Article(id=1239148841568620698, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239148837584040649, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024.03.18, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1694620800000, receivedDateStr=2023-09-14, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773365706101, onlineDateStr=2026-03-13, pubDate=1711814400000, pubDateStr=2024-03-31, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773365706101, onlineIssueDateStr=2026-03-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773365706101, creator=13701087609, updateTime=1773365706101, updator=13701087609, issue=Issue{id=1239148837584040649, tenantId=1146029695717560320, journalId=1205117023404326918, year='2024', volume='44', issue='3', pageStart='373', pageEnd='552', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773365705152, creator=13701087609, updateTime=1773367146269, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1239154882125550205, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239148837584040649, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1239154882125550206, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239148837584040649, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=522, endPage=531, ext={EN=ArticleExt(id=1239148841929330860, articleId=1239148841568620698, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Quality evaluation of Jinteng Qingbi granules based on UHPLC multi-index component determination and principal component analysis*, columnId=1239148841803501731, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Quality Control, runingTitle=null, highlight=null, articleAbstract=
Objective:

To establish a UHPLC method for simultaneously determining sinomenine,neochlorogenic acid,magnolflorine,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,harpagoside,ligustilide and ammonium glycyrrhizate in Jinteng Qingbi granules,and evaluate the quality of Jinteng Qingbi granules combined with principal component analysis.

Methods:

UHPLC wavelength switching method was employed in the study. The chromatographic separation was performed on a Waters XSelect® CSH C18 column (150 mm×4.6 mm,2.5 μm) using methanol-acetonitrile (1:1) as mobile phase A and 0.2% phosphoric acid aqueous solution as mobile phase B in a gradient mode at 25 ℃. The flow rate was 1.0 mL·min-1,and the UV detection wavelength was chosen at 218 nm for sinomenine during 0-17 min,326 nm for neochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C during 17-25 min and 34-98nm,263 nm for magnolflorine,harpagoside,ligustilide and ammonium glycyrrhizate during 25-34 min and 98-125nm,respectively. Furthermore,multiple statistical analysis was conducted on the contents of 11 components in 20 batches of Jinteng Qingbi granules using SPSS27.0 software.

Resluts:

Satisfactory linearities of sinomenine,neochlorogenic acid,magnolflorine,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,harpagoside,ligustilide and ammonium glycyrrhizate were in the ranges of 14.36-143.61 μg·mL-1 (r=0.999 9),7.71-77.15 μg·mL-1 (r=0.999 8),9.18-91.83 μg·mL-1 (r=0.999 7),10.71-107.07 μg·mL-1 (r=0.999 8),12.88-128.80 μg·mL-1 (r=0.999 8),5.20-51.95 μg·mL-1 (r=0.999 7),5.18-51.84 μg·mL-1 (r=0.999 8),5.40-53.95 μg·mL-1 (r=0.999 8),2.62-26.16 μg·mL-1 (r=0.999 9),6.31-63.06 μg·mL-1 (r=0.999 9) and 11.13-111.26 μg·mL-1 (r=0.997 6),respectively. The average recoveries (n=6) were 98.3%,98.3%,98.5%,98.9%,99.2%,101.0%,98.1%,97.1%,96.8%,98.0% and 98.7%,respectively,with RSDs less than 3.0%. The contents ranges of sinomenine and other 10 components of 20 batches of Jinteng Qingbi granules samples were 2.206-2.704,1.071-1.403,2.096-2.487,1.321-1.724,2.241-2.612,0.605-0.749,0.363-0.412,0.835-1.020,0.151-0.191,0.791-1.188 and 1.008-1.363 mg·g-1,respectively. The results of principal component analysis showed that the quality differences between batches of continuously produced Jinteng Qingbi granules were relatively smaller,and the comprehensive quality of samples S1,S5 and S7 was relatively better.

Conclusion:

The established UHPLC method of multi-index component determination is simple,accurate and stable. Combined with principal component analysis,it can be used for quality evaluation of Jinteng Qingbi granules comprehensively.

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目的:

建立同时测定金藤清痹颗粒中青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分含量的UHPLC法,并结合主成分分析对制剂进行质量评价。

方法:

采用UHPLC波长切换法,色谱柱为Waters XSelect® CSH C18柱(150 mm×4.6 mm,2.5 μm),以甲醇-乙腈(1:1)为流动相A,0.2%磷酸水溶液为流动相B,梯度洗脱,流速1.0 mL·min-1,柱温25 ℃,检测波长218 nm(0~17 min时,检测青藤碱)、326 nm (17~25 min和34~98 min时,检测新绿原酸、隐绿原酸、绿原酸和异绿原酸A、异绿原酸B、异绿原酸C)、263 nm(25~34 min和98~125 min时,检测木兰花碱、哈巴俄苷、藁本内酯、甘草酸铵)。利用SPSS27.0软件对20批金藤清痹颗粒中成分含量进行多元统计分析。

结果:

青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分的质量浓度分别在14.36~143.61 μg·mL-1r=0.999 9)、7.71~77.15 μg·mL-1r=0.999 8)、9.18~91.83 μg·mL-1r=0.999 7)、10.71~107.07 μg·mL-1r=0.999 8)、12.88~128.80 μg·mL-1r=0.999 8)、5.20~51.95 μg·mL-1r=0.999 7)、5.18~51.84 μg·mL-1r=0.999 8)、5.40~53.95 μg·mL-1r=0.999 8)、2.62~26.16 μg·mL-1r=0.999 9)、6.31~63.06 μg·mL-1r=0.999 9)和11.13~111.26 μg·mL-1r=0.997 6)范围内与峰面积的线性关系良好;平均加样回收率(n=6)分别为98.3%、98.3%、98.5%、98.9%、99.2%、101.0%、98.1%、97.1%、96.8%、98.0%和98.7%,RSD均小于3.0%。20批金藤清痹颗粒样品中,上述青藤碱等11个成分的含量测定结果(n=6)依次为2.206~2.704、1.071~1.403、2.096~2.487、1.321~1.724、2.241~2.612、0.605~0.749、0.363~0.412、0.835~1.020、0.151~0.191、0.791~1.188和1.008~1.363 mg·g-1。主成分分析结果显示,连续生产的金藤清痹颗粒批次间质量差异较小,且以样品S1、S5和S7的综合质量相对更好。

结论:

