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Article(id=1193877803474055309, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1193877800143777917, articleNumber=1001-2494(2025)02-0172-08, orderNo=null, doi=10.11669/cpj.2025.02.009, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1712073600000, receivedDateStr=2024-04-03, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1762572249524, onlineDateStr=2025-11-08, pubDate=1737475200000, pubDateStr=2025-01-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1762572249524, onlineIssueDateStr=2025-11-08, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1762572249524, creator=13701087609, updateTime=1762572249524, updator=13701087609, issue=Issue{id=1193877800143777917, tenantId=1146029695717560320, journalId=1190317699101192196, year='2025', volume='60', issue='2', pageStart='109', pageEnd='206', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=0, articleOrder=1, issueType=-1, specialIssue=null, createTime=1762572248731, creator=13701087609, updateTime=1762584852274, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1193930663289123481, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1193877800143777917, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1193930663289123482, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1193877800143777917, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=172, endPage=179, ext={EN=ArticleExt(id=1193877803679576209, articleId=1193877803474055309, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Simultaneous Determination of 9 Components in Xiaoxianxiong Decoction Based on QAMS Method, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of hypoxanthine, vanillic acid, magnolithine, demethyleneberberine, Columbamine, Jatrorrhizine hydrochloride, coptisine hydrochloride, palmatine hydrochloride and berberine hydrochloride in Xiaoxianxiong decoction. METHODS The separation was performed on a Spursil C18 column (4.6 mm×250 mm, 5 μm) with acetonitrile and 0.05 mol·L-1 potassium dihydrogen phosphate solution plus phosphoric acid adjusted to pH=4.0 as the mobile phases. Detection wavelength: 0-20 min: 290 nm; 20-45 min: 300 nm, column temperature 30 ℃, injection volume 10 μL. Using berberine hydrochloride as the internal reference material, the relative correction factors of the remaining eight components were calculated and the content of each component was calculated, and the results were compared with those of the external standard method (ESM) and the QAMS method. The differences between the results of the external standard method (ESM) and QAMS method were compared. RESULTS The nine components of Xiaoxianxiong decoction showed a good linear relationship with peak area in a certain concentration range (r>0.998 0), with the average spiked recoveries of 94.36%-105.57% and the RSD was 0.52%-3.48%, and the results of component contents determined by QAMS and ESM were not statistically significant. CONCLUSION The results of the established QAMS for Xiaoxianxiong decoction are accurate and reliable, and the method is simple and easy to be applied to the quality control of Xiaoxianxiong Decoction.

, correspAuthors=Xia 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=Yuan LIU, Zhiyuan NING, Chenxi LU, Zhiyong CHEN, Hui REN, Feifei REN, Xia DU), CN=ArticleExt(id=1193877893114720564, articleId=1193877803474055309, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=基于一测多评法同时测定经典名方小陷胸汤中9种成分含量, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 建立一测多评法(quantitative analysis of multi-components by single marker, QAMS)同时测定小陷胸汤中次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的含量。方法 本实验采用色谱柱Spursil C18(4.6 mm×250 mm,5 μm),以乙腈-0.05 mol·L-1磷酸二氢钾溶液加磷酸调pH=4.0为流动相,检测波长: 0~20 min:290 nm;20~45 min:300 nm,柱温30 ℃,进样量10 μL。以盐酸小檗碱为内参物,分别计算其余8个成分的相对校正因子并计算各成分含量,比较外标法(external standard method, ESM)和QAMS法结果的差异。结果 小陷胸汤9个成分在一定浓度范围与峰面积呈良好的线性关系(r>0.998 0),平均加样回收率为94.36%~105.57%,相对标准偏差(relative standard deviation,RSD)为0.52%~3.48%,QAMS法与ESM法测定的结果无显著性差异。结论 本研究建立的小陷胸汤QAMS法结果准确可靠,该方法简便易行,可用于小陷胸汤的质量控制。

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*杜霞,女,博士,副研究员,硕士生导师 研究方向:中药分析,计算化学
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刘园,女,硕士,研究实习员 研究方向:中药分析;

宁致远,男,硕士 研究方向:中药分析。刘园与宁致远为共同第一作者

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1-Hypoxanthine; 2-Vanillic acid; 3-Magnolithine; 4-Demethyleneberberine; 5-Columbamine; 6-Jatrorrhizine hydrochloride; 7-Coptisine hydrochloride; 8-Palmatine hydrochloride; 9-Berberine hydrochlorid.

, figureFileSmall=W9dWLYD1t5bjHcM2Iu8Tsw==, figureFileBig=j/umDzg2lXGJMFKXxAXldw==, tableContent=null), ArticleFig(id=1193928848141156969, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=图1, caption=小陷胸汤供试品和混合对照品的色谱图

1-次黄嘌呤;2-香草酸;3-木兰花碱;4-去亚甲基小檗碱;5-非洲防己碱;6-盐酸药根碱;7-盐酸黄连碱;8-盐酸巴马汀;9-盐酸小檗碱。

, figureFileSmall=W9dWLYD1t5bjHcM2Iu8Tsw==, figureFileBig=j/umDzg2lXGJMFKXxAXldw==, tableContent=null), ArticleFig(id=1193928848225043050, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Fig.2, caption=Chromatograms of reference product (A) and Xiaoxianxiong Decoction by the modern decocting method (B) and the traditional decocting method (C)

1-Hypoxanthine;2-Vanillic acid;3-Magnolithine;4-Demethyleneberberine;5-Columbamine;6-Jatrorrhizine hydrochloride;7-Coptisine hydrochloride;8-Palmatine hydrochloride;9-Berberine hydrochloride.

