoic acid A)Y=1 349.3X+165.50.772 7~23.179 50.999 9
多孔菌酸C(polyporenic acid C)Y=1 692.0X+0.90.098 8~2.964 01.000
3-表去氢土莫酸(3-epidehydrotumulosic acid)Y=827.1X+2.40.040 0~1.200 80.999 9
3-O-乙酰-16α-羟基松苓新酸(3-O-acetyl-16alpha-hydroxydehydrotrametenolic acid)Y=1 549.1X+3.50.028 5~0.855 81.000
去氢茯苓酸(dehydropachymic acid)Y=1 214.6X+5.40.071 7~2.149 51.000
茯苓酸(pachymic acid)Y=680.51X+51.80.103 1~3.091 50.999 6
松苓新酸(dehydrotrametenolic acid)Y=2 166.3X+63.80.453 7~13.609 51.000
栓菌酸(trametenolic acid)Y=901.3X-25.00.172 6~5.176 50.999 8
去氢齿孔酸(dehydroeburicoic acid)Y=2 017.1X+39.80.3567~10.701 01.000
), ArticleFig(id=1239173822914556032, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=表1, caption=

11个成分的回归方程、线性范围

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
线性范围
(linear range)/μg
r
茯苓酸B(poricoic acid B)Y=1 681.9X+15.10.184 7~5.541 01.000
去氢土莫酸(dehydrotumulosic acid)Y=1 716.3X-14.80.077 9~2.337 00.999 7
茯苓酸A(poricoic acid A)Y=1 349.3X+165.50.772 7~23.179 50.999 9
多孔菌酸C(polyporenic acid C)Y=1 692.0X+0.90.098 8~2.964 01.000
3-表去氢土莫酸(3-epidehydrotumulosic acid)Y=827.1X+2.40.040 0~1.200 80.999 9
3-O-乙酰-16α-羟基松苓新酸(3-O-acetyl-16alpha-hydroxydehydrotrametenolic acid)Y=1 549.1X+3.50.028 5~0.855 81.000
去氢茯苓酸(dehydropachymic acid)Y=1 214.6X+5.40.071 7~2.149 51.000
茯苓酸(pachymic acid)Y=680.51X+51.80.103 1~3.091 50.999 6
松苓新酸(dehydrotrametenolic acid)Y=2 166.3X+63.80.453 7~13.609 51.000
栓菌酸(trametenolic acid)Y=901.3X-25.00.172 6~5.176 50.999 8
去氢齿孔酸(dehydroeburicoic acid)Y=2 017.1X+39.80.3567~10.701 01.000
), ArticleFig(id=1239173823002636420, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Tab. 2, caption=

The recoveries of 11 components

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
样品含量
(content)/mg
加入量
(added)/mg
测得量
(measured)/mg
回收率
(recovery)/%
平均回收率
(average recovery)/%
RSD/%
茯苓酸B(poricoic acid B)1.076 11.108 22.200 7101.5103.51.7
1.075 21.108 22.251 8106.4
1.075 21.108 22.233 2104.6
1.076 11.108 22.211 5102.5
1.075 71.108 22.212 9102.7
1.076 11.108 22.221 7103.5
去氢土莫酸(dehydrotumulosic acid)0.704 80.467 41.198 8103.8105.51.6
0.704 20.467 41.227 0107.9
0.704 20.467 41.203 0104.5
0.704 80.467 41.204 5104.6
0.704 50.467 41.223 4107.3
0.704 80.467 41.207 4105.0
茯苓酸A(poricoic acid A)5.790 84.635 910.570 8102.5102.01.8
5.786 24.635 910.666 1104.2
5.786 24.635 910.619 0103.4
5.790 84.635 910.514 0101.5
5.78854.635 910.364 999.0
5.790 84.635 910.521 2101.6
多孔菌酸C(polyporenic acid C)0.630 80.592 81.239 1102.5102.71.0
0.630 30.592 81.245 7103.6
0.630 30.592 81.247 3103.8
0.630 80.592 81.231 9101.3
0.630 60.592 81.235 2101.9
0.630 80.592 81.244 7103.3
3-表去氢土莫酸(3-epidehydrotumulosic acid)0.242 00.240 20.475 997.499.62.3
0.241 80.240 20.488 9102.9
0.241 80.240 20.484 8101.2
0.242 00.240 20.474 296.7
0.241 90.240 20.482 4100.1
0.242 00.240 20.481 199.5
3-O-乙酰-16α-羟基松苓新酸(3-O-acetyl-16 alpha-hydroxydehydrotrametenolic acid)0.187 10.171 20.351 696.497.52.9
0.187 00.171 20.350 595.9
0.187 00.171 20.355 398.5
0.187 10.171 20.349 595.3
0.187 10.171 20.350 595.9
0.187 10.171 20.363 3102.7
去氢茯苓酸(dehydropachymic acid)0.413 30.429 90.837 598.695.41.9
0.413 00.429 90.825 695.8
0.413 00.429 90.823 695.3
0.413 30.429 90.815 193.2
0.413 20.429 90.818 794.1
0.413 30.429 90.823 395.2
茯苓酸(pachymic acid)0.564 20.618 31.154 295.095.72.7
0.563 80.618 31.140 292.6
0.563 80.618 31.158 295.8
0.564 20.618 31.146 293.6
0.564 00.618 31.169 897.8
0.564 20.618 31.179 499.4
松苓新酸(dehydrotrametenolic acid)2.887 92.721 95.659 5101.798.63.1
2.885 62.721 95.523 397.1
2.885 62.721 95.545 997.9
2.887 92.721 95.482 095.6
2.886 72.721 95.497 496.2
2.887 92.721 95.695 0103.0
栓菌酸(trametenolic acid)1.049 51.035 32.161 8107.3103.82.3
1.048 71.035 32.107 2102.2
1.048 71.035 32.134 0104.8
1.049 51.035 32.104 2101.9
1.049 11.035 32.099 3101.4
1.049 51.035 32.141 5105.4
去氢齿孔酸(dehydroeburicoic acid)2.455 12.140 24.526 897.296.21.8
2.453 12.140 24.534 797.6
2.453 12.140 24.529 297.4
2.455 12.140 24.451 194.1
2.454 12.140 24.439 593.7
2.455 12.140 24.522 497.0
), ArticleFig(id=1239173823124271242, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=表2, caption=