建立的UHPLC多指标成分含量测定方法简便、准确、稳定,结合主成分分析,可全面地评价金藤清痹颗粒的产品质量。

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China J Chin Mater Med202247 (13):3530, articleTitle=Quality evaluation of Ginseng Radix et Rhizoma based on UPLC characteristic chromatogram and quantitative analysis of multi-components by single marker (QAMS), refAbstract=null)], funds=[Fund(id=1239171020813300406, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, awardId=2021CXGC010508, language=CN, fundingSource=*山东省重点研发计划(重大科技创新工程)项目(2021CXGC010508), fundOrder=null, country=null), Fund(id=1239171020918158008, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, awardId=鲁动能办〔2021〕23号, language=CN, fundingSource=山东省新旧动能转换重大产业攻关项目(鲁动能办〔2021〕23号), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1239171012001067286, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, xref=1., ext=[AuthorCompanyExt(id=1239171012005261591, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, companyId=1239171012001067286, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 276006,China), AuthorCompanyExt(id=1239171012013650200, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, companyId=1239171012001067286, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.鲁南制药集团股份有限公司 经方与现代中药融合创新全国重点实验室,临沂276006)]), AuthorCompany(id=1239171012101730587, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, xref=2., ext=[AuthorCompanyExt(id=1239171012110119196, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, companyId=1239171012101730587, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.Shandong Engineering Research Center of Generic Manufacture Technology of Traditional Chinese Medicine,Lunan Hope Pharmaceutical Co.,Ltd.,Linyi 276006,China), AuthorCompanyExt(id=1239171012114313502, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, companyId=1239171012101730587, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.鲁南厚普制药有限公司 中药制药共性技术山东省工程研究中心,临沂 276006)])], figs=[ArticleFig(id=1239171017797595751, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Fig.1, caption=UHPLC chromatograms of mixed reference substance (A),sample (B),negative sample without Sinomenii Caulis (C),negative sample without Lonicerae Japonicae Flos (D),negative sample without Angelicae Sinensis Radix (E),negative sample without Glycyrrhizae Radix et Rhizoma (F) and negative sample without Scrophulariae Radix (G), figureFileSmall=xKAOwcl8i+6RTul4CQ4wTQ==, figureFileBig=eXPnl8opqOn1kb1pRRWRCg==, tableContent=null), ArticleFig(id=1239171017864704621, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=图1 , caption=混合对照品(A)、样品(B)、缺青风藤阴性样品(C)、缺金银花阴性样品(D)、缺当归阴性样品(E)、缺甘草阴性样品(F)和缺玄参阴性样品(G)的色谱图

1.青藤碱(sinomenine) 2.新绿原酸(neochlorogenic acid) 3.木兰花碱(magnolflorine) 4.隐绿原酸(cryptochlorogenic acid) 5.绿原酸(chlorogenic acid) 6.异绿原酸B(isochlorogenic acid B) 7.异绿原酸A(isochlorogenic acid A) 8.异绿原酸C(isochlorogenic acid C) 9.哈巴俄苷(harpagoside) 10.藁本内酯(ligustilide) 11.甘草酸铵(ammonium glycyrrhizate)

, figureFileSmall=xKAOwcl8i+6RTul4CQ4wTQ==, figureFileBig=eXPnl8opqOn1kb1pRRWRCg==, tableContent=null), ArticleFig(id=1239171017986339443, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Fig.2, caption=Scatter plot of principal component scores, figureFileSmall=078gsuG8ZqY+Yi512QWaVQ==, figureFileBig=u150la0TImnjiUn04qRK4A==, tableContent=null), ArticleFig(id=1239171018087002744, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=图2 , caption=主成分得分散点图, figureFileSmall=078gsuG8ZqY+Yi512QWaVQ==, figureFileBig=u150la0TImnjiUn04qRK4A==, tableContent=null), ArticleFig(id=1239171018183471741, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Tab.1, caption=

The linear regression equations,correlation coefficients and linear ranges of eleven components

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
r线性范围
(linear range)/(μg·mL-1
青藤碱(sinomenine) Y=8.790×10-2X-8.438×10-20.999 914.36~143.61
新绿原酸(neochlorogenic acid) Y=1.740×10-1X-1.157×10-10.999 87.71~77.15
木兰花碱(magnolflorine) Y=9.129×10-2X+2.428×10-20.999 79.18~91.83
隐绿原酸(cryptochlorogenic acid) Y=1.521×10-1X-1.959×10-10.999 810.71~107.07
绿原酸(chlorogenic acid) Y=1.765×10-1X-2.163×10-10.999 812.88~128.80
异绿原酸B(isochlorogenic acid B) Y=1.707×10-1X-1.826×10-10.999 75.20~51.95
异绿原酸A(isochlorogenic acid A) Y=2.088×10-1X-2.127×10-10.999 85.18~51.84
异绿原酸C(isochlorogenic acid C) Y=1.951×10-1X-2.342×10-10.999 85.40~53.95
哈巴俄苷(harpagoside) Y=1.092×10-1X-1.288×10-20.999 92.62~26.16
藁本内酯(ligustilide) Y=3.193×10-2X-1.273×10-30.999 96.31~63.06
甘草酸铵(ammonium glycyrrhizate) Y=3.646×10-2X-1.103×10-10.997 611.13~111.26
), ArticleFig(id=1239171018288329345, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=表1, caption=

11个成分的线性回归方程、相关系数及线性范围

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
r线性范围
(linear range)/(μg·mL-1
青藤碱(sinomenine) Y=8.790×10-2X-8.438×10-20.999 914.36~143.61
新绿原酸(neochlorogenic acid) Y=1.740×10-1X-1.157×10-10.999 87.71~77.15
木兰花碱(magnolflorine) Y=9.129×10-2X+2.428×10-20.999 79.18~91.83
隐绿原酸(cryptochlorogenic acid) Y=1.521×10-1X-1.959×10-10.999 810.71~107.07
绿原酸(chlorogenic acid) Y=1.765×10-1X-2.163×10-10.999 812.88~128.80
异绿原酸B(isochlorogenic acid B) Y=1.707×10-1X-1.826×10-10.999 75.20~51.95
异绿原酸A(isochlorogenic acid A) Y=2.088×10-1X-2.127×10-10.999 85.18~51.84
异绿原酸C(isochlorogenic acid C) Y=1.951×10-1X-2.342×10-10.999 85.40~53.95
哈巴俄苷(harpagoside) Y=1.092×10-1X-1.288×10-20.999 92.62~26.16
藁本内酯(ligustilide) Y=3.193×10-2X-1.273×10-30.999 96.31~63.06
甘草酸铵(ammonium glycyrrhizate) Y=3.646×10-2X-1.103×10-10.997 611.13~111.26
), ArticleFig(id=1239171018397381254, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Tab.2, caption=