, figureFileSmall=tJ6xzYQ7S2oav/+dH34jCg==, figureFileBig=atc+QNMJjAJCkQsG9EG/yw==, tableContent=null), ArticleFig(id=1193928848304734827, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=图2, caption=对照品(A)及小陷胸汤现代煎煮方法(B)与传统代煎煮法(C)的色谱图

1-次黄嘌呤;2-香草酸;3-木兰花碱;4-去亚甲基小檗碱;5-非洲防己碱;6-盐酸药根碱;7-盐酸黄连碱;8-盐酸巴马汀;9-盐酸小檗碱。

, figureFileSmall=tJ6xzYQ7S2oav/+dH34jCg==, figureFileBig=atc+QNMJjAJCkQsG9EG/yw==, tableContent=null), ArticleFig(id=1193928848367649388, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.1, caption=

Linear regression equations of 9 components in Xiaoxianxiong Decoction

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Regression equation r Linear range/
μg·mL-1
Hypoxanthine Y=0.295 4X+6.019 8 r=0.999 6 52.56 -525.6
Vanillic acid Y=17.259X-31.441 r=0.998 0 3.92 -39.2
Magnolithine Y=9.635 5X+0.896 7 r=0.999 9 3.6 -36
Demethyleneberberine Y=15.644X-3.302 5 r=0.999 9 1.456 -4.56
Columbamine Y=18.515X+2.755 r=0.999 9 3.834 -38.34
Jatrorrhizine hydrochloride Y=17.074X+6.610 7 r=0.999 9 10.274 -102.74
Coptisine hydrochloride Y=17.074X+6.610 7 r=0.999 9 11.664 -116.64
Palmatine hydrochloride Y=11.221X+4.314 r=0.999 9 9.504 -95.04
Berberine hydrochloride Y=15.165X+33.413 r=0.999 9 32.208 -322.08
), ArticleFig(id=1193928848434758253, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表1, caption=

小陷胸汤中9个成分的线性回归方程

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Regression equation r Linear range/
μg·mL-1
Hypoxanthine Y=0.295 4X+6.019 8 r=0.999 6 52.56 -525.6
Vanillic acid Y=17.259X-31.441 r=0.998 0 3.92 -39.2
Magnolithine Y=9.635 5X+0.896 7 r=0.999 9 3.6 -36
Demethyleneberberine Y=15.644X-3.302 5 r=0.999 9 1.456 -4.56
Columbamine Y=18.515X+2.755 r=0.999 9 3.834 -38.34
Jatrorrhizine hydrochloride Y=17.074X+6.610 7 r=0.999 9 10.274 -102.74
Coptisine hydrochloride Y=17.074X+6.610 7 r=0.999 9 11.664 -116.64
Palmatine hydrochloride Y=11.221X+4.314 r=0.999 9 9.504 -95.04
Berberine hydrochloride Y=15.165X+33.413 r=0.999 9 32.208 -322.08
), ArticleFig(id=1193928848522838638, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.2, caption=

Experimental results of sample recovery of Xiaoxianxiong Decoction. n=6

, figureFileSmall=null, figureFileBig=null, tableContent=
Component V(Sample)/mL m(Sample)/μg m(Added)/μg m(Measured)/μg Recovery/% Average recovery/% RSD/%
Hypoxanthine 1.5 482.96 482.97 972.33 101.32 98.20 2.83
1.5 482.85 482.97 952.08 97.15
1.5 484.03 482.97 974.66 101.59
1.5 483.19 482.97 955.60 97.81
1.5 449.68 482.97 905.41 94.36
1.5 449.67 482.97 918.02 96.97
Vanillic acid 1.5 84.27 84.26 165.72 96.66 100.32 2.24
1.5 84.24 84.26 170.59 102.48
1.5 84.29 84.26 168.22 99.60
1.5 84.31 84.26 168.97 100.47
1.5 83.59 84.26 167.76 99.89
1.5 83.53 84.26 170.20 102.85
Magnolithine 1.5 76.58 76.56 151.54 97.90 99.57 1.41
1.5 76.79 76.56 152.31 98.64
1.5 75.62 76.56 152.28 100.13
1.5 76.49 76.56 152.08 98.74
1.5 75.79 76.56 153.70 101.77
1.5 75.37 76.56 152.10 100.23
Desmethyleneberberine 1.5 39.09 39.09 78.99 102.08 102.01 3.48
1.5 38.79 39.09 79.28 103.59
1.5 39.54 39.09 77.17 96.28
1.5 40.02 39.09 78.93 99.55
1.5 41.57 39.09 82.84 105.59
1.5 41.13 39.09 82.16 104.95
Columbamine 1.5 58.99 59.04 116.50 97.40 101.53 3.14
1.5 59.28 59.04 117.33 98.31
1.5 58.79 59.04 118.69 101.45
1.5 58.97 59.04 119.43 102.40
1.5 60.49 59.04 121.93 104.05
1.5 60.61 59.04 122.94 105.57
Jatrorrhizine hydrochloride 1.5 189.16 189.19 381.51 101.67 100.29 0.98
1.5 189.47 189.19 377.67 99.48
1.5 189.65 189.19 379.09 100.13
1.5 188.98 189.19 376.97 99.36
1.5 189.73 189.19 378.46 99.76
1.5 189.83 189.19 381.52 101.32
Coptisine hydrochloride 1.5 290.99 291.02 584.10 100.72 99.89 1.24
1.5 291.47 291.02 587.35 101.67
1.5 289.94 291.02 580.62 99.89
1.5 290.87 291.02 577.83 98.60
1.5 290.49 291.02 581.64 100.05
1.5 290.12 291.02 576.58 98.43
Palmatine hydrochloride 1.5 201.73 201.74 407.40 101.95 100.44 1.42
1.5 201.58 201.74 400.05 98.38
1.5 201.97 201.74 402.98 99.63
1.5 200.99 201.74 405.62 101.43
1.5 201.96 201.74 406.98 101.62
1.5 202.11 201.74 403.15 99.65
Berberine hydrochloride 1.5 902.75 902.76 1 806.06 100.06 99.82 0.52
1.5 901.98 902.76 1 797.46 99.19
1.5 902.69 902.76 1 811.16 100.63
1.5 902.58 902.76 1 805.66 100.04
1.5 903.64 902.76 1 801.57 99.46
1.5 903.48 902.76 1 801.94 99.52
), ArticleFig(id=1193928848598336111, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表2, caption=