11个成分加样回收率试验(n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
样品含量
(content)/mg
加入量
(added)/mg
测得量
(measured)/mg
回收率
(recovery)/%
平均回收率
(average recovery)/%
RSD/%
茯苓酸B(poricoic acid B)1.076 11.108 22.200 7101.5103.51.7
1.075 21.108 22.251 8106.4
1.075 21.108 22.233 2104.6
1.076 11.108 22.211 5102.5
1.075 71.108 22.212 9102.7
1.076 11.108 22.221 7103.5
去氢土莫酸(dehydrotumulosic acid)0.704 80.467 41.198 8103.8105.51.6
0.704 20.467 41.227 0107.9
0.704 20.467 41.203 0104.5
0.704 80.467 41.204 5104.6
0.704 50.467 41.223 4107.3
0.704 80.467 41.207 4105.0
茯苓酸A(poricoic acid A)5.790 84.635 910.570 8102.5102.01.8
5.786 24.635 910.666 1104.2
5.786 24.635 910.619 0103.4
5.790 84.635 910.514 0101.5
5.78854.635 910.364 999.0
5.790 84.635 910.521 2101.6
多孔菌酸C(polyporenic acid C)0.630 80.592 81.239 1102.5102.71.0
0.630 30.592 81.245 7103.6
0.630 30.592 81.247 3103.8
0.630 80.592 81.231 9101.3
0.630 60.592 81.235 2101.9
0.630 80.592 81.244 7103.3
3-表去氢土莫酸(3-epidehydrotumulosic acid)0.242 00.240 20.475 997.499.62.3
0.241 80.240 20.488 9102.9
0.241 80.240 20.484 8101.2
0.242 00.240 20.474 296.7
0.241 90.240 20.482 4100.1
0.242 00.240 20.481 199.5
3-O-乙酰-16α-羟基松苓新酸(3-O-acetyl-16 alpha-hydroxydehydrotrametenolic acid)0.187 10.171 20.351 696.497.52.9
0.187 00.171 20.350 595.9
0.187 00.171 20.355 398.5
0.187 10.171 20.349 595.3
0.187 10.171 20.350 595.9
0.187 10.171 20.363 3102.7
去氢茯苓酸(dehydropachymic acid)0.413 30.429 90.837 598.695.41.9
0.413 00.429 90.825 695.8
0.413 00.429 90.823 695.3
0.413 30.429 90.815 193.2
0.413 20.429 90.818 794.1
0.413 30.429 90.823 395.2
茯苓酸(pachymic acid)0.564 20.618 31.154 295.095.72.7
0.563 80.618 31.140 292.6
0.563 80.618 31.158 295.8
0.564 20.618 31.146 293.6
0.564 00.618 31.169 897.8
0.564 20.618 31.179 499.4
松苓新酸(dehydrotrametenolic acid)2.887 92.721 95.659 5101.798.63.1
2.885 62.721 95.523 397.1
2.885 62.721 95.545 997.9
2.887 92.721 95.482 095.6
2.886 72.721 95.497 496.2
2.887 92.721 95.695 0103.0
栓菌酸(trametenolic acid)1.049 51.035 32.161 8107.3103.82.3
1.048 71.035 32.107 2102.2
1.048 71.035 32.134 0104.8
1.049 51.035 32.104 2101.9
1.049 11.035 32.099 3101.4
1.049 51.035 32.141 5105.4
去氢齿孔酸(dehydroeburicoic acid)2.455 12.140 24.526 897.296.21.8
2.453 12.140 24.534 797.6
2.453 12.140 24.529 297.4
2.455 12.140 24.451 194.1
2.454 12.140 24.439 593.7
2.455 12.140 24.522 497.0
), ArticleFig(id=1239173823237517454, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=EN, label=Tab. 3, caption=