Results of recovery test

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
取样量
(weight)/g
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/%
青藤碱(sinomenine)0.500 01.296 01.029 42.303 0 97.8299.41.2
0.500 11.296 31.029 42.326 8 100.11
0.503 81.305 81.029 42.320 0 98.52
0.504 61.307 91.029 42.334 8 99.76
0.505 31.309 71.029 42.330 3 99.15
0.504 81.308 41.029 42.346 5 100.85
新绿原酸(neochlorogenic acid)0.500 00.691 50.593 21.281 5 99.4699.50.60
0.500 10.691 60.593 21.280 5 99.28
0.503 80.696 80.593 21.284 8 99.12
0.504 60.697 90.593 21.287 3 99.36
0.505 30.698 80.593 21.295 8 100.64
0.504 80.698 10.593 21.285 3 98.99
木兰花碱(magnolflorine)0.500 01.131 01.262 12.381 5 99.0998.70.27
0.500 11.131 21.262 12.376 3 98.65
0.503 81.139 61.262 12.382 8 98.50
0.504 61.141 41.262 12.387 0 98.69
0.505 31.143 01.262 12.384 5 98.37
0.504 81.141 91.262 12.389 8 98.87
隐绿原酸(cryptochlorogenic acid)0.500 00.859 50.840 71.699 0 99.8699.00.72
0.500 10.859 70.840 71.698 0 99.71
0.503 80.866 00.840 71.695 5 98.67
0.504 60.867 40.840 71.700 3 99.07
0.505 30.868 60.840 71.698 8 98.75
0.504 80.867 80.840 71.691 3 97.95
绿原酸(chlorogenic acid)0.500 01.305 51.137 42.439 5 99.7099.60.67
0.500 11.305 81.137 42.449 0 100.51
0.503 81.315 41.137 42.449 5 99.71
0.504 61.317 51.137 42.453 8 99.90
0.505 31.319 31.137 42.443 0 98.80
0.504 81.318 01.137 42.442 0 98.82
异绿原酸B(isochlorogenic acid B)0.500 00.364 00.370 30.732 0 99.3899.41.0
0.500 10.364 10.370 30.730 0 98.81
0.503 80.366 80.370 30.735 0 99.43
0.504 60.367 30.370 30.740 0 100.65
0.505 30.367 90.370 30.739 5 100.35
0.504 80.367 50.370 30.730 3 97.97
异绿原酸A(isochlorogenic acid A)0.500 00.204 00.162 20.366 5 100.1898.11.3
0.500 10.204 00.162 20.362 3 97.60
0.503 80.205 60.162 20.365 3 98.46
0.504 60.205 90.162 20.362 5 96.5
0.505 30.206 20.162 20.365 3 98.09
0.504 80.206 00.162 20.365 0 98.03
异绿原酸C(isochlorogenic acid C)0.500 00.510 00.471 10.976 0 98.9297.90.94
0.500 10.510 10.471 10.971 0 97.83
0.503 80.513 90.471 10.977 8 98.47
0.504 60.514 70.471 10.975 3 97.77
0.505 30.515 40.471 10.978 8 98.37
0.504 80.514 90.471 10.968 5 96.29
哈巴俄苷(harpagoside)0.500 00.080 00.080 60.159 8 99.0198.90.66
0.500 10.080 00.080 60.160 0 99.26
0.503 80.080 60.080 60.160 3 98.88
0.504 60.080 70.080 60.159 8 98.14
0.505 30.080 80.080 60.160 0 98.26
0.504 80.080 80.080 60.161 3 99.88
藁本内酯(ligustilide)0.500 00.600 00.568 81.163 5 99.0798.40.86
0.500 10.600 10.568 81.165 8 99.45
0.503 80.604 60.568 81.166 8 98.84
0.504 60.605 50.568 81.163 5 98.10
0.505 30.606 40.568 81.159 5 97.24
0.504 80.605 80.568 81.162 0 97.78
甘草酸铵(ammonium glycyrrhizate)0.500 00.579 50.429 61.004 8 99.0098.50.50
0.500 10.579 60.429 61.002 3 98.39
0.503 80.583 90.429 61.003 0 97.56
0.504 60.584 80.429 61.008 8 98.70
0.505 30.585 60.429 61.008 8 98.51
0.504 80.585 10.429 61.008 5 98.56
), ArticleFig(id=1239171018548376205, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=表2, caption=

加样回收率试验结果(n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
取样量
(weight)/g
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/%
青藤碱(sinomenine)0.500 01.296 01.029 42.303 0 97.8299.41.2
0.500 11.296 31.029 42.326 8 100.11
0.503 81.305 81.029 42.320 0 98.52
0.504 61.307 91.029 42.334 8 99.76
0.505 31.309 71.029 42.330 3 99.15
0.504 81.308 41.029 42.346 5 100.85
新绿原酸(neochlorogenic acid)0.500 00.691 50.593 21.281 5 99.4699.50.60
0.500 10.691 60.593 21.280 5 99.28
0.503 80.696 80.593 21.284 8 99.12
0.504 60.697 90.593 21.287 3 99.36
0.505 30.698 80.593 21.295 8 100.64
0.504 80.698 10.593 21.285 3 98.99
木兰花碱(magnolflorine)0.500 01.131 01.262 12.381 5 99.0998.70.27
0.500 11.131 21.262 12.376 3 98.65
0.503 81.139 61.262 12.382 8 98.50
0.504 61.141 41.262 12.387 0 98.69
0.505 31.143 01.262 12.384 5 98.37
0.504 81.141 91.262 12.389 8 98.87
隐绿原酸(cryptochlorogenic acid)0.500 00.859 50.840 71.699 0 99.8699.00.72
0.500 10.859 70.840 71.698 0 99.71
0.503 80.866 00.840 71.695 5 98.67
0.504 60.867 40.840 71.700 3 99.07
0.505 30.868 60.840 71.698 8 98.75
0.504 80.867 80.840 71.691 3 97.95
绿原酸(chlorogenic acid)0.500 01.305 51.137 42.439 5 99.7099.60.67
0.500 11.305 81.137 42.449 0 100.51
0.503 81.315 41.137 42.449 5 99.71
0.504 61.317 51.137 42.453 8 99.90
0.505 31.319 31.137 42.443 0 98.80
0.504 81.318 01.137 42.442 0 98.82
异绿原酸B(isochlorogenic acid B)0.500 00.364 00.370 30.732 0 99.3899.41.0
0.500 10.364 10.370 30.730 0 98.81
0.503 80.366 80.370 30.735 0 99.43
0.504 60.367 30.370 30.740 0 100.65
0.505 30.367 90.370 30.739 5 100.35
0.504 80.367 50.370 30.730 3 97.97
异绿原酸A(isochlorogenic acid A)0.500 00.204 00.162 20.366 5 100.1898.11.3
0.500 10.204 00.162 20.362 3 97.60
0.503 80.205 60.162 20.365 3 98.46
0.504 60.205 90.162 20.362 5 96.5
0.505 30.206 20.162 20.365 3 98.09
0.504 80.206 00.162 20.365 0 98.03
异绿原酸C(isochlorogenic acid C)0.500 00.510 00.471 10.976 0 98.9297.90.94
0.500 10.510 10.471 10.971 0 97.83
0.503 80.513 90.471 10.977 8 98.47
0.504 60.514 70.471 10.975 3 97.77
0.505 30.515 40.471 10.978 8 98.37
0.504 80.514 90.471 10.968 5 96.29
哈巴俄苷(harpagoside)0.500 00.080 00.080 60.159 8 99.0198.90.66
0.500 10.080 00.080 60.160 0 99.26
0.503 80.080 60.080 60.160 3 98.88
0.504 60.080 70.080 60.159 8 98.14
0.505 30.080 80.080 60.160 0 98.26
0.504 80.080 80.080 60.161 3 99.88
藁本内酯(ligustilide)0.500 00.600 00.568 81.163 5 99.0798.40.86
0.500 10.600 10.568 81.165 8 99.45
0.503 80.604 60.568 81.166 8 98.84
0.504 60.605 50.568 81.163 5 98.10
0.505 30.606 40.568 81.159 5 97.24
0.504 80.605 80.568 81.162 0 97.78
甘草酸铵(ammonium glycyrrhizate)0.500 00.579 50.429 61.004 8 99.0098.50.50
0.500 10.579 60.429 61.002 3 98.39
0.503 80.583 90.429 61.003 0 97.56
0.504 60.584 80.429 61.008 8 98.70
0.505 30.585 60.429 61.008 8 98.51
0.504 80.585 10.429 61.008 5 98.56
), ArticleFig(id=1239171018682593940, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Tab.3, caption=