小陷胸汤加样回收实验结果。n=6

, figureFileSmall=null, figureFileBig=null, tableContent=
Component V(Sample)/mL m(Sample)/μg m(Added)/μg m(Measured)/μg Recovery/% Average recovery/% RSD/%
Hypoxanthine 1.5 482.96 482.97 972.33 101.32 98.20 2.83
1.5 482.85 482.97 952.08 97.15
1.5 484.03 482.97 974.66 101.59
1.5 483.19 482.97 955.60 97.81
1.5 449.68 482.97 905.41 94.36
1.5 449.67 482.97 918.02 96.97
Vanillic acid 1.5 84.27 84.26 165.72 96.66 100.32 2.24
1.5 84.24 84.26 170.59 102.48
1.5 84.29 84.26 168.22 99.60
1.5 84.31 84.26 168.97 100.47
1.5 83.59 84.26 167.76 99.89
1.5 83.53 84.26 170.20 102.85
Magnolithine 1.5 76.58 76.56 151.54 97.90 99.57 1.41
1.5 76.79 76.56 152.31 98.64
1.5 75.62 76.56 152.28 100.13
1.5 76.49 76.56 152.08 98.74
1.5 75.79 76.56 153.70 101.77
1.5 75.37 76.56 152.10 100.23
Desmethyleneberberine 1.5 39.09 39.09 78.99 102.08 102.01 3.48
1.5 38.79 39.09 79.28 103.59
1.5 39.54 39.09 77.17 96.28
1.5 40.02 39.09 78.93 99.55
1.5 41.57 39.09 82.84 105.59
1.5 41.13 39.09 82.16 104.95
Columbamine 1.5 58.99 59.04 116.50 97.40 101.53 3.14
1.5 59.28 59.04 117.33 98.31
1.5 58.79 59.04 118.69 101.45
1.5 58.97 59.04 119.43 102.40
1.5 60.49 59.04 121.93 104.05
1.5 60.61 59.04 122.94 105.57
Jatrorrhizine hydrochloride 1.5 189.16 189.19 381.51 101.67 100.29 0.98
1.5 189.47 189.19 377.67 99.48
1.5 189.65 189.19 379.09 100.13
1.5 188.98 189.19 376.97 99.36
1.5 189.73 189.19 378.46 99.76
1.5 189.83 189.19 381.52 101.32
Coptisine hydrochloride 1.5 290.99 291.02 584.10 100.72 99.89 1.24
1.5 291.47 291.02 587.35 101.67
1.5 289.94 291.02 580.62 99.89
1.5 290.87 291.02 577.83 98.60
1.5 290.49 291.02 581.64 100.05
1.5 290.12 291.02 576.58 98.43
Palmatine hydrochloride 1.5 201.73 201.74 407.40 101.95 100.44 1.42
1.5 201.58 201.74 400.05 98.38
1.5 201.97 201.74 402.98 99.63
1.5 200.99 201.74 405.62 101.43
1.5 201.96 201.74 406.98 101.62
1.5 202.11 201.74 403.15 99.65
Berberine hydrochloride 1.5 902.75 902.76 1 806.06 100.06 99.82 0.52
1.5 901.98 902.76 1 797.46 99.19
1.5 902.69 902.76 1 811.16 100.63
1.5 902.58 902.76 1 805.66 100.04
1.5 903.64 902.76 1 801.57 99.46
1.5 903.48 902.76 1 801.94 99.52
), ArticleFig(id=1193928848669639280, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.3, caption=

Calculation results of 8 components relative correction factors with berberine hydrochloride as the internal reference

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Hypoxanthine Vanillic acid Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
1 49.631 9 0.929 1 1.580 5 0.991 9 0.821 2 0.891 0 1.355 4 0.809 8
2 49.625 7 0.923 8 1.582 0 0.993 6 0.822 4 0.891 3 1.357 0 0.811 6
3 49.098 8 0.965 7 1.581 3 0.997 5 0.821 4 0.890 6 1.357 7 0.812 9
4 46.832 5 0.955 7 1.601 3 1.040 5 0.829 9 0.900 5 1.376 0 0.824 0
5 47.542 5 0.934 5 1.606 6 1.016 4 0.830 0 0.901 4 1.373 7 0.827 3
6 46.422 3 0.963 2 1.621 6 1.002 8 0.835 0 0.906 2 1.372 4 0.829 4
Mean value 48.192 4 0.945 3 1.595 5 1.007 1 0.826 7 0.896 8 1.365 3 0.819 2
RSD/% 2.98 1.94 1.07 1.84 0.70 0.75 0.70 1.06
), ArticleFig(id=1193928848745136753, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表3, caption=

以盐酸小檗碱为内参物的8个成分相对校正因子计算结果

, figureFileSmall=null, figureFileBig=null, tableContent=
Component Hypoxanthine Vanillic acid Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
1 49.631 9 0.929 1 1.580 5 0.991 9 0.821 2 0.891 0 1.355 4 0.809 8
2 49.625 7 0.923 8 1.582 0 0.993 6 0.822 4 0.891 3 1.357 0 0.811 6
3 49.098 8 0.965 7 1.581 3 0.997 5 0.821 4 0.890 6 1.357 7 0.812 9
4 46.832 5 0.955 7 1.601 3 1.040 5 0.829 9 0.900 5 1.376 0 0.824 0
5 47.542 5 0.934 5 1.606 6 1.016 4 0.830 0 0.901 4 1.373 7 0.827 3
6 46.422 3 0.963 2 1.621 6 1.002 8 0.835 0 0.906 2 1.372 4 0.829 4
Mean value 48.192 4 0.945 3 1.595 5 1.007 1 0.826 7 0.896 8 1.365 3 0.819 2
RSD/% 2.98 1.94 1.07 1.84 0.70 0.75 0.70 1.06
), ArticleFig(id=1193928848837411442, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.4, caption=

Effects of different instruments on relative correction factors

, figureFileSmall=null, figureFileBig=null, tableContent=
Instrument Chromatogr-
aphic column		 Relative correction factor
Hypoxanthine Vanillic
acid		 Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
Agilent 1260 Dikma 50.013 6 0.939 1 1.581 7 0.994 7 0.821 9 0.891 7 1.357 3 0.811 2
Agilent 49.962 9 0.963 5 1.582 0 0.996 7 0.821 9 0.891 7 1.357 3 0.811 3
Agilent DEAEQ14098 Dikma 49.625 7 0.923 8 1.582 0 0.993 6 0.822 4 0.891 3 1.357 0 0.811 6
Agilent 49.475 8 0.964 9 1.582 3 0.996 6 0.821 9 0.891 6 1.357 1 0.811 2
Mean value 49.769 5 0.947 8 1.582 0 0.995 4 0.822 0 0.891 6 1.357 2 0.811 3
RSD/% 0.52 2.10 0.02 0.15 0.03 0.02 0.01 0.02
), ArticleFig(id=1193928848908714611, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表4, caption=