Contents of 11 triterpenoid components in Poriae Cutis

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(No.)
含量(content)/%
茯苓酸B
(poricoic acid B)
去氢土莫酸
(dehydrotumulosic acid)
茯苓酸A
(poricoic acid A)
多孔菌酸C
(polyporenic acid C)
3-表去氢土莫酸
(3-epidehydrotumulosic acid)
3-O-乙酰-16α-羟基松苓新酸
(3-O-acetyl-16 alpha-hydroxydehydrotrametenolic acid)
去氢茯苓酸
(dehydropachymic acid)
茯苓酸
(pachymic acid)
松苓新酸
(dehydrotrametenolic acid)
栓菌酸
(trametenolic acid)
去氢齿孔酸
(dehydroeburicoic acid)
总和
(sum)
P-10.2160.1020.7740.0990.0560.0270.0790.1690.3960.2080.2582.384
P-20.6210.1872.3510.1730.0730.0630.1440.2410.8410.4390.6115.745
P-30.6240.1741.5160.1590.0670.0600.1220.2490.8760.4940.4174.759
P-40.2210.1381.0140.1180.0460.0430.1000.1610.5260.2970.4393.103
P-50.3300.2481.4890.1760.0660.0630.1780.2220.5870.3810.7734.513
P-60.2550.2041.4120.1500.0540.0550.1440.1910.6560.4360.6904.245
P-70.2250.1350.9610.1130.0450.0450.1020.1590.5160.3070.4223.029
P-80.1980.1331.0750.1280.0410.0340.0770.1430.5420.3040.4593.133
P-90.2420.1991.3660.1440.0540.0530.1410.2160.6060.4190.6584.096
P-100.1870.1240.8810.1020.0430.0350.0920.1400.4280.2570.3792.668
P-110.2690.2161.5050.1530.0570.0590.1520.2180.6920.4650.7354.520
P-120.2490.2051.4590.1440.0480.0560.1430.1660.6510.4060.7164.244
P-130.2720.1991.4820.1410.0560.0490.1490.1870.6430.4280.6984.304
P-140.2570.2101.5160.1470.0550.0540.1500.2010.6590.4270.7594.435
P-150.2480.1991.5010.1440.0570.0570.1510.1810.6800.4450.7494.411
P-160.2560.1951.4960.1350.0480.0500.1470.1950.6470.3960.7374.301
P-170.2050.2151.3530.1580.0620.0500.1490.1830.5500.3570.6463.927
P-180.2620.2041.3320.1460.0540.0600.1460.1850.6770.4400.6704.177
P-190.2210.1350.9220.1130.0410.0360.0840.1400.5000.3050.3532.850
P-200.2530.1351.0780.1200.0460.0450.0940.1460.6210.3700.3753.283
P-210.1870.1310.9040.1080.0440.0380.0890.1280.5140.3360.3402.818
), ArticleFig(id=1239173823489175701, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173813817111235, language=CN, label=表3, caption=