Contents of eleven components in Jinteng Qingbi granules

, figureFileSmall=null, figureFileBig=null, tableContent=
样品编号
(sample No.)
含量(content)/(mg·g-1
青藤碱
(sinomenine)
新绿原酸
(neochloro-genic acid)
木兰花碱
(magnolflorine)
隐绿原酸
(cryptochloro-genic acid)
绿原酸
(chloroge-nic acid)
异绿原酸B
(isochloroge- nic acid B)
异绿原酸A
(isochloroge- nic acid A)
异绿原酸C
(isochloroge-nic acid C)
哈巴俄苷
(harpagoside)
藁本内酯
(ligustilide)
甘草酸铵
(ammonium glycyrrhizate)
S12.5921.3832.2621.7192.6120.7280.4081.0200.1601.2001.159
S22.5011.3092.1411.5982.4470.6750.3910.9450.1551.1371.111
S32.4761.3182.1441.6052.4090.6930.3930.9520.1541.1221.097
S42.4671.3132.1581.6062.4130.7010.3930.9570.1521.1141.093
S52.5831.3622.2511.6822.5200.7260.4101.0000.1591.1661.133
S62.5881.3722.2431.6282.5180.7340.3960.9950.1561.1711.130
S72.5231.3652.1961.6692.5420.7260.4060.9860.1801.1631.124
S82.4361.2942.1091.6112.4390.7040.3950.9490.1651.1131.090
S92.5491.2262.4101.5282.5200.6900.4030.9500.1840.8841.331
S102.3891.1512.2811.3862.3760.6370.3810.9000.1760.8331.255
S112.4581.1812.3401.4232.4400.6470.3880.9220.1790.8541.285
S122.6161.2552.4871.5142.5940.6990.4120.9820.1910.9021.363
S132.2191.0952.1461.3212.2490.6160.3630.8460.1820.7951.206
S142.2061.0712.0961.3542.2410.6050.3640.8350.1790.7911.187
S152.6201.2592.2231.6092.5000.6940.3970.9470.1710.8271.043
S162.7041.3192.3181.6622.6010.7190.3990.9890.1630.8691.074
S172.6191.2562.2461.5422.4480.6820.3900.9490.1550.8251.029
S182.5591.2362.2001.5052.3920.6840.3800.9280.1510.8081.008
S192.6041.2572.2401.5842.4370.6840.3880.9440.1550.8171.026
S202.5691.2422.2151.5142.4070.6700.3830.9350.1540.7981.015
), ArticleFig(id=1239171018846171800, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=表3, caption=

金藤清痹颗粒中11个成分的含量(n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品编号
(sample No.)
含量(content)/(mg·g-1
青藤碱
(sinomenine)
新绿原酸
(neochloro-genic acid)
木兰花碱
(magnolflorine)
隐绿原酸
(cryptochloro-genic acid)
绿原酸
(chloroge-nic acid)
异绿原酸B
(isochloroge- nic acid B)
异绿原酸A
(isochloroge- nic acid A)
异绿原酸C
(isochloroge-nic acid C)
哈巴俄苷
(harpagoside)
藁本内酯
(ligustilide)
甘草酸铵
(ammonium glycyrrhizate)
S12.5921.3832.2621.7192.6120.7280.4081.0200.1601.2001.159
S22.5011.3092.1411.5982.4470.6750.3910.9450.1551.1371.111
S32.4761.3182.1441.6052.4090.6930.3930.9520.1541.1221.097
S42.4671.3132.1581.6062.4130.7010.3930.9570.1521.1141.093
S52.5831.3622.2511.6822.5200.7260.4101.0000.1591.1661.133
S62.5881.3722.2431.6282.5180.7340.3960.9950.1561.1711.130
S72.5231.3652.1961.6692.5420.7260.4060.9860.1801.1631.124
S82.4361.2942.1091.6112.4390.7040.3950.9490.1651.1131.090
S92.5491.2262.4101.5282.5200.6900.4030.9500.1840.8841.331
S102.3891.1512.2811.3862.3760.6370.3810.9000.1760.8331.255
S112.4581.1812.3401.4232.4400.6470.3880.9220.1790.8541.285
S122.6161.2552.4871.5142.5940.6990.4120.9820.1910.9021.363
S132.2191.0952.1461.3212.2490.6160.3630.8460.1820.7951.206
S142.2061.0712.0961.3542.2410.6050.3640.8350.1790.7911.187
S152.6201.2592.2231.6092.5000.6940.3970.9470.1710.8271.043
S162.7041.3192.3181.6622.6010.7190.3990.9890.1630.8691.074
S172.6191.2562.2461.5422.4480.6820.3900.9490.1550.8251.029
S182.5591.2362.2001.5052.3920.6840.3800.9280.1510.8081.008
S192.6041.2572.2401.5842.4370.6840.3880.9440.1550.8171.026
S202.5691.2422.2151.5142.4070.6700.3830.9350.1540.7981.015
), ArticleFig(id=1239171018972000927, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Tab.4, caption=

Eigenvalue,variance contribution,accumulative contribution and eigenvector of three principal components

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
主成分(principal component)
F1 F2 F3
青藤碱(sinomenine)0.318 40.061 8-0.487 5
新绿原酸(neochlorogenic acid)0.367 3-0.127 30.136 5
木兰花碱(magnolflorine)0.110 50.540 5-0.332 6
隐绿原酸(cryptochlorogenic acid)0.363 9-0.146 70.080 6
绿原酸(chlorogenic acid)0.345 50.240 3-0.098 0
异绿原酸B(isochlorogenic acid B)0.373 80.003 10.066 9
异绿原酸A(isochlorogenic acid A)0.352 40.192 20.081 6
异绿原酸C(isochlorogenic acid C)0.378 80.060 50.005 5
哈巴俄苷(harpagoside)-0.136 80.504 30.300 6
藁本内酯(ligustilide)0.255 3-0.134 80.635 0
甘草酸铵(ammonium glycyrrhizate)-0.092 90.543 60.332 6
特征值(eigenvalue)6.8452.5671.191
方差贡献率(variance contribution)/%62.23123.34010.823
累计方差贡献率(accumulative contribution)/%62.23185.57196.394
), ArticleFig(id=1239171019085247140, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=表4, caption=

