Article(id=1239173816769892936, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239173808419033435, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024.02.05, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1673452800000, receivedDateStr=2023-01-12, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773371660654, onlineDateStr=2026-03-13, pubDate=1709136000000, pubDateStr=2024-02-29, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773371660654, onlineIssueDateStr=2026-03-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773371660654, creator=13701087609, updateTime=1773371660654, 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=233, endPage=241, ext={EN=ArticleExt(id=1239173817491313234, articleId=1239173816769892936, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Study on UPLC fingerprint and simultaneous determination of 5 components in Frucius Aceris Fabri*, columnId=1239148838318043851, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Ingredient Analys, runingTitle=null, highlight=null, articleAbstract=
Objective:

To establish a method of UPLC fingerprint and quantitative analysis of gallic acid, protocatechuic acid, p-hydroxycinnamic acid, myricitrin and quercitrin for Frucius Aceris Fabri, and to provide a reference for the quality control of Frucius Aceris Fabri.

Methods:

The determination was performed on a Waters CORTECS UPLC T3 column (150 mm×2.1 mm, 1.6 μm), with mobile phase consisting of acetonitrile -0.1% phosphoric acid by gradually elution at a flow rate of 0.20 mL·min-1. The column temperature was 30 ℃, and detection wavelength was set at 300 nm. The quality of 10 batches of Frucius Aceris Fabri was evaluated by similarity analysis, CA and TOPSIS analysis of fingerprints.

Results:

The UPLC fingerprint of Frucius Aceris Fabri was established, and 14 peaks were selected as the characteristic fingerprint peaks. Five chemical components were identified, which were gallic acid, protocatechuic acid, p-hydroxycinnamic acid, myricitrin and quercitrin. And a quantitative method for the determination of the 5 chemical components was established. Good similarities were found in the established fingerprint through similarity analysis. CA and TOPSIS analysis showed that 10 batches of Frucius Aceris Fabri samples could be clustered into 3 groups. The sample S9 from Jiangxi was classified as class Ⅰ with best quality. The samples from Guangxi and Guangdong were classified as Class Ⅱ with medium quality. The samples S1, S4 and S6 from Jiangxi were classified as Ⅲ with worst quality. The linear relationship of the 5 chemical components was good, and the r values were all above 0.999. The contents of the 5 chemical components in 10 batches of samples were 0.236-0.356 mg·g-1, 0.118-0.398 mg·g-1, 0.108-0.141 mg·g-1, 0.146-0.222 mg·g-1 and 0.046-0.104 mg·g-1, respectively.

Conclusion:

The established UPLC fingerprint and quantitative analysis methods of Frucius Aceris Fabri are precise and stable, which can be used for evaluating and controlling the quality of Frucius Aceris Fabri.

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目的:

建立中药蝴蝶果超高效液相色谱(UPLC)指纹图谱及没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷5个成分的定量分析方法,为中药蝴蝶果质量控制提供依据。

方法:

采用Waters CORTECS UPLC T3色谱柱(150 mm×2.1 mm,1.6 μm),以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,柱温30 ℃,流速0.20 mL·min-1,检测波长300 nm,进样量为1 μL;采用“中药色谱指纹图谱相似度评价系统”生成对照指纹图谱并进行相似度分析,结合聚类分析、TOPSIS多元统计分析法进行评价。

结果:

建立了中药蝴蝶果UPLC指纹图谱,共标示了14个共有峰,指认了其中没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷5个成分,并建立了该5个成分的定量分析方法。10批次中药蝴蝶果的指纹图谱相似度均在0.9以上,其中来源于江西的样品S9聚为Ⅰ类,质量最佳;来源于广西和广东的样品聚为Ⅱ类,质量次之;来源于江西的样品S1、S4、S6聚为Ⅲ类,质量最差。10批样品中上述5个成分的含量分别为0.236~0.356、0.118~0.398、0.108~0.141、0.146~0.222、0.046~0.104 mg·g-1

结论:

所建立的蝴蝶果指纹图谱及含量测定分析方法精密度高,稳定性好,可用于中药蝴蝶果的质量控制及评价。

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1.没食子酸(gallic acid) 3.原儿茶酸(protocatechuic acid) 7.对羟基肉桂酸(p-hydroxycinnamic acid) 11.杨梅苷(myricitrin) 14.槲皮苷(quercitrin)