茯苓皮样品中11个三萜类成分的含量

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(No.)
含量(content)/%
茯苓酸B
(poricoic acid B)
去氢土莫酸
(dehydrotumulosic acid)
茯苓酸A
(poricoic acid A)
多孔菌酸C
(polyporenic acid C)
3-表去氢土莫酸
(3-epidehydrotumulosic acid)
3-O-乙酰-16α-羟基松苓新酸
(3-O-acetyl-16 alpha-hydroxydehydrotrametenolic acid)
去氢茯苓酸
(dehydropachymic acid)
茯苓酸
(pachymic acid)
松苓新酸
(dehydrotrametenolic acid)
栓菌酸
(trametenolic acid)
去氢齿孔酸
(dehydroeburicoic acid)
总和
(sum)
P-10.2160.1020.7740.0990.0560.0270.0790.1690.3960.2080.2582.384
P-20.6210.1872.3510.1730.0730.0630.1440.2410.8410.4390.6115.745
P-30.6240.1741.5160.1590.0670.0600.1220.2490.8760.4940.4174.759
P-40.2210.1381.0140.1180.0460.0430.1000.1610.5260.2970.4393.103
P-50.3300.2481.4890.1760.0660.0630.1780.2220.5870.3810.7734.513
P-60.2550.2041.4120.1500.0540.0550.1440.1910.6560.4360.6904.245
P-70.2250.1350.9610.1130.0450.0450.1020.1590.5160.3070.4223.029
P-80.1980.1331.0750.1280.0410.0340.0770.1430.5420.3040.4593.133
P-90.2420.1991.3660.1440.0540.0530.1410.2160.6060.4190.6584.096
P-100.1870.1240.8810.1020.0430.0350.0920.1400.4280.2570.3792.668
P-110.2690.2161.5050.1530.0570.0590.1520.2180.6920.4650.7354.520
P-120.2490.2051.4590.1440.0480.0560.1430.1660.6510.4060.7164.244
P-130.2720.1991.4820.1410.0560.0490.1490.1870.6430.4280.6984.304
P-140.2570.2101.5160.1470.0550.0540.1500.2010.6590.4270.7594.435
P-150.2480.1991.5010.1440.0570.0570.1510.1810.6800.4450.7494.411
P-160.2560.1951.4960.1350.0480.0500.1470.1950.6470.3960.7374.301
P-170.2050.2151.3530.1580.0620.0500.1490.1830.5500.3570.6463.927
P-180.2620.2041.3320.1460.0540.0600.1460.1850.6770.4400.6704.177
P-190.2210.1350.9220.1130.0410.0360.0840.1400.5000.3050.3532.850
P-200.2530.1351.0780.1200.0460.0450.0940.1460.6210.3700.3753.283
P-210.1870.1310.9040.1080.0440.0380.0890.1280.5140.3360.3402.818
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基于波长转换RP-HPLC法的茯苓皮多成分含量测定及特征图谱方法研究*
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徐凡 1 , 袁杰 2, 3 , 蒲婧哲 2, 3 , 胡冲 2, 3 , 张亚中 2, 3, 4, ** , 刘守金 1
药物分析杂志 | 成分分析 2024,44(2): 214-223
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药物分析杂志 | 成分分析 2024, 44(2): 214-223
基于波长转换RP-HPLC法的茯苓皮多成分含量测定及特征图谱方法研究*
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徐凡1 , 袁杰2, 3, 蒲婧哲2, 3, 胡冲2, 3, 张亚中2, 3, 4, ** , 刘守金1
作者信息
  • 1.安徽中医药大学药学院,合肥 230012
  • 2.安徽省食品药品检验研究院,合肥 230051
  • 3.国家药品监督管理局中药质量研究与评价重点实验室,合肥 230051
  • 4.中药饮片制造新技术安徽省重点实验室,合肥 230012
  • Tel:15055638554;E-mail:

通讯作者:

**Tel:15055638554;E-mail:
Determination and characterization of Poriae Cutis multi-component content based on wavelength conversion RP-HPLC method*
Fan XU1 , Jie YUAN2, 3, Jing-zhe PU2, 3, Chong HU2, 3, Ya-zhong ZHANG2, 3, 4, ** , Shou-jin LIU1
Affiliations
  • 1.School of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230012, China
  • 2.Anhui Institute for Food and Drug Control, Hefei 230051, China
  • 3.State Drug Administration Key Laboratory of Quality Research and Evaluation of Chinese Medicine, Hefei 230051, China
  • 4.Anhui Province Key Laboratory of Traditional Chinese Medicine Decoction Pieces of New Manufacturing Technology, Hefei 230012, China
出版时间: 2024-02-29 doi: 10.16155/j.0254-1793.2024.02.03
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目的:

本文通过建立茯苓皮的双波长切换HPLC特征图谱方法,同时建立茯苓皮中11个三萜类成分含量的测定方法,旨在为茯苓皮全面的质量评价提供定性和定量方法。

方法:

采用Agilent 5 HC-C18(2)色谱柱(250 mm×4.6 mm,5 μm),流动相为乙腈(含3%四氢呋喃)(A)和0.1%磷酸水溶液(B),梯度洗脱,流速1 mL·min-1,柱温35 ℃,进样体积20 μL,检测波长210、243 nm。

结果:

所建立的特征图谱能有效的识别18个共有峰,精密度、重复性、稳定性(48 h)试验的RSD均<3.7%(n=6);11个待测化学成分能够达到良好分离,在考察质量范围内线性关系良好(r均≥0.999 6),平均回收率95.4%~105.5%,RSD 1.0%~3.1%,精密度、重复性、稳定性(48 h)试验的RSD均≤3.0%(n=6);相似度分析结果显示,大部分产地的茯苓皮药材的相似度很高;含量测定结果表明,11个三萜类成分中茯苓酸A在所有批次茯苓皮中占比最高,另外茯苓酸A、松苓新酸、茯苓酸B、去氢齿孔酸、栓菌酸5个成分的含量波动相对较大,其他成分波动较平缓;不同产地茯苓皮样品无明显地域上的差异。

结论:

建立了茯苓皮特征图谱和多成分含量测定方法,为茯苓皮的质量控制奠定了基础。

茯苓皮  /  三萜  /  特征图谱  /  含量测定  /  波长转换
Objective:

To comprehensively evaluate the quality of Poriae Cutis, and to establish a dual wavelength switching HPLC method for comparing the characteristic spectra of Poriae Cutis and studying the content of 11 triterpenoid components, to provide reference for the qualitative and quantitative research of Poriae Cutis.