3个主成分的特征值、方差贡献率、累计方差贡献率及其特征向量

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
主成分(principal component)
F1 F2 F3
青藤碱(sinomenine)0.318 40.061 8-0.487 5
新绿原酸(neochlorogenic acid)0.367 3-0.127 30.136 5
木兰花碱(magnolflorine)0.110 50.540 5-0.332 6
隐绿原酸(cryptochlorogenic acid)0.363 9-0.146 70.080 6
绿原酸(chlorogenic acid)0.345 50.240 3-0.098 0
异绿原酸B(isochlorogenic acid B)0.373 80.003 10.066 9
异绿原酸A(isochlorogenic acid A)0.352 40.192 20.081 6
异绿原酸C(isochlorogenic acid C)0.378 80.060 50.005 5
哈巴俄苷(harpagoside)-0.136 80.504 30.300 6
藁本内酯(ligustilide)0.255 3-0.134 80.635 0
甘草酸铵(ammonium glycyrrhizate)-0.092 90.543 60.332 6
特征值(eigenvalue)6.8452.5671.191
方差贡献率(variance contribution)/%62.23123.34010.823
累计方差贡献率(accumulative contribution)/%62.23185.57196.394
), ArticleFig(id=1239171019177521833, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=EN, label=Tab.5, caption=

Overall evaluation score for the quality of 20 batches of Jinteng Qingbi granules

, figureFileSmall=null, figureFileBig=null, tableContent=
批号
(sample No.)
得分(score)综合得分
(overall score)
综合排名
(overall ranking)
F1 F2 F3
S13.960.250.832.611
S20.28-1.200.970.0012
S30.53-1.720.810.0111
S40.66-1.630.710.1010
S52.91-0.140.761.862
S62.48-0.280.681.555
S72.490.291.521.783
S80.60-1.331.210.199
S90.102.94-0.160.737
S10-3.121.460.18-1.5818
S11-1.932.13-0.07-0.7115
S121.124.26-0.281.664
S13-5.69-0.040.80-3.4619
S14-6.09-0.460.89-3.8020
S150.66-0.11-1.020.278
S162.490.37-1.541.476
S170.17-0.9-1.67-0.2913
S18-0.88-1.51-1.47-1.0617
S190.10-1.05-1.56-0.3514
S20-0.85-1.30-1.61-1.0116
), ArticleFig(id=1239171019282379438, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239148841568620698, language=CN, label=表5, caption=

20批金藤清痹颗粒质量的综合评价得分

, figureFileSmall=null, figureFileBig=null, tableContent=
批号
(sample No.)
得分(score)综合得分
(overall score)
综合排名
(overall ranking)
F1 F2 F3
S13.960.250.832.611
S20.28-1.200.970.0012
S30.53-1.720.810.0111
S40.66-1.630.710.1010
S52.91-0.140.761.862
S62.48-0.280.681.555
S72.490.291.521.783
S80.60-1.331.210.199
S90.102.94-0.160.737
S10-3.121.460.18-1.5818
S11-1.932.13-0.07-0.7115
S121.124.26-0.281.664
S13-5.69-0.040.80-3.4619
S14-6.09-0.460.89-3.8020
S150.66-0.11-1.020.278
S162.490.37-1.541.476
S170.17-0.9-1.67-0.2913
S18-0.88-1.51-1.47-1.0617
S190.10-1.05-1.56-0.3514
S20-0.85-1.30-1.61-1.0116
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基于UHPLC多成分测定与主成分分析的金藤清痹颗粒质量评价*
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邓丽华 1, 2 , 许克宁 1, 2 , 袁晓梅 1, 2 , 丁兵 1 , 王丽苹 1, 2 , 徐丽 1, 2 , 杨本官 1 , 刘源慧 2 , 范建伟 1, 2, **
药物分析杂志 | 质量分析 2024,44(3): 522-531
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药物分析杂志 | 质量分析 2024, 44(3): 522-531
基于UHPLC多成分测定与主成分分析的金藤清痹颗粒质量评价*
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邓丽华1, 2 , 许克宁1, 2, 袁晓梅1, 2, 丁兵1, 王丽苹1, 2, 徐丽1, 2, 杨本官1, 刘源慧2, 范建伟1, 2, **
作者信息
  • 1.鲁南制药集团股份有限公司 经方与现代中药融合创新全国重点实验室,临沂276006
  • 2.鲁南厚普制药有限公司 中药制药共性技术山东省工程研究中心,临沂 276006
  • Tel:13754746480;E-mail:

通讯作者:

**Tel:(0539)8336639;E-mail:
Quality evaluation of Jinteng Qingbi granules based on UHPLC multi-index component determination and principal component analysis*
Li-hua DENG1, 2 , Ke-ning XU1, 2, Xiao-mei YUAN1, 2, Bing DING1, Li-ping WANG1, 2, Li XU1, 2, Ben-guan YANG1, Yuan-hui LIU2, Jian-wei FAN1, 2, **
Affiliations
  • 1.State Key Laboratory of Integration and Innovation of Classic Formula and Modern Chinese Medicine,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 276006,China
  • 2.Shandong Engineering Research Center of Generic Manufacture Technology of Traditional Chinese Medicine,Lunan Hope Pharmaceutical Co.,Ltd.,Linyi 276006,China
出版时间: 2024-03-31 doi: 10.16155/j.0254-1793.2024.03.18
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目的:

建立同时测定金藤清痹颗粒中青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分含量的UHPLC法,并结合主成分分析对制剂进行质量评价。

方法:

采用UHPLC波长切换法,色谱柱为Waters XSelect® CSH C18柱(150 mm×4.6 mm,2.5 μm),以甲醇-乙腈(1:1)为流动相A,0.2%磷酸水溶液为流动相B,梯度洗脱,流速1.0 mL·min-1,柱温25 ℃,检测波长218 nm(0~17 min时,检测青藤碱)、326 nm (17~25 min和34~98 min时,检测新绿原酸、隐绿原酸、绿原酸和异绿原酸A、异绿原酸B、异绿原酸C)、263 nm(25~34 min和98~125 min时,检测木兰花碱、哈巴俄苷、藁本内酯、甘草酸铵)。利用SPSS27.0软件对20批金藤清痹颗粒中成分含量进行多元统计分析。

结果:

青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分的质量浓度分别在14.36~143.61 μg·mL-1r=0.999 9)、7.71~77.15 μg·mL-1r=0.999 8)、9.18~91.83 μg·mL-1r=0.999 7)、10.71~107.07 μg·mL-1r=0.999 8)、12.88~128.80 μg·mL-1r=0.999 8)、5.20~51.95 μg·mL-1r=0.999 7)、5.18~51.84 μg·mL-1r=0.999 8)、5.40~53.95 μg·mL-1r=0.999 8)、2.62~26.16 μg·mL-1r=0.999 9)、6.31~63.06 μg·mL-1r=0.999 9)和11.13~111.26 μg·mL-1r=0.997 6)范围内与峰面积的线性关系良好;平均加样回收率(n=6)分别为98.3%、98.3%、98.5%、98.9%、99.2%、101.0%、98.1%、97.1%、96.8%、98.0%和98.7%,RSD均小于3.0%。20批金藤清痹颗粒样品中,上述青藤碱等11个成分的含量测定结果(n=6)依次为2.206~2.704、1.071~1.403、2.096~2.487、1.321~1.724、2.241~2.612、0.605~0.749、0.363~0.412、0.835~1.020、0.151~0.191、0.791~1.188和1.008~1.363 mg·g-1。主成分分析结果显示,连续生产的金藤清痹颗粒批次间质量差异较小,且以样品S1、S5和S7的综合质量相对更好。