, figureFileSmall=Rwvqn25/HnBHTCndTVvdTQ==, figureFileBig=RZkOcdktbfC9xqdc/iPYhA==, tableContent=null), ArticleFig(id=1239173824789402553, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Fig.3, caption=Cluster analysis tree of 10 batches of Frucius Aceris Fabri, figureFileSmall=86D0Brh+AxOsGKmx+5cexQ==, figureFileBig=OLneoqjQJw0ZpZyxl17JCQ==, tableContent=null), ArticleFig(id=1239173824894260157, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=图3, caption=10批蝴蝶果药材聚类分析树状图, figureFileSmall=86D0Brh+AxOsGKmx+5cexQ==, figureFileBig=OLneoqjQJw0ZpZyxl17JCQ==, tableContent=null), ArticleFig(id=1239173826391626689, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Fig.4, caption=Chromatograms of content determination of 5 components in Frucius Aceris Fabri, figureFileSmall=GpFxEhCoa3C7xcTOQxRpSg==, figureFileBig=H2dbhKCmiDSeGQpeZ2Y83g==, tableContent=null), ArticleFig(id=1239173826492289989, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=图4, caption=蝴蝶果药材5个成分含量测定色谱图

1.没食子酸(gallic acid) 2.原儿茶酸(protocatechuic acid) 3.对羟基肉桂酸(p-hydroxycinnamic acid) 4.杨梅苷(myricitrin) 5.槲皮苷(quercitrin)

, figureFileSmall=GpFxEhCoa3C7xcTOQxRpSg==, figureFileBig=H2dbhKCmiDSeGQpeZ2Y83g==, tableContent=null), ArticleFig(id=1239173826597147595, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Tab.1, caption=

Samples information of Frucius Aceris Fabri

, figureFileSmall=null, figureFileBig=null, tableContent=
样品编号
(sample No.)
批号
(batch No.)
产地
(habitat)
样品编号
(sample No.)
批号
(batch No.)
产地
(habitat)
S12103217江西(Jiangxi)S62104022江西(Jiangxi)
S22103218广西(Guangxi)S72104201广东(Guangdong)
S32103219广东(Guangdong)S82104202广东(Guangdong)
S42103202江西(Jiangxi)S92105055江西(Jiangxi)
S52103220广西(Guangxi)S102105057广西(Guangxi)
), ArticleFig(id=1239173826706199501, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=表1, caption=

蝴蝶果药材样品信息

, figureFileSmall=null, figureFileBig=null, tableContent=
样品编号
(sample No.)
批号
(batch No.)
产地
(habitat)
样品编号
(sample No.)
批号
(batch No.)
产地
(habitat)
S12103217江西(Jiangxi)S62104022江西(Jiangxi)
S22103218广西(Guangxi)S72104201广东(Guangdong)
S32103219广东(Guangdong)S82104202广东(Guangdong)
S42103202江西(Jiangxi)S92105055江西(Jiangxi)
S52103220广西(Guangxi)S102105057广西(Guangxi)
), ArticleFig(id=1239173826815251408, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Tab.2, caption=

RRT of 10 batches of Frucius Aceris Fabri

, figureFileSmall=null, figureFileBig=null, tableContent=
共有峰号
(common peak No.)
RRTRSD/%
S1S2S3S4S5S6S7S8S9S10
10.570.570.570.570.580.570.570.570.570.570.29
20.830.830.830.830.830.830.830.830.830.830.13
31.001.001.001.001.001.001.001.001.001.000.00
41.911.911.921.911.911.921.931.911.911.920.37
51.961.951.961.951.951.961.971.951.951.960.35
62.182.182.192.182.172.192.202.182.182.190.35
72.232.232.242.232.222.242.252.222.232.240.35
82.272.262.272.262.252.282.282.262.272.280.37
92.362.352.362.352.342.372.372.352.362.360.36
102.492.492.502.492.482.502.512.492.502.500.33
112.642.632.642.632.622.642.652.622.632.640.39
122.702.692.702.692.682.712.712.682.692.700.40
133.143.133.153.133.113.153.163.113.143.150.49
143.273.253.273.253.243.283.283.243.263.270.49
), ArticleFig(id=1239173826903331798, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=表2, caption=