Methods:

Agilent 5 HC-C18(2) column(250 mm×4.6 mm, 5 μm) was adopted. Acetonitrile solution (contain 3% tetrahydrofuran) (A) and 0.1% formic acid aqueous solution (B) were used as the mobile phase with gradient elution at a flow rate of 1.0 mL·min-1. The column temperature was 30 ℃ and the injection volume was 20 μL. The detection wavelengths were 210 and 243 nm.

Results:

The feature profiles developed were effective in identifying the 18 shared peaks. RSD for precision, repeatability and stability (48 h) tests were all less than 3.72%(n=6). The 11 chemical components to be measured were well separated, with good linearity in the mass range examined (all r ≥ 0.999 6). The average recovery rate was 95.4%-105.5%, and the RSD was 1.0%-3.1%. The RSDs of precision, repeatability, and stability (48 h) tests were all less than or equal to 3.0%(n=6). The results of similarity analysis showed that most of the origins of Poriae Cutis were very similar to each other. The results of content determination showed that among the 11 triterpenoid constituents, poricoic acid A accounted for the highest percentage in all batches of Poriae Cutis. In addition, the content of five components, poricoic acid A, dehydrotrametenolic acid, poricoic acid B, dehydroeburicoic acid and trametenolic acid, fluctuated relatively more, while the other components fluctuated more gently. No significant geographic variation in samples from different origins.

Conclusion:

A method for the determination of Poriae Cutis characteristics and multi-component content was established, which laid the foundation for quality control of Poriae Cutis.