不同仪器色谱柱对相对校正因子的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
Instrument Chromatogr-
aphic column		 Relative correction factor
Hypoxanthine Vanillic
acid		 Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
Agilent 1260 Dikma 50.013 6 0.939 1 1.581 7 0.994 7 0.821 9 0.891 7 1.357 3 0.811 2
Agilent 49.962 9 0.963 5 1.582 0 0.996 7 0.821 9 0.891 7 1.357 3 0.811 3
Agilent DEAEQ14098 Dikma 49.625 7 0.923 8 1.582 0 0.993 6 0.822 4 0.891 3 1.357 0 0.811 6
Agilent 49.475 8 0.964 9 1.582 3 0.996 6 0.821 9 0.891 6 1.357 1 0.811 2
Mean value 49.769 5 0.947 8 1.582 0 0.995 4 0.822 0 0.891 6 1.357 2 0.811 3
RSD/% 0.52 2.10 0.02 0.15 0.03 0.02 0.01 0.02
), ArticleFig(id=1193928848971629172, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.5, caption=

Effects of different column temperatures on relative correction factors

, figureFileSmall=null, figureFileBig=null, tableContent=
T/℃ Relative correction factor
Hypoxanthine Vanillic
acid		 Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
25 49.420 0 0.940 1 1.581 8 0.994 3 0.821 9 0.891 7 1.357 2 0.811 2
30 49.625 7 0.923 8 1.582 0 0.993 6 0.822 4 0.891 3 1.357 0 0.811 6
35 49.236 4 0.939 9 1.581 7 0.994 5 0.821 9 0.891 6 1.357 1 0.811 2
Mean value 49.427 4 0.934 6 1.581 8 0.994 1 0.822 1 0.891 5 1.357 1 0.811 3
RSD/% 0.39 1.00 0.01 0.05 0.03 0.02 0.01 0.02
), ArticleFig(id=1193928849047126645, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表5, caption=

不同柱温对相对校正因子的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
T/℃ Relative correction factor
Hypoxanthine Vanillic
acid		 Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
25 49.420 0 0.940 1 1.581 8 0.994 3 0.821 9 0.891 7 1.357 2 0.811 2
30 49.625 7 0.923 8 1.582 0 0.993 6 0.822 4 0.891 3 1.357 0 0.811 6
35 49.236 4 0.939 9 1.581 7 0.994 5 0.821 9 0.891 6 1.357 1 0.811 2
Mean value 49.427 4 0.934 6 1.581 8 0.994 1 0.822 1 0.891 5 1.357 1 0.811 3
RSD/% 0.39 1.00 0.01 0.05 0.03 0.02 0.01 0.02
), ArticleFig(id=1193928849101652598, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.6, caption=

Relative retention times measured by different instruments and columns. s

, figureFileSmall=null, figureFileBig=null, tableContent=
Instrument Chromatographic
column		 Hypoxanthine Vanillic
acid		 Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
Agilent 1260 Dikma 0.150 0 0.590 9 0.732 4 0.834 6 0.883 9 0.895 7 0.915 3 0.972 7
Agilent 0.153 4 0.575 2 0.696 2 0.800 6 0.860 5 0.871 6 0.898 5 0.962 8
Agilent DEAEQ14098 Dikma 0.153 4 0.610 9 0.733 8 0.838 0 0.887 9 0.898 2 0.916 3 0.974 9
Agilent 0.163 2 0.582 0 0.701 0 0.805 4 0.864 9 0.875 6 0.900 7 0.965 7
Mean value 0.155 0 0.589 7 0.715 9 0.819 6 0.874 3 0.885 3 0.907 7 0.969 0
RSD/% 3.68 2.62 2.80 2.37 1.56 1.54 1.04 0.59
), ArticleFig(id=1193928849156178551, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表6, caption=

不同仪器和色谱柱测得的相对保留时间。s

, figureFileSmall=null, figureFileBig=null, tableContent=
Instrument Chromatographic
column		 Hypoxanthine Vanillic
acid		 Magnolithine Demethyleneberberine Columbamine Jatrorrhizine
hydrochloride		 Coptisine
hydrochloride		 Palmatine
hydrochloride
Agilent 1260 Dikma 0.150 0 0.590 9 0.732 4 0.834 6 0.883 9 0.895 7 0.915 3 0.972 7
Agilent 0.153 4 0.575 2 0.696 2 0.800 6 0.860 5 0.871 6 0.898 5 0.962 8
Agilent DEAEQ14098 Dikma 0.153 4 0.610 9 0.733 8 0.838 0 0.887 9 0.898 2 0.916 3 0.974 9
Agilent 0.163 2 0.582 0 0.701 0 0.805 4 0.864 9 0.875 6 0.900 7 0.965 7
Mean value 0.155 0 0.589 7 0.715 9 0.819 6 0.874 3 0.885 3 0.907 7 0.969 0
RSD/% 3.68 2.62 2.80 2.37 1.56 1.54 1.04 0.59
), ArticleFig(id=1193928849214898808, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.7, caption=

Determination of 9 components in Xiaoxianxiong Decoction by QAMS and ESM. μg·mL-1

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Berberine Hypoxanthine Vanillic acid Magnolithine Demethyleneberberine
ESM ESM QAMS ESM QAMS ESM QAMS ESM QAMS
1 182.133 333.313 327.160 15.907 15.571 15.246 15.383 8.204 8.284
2 183.768 340.563 334.276 16.251 15.908 14.996 15.131 8.131 8.210
3 181.545 319.419 313.522 15.213 14.892 15.140 15.276 7.940 8.016
4 184.292 342.149 335.832 15.775 15.442 15.259 15.396 8.187 8.266
5 181.746 343.408 337.068 15.206 14.885 14.565 14.697 8.169 8.248
6 181.936 294.641 289.202 16.260 15.917 14.590 14.721 8.182 8.261
No. Columbamine Jatrorrhizine hydrochloride Coptisine hydrochloride Palmatine hydrochloride
ESM QAMS ESM QAMS ESM QAMS ESM QAMS
1 11.696 11.771 38.086 38.350 58.560 58.890 40.634 40.949
2 12.309 12.388 38.285 38.550 58.423 58.752 40.577 40.892
3 12.197 12.275 37.906 38.169 57.638 57.962 40.235 40.547
4 12.196 12.274 38.492 38.759 58.557 58.886 40.656 40.971
5 12.285 12.364 38.774 39.043 59.280 59.614 41.034 41.353
6 11.932 12.009 38.671 38.940 58.631 58.961 40.721 41.037
), ArticleFig(id=1193928849286201977, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表7, caption=