结论:

建立的UHPLC多指标成分含量测定方法简便、准确、稳定,结合主成分分析,可全面地评价金藤清痹颗粒的产品质量。

金藤清痹颗粒  /  超高效液相色谱  /  波长切换  /  含量测定  /  主成分分析  /  质量评价
Objective:

To establish a UHPLC method for simultaneously determining sinomenine,neochlorogenic acid,magnolflorine,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,harpagoside,ligustilide and ammonium glycyrrhizate in Jinteng Qingbi granules,and evaluate the quality of Jinteng Qingbi granules combined with principal component analysis.

Methods:

UHPLC wavelength switching method was employed in the study. The chromatographic separation was performed on a Waters XSelect® CSH C18 column (150 mm×4.6 mm,2.5 μm) using methanol-acetonitrile (1:1) as mobile phase A and 0.2% phosphoric acid aqueous solution as mobile phase B in a gradient mode at 25 ℃. The flow rate was 1.0 mL·min-1,and the UV detection wavelength was chosen at 218 nm for sinomenine during 0-17 min,326 nm for neochlorogenic acid,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C during 17-25 min and 34-98nm,263 nm for magnolflorine,harpagoside,ligustilide and ammonium glycyrrhizate during 25-34 min and 98-125nm,respectively. Furthermore,multiple statistical analysis was conducted on the contents of 11 components in 20 batches of Jinteng Qingbi granules using SPSS27.0 software.

Resluts:

Satisfactory linearities of sinomenine,neochlorogenic acid,magnolflorine,cryptochlorogenic acid,chlorogenic acid,isochlorogenic acid B,isochlorogenic acid A,isochlorogenic acid C,harpagoside,ligustilide and ammonium glycyrrhizate were in the ranges of 14.36-143.61 μg·mL-1 (r=0.999 9),7.71-77.15 μg·mL-1 (r=0.999 8),9.18-91.83 μg·mL-1 (r=0.999 7),10.71-107.07 μg·mL-1 (r=0.999 8),12.88-128.80 μg·mL-1 (r=0.999 8),5.20-51.95 μg·mL-1 (r=0.999 7),5.18-51.84 μg·mL-1 (r=0.999 8),5.40-53.95 μg·mL-1 (r=0.999 8),2.62-26.16 μg·mL-1 (r=0.999 9),6.31-63.06 μg·mL-1 (r=0.999 9) and 11.13-111.26 μg·mL-1 (r=0.997 6),respectively. The average recoveries (n=6) were 98.3%,98.3%,98.5%,98.9%,99.2%,101.0%,98.1%,97.1%,96.8%,98.0% and 98.7%,respectively,with RSDs less than 3.0%. The contents ranges of sinomenine and other 10 components of 20 batches of Jinteng Qingbi granules samples were 2.206-2.704,1.071-1.403,2.096-2.487,1.321-1.724,2.241-2.612,0.605-0.749,0.363-0.412,0.835-1.020,0.151-0.191,0.791-1.188 and 1.008-1.363 mg·g-1,respectively. The results of principal component analysis showed that the quality differences between batches of continuously produced Jinteng Qingbi granules were relatively smaller,and the comprehensive quality of samples S1,S5 and S7 was relatively better.

Conclusion:

The established UHPLC method of multi-index component determination is simple,accurate and stable. Combined with principal component analysis,it can be used for quality evaluation of Jinteng Qingbi granules comprehensively.