Poriae Cutis  /  triterpenoids  /  specific chromatograms  /  content determination  /  wavelength conversion
徐凡, 袁杰, 蒲婧哲, 胡冲, 张亚中, 刘守金. 基于波长转换RP-HPLC法的茯苓皮多成分含量测定及特征图谱方法研究*. 药物分析杂志, 2024 , 44 (2) : 214 -223 . DOI: 10.16155/j.0254-1793.2024.02.03
Fan XU, Jie YUAN, Jing-zhe PU, Chong HU, Ya-zhong ZHANG, Shou-jin LIU. Determination and characterization of Poriae Cutis multi-component content based on wavelength conversion RP-HPLC method*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (2) : 214 -223 . DOI: 10.16155/j.0254-1793.2024.02.03
茯苓皮为多孔菌科真菌茯苓Poria cocos (Schw.) Wolf菌核的干燥外皮,具有利水消肿的功效,用于水肿,小便不利[1]。在东汉之前,茯苓一直是以整体入药,在东汉时期才有了茯苓和茯苓皮之分,茯苓皮开始单独入药,东汉的《中藏经》卷六[2]首次记载了以茯苓皮入药的“五皮散”。而茯苓皮作为独立药材的记载最早见于明代《本草纲目》[3],2020年版《中华人民共和国药典》(以下简称《中国药典》)中茯苓皮也已经单独记载。目前市场流通茯苓皮主要来源于安徽、湖北地区,但报道显示以云南产质量最佳[4]
茯苓皮中主要含有多糖类和三萜类化学成分,其中三萜类成分已被证实具有利尿、抗氧化、抗炎、抗肿瘤、降血脂、抗癫痫、美白等药理作用[5-11]。目前学者对茯苓中三萜类成分的研究已经较为深入,对茯苓皮中三萜类成分的研究较少,近10年来多见对茯苓皮中总三萜或1~5个三萜类成分的测定[12-14],且2020年版《中国药典》中未对茯苓皮进行含量测定的规定,特异性不足,课题组前期已完成茯苓皮基础检查项如水分、总灰分等的测定,有一定实验基础。因此,本研究通过建立茯苓皮特征图谱,以及多成分含量测定的方法,为后期实验提供帮助,为茯苓皮的鉴别及质量控制提供参考。
Agilent Technologies 1260高效液相色谱仪[安捷伦科技(中国)有限公司];ML204型万分之一电子天平、XP26型十万分之一电子天平(托利多仪器上海有限公司);Elmasonic S 30(H)超声波清洗器(德祥科技有限公司);116B摇摆式六两装高速中药粉碎机(永历制药机械有限公司);5HC-C18(2)色谱柱(250 mm×4.6 mm,5 μm)(Agilent公司)。
对照品茯苓酸B(批号D28GB172711,纯度以95%计)、去氢土莫酸(批号A19GB158345,纯度以95%计)、茯苓酸A(批号J08GB154007,纯度以96%计)、多孔菌酸C(批号P08J11L107514,纯度以95%计)、3-表去氢土莫酸(批号A13HB180294,纯度以95%计)、3-O-乙酰-16α-羟基松苓新酸(批号P08J11S107846,纯度以95%计)、去氢茯苓酸(批号A17GB144422,纯度以98%计)、茯苓酸(批号J02GB150406,纯度以97%计)、松苓新酸(批号P24J10S80630,纯度以98%计)、栓菌酸(批号A14HB180295,纯度以95%计)、去氢齿孔酸(批号A14HB180296,纯度以95%计)均购自上海源叶生物科技有限公司。甲醇、乙腈、四氢呋喃为色谱纯[赛默飞世尔科技(中国)有限公司],磷酸为分析醇(国药集团化学试剂有限公司),水为纯净水。
21批茯苓皮药材于安徽、云南、广西、贵州及湖北产地收集,经安徽省食品药品检验研究院张亚中主任中药师鉴定多孔菌科真菌茯苓Poria cocos (Schw.) Wolf菌核的干燥外皮,产地及编号分别为安徽省六安市金寨县(P-1~P-4)、安庆市宿松县(P-5)、岳西县(P-6、P-7),云南省楚雄彝族自治州双柏县(P-8)、保山市腾冲县(P-9、P-10),广西壮族自治区玉林市(P-11~P-13),贵州省黔东南苗族侗族自治州黎平县(P-14~P-16),湖北省宜昌市夷陵区(P-17)、黄冈市罗田县(P-18~P-21)。
采用Agilent 5 HC-C18(2)色谱柱(250 mm×4.6 mm,5 μm),流动相为乙腈(含3%四氢呋喃)(A)和0.1%磷酸水溶液(B),梯度洗脱(0~10 min,45%A;10~15 min,45%A→52%A;15~40 min,52%A;40~50 min,52%A→70%A;50~65 min,70%A→75%A;65~71 min,75%A;71~74 min,75%A→80%A;74~95 min,80%A),流速1 mL·min-1,柱温35 ℃,进样体积20 μL,检测波长243 nm和210 nm(62~64.5 min、67~68.8 min、80~83 min),理论塔板数按茯苓酸B计算应≥8 000。
取各对照品适量,精密称量,加甲醇制成每1 mL含茯苓酸B 0.184 7 mg、去氢土莫酸0.077 9 mg、茯苓酸A 0.772 7 mg、多孔菌酸C 0.098 8 mg、3-表去氢土莫酸0.040 0 mg、3-O-乙酰-16α-羟基松苓新酸0.028 5 mg、去氢茯苓酸0.071 7 mg、茯苓酸0.103 1 mg、松苓新酸0.453 7 mg、栓菌酸0.172 6 mg、去氢齿孔酸0.356 7 mg的混合对照品溶液。