一测多评法(QAMS)和外标法(ESM)测定小陷胸汤中9个成分的含量。μg·mL-1

, figureFileSmall=null, figureFileBig=null, tableContent=
No. Berberine Hypoxanthine Vanillic acid Magnolithine Demethyleneberberine
ESM ESM QAMS ESM QAMS ESM QAMS ESM QAMS
1 182.133 333.313 327.160 15.907 15.571 15.246 15.383 8.204 8.284
2 183.768 340.563 334.276 16.251 15.908 14.996 15.131 8.131 8.210
3 181.545 319.419 313.522 15.213 14.892 15.140 15.276 7.940 8.016
4 184.292 342.149 335.832 15.775 15.442 15.259 15.396 8.187 8.266
5 181.746 343.408 337.068 15.206 14.885 14.565 14.697 8.169 8.248
6 181.936 294.641 289.202 16.260 15.917 14.590 14.721 8.182 8.261
No. Columbamine Jatrorrhizine hydrochloride Coptisine hydrochloride Palmatine hydrochloride
ESM QAMS ESM QAMS ESM QAMS ESM QAMS
1 11.696 11.771 38.086 38.350 58.560 58.890 40.634 40.949
2 12.309 12.388 38.285 38.550 58.423 58.752 40.577 40.892
3 12.197 12.275 37.906 38.169 57.638 57.962 40.235 40.547
4 12.196 12.274 38.492 38.759 58.557 58.886 40.656 40.971
5 12.285 12.364 38.774 39.043 59.280 59.614 41.034 41.353
6 11.932 12.009 38.671 38.940 58.631 58.961 40.721 41.037
), ArticleFig(id=1193928849370088058, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=EN, label=Tab.8, caption=

Determination of 9 components in Xiaoxianxiong Decoction by QAMS and ESM. μg·mL-1

, figureFileSmall=null, figureFileBig=null, tableContent=
Component ESM QAMS
Berberine hydrochloride 237.219 9 237.219 9
Hypoxanthine 552.828 539.433
Vanillic acid 17.834 17.309
Magnolithine 17.969 18.022
Demethyleneberberine 6.976 7.398
Columbamine 12.044 12.817
Jatrorrhizine hydrochloride 37.918 38.614
Coptisine hydrochloride 80.134 81.050
Palmatine hydrochloride 46.010 47.017
), ArticleFig(id=1193928849424614011, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1193877803474055309, language=CN, label=表8, caption=

现代煎煮方法QAMS法和ESM法测定小陷胸汤中9个成分的含量。μg·mL-1

, figureFileSmall=null, figureFileBig=null, tableContent=
Component ESM QAMS
Berberine hydrochloride 237.219 9 237.219 9
Hypoxanthine 552.828 539.433
Vanillic acid 17.834 17.309
Magnolithine 17.969 18.022
Demethyleneberberine 6.976 7.398
Columbamine 12.044 12.817
Jatrorrhizine hydrochloride 37.918 38.614
Coptisine hydrochloride 80.134 81.050
Palmatine hydrochloride 46.010 47.017
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基于一测多评法同时测定经典名方小陷胸汤中9种成分含量
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刘园 1 , 宁致远 2 , 陆晨曦 1 , 陈志永 1 , 任慧 1 , 任菲菲 1 , 杜霞 1, *
中国药学杂志 | 论著 2025,60(2): 172-179
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中国药学杂志 | 论著 2025, 60(2): 172-179
基于一测多评法同时测定经典名方小陷胸汤中9种成分含量
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刘园1, 宁致远2, 陆晨曦1, 陈志永1, 任慧1, 任菲菲1, 杜霞1, *
作者信息
  • 1 陕西省中医药研究院 中药研究所, 西安 710061
  • 2 澳门科技大学中药质量研究国家重点实验室, 澳门 999078

    • 刘园,女,硕士,研究实习员 研究方向:中药分析;

    宁致远,男,硕士 研究方向:中药分析。刘园与宁致远为共同第一作者

通讯作者:

*杜霞,女,博士,副研究员,硕士生导师 研究方向:中药分析,计算化学
Simultaneous Determination of 9 Components in Xiaoxianxiong Decoction Based on QAMS Method
Yuan LIU1, Zhiyuan NING2, Chenxi LU1, Zhiyong CHEN1, Hui REN1, Feifei REN1, Xia DU1, *
Affiliations
  • 1 Institute of traditional Chinese Medicine, Shaanxi Academy of Traditional Chinese Medicine, Xi'an 710061, China
  • 2 State Key Laboratory for Quality Research of Chinese Medicines, Macau University of Science and Technology, Macao 999078,China
出版时间: 2025-01-22 doi: 10.11669/cpj.2025.02.009
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摘要
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目的 建立一测多评法(quantitative analysis of multi-components by single marker, QAMS)同时测定小陷胸汤中次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的含量。方法 本实验采用色谱柱Spursil C18(4.6 mm×250 mm,5 μm),以乙腈-0.05 mol·L-1磷酸二氢钾溶液加磷酸调pH=4.0为流动相,检测波长: 0~20 min:290 nm;20~45 min:300 nm,柱温30 ℃,进样量10 μL。以盐酸小檗碱为内参物,分别计算其余8个成分的相对校正因子并计算各成分含量,比较外标法(external standard method, ESM)和QAMS法结果的差异。结果 小陷胸汤9个成分在一定浓度范围与峰面积呈良好的线性关系(r>0.998 0),平均加样回收率为94.36%~105.57%,相对标准偏差(relative standard deviation,RSD)为0.52%~3.48%,QAMS法与ESM法测定的结果无显著性差异。结论 本研究建立的小陷胸汤QAMS法结果准确可靠,该方法简便易行,可用于小陷胸汤的质量控制。