Jinteng Qingbi granules  /  UHPLC  /  wavelength switching  /  content determination  /  principal component analysis  /  quality evaluation
邓丽华, 许克宁, 袁晓梅, 丁兵, 王丽苹, 徐丽, 杨本官, 刘源慧, 范建伟. 基于UHPLC多成分测定与主成分分析的金藤清痹颗粒质量评价*. 药物分析杂志, 2024 , 44 (3) : 522 -531 . DOI: 10.16155/j.0254-1793.2024.03.18
Li-hua DENG, Ke-ning XU, Xiao-mei YUAN, Bing DING, Li-ping WANG, Li XU, Ben-guan YANG, Yuan-hui LIU, Jian-wei FAN. Quality evaluation of Jinteng Qingbi granules based on UHPLC multi-index component determination and principal component analysis*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (3) : 522 -531 . DOI: 10.16155/j.0254-1793.2024.03.18
类风湿性关节炎(rheumatoid arthritis,RA)是一种以慢性进展性滑膜炎症为特征的自身免疫性疾病,其临床发病率约0.5%~1.0%,既严重影响患者生活质量,又为社会带来极大的经济负担[1]。目前,临床常会选用非甾体抗炎药、糖皮质激素、抗风湿药等对症治疗,但上述药物易引起脱发、口腔炎、肝毒性等不良反应,极大限制了对RA患者的治疗[2]。RA属于中医学“痹证”范畴,又称“尪痹”“顽痹”“鹤膝风”等,为中医优势病种[3],中医药治疗RA具有独特优势和良好疗效。金藤清痹颗粒源于清·鲍相璈《验方新编》中“四妙勇安汤”,由金银花、青风藤、白花蛇舌草、玄参、白芍、生地黄、山慈菇、鹿衔草、当归、甘草、蜈蚣等11味药物组成,具有清热解毒、活血消肿、通痹止痛的功效,用于RA活动期的治疗[4]。现代临床研究表明,金藤清痹颗粒可明显改善活动期RA患者的临床症状、体征及各项生化指标,疗效确切[5]
金藤清痹颗粒现行质量标准[6]仅以青藤碱、绿原酸2个成分作为定量控制指标,迄今未见有金藤清痹颗粒多成分同时含量测定的文献报道。中药化学成分的复杂性决定了以多指标成分的同时测定进行质量控制更加全面、科学、合理[7-8]。为进一步评价金藤清痹颗粒产品质量,本研究采用UHPLC波长切换法对金藤清痹颗粒中青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个指标成分含量进行同时测定,并结合主成分分析,为全面、客观、有效地评价金藤清痹颗粒的质量提供科学参考。
Thermo Vanquish Flex超高效液相色谱系统(配备DAD检测器、Chromeleon 7.2色谱工作站,Thermo Fisher Scientific公司);Waters XSelect® CSH C18色谱柱(150 mm×4.6 mm,2.5 μm;Waters公司);XS204万分之一电子分析天平、XS105DU十万分之一电子分析天平(Mettler-Toledo公司);JY-15超声波清洗机(鼎泰(湖北)生化科技设备制造有限公司)。
对照品青藤碱(批号110774-201808,纯度94.6%)、绿原酸(批号110753-201817,纯度96.8%)、哈巴俄苷(批号111730-201709,纯度95.9%)、甘草酸铵(批号110731-202021,纯度96.2%)均购自中国食品药品检定研究院,新绿原酸(批号MUST-21030108,纯度99.67%)、隐绿原酸(批号MUST-21082610,纯度99.88%)、异绿原酸B(批号MUST-21030602,纯度99.05%)、异绿原酸A(批号MUST-21102611,纯度98.46%)、异绿原酸C(批号MUST-21081010,纯度99.77%)均购自成都曼思特生物科技有限公司,木兰花碱(批号A31HB193420,纯度98.0%)购自上海源叶生物科技有限公司;藁本内酯(批号070017-201912,纯度98.0%)购自上海鸿永生物科技有限公司。甲醇、乙腈为色谱纯(Merck公司),其他试剂均为分析纯,水为自制超纯水。20批金藤清痹颗粒样品(规格:每袋10 g;批号分别为28200081、28210001、28210011、28210021、28210031、28210041、28210051、28210061、28210071、28210081、28210091、28210101、28210111、28210121、28210131、28210141、28210151、28210161、28210171、28210181、28210191,编号依次为S1~S20)由鲁南厚普制药有限公司提供。
取金藤清痹颗粒适量,研细,精密称取1 g,置25 mL量瓶中,加入40%甲醇水溶液20 mL,超声处理(功率300 W,频率40 kHz)30 min,取出,放冷,定容至刻度,摇匀,用微孔滤膜(Φ 0.22 μm)滤过,取续滤液,即得。
精密称取“1.2”项下各对照品适量,加40%甲醇水溶液制成青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵的质量浓度分别为143.61、77.15、91.83、107.07、128.75、51.95、51.84、53.95、26.16、63.06和111.26 μg·mL-1的混合对照品溶液。
参考金藤清痹颗粒处方与制法[6],分别制备缺青风藤、缺金银花、缺当归、缺甘草、缺玄参的5种阴性样品,“2.1”项下方法,分别制得相应阴性对照溶液。
色谱柱:Waters XSelect® CSH C18柱(150 mm×4.6 mm,2.5 μm);流动相:甲醇-乙腈(1:1)为流动相A,0.2%磷酸水溶液为流动相B,梯度洗脱(0~13 min,3%A→7%A;13~45 min,7%A→13%A;45~52 min,13%A→20%A;52~85 min,20%A→25%A;85~95 min,25%A →35%A;95~102 min,35%A→55%A;102~120 min,55%A→60%A;120~125 min,60%A→70%A);流速:1.0 mL·min-1;柱温:25 ℃;检测波长:218 nm(0~17 min时,检测青藤碱)、326 nm(17~25 min和34~98 min时,检测新绿原酸、隐绿原酸、绿原酸和异绿原酸A、异绿原酸B、异绿原酸C)、263 nm(25~34 min和98~125 min时,检测木兰花碱、哈巴俄苷、藁本内酯、甘草酸铵);进样量:5 μL。
分别精密吸取混合对照品溶液、供试品溶液及各阴性对照溶液,按“2.4”项下色谱条件进样分析。样品中青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯、甘草酸铵各峰的分离度和对称性良好,其分离度均大于1.5,对称因子均在0.90~1.10;对照品溶液及样品溶液目标峰的保留时间一致,且阴性样品无干扰。色谱图见图1
精密吸取“2.2”项下混合对照品溶液1、2、3、5、8、10 mL,分别置于10 mL量瓶中,以40%甲醇水溶液稀释至刻度,摇匀,即得系列浓度的混合对照品溶液。精密吸取上述系列混合对照品溶液,按“2.4”项下色谱条件进样测定。以峰面积(Y)对质量浓度(X)进行线性回归,得青藤碱等11个成分的回归方程及其线性范围,见表1
取“2.2”项下混合对照品溶液,按“2.4”项下色谱条件进样,连续测定6次,计算相应峰面积及其RSD。结果青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵峰面积的RSD分别为0.71%、0.60%、0.60%、0.76%、0.83%、1.1%、1.6%、0.75%、0.67%、0.71%和0.82%,表明仪器精密度良好。
精密称取同一批样品(批号28200081,编号S1)6份,按“2.1”项下方法制备供试品溶液,按“2.4”项下色谱条件进样,记录相应峰面积。计算青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵的平均含量(n=6)分别为2.592、1.383、2.262、1.719、2.611、0.728、0.408、1.020、0.160、1.200和1.159 mg·g-1,其RSD依次为0.22%、0.41%、0.23%、0.58%、0.52%、0.87%、1.4%、1.7%、0.26%、0.77%和0.65%,表明该法重复性良好。
室温条件下取S1号样品,按“2.1”项下方法制备供试品溶液,并按“2.4”项下色谱条件分别于制备后0、3、6、9、12、24 h进样测定,记录相应峰面积。结果青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵峰面积的RSD分别为0.30%、0.25%、0.36%、0.32%、0.29%、0.68%、0.69%、0.56%、1.6%、0.69%和0.79%,表明供试品溶液室温下24 h内稳定。