10批蝴蝶果药材共有峰的RRT

, figureFileSmall=null, figureFileBig=null, tableContent=
共有峰号
(common peak No.)
RRTRSD/%
S1S2S3S4S5S6S7S8S9S10
10.570.570.570.570.580.570.570.570.570.570.29
20.830.830.830.830.830.830.830.830.830.830.13
31.001.001.001.001.001.001.001.001.001.000.00
41.911.911.921.911.911.921.931.911.911.920.37
51.961.951.961.951.951.961.971.951.951.960.35
62.182.182.192.182.172.192.202.182.182.190.35
72.232.232.242.232.222.242.252.222.232.240.35
82.272.262.272.262.252.282.282.262.272.280.37
92.362.352.362.352.342.372.372.352.362.360.36
102.492.492.502.492.482.502.512.492.502.500.33
112.642.632.642.632.622.642.652.622.632.640.39
122.702.692.702.692.682.712.712.682.692.700.40
133.143.133.153.133.113.153.163.113.143.150.49
143.273.253.273.253.243.283.283.243.263.270.49
), ArticleFig(id=1239173827024966616, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Tab.3, caption=

RPA of 10 batches of Frucius Aceris Fabri

, figureFileSmall=null, figureFileBig=null, tableContent=
共有峰号
(common peak No.)
RPA
S1S2S3S4S5S6S7S8S9S10
10.7400.9610.8770.6750.5190.9081.2721.4921.2381.043
20.4980.4120.3680.4520.2940.4120.7380.8520.5780.590
31.0001.0001.0001.0001.0001.0001.0001.0001.0001.000
40.6730.7410.9790.6520.5710.8901.0191.6991.4261.219
51.0331.8633.8201.0091.5831.4313.3743.9656.0962.699
62.6883.0194.6492.7682.6993.3734.2715.8224.8724.786
72.0301.9782.6081.9211.4102.6072.8914.5473.2003.249
81.3591.2461.0821.5590.5201.4731.0292.4121.9201.786
91.9711.9793.3971.9711.8552.3632.7704.0243.1193.231
101.7561.7902.6961.6991.4052.2382.4624.2542.8483.040
110.4940.5510.6980.5060.4350.5030.8420.9850.9410.693
120.2940.3020.5150.3010.2780.3200.4550.5650.5940.518
132.0321.7302.8502.1041.5642.0432.4052.3142.2522.505
140.1360.2800.2470.1430.1600.1840.3440.4150.3800.259
), ArticleFig(id=1239173827121435613, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=表3, caption=

10批蝴蝶果药材共有峰的RPA

, figureFileSmall=null, figureFileBig=null, tableContent=
共有峰号
(common peak No.)
RPA
S1S2S3S4S5S6S7S8S9S10
10.7400.9610.8770.6750.5190.9081.2721.4921.2381.043
20.4980.4120.3680.4520.2940.4120.7380.8520.5780.590
31.0001.0001.0001.0001.0001.0001.0001.0001.0001.000
40.6730.7410.9790.6520.5710.8901.0191.6991.4261.219
51.0331.8633.8201.0091.5831.4313.3743.9656.0962.699
62.6883.0194.6492.7682.6993.3734.2715.8224.8724.786
72.0301.9782.6081.9211.4102.6072.8914.5473.2003.249
81.3591.2461.0821.5590.5201.4731.0292.4121.9201.786
91.9711.9793.3971.9711.8552.3632.7704.0243.1193.231
101.7561.7902.6961.6991.4052.2382.4624.2542.8483.040
110.4940.5510.6980.5060.4350.5030.8420.9850.9410.693
120.2940.3020.5150.3010.2780.3200.4550.5650.5940.518
132.0321.7302.8502.1041.5642.0432.4052.3142.2522.505
140.1360.2800.2470.1430.1600.1840.3440.4150.3800.259
), ArticleFig(id=1239173827238876128, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Tab.4, caption=

TOPSIS evaluation results of 10 batches of Frucius Aceris Fabri

, figureFileSmall=null, figureFileBig=null, tableContent=
样品序号
(sample No.)
正理想解距离D+
(perfect ideal solution distance D+)
负理想解距离D-
(negative ideal solution distance D-)
相对接近度C
(relative proximity C)
排序结果
(sort result)
S10.1450.0850.3699
S20.1130.0750.3996
S30.0940.1060.532
S40.1470.0910.3838
S50.1170.0880.4284
S60.1350.0740.35510
S70.1010.0890.4683
S80.1160.0830.4175
S90.0930.1430.6071
S100.1150.0730.3887
), ArticleFig(id=1239173827352122342, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=表4, caption=