小陷胸汤  /  质量分析  /  一测多评法  /  高效液相色谱法

OBJECTIVE To establish a quantitative analysis of multi-components by single marker (QAMS) for the simultaneous determination of hypoxanthine, vanillic acid, magnolithine, demethyleneberberine, Columbamine, Jatrorrhizine hydrochloride, coptisine hydrochloride, palmatine hydrochloride and berberine hydrochloride in Xiaoxianxiong decoction. METHODS The separation was performed on a Spursil C18 column (4.6 mm×250 mm, 5 μm) with acetonitrile and 0.05 mol·L-1 potassium dihydrogen phosphate solution plus phosphoric acid adjusted to pH=4.0 as the mobile phases. Detection wavelength: 0-20 min: 290 nm; 20-45 min: 300 nm, column temperature 30 ℃, injection volume 10 μL. Using berberine hydrochloride as the internal reference material, the relative correction factors of the remaining eight components were calculated and the content of each component was calculated, and the results were compared with those of the external standard method (ESM) and the QAMS method. The differences between the results of the external standard method (ESM) and QAMS method were compared. RESULTS The nine components of Xiaoxianxiong decoction showed a good linear relationship with peak area in a certain concentration range (r>0.998 0), with the average spiked recoveries of 94.36%-105.57% and the RSD was 0.52%-3.48%, and the results of component contents determined by QAMS and ESM were not statistically significant. CONCLUSION The results of the established QAMS for Xiaoxianxiong decoction are accurate and reliable, and the method is simple and easy to be applied to the quality control of Xiaoxianxiong Decoction.