取茯苓皮粉末(过4号筛)约1 g,精密称量,置具塞锥形瓶中,精密加甲醇25 mL,密塞,称量,超声处理(功率100 W,频率40 kHz)30 min,取出,冷却,称量,用甲醇补足失去的量,摇匀,静置,取上清液,用0.22 μm微孔滤膜滤过,取续滤液,即得。
通过21批茯苓皮药材的HPLC色谱图,建立茯苓皮的特征图谱,对茯苓皮中化学成分进行表征,为茯苓皮的鉴别和质量控制提供支持。
取茯苓皮粉末(过4号筛)约1 g,精密称定,按“2.3”项下方法制备供试品溶液,按“2.1”项色谱条件进样,记录HPLC色谱图。采用国家药典委员会的中药色谱指纹图谱相似度评价系统软件(2012.1版)进行评价,设定P-1样品图谱为参照图谱,并将其余样品的色谱峰与P-1样品的色谱峰进行自动匹配,生成对照特征图谱,对21批茯苓皮药材特征图谱中出峰时间基本一致、峰面积较大的18个共有特征峰进行相似度分析。生成的茯苓皮药材对照特征图谱见图1,21批茯苓皮药材HPLC叠加图见图2
通过《中药色谱指纹图谱相似度评价系统》对21批茯苓皮相似度进行评估,结果的数值越大,相似度越高。结果显示,除P-8样品外,其余20批样品与对照特征图谱的相似度均>0.9,说明大部分产地的茯苓皮中三萜类成分具有较高的相似度。
通过中药色谱指纹图谱相似度评价系统软件对21批茯苓皮进行共有峰标识,根据各峰的峰面积、峰形和分离度,选择18个共有峰作为特征峰,并与参照物图谱进行比对,最终确定3号峰为茯苓酸B,4号峰为去氢土莫酸,6号峰为茯苓酸A,7号峰为多孔菌酸C,8号峰为3-表去氢土莫酸,9号峰为3-O-乙酰-16α-羟基松苓新酸,11号峰为去氢茯苓酸,12号峰为茯苓酸,15号峰为松苓新酸,16号峰为栓菌酸,17号峰为去氢齿孔酸。
精密吸取供试品溶液20 μL,按“2.1”项下色谱条件重复进样6次,记录色谱图。以峰面积较大的茯苓酸A峰作为参照峰,计算共有峰的相对保留时间及相对峰面积。共有峰的相对保留时间RSD均<1.0%,相对峰面积RSD均<2.4%,结果表明仪器精密度良好。
精密吸取同一茯苓皮(P-8)供试品溶液20 μL,分别于0、2、4、6、8、10、12、24、36、48 h按“2.1”项下色谱条件测定,记录色谱图。以茯苓酸A峰为参照峰,计算共有峰的相对保留时间及相对峰面积。共有峰相对保留时间的RSD均<1.0%,相对峰面积的RSD均<2.4%,结果表明供试品溶液在48 h内稳定性良好。
取同一批茯苓皮供试品(P-8)6份,精密称定,按“2.3”项下方法分别制备供试品溶液,精密吸取供试品溶液20 μL,分别进样测定,记录色谱图。以茯苓酸A峰为参照峰,计算共有峰的相对保留时间及相对峰面积。共有峰相对保留时间的RSD均<1.0%,相对峰面积的RSD均<3.7%,结果表明该方法重复性好。
在特征图谱研究的基础上,通过特征图谱色谱条件、供试品溶液的和对照品溶液的制备,测定其中11个化学成分的含量,从定量的角度对茯苓皮的质量进行控制,和特征图谱形成全面控制茯苓皮的质量控制体系。
精密吸取混合对照品溶液、供试品溶液及纯溶剂溶液(甲醇,阴性对照),在相同的色谱条件下,测定并记录色谱图,结果表明阴性对照无干扰,专属性强。茯苓皮样品与混合对照品的色谱图见图3
取混合对照品溶液适量,以进样量1、5、10、15、20、30 μL进样,按“2.1”项下色谱条件进行测定。以待测成分的进样质量(X)为横坐标,待测成分峰面积(Y)为纵坐标,绘制标准曲线,计算回归方程及相关系数,结果见表1
取混合对照品溶液,按“2.1”项下色谱条件进行分析,连续进样6次,记录待测成分峰面积,计算RSD。11个成分峰面积的RSD均<2.0%,结果表明仪器精密度良好。
取同一份供试品(P-8)溶液,分别于0、2、4、6、8、10、12、24、36、48 h进行分析,记录待测成分峰面积,计算含量及相应RSD。11个成分含量的RSD均<2.5%,结果表明供试品溶液在48 h内基本稳定。
取同一批茯苓皮供试品(P-8)6份,精密称定,按“2.3”项下方法制备供试品溶液,按“2.1”项下色谱条件测定,记录峰面积,计算含量及相应RSD。11个成分含量为0.214 8%、0.132 3%、1.155 8%、0.129 4%、0.048 3%、0.037 8%、0.082 5%、0.121 5%、0.578 9%、0.197 3%、0.490 0%,RSD为2.6%、1.4%、2.9%、2.1%、1.7%、2.3%、1.2%、3.0%、2.2%、2.1%、2.8%,RSD均≤3.0%,说明本方法的重复性良好。
取已知含量的茯苓皮粉末(过4号筛)约0.5 g,共6份,精密称定,分别加入一定量混合对照品溶液,按照“2.3”项下方法制备供试品溶液,进行分析,计算各待测成分的平均回收率及RSD,见表2。11个待测成分的平均回收率在95.4%~105.5%,RSD均≤3.1%,表明本方法的准确度良好。
分别取21批茯苓皮样品(P1~P21),按照“2.3”项下方法制备供试品溶液,再按“2.1”项下色谱条件进行测定,用外标法计算样品含量,结果见表3
将21批茯苓样品中11个三萜类成分的含量测定结果导入SPSS 21.0软件中,采用聚类分析法结合平方欧氏距离(d)进行分析,生成21批不同产地茯苓中三萜类成分含量的聚类分析树状图,详见图4。同时将数据导入SIMCA-P 14.0软件进行主成分分析(PCA),以21批样品中11个三萜类成分的含量为变量,建立PCA模型,结果见图5
建立的茯苓皮特征图谱精密度、稳定性、重复性均符合2020年版《中国药典》规定,所建立的特征图谱标定大部分共有峰,11个三萜类成分分离度好。建立的含量测定方法专属性强,准确可靠。相似度分析结果显示,大部分产地的茯苓皮药材中11个三萜类成分的相似度很高。从表3中11个三萜类成分的含量测定结果可以看到,茯苓酸A在所有批次茯苓皮中占比最高,平均含量>1%。除个别样品外,茯苓皮中11个成分的含量变化呈现正相关趋势,猜测与其结构相似,在植物体内的积累规律一致有关。11个成分中茯苓酸A、松苓新酸、茯苓酸B、去氢齿孔酸、栓菌酸的含量波动变化较大,表明不同生态环境可能对以上5个成分的积累影响更大。