取含量已知的S1号样品细粉6份,每份0.5 g,精密称定,分别置于25 mL量瓶中,每份加入混合对照品储备液10 mL(每1 mL分别含青藤碱102.94 μg、新绿原酸59.32 μg、木兰花碱126.21 μg、隐绿原酸84.07 μg、绿原酸113.74 μg、异绿原酸B 37.03 μg、异绿原酸A 16.22 μg、异绿原酸C 47.11 μg、哈巴俄苷8.06 μg、藁本内酯56.88 μg和甘草酸铵42.96 μg),按“2.1”项下方法制备加样供试溶液,并按“2.4”项下色谱条件进样,计算各成分回收率及其RSD,结果见表2
取20批金藤清痹颗粒样品(编号S1~S20),分别按“2.1”项下方法制备供试品溶液,每批样品平行3份,并按“2.4”项下色谱条件进样测定,计算各批样品中青藤碱等11个成分的含量,结果见表3
将“2.6”项下样品含量测定结果的标准化得分作为变量,导入SPSS 27.0软件进行主成分分析,并以特征值>1为判定标准,按方差贡献率大小提取出3个主成分,依次记作F1F2F3,且三者的累计方差贡献率达96.394%,表明该3个主成分基本反映了11个指标的全部信息,可对样品降维分析,结果见表4。在主成分F1中,青藤碱、新绿原酸、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A和异绿原酸C的特征向量的绝对值均大于其他变量,表明F1主要反映了青风藤、金银花2味药材的质量信息;同理,F2主要反映了玄参、甘草和青风藤3味药材的质量信息,F3反映了当归、青风藤和玄参的质量信息。
依据各指标成分的特征向量,分别求得3个主成分F1~F3的函数表达式:Y1=0.318 4X1+0.367 3X2+0.110 5X3+0.363 9X4+0.345 5X5+0.373 8X6+0.352 4X7+0.378 8X8-0.136 8X9+0.255 3X10-0.092 9X11Y2=0.061 8X1-0.127 3X2+0.540 5X3-0.146 7X4+0.240 3X5+0.003 1X6+0.192 2X7+0.060 5X8+0.504 3X9-0.134 8X10+0.543 6X11Y3=-0.487 5X1+0.136 5X2-0.332 6X3+0.080 6X4-0.098 0X5+0.066 9X6+0.081 6X7+0.005 5X8+0.300 6X9+0.635 0X10+0.332 6X11;式中,Y1Y2Y3分别代表3个主成分F1F2F3的得分,X1X2~Xn依次表示青藤碱、新绿原酸、木兰花碱等11个成分含量的标准化数据。
将20批金藤清痹颗粒样品(S1~S20)中青藤碱等11个成分含量的标准化数据,依次代入上述函数表达式中,得到主成分得分,并导入SPSS 27.0软件绘制出主成分得分散点图,见图2。图中样品点的空间距离越近,表示样品的化学成分更为接近,其中,S1~S8、S15~S20、S9~S12、S13~S14分别聚为4个居群。分别以各主成分对应的方差贡献率作为权重,对主成分得分及其相应的权重进行线性加权,构建金藤清痹颗粒评价函数:Y=0.622 3 Y1+0.233 4Y2+0.108 2Y3,计算各样品的综合评价得分,分值越高表示该样品整体质量越好[9-10]。结果表明,20批金藤清痹颗粒样品中,以S1、S5、S7的整体质量更好,结果见表5
为了对金藤清痹颗粒进行全面质量评控,本研究前期针对该制剂药味组成,结合现行《中华人民共和国药典》及相关文献报道,选择君药金银花中的绿原酸、隐绿原酸、新绿原酸、异绿原酸A、异绿原酸B、异绿原酸C、木犀草苷[11-12]和青风藤中的青藤碱、木兰花碱[13],臣药当归中的阿魏酸、藁本内酯[14]和白芍中的芍药苷,佐药玄参中的哈巴苷、哈巴俄苷[15]及使药甘草中的甘草苷、甘草酸铵,一并作为候选目标成分进行定量分析方法学研究。结果发现,金藤清痹颗粒中木犀草苷、阿魏酸、芍药苷、哈巴苷及甘草苷5个成分的含量相对较低,甚者未能检出。为确保定量分析结果准确性,本研究最终选定绿原酸、隐绿原酸、新绿原酸、异绿原酸A、异绿原酸B、异绿原酸C、青藤碱、木兰花碱、藁本内酯、哈巴俄苷、甘草酸铵等11个成分作为待测成分,藉此对金藤清痹颗粒进行多指标成分定量控制。
上述绿原酸等11个待测目标成分的结构多样,包含有机酸类(绿原酸、隐绿原酸、新绿原酸、异绿原酸A、异绿原酸B、异绿原酸C),生物碱类(青藤碱、木兰花碱),香豆素类(藁本内酯),三萜皂苷类(甘草酸铵)及环烯醚萜苷类(哈巴俄苷)。为实现对绿原酸等11个成分的同时测定,利用DAD检测器在200~400 nm范围内进行全波长扫描,并综合考虑各成分的紫外吸收及相互干扰情况,最终确定采用波长切换法在218 nm下检测青藤碱,326 nm下检测新绿原酸、隐绿原酸、绿原酸、异绿原酸A、异绿原酸B和异绿原酸C,263 nm下检测木兰花碱、哈巴俄苷、藁本内酯和甘草酸铵。
分别考察了Waters CORTECS UPLC T3、Waters Acquity UPLC HSS T3、Waters UPLC BEH Shield PR18、Waters XSelect® CSH C18和Thermo GOLDTM C18 5款色谱柱的适用性,结果相同条件下Waters XSelect® CSH C18柱能使11个成分达到基线分离,其峰形和分离度更佳,柱效更高,成为本研究待测成分分离分析的最佳选择。同时,比较了甲醇-0.1%甲酸水溶液、乙腈-0.1%甲酸水溶液、甲醇-乙腈(1:1)-0.1%甲酸水溶液、甲醇-乙腈(1:1)-0.2%甲酸水溶液、甲醇-乙腈(1:1)-0.1%磷酸水溶液和甲醇-乙腈(1:1)-0.2%磷酸水溶液6种流动相系统,结果以甲醇-乙腈(1:1)为有机相、0.2%磷酸水溶液为水相时,更利于待测目标成分的分离,基线噪音最小,各色谱峰分离度、对称性更佳,故选用甲醇-乙腈(1:1)-0.2%磷酸水溶液为流动相系统进行梯度洗脱。
针对超声提取和回流提取2种提取方法,以20%甲醇水溶液、40%甲醇水溶液、70%甲醇水溶液和甲醇为提取溶剂分别进行了考察。结果表明,上述2种提取方法及4种提取溶剂对待测目标成分的提取效果无明显差别,为降低流动相与提取溶剂之间因极性差异所造成的溶剂效应,最终确定以40%甲醇水溶液为提取溶剂,进行超声提取。
作为常用的复杂数据统计分析方法,主成分分析利用降维思想,将多个相互关联的多变量通过线性变换,在保留原始数据大部分信息的前提下,选择少数几个具有代表性的综合指标(主成分)代替多个原始变量,可以最大限度地保留样本的原始信息并对样本做出整体性评价[16-18]。本研究中,主成分分析结果表明金藤清痹颗粒的制剂工艺稳定性好,连续生产批次之间的整体质量更为接近,而非连续生产批次之间的质量差异,更多地体现出药材在不同投料时间阶段内的质量波动性。
本研究采用波长切换技术,建立了UHPLC同时测定金藤清痹颗粒中青藤碱、新绿原酸、木兰花碱、隐绿原酸、绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、哈巴俄苷、藁本内酯和甘草酸铵11个成分的方法。该法简便、准确、稳定,通过结合多元统计分析,可用于金藤清痹颗粒的质量评价。
  • *山东省重点研发计划(重大科技创新工程)项目(2021CXGC010508)
  • 山东省新旧动能转换重大产业攻关项目(鲁动能办〔2021〕23号)
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2024年第44卷第3期
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doi: 10.16155/j.0254-1793.2024.03.18
  • 接收时间:2023-09-14
  • 首发时间:2026-03-13
  • 出版时间:2024-03-31
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  • 收稿日期:2023-09-14
基金
*山东省重点研发计划(重大科技创新工程)项目(2021CXGC010508)
山东省新旧动能转换重大产业攻关项目(鲁动能办〔2021〕23号)
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    1.鲁南制药集团股份有限公司 经方与现代中药融合创新全国重点实验室,临沂276006
    2.鲁南厚普制药有限公司 中药制药共性技术山东省工程研究中心,临沂 276006

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https://castjournals.cast.org.cn/joweb/ywfxzz/CN/10.16155/j.0254-1793.2024.03.18
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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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