Xiaoxianxiong decoction  /  quantitative analysis  /  multi-components by single marker  /  HPLC
刘园, 宁致远, 陆晨曦, 陈志永, 任慧, 任菲菲, 杜霞. 基于一测多评法同时测定经典名方小陷胸汤中9种成分含量. 中国药学杂志, 2025 , 60 (2) : 172 -179 . DOI: 10.11669/cpj.2025.02.009
Yuan LIU, Zhiyuan NING, Chenxi LU, Zhiyong CHEN, Hui REN, Feifei REN, Xia DU. Simultaneous Determination of 9 Components in Xiaoxianxiong Decoction Based on QAMS Method[J]. Chinese Pharmaceutical Journal, 2025 , 60 (2) : 172 -179 . DOI: 10.11669/cpj.2025.02.009
正文
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经典名方小陷胸汤取自张仲景《伤寒论·辨太阳病脉证并治》,是以瓜蒌、半夏和黄连组成的方剂。具有清热化痰,开胸散结之功效[1]。方中瓜蒌为君药,甘寒而润,有清热化痰、开胸散结的功效,在该方熬制时瓜蒌应先煮,是为了“以缓治上”,疏通胸膈之痹,可以清肺热,润肺燥,主治痰热咳喘[2-3];半夏为臣药,可化痰散结,降逆止呕,利咽喉,消肿止痛[2];黄连同为臣药,可清热燥湿,泻火解毒[4]。在临床上主要用于治疗冠心病、糖尿病、胃炎、肺炎、肋间神经痛、胰腺炎等病[4-6]。
目前,对于小陷胸汤的研究多集中在临床药效及作用机制,关于小陷胸汤的成分分析和质量控制报道较少。临床常用小陷胸汤汤剂加减方,由于煎煮方法存在差异,难以实现对成分的有效控制,进而影响疗效。中药因其多组分、多靶点、多通路的特点,不同组分间相互协同发挥药效,单一成分含量测定无法全面评估其质量[7],一测多评法(quantitative analysis of multi-components by single marker, QAMS)因具有只测定1个对照品便可评价多个成分的优势,已成为一种适合中药及其复方制剂的多指标质量评价模式[8],它以性质稳定、价廉易得的对照品为内标物,同时测定其他多种成分的含量,解决了中药对照品昂贵、稀缺、管理困难等问题,是其他分析方法的扩展与延伸[7]。因为小陷胸汤主治为“小结胸病,正在心下,按之则痛,脉浮滑者,小陷胸汤主之。”临床主要用来治疗冠心病、动脉粥样硬化、心律失常、心衰、心绞痛等心血管疾病,其主要药效成分有次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱等。本实验采用QAMS法测定上述9个药效成分的含量,并将一测多评法结果与外标法进行比较,探讨小陷胸汤各成分QAMS法评价的可行性,为小陷胸汤的研发及质量控制奠定基础。
1 仪器与试药
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1.1 仪器
Agilent 1260型高效液相色谱仪、色谱柱Spursil C18(4.6 mm×250 mm,5 μm)及Agilent 5 TC-C18(4.6 mm×250 mm,5 μm)(安捷伦公司);分析天平(赛多利斯科学仪器有限公司);水浴锅(北京科伟永兴仪器有限公司);超声仪(宁波新艺超声设备有限公司)。
1.2 试剂与材料
对照品次黄嘌呤(批号:CHB201107,纯度HPLC≥98%)、香草酸(批号:110776-201503,纯度HPLC≥99.8%)、木兰花碱(批号:CHB201113,纯度HPLC≥98%)、去亚甲基小檗碱(批号:CHB210114,纯度HPLC≥98%)、非洲防己碱(批号:CHB201126,纯度HPLC≥98%)(中国食品药品检定研究院);盐酸药根碱(批号:110733-201609,纯度HPLC≥89.5%)、盐酸黄连碱(批号:112026-201802,纯度HPLC≥94.0%)、盐酸巴马汀(批号:110732-201913,纯度HPLC≥85.7%)、盐酸小檗碱(批号:110713-202015,纯度HPLC≥85.9%)(成都克洛玛生物科技有限公司);水为娃哈哈纯水;乙腈、甲醇为色谱纯;磷酸二氢钾、磷酸及其余试剂均为分析纯。
瓜蒌、半夏、黄连三味药材均购于陕西兴盛德药业有限公司,经由陕西省中医药研究院孟雪老师鉴定为正品。瓜蒌表面呈橙红色和橙黄色,表面较光滑,质脆,易破开,内表面黄白色,有红黄色丝络,果瓤橙黄色,黏稠,与多数种子黏结成团。具焦糖气,味微酸、甜。炮制方法为切块。
清半夏为半夏的炮制加工品。外观为椭圆形、类圆形和不规则状。质脆,易折断,断面略呈粉性或角质样。气微,味微涩、微有麻舌感。
黄连(味连)集聚成簇,呈弯曲状,形如鸡爪,单枝根茎长3~6 cm,表面呈黄褐色,粗糙,有不规则结节状隆起、须根及须根残基,顶端常留有残余的茎或叶柄。气微,味极苦。
2 方法与结果
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2.1 色谱条件
使用Agilent 1260型高效液相色谱仪,色谱柱为Spursil C18(4.6 mm×250 mm, 5 μm),乙腈(A)-0.05 mol·L-1磷酸二氢钾(B)(加磷酸调节pH=4.0)为流动相;梯度洗脱程序为0~10 min,2%~10%A;10~17 min,10%~15%A;17~23 min,15%~37%A;23~27 min,37%~40%A;27~37 min,40%A;37~45 min,40%~2%A,检测波长: 0~20 min:290 nm;20~45 min:300 nm,初始流速0.8 mL·min-1,在27~37 min流速切为0.6 mL·min-1,37 min之后恢复到0.8 mL·min-1,柱温30 ℃,进样量10 μL。
2.2 供试品溶液的制备
考证小陷胸汤古籍记载结合现代临床安全用量,折算现代计量单位为瓜蒌30 g、半夏12 g、黄连6 g。先取瓜蒌置于煎药器中,加入1 200 mL去离子水,浸泡30 min,先武火后文火煎煮至600 mL,过滤去渣。分别将黄连和半夏加入瓜蒌煎煮药液中,继续先武火后文火煎煮至400 mL,过滤去渣。水煎液保存在4 ℃冰箱以供后续制备。取3 mL水煎液于电热恒温水浴锅中蒸干,用体积分数20%甲醇10 mL溶于已蒸干的蒸发皿中,取上清液,以0.45 μm微孔滤膜过滤,即为供试品溶液。放在4 ℃冰箱保存备用。
现代煎煮方法参照《医疗机构中药煎药室管理规范》[9]。用水量一般以浸过药面2~5 cm为宜,每剂药煎煮2次,药物煮沸后再煎煮20~30 min,第2煎的煎煮时间应比第1煎的时间略缩短。即瓜蒌30 g、半夏12 g、黄连6 g,先取瓜蒌置于煎药器中,加入500 mL去离子水,浸泡30 min,先武火后文火煎煮30 min,过滤去渣。将黄连和半夏加入瓜蒌煎煮药液中,继续先武火后文火煎煮25 min,过滤去渣。水煎液保存在4 ℃冰箱以供后续制备。取3 mL水煎液于电热恒温水浴锅中蒸干,用体积分数20%甲醇10 mL溶于已蒸干的蒸发皿中,取上清液,以0.45 μm微孔滤膜过滤,即为供试品溶液。放在4 ℃冰箱保存备用。
2.3 混合对照品溶液的制备
分别称取次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱配制成质量浓度分别为526、39、36、15、38、103、117、95、322 μg·mL-1的混合标准品,将上述混合对照品溶液稀释成6个不同质量浓度,分别为原标准品浓度的0.8、0.6、0.4、0.2、0.1倍,并放于4 ℃保存,备用。
2.4 系统适应性试验
取混合对照品溶液,供试品溶液10 μL,按“2.1”项下的色谱条件进样,结果见图1。
2.5 方法学考察
2.5.1 线性关系考察
分别精密吸取“2.3”项下6个不同质量浓度的混合对照品溶液各10 μL,按“2.1”项下色谱条件测定,以对照品质量浓度为横坐标(X),峰面积为纵坐标(Y),绘制次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的标准曲线,结果表明,9个成分质量浓度在相应的范围内线性关系良好。结果见表1。
2.5.2 精密度实验
取本品适量,按“2.1”项下制备供试品溶液,在“2.1”项色谱条件下进样测定6次,测得次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱峰面积相对标准偏差(RSD)分别为3.37%、3.11%、2.10%、1.94%、3.23%、0.33%、0.65%、0.37%、0.33%,表明该仪器在实验中具有良好的精密度。
2.5.3 重复性试验
采用同一水煎液,按照“2.1”中所述的方法平行配制6份样品,测定6次,测得次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的峰面积的RSD分别为3.13%、3.06%、2.48%、2.18%、2.27%、1.03%、1.38%、0.20%、0.64%,说明此法具有较好的重复性。
2.5.4 稳定性试验