聚类分析结果表明,21批样品按d=4可分为4类:样品P-5、P-6、P-9、P-11~P-18聚为Ⅰ类,特征为11个成分整体含量中上,总含量在3.927%~4.520%;样品P-3单独聚为Ⅱ类,特征为11个成分总含量较高,其中茯苓酸B含量>0.624%;样品P-1、P-4、P-7、P-8、P10、P-19~P-21聚为Ⅲ类,特征为11个成分整体含量中等偏低,总含量在2.384%~3.283%;样品P-2单独聚为Ⅳ类,特征为11个成分整体含量偏高,总含量达到5.745%。由含量测定结果可见,21批茯苓皮样品中11个成分的总含量整体差异较大,以不同省份划分产地区域,同一聚类中样品所属区域较分散,同一区域内不同产地样品分布在不同聚类中,未体现出显著的区域性特征。主成分分析建立的模型拟合优度R2为0.949,预测优度Q2为0.833,均>0.5,模型良好可靠,为可用模型,且主成分分析结果与聚类分析结果一致。
采用DAD检测器进行全波长扫描,峰10、12、16在201 nm有最大吸收,其余共有峰在243 nm有最大吸收,观察色谱图发现在201 nm有较明显的端吸收,影响茯苓酸、栓菌酸色谱峰峰型对称性。比较不同检测波长(201、205、210 nm)色谱图,发现选取210 nm作为检测波长时,峰10、12、16峰形较好,基线较平稳,与201 nm峰面积无较大差异,故选择210、243 nm进行双波长检测。
本研究考察了乙腈-0.05%磷酸水溶液、乙腈-0.1%磷酸水溶液、乙腈-0.5%磷酸水溶液,观察色谱图发现乙腈-0.1%磷酸水溶液与乙腈-0.5%磷酸水溶液的分离效果差别不大,考虑pH对色谱柱柱效的影响,故选择乙腈-0.1%磷酸水溶液。个别色谱峰有肩峰较难分离,查阅文献发现在乙腈中加入3%的四氢呋喃[15]可以使分离效果更好,通过比较色谱峰分离度及峰形,最终确定以乙腈(3%的四氢呋喃)-0.1%磷酸水溶液为流动相。
考察了Welch Ultimate Plus C18色谱柱(250 mm×4.6 mm,5 μm)、Agilent Eclipse XDB-C18色谱柱(250 mm×4.6 mm,5 μm)、Agilent 5HC-C18(2)色谱柱(250 mm×4.6 mm,5 μm)对茯苓皮中11个成分分离度的影响。3-O-乙酰-16α-羟基松苓新酸与旁边的干扰峰较难分离,采用Agilent 5HC-C18(2)色谱柱(250 mm×4.6 mm,5 μm)可以达到很好的分离效果。
分别取茯苓皮样品(P-8)粉末约1 g,置具塞锥形瓶中,考察不同提取方式(超声提取、热回流提取)、不同提取时间(15、30、60、90 min)、不同提取溶剂(50%甲醇、甲醇、无水乙醇、95%乙醇)以及不同溶剂用量(15、25、50 mL),最终选择25 mL甲醇超声处理30 min为前处理方法。
茯苓皮利尿作用较茯苓更强,研究发现茯苓皮中总三萜的含量明显高于茯苓,且已证实四环三萜类化合物是茯苓皮利尿作用的主要活性成分[5,14],故茯苓皮中三萜类成分的高低影响着茯苓皮利尿效果,制定一种可以准确定量茯苓皮中三萜类成分的方法很有必要。本文建立的茯苓皮特征图谱共得到18个共有峰,其中11个峰得到对照品标定。各特征峰分离度较好,较为全面的反映了茯苓皮药材的特异性和整体性信息,可以用于茯苓皮的定性鉴别。
建立的茯苓皮含量测定方法专属性强,准确可靠,分析含量测定结果发现,茯苓皮中11个三萜类成分的含量变化呈现正相关趋势,猜测与其结构相似,在植物体内的积累规律一致有关,其中茯苓酸A、松苓新酸、茯苓酸B、去氢齿孔酸、栓菌酸5个成分的含量波动相对较大,其他成分波动较平缓,猜测外界生态因子可能主要对以上5个成分的积累影响更大;对不同产地21批茯苓皮药材中11个三萜类成分含量测定结果的分析显示,茯苓皮样品无明显地域上的差异。
综上所述,本研究建立的特征图谱可以在对照品缺乏的情况下,用于茯苓皮的定性鉴别。建立的含量测定方法可以同时用于茯苓皮中多个三萜类成分的含量测定,为茯苓皮全面的质量控制提供帮助。后期有必要扩大不同产地样品量,进一步分析产地是否对茯苓皮三萜类成分积累有影响,为全面掌握茯苓皮药材的品质提供帮助。
  • *国家重点研发计划项目-中药材净切制关键技术与智能设备研究及应用(2019YFC1711500)
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2024年第44卷第2期
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doi: 10.16155/j.0254-1793.2024.02.03
  • 首发时间:2026-03-13
  • 出版时间:2024-02-29
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  • 修回日期:2023-11-22
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*国家重点研发计划项目-中药材净切制关键技术与智能设备研究及应用(2019YFC1711500)
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    1.安徽中医药大学药学院,合肥 230012
    2.安徽省食品药品检验研究院,合肥 230051
    3.国家药品监督管理局中药质量研究与评价重点实验室,合肥 230051
    4.中药饮片制造新技术安徽省重点实验室,合肥 230012

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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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