精密提取10 μL标准溶液,根据“2.1”中规定的色谱条件,在配制后的第0、2、4、6、8、12小时进样,次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱峰面积的RSD分别为3.03%、2.98%、1.77%、1.22%、1.98%、0.88%、0.90%、0.63%、0.59%,表示该试样具有较好的稳定性。
2.5.5 加样回收率实验
精密吸取已知含量的小陷胸汤供试品各6份,每份量取1.5 mL,分别精密加入次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的对照品储备液,按照“2.1”项方法制备加样回收供试溶液,按照“2.1”项色谱条件进样测定,计算加样回收率,各成分RSD值分别为2.83%、2.24%、1.41%、3.48%、3.14%、0.98%、1.24%、1.42%、0.52%,结果见表2,表明方法的准确性良好。
2.6 相对校正因子的测定
精密吸取“2.3”项下的混合对照品溶液,按照“2.1”项的色谱条件,连续进样6次,测定峰面积,以盐酸小檗碱为内参物,分别测定计算次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀的相对校正因子(公式1)。
fs/i=fs/fi=(As×ρi)/(Ai×ρs)
式中,As为盐酸小檗碱的峰面积,ρi为待测组分i的质量浓度,ρs为盐酸小檗碱的质量浓度,Ai为待测组分i的峰面积。结果见表3。
2.7 相对校正因子的耐用性考察
2.7.1 不同色谱柱和仪器对相对校正因子的影响
采用Agilent 1260型高效液相色谱系统和Agilent DEAEQ14098系统分别考察了Spursil C18(4.6 mm×250 mm,5 μm)和Agilent 5 TC-C18(4.6 mm×250 mm,5 μm)色谱柱,以盐酸小檗碱为内参物,测定不同色谱柱和仪器对各待测成分的相对校正因子的影响,结果见表4。结果表明,次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀与内参物盐酸小檗碱在不同仪器色谱柱下重现性良好,RSD在0.01%~2.10%之间。
2.7.2 不同柱温对测定小陷胸汤中8个成分的相对校正因子的影响
采用“2.1”项下的色谱条件,以盐酸小檗碱为内参物,考察柱温分别为25、30、35 ℃时对相对校正因子的影响。结果见表5,次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀与内参物盐酸小檗碱在不同柱温下相对校正因子的适用性良好。
2.7.3 待测成分色谱峰的定位
待测成分色谱峰的定位,采用相对保留时间定位法,已知内标峰的保留时间,根据8个待测成分的相对保留时间,即可对其进行定位。选择以盐酸小檗碱的保留时间相对值作为待测成分定位标准,计算不同高效液相色谱仪和不同色谱柱的相对保留时间结果在不同高效液相色谱仪和色谱柱条件下,测得各待测成分相对保留时间RSD均小于5.00%。见表6。
2.8 传统煎煮方法一测多评法与外标法检测结果的比较
采用外标法(ESM)分别对6批不同小陷胸汤样品中次黄嘌呤、香草酸、木兰花碱、去亚甲基小檗碱、非洲防己碱、盐酸药根碱、盐酸黄连碱、盐酸巴马汀进行含量测定,并与一测多评法计算的结果进行比较,验证一测多评法用于小陷胸汤中9个成分测定的准确度和可靠性, 2种方法测定结果见表7。用SPSS 27.0和GraphPad Prism 8.4.2进行数据分析,采用可以适用于该数据的非参数检验进行分析,P>0.05,表明两种方法测定结果无显著性差异。本研究建立的一测多评法具有较好的可信度。
2.9 现代煎煮方法一测多评法与外标法检测结果的比较
由图2比对可以看出,小陷胸汤现代煎煮方法和传统煎煮方法图谱相似,并将现代煎煮方法各成分按照ESM法与QAMS法分别进行计算(表8),用SPSS 27.0和GraphPad Prism 8.4.2进行数据分析,采用非参数检验进行分析,结果同样为P > 0.05,现代煎煮方法适用于所建立的一测多评法。
3 讨论
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3.1 供试品提取方法的选择
本实验首先对冻干粉和传统汤剂两种制样方法进行了考察,考虑到将样品制成冻干粉具有稳定性好、易于保存,不易被污染等优点,本实验首先尝试冻干粉,但是图谱出峰很少,于是换用汤剂,发现汤剂出峰数量明显优于冻干粉。接下来对小陷胸汤提取溶剂进行了考察,包括体积分数20%、50%、70%甲醇、盐酸甲醇(1∶100)等提取方法[10],通过对比分析,使用体积分数20%甲醇时出峰数量较多,各成分响应较好,基线平稳,且查询《中国药典》2020年版药典中瓜蒌、半夏、黄连单药材提取方法进行对比,仍旧是体积分数20%甲醇提取的汤剂出峰数量更多,峰型更好。最终采用体积分数20%甲醇提取方法制备样品。
3.2 液相色谱条件的考察
流动相分别比较了甲醇-0.1%磷酸水、乙腈-0.1%甲酸水、乙腈-0.1%磷酸水、乙腈-0.05 mol·L-1磷酸二氢钾等、pH值分别考察了2、3、4、5、6,经高效液相色谱仪的DAD检测器扫描190~400 nm范围内的光谱图,结果显示,9个成分在280~300 nm之间的吸收较好,乙腈-0.05 mol·L-1磷酸二氢钾为流动相时分离效果最好,灵敏度较高,出峰数量较多。最终选择流动相为乙腈-0.05 mol·L-1磷酸二氢钾,PH为4,波长为290 nm切300 nm的液相色谱条件。
3.3 QAMS法的准确性与含量测定结果
本实验通过考察不同仪器、不同色谱柱、不同柱温对相对校正因子的影响,计算得出RSD均在5%以下,证明了小陷胸汤QAMS法的耐用性,并对不同批次的小陷胸汤样品分别采用ESM法和QAMS法计算9个成分的含量,结果接近,两种方法没有显著性差异,证明建立的QAMS法准确性和可靠性较好。
3.4 内参物选择
小陷胸汤在心血管疾病中应用广泛,可用于治疗各类冠心病、心绞痛、降脂、抗动脉粥样硬化等。经过对照品和指纹图谱比对,小陷胸汤中生物碱居多,ESM法计算得出小陷胸汤中盐酸小檗碱含量最高,通过查阅文献,小檗碱对心血管有保护作用,盐酸小檗碱可以抑制血管内皮通透性增加[11]、调节自噬、促进细胞增殖和抑制细胞凋亡,从而降低血脂水平,改善内膜增生,拮抗颈动脉脂质堆积改善动脉粥样硬化[12]、参与肠道菌群的调节发挥治疗心血管的作用;除此之外还有强大的抗菌作用、抗炎作用、免疫调节等[13]。小檗碱药理作用广泛,含量较高,稳定性好,廉价易得,故选取其为内参物。
此外,本研究中指认出的其余含量较高的成分中,盐酸巴马汀可以抑制纤维细胞活化延缓细胞纤维化[14],还可以减轻炎症反应、氧化损伤和心肌细胞凋亡[15];盐酸黄连碱能通过抑制自噬保护心肌细胞免受缺氧/复氧诱导的损伤[16];药根碱可以通过蛋白激酶B/AMP依赖蛋白激酶/内皮型一氧化氮合酶(Akt/AMPK/e NOS)信号通路改善血管内皮功能紊乱[17],并且具有良好的调节血脂活性[17],木兰花碱也具有抗炎、抗菌、免疫调节等多项药理活性。香草酸可以维持血管稳态,减少细胞凋亡[18]。次黄嘌呤可以降低小鼠血脂四项,降低脏器脂肪,提高超氧化物歧化酶(SOD)酶活性[19]。
4 结论
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本研究建立的QAMS法同时测定了小陷胸汤9个成分的含量的方法准确、稳定、重复性好,在节省成本的同时,可以更全面地反映小陷胸汤内在质量。
基金
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  • 国家自然科学基金青年科学基金项目资助(82204711)
  • 陕西省中医药管理局“双链融合”中青年科研创新团队项目资助(2022-SLRH-YQ-003)
  • 陕西省中医药管理局“医研校企”中医药传承创新平台项目资助
文献
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2025年第60卷第2期
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文章信息
doi: 10.11669/cpj.2025.02.009
  • 接收时间：2024-04-03
  • 首发时间：2025-11-08
  • 出版时间：2025-01-22
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  • 收稿日期：2024-04-03
基金
国家自然科学基金青年科学基金项目资助(82204711)
陕西省中医药管理局“双链融合”中青年科研创新团队项目资助(2022-SLRH-YQ-003)
陕西省中医药管理局“医研校企”中医药传承创新平台项目资助
作者信息
    1 陕西省中医药研究院 中药研究所, 西安 710061
    2 澳门科技大学中药质量研究国家重点实验室, 澳门 999078

通讯作者:

*杜霞,女,博士,副研究员,硕士生导师 研究方向:中药分析,计算化学
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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