10批蝴蝶果药材TOPSIS评价计算结果

, figureFileSmall=null, figureFileBig=null, tableContent=
样品序号
(sample No.)
正理想解距离D+
(perfect ideal solution distance D+)
负理想解距离D-
(negative ideal solution distance D-)
相对接近度C
(relative proximity C)
排序结果
(sort result)
S10.1450.0850.3699
S20.1130.0750.3996
S30.0940.1060.532
S40.1470.0910.3838
S50.1170.0880.4284
S60.1350.0740.35510
S70.1010.0890.4683
S80.1160.0830.4175
S90.0930.1430.6071
S100.1150.0730.3887
), ArticleFig(id=1239173827440202729, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Tab.5, caption=

Results of the linear relationship of 5 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
LOQ/(μg·mL-1LOD/(μg·mL-1
没食子酸(gallic acid)Y=6.42×103X+9.00×1030.999 96.602~66.0230.1320.039 6
原儿茶酸(protocatechuic acid)Y=8.74×103X+1.42×1030.999 86.980~69.7970.1400.041 9
对羟基肉桂酸(p-hydroxycinnamic acid)Y=3.40×104X+4.55×1040.999 86.152~61.5230.1230.036 9
杨梅苷(myricitrin)Y=6.26×103X+6.01×1030.999 74.127~41.2720.0830.024 8
槲皮苷(quercitrin)Y=6.64×103X+3.91×1030.999 82.640~26.4040.0530.015 8
), ArticleFig(id=1239173827549254639, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=表5, caption=

5个成分线性关系结果

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
r线性范围
(linear range)/(μg·mL-1
LOQ/(μg·mL-1LOD/(μg·mL-1
没食子酸(gallic acid)Y=6.42×103X+9.00×1030.999 96.602~66.0230.1320.039 6
原儿茶酸(protocatechuic acid)Y=8.74×103X+1.42×1030.999 86.980~69.7970.1400.041 9
对羟基肉桂酸(p-hydroxycinnamic acid)Y=3.40×104X+4.55×1040.999 86.152~61.5230.1230.036 9
杨梅苷(myricitrin)Y=6.26×103X+6.01×1030.999 74.127~41.2720.0830.024 8
槲皮苷(quercitrin)Y=6.64×103X+3.91×1030.999 82.640~26.4040.0530.015 8
), ArticleFig(id=1239173827633140722, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=EN, label=Tab.6, caption=

Results of content determination of 5 components in Frucius Aceris Fabri

, figureFileSmall=null, figureFileBig=null, tableContent=
样品序号
(sample No.)
含量(content)/(mg·g-1
没食子酸
(gallic acid)
原儿茶酸
(protocatechuic acid)
对羟基肉桂酸
p-hydroxycinnamic acid)
杨梅苷
(myricitrin)
槲皮苷
(quercitrin)
S10.2510.2310.1110.1590.046
S20.3560.2500.1160.1930.104
S30.2680.2050.1310.1870.078
S40.2360.2390.1080.1690.049
S50.2880.3980.1280.2120.091
S60.2860.2090.1340.1460.058
S70.3510.1820.1320.2040.096
S80.2810.1180.1410.1750.079
S90.3020.1590.1280.2220.088
S100.2750.1720.1410.1730.070
平均值(average)0.2900.2160.1270.1840.076
RSD/%13.334.99.213.026.2
), ArticleFig(id=1239173827721221110, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239173816769892936, language=CN, label=表6, caption=

10批蝴蝶果药材5个成分含量测定结果(n=2)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品序号
(sample No.)
含量(content)/(mg·g-1
没食子酸
(gallic acid)
原儿茶酸
(protocatechuic acid)
对羟基肉桂酸
p-hydroxycinnamic acid)
杨梅苷
(myricitrin)
槲皮苷
(quercitrin)
S10.2510.2310.1110.1590.046
S20.3560.2500.1160.1930.104
S30.2680.2050.1310.1870.078
S40.2360.2390.1080.1690.049
S50.2880.3980.1280.2120.091
S60.2860.2090.1340.1460.058
S70.3510.1820.1320.2040.096
S80.2810.1180.1410.1750.079
S90.3020.1590.1280.2220.088
S100.2750.1720.1410.1730.070
平均值(average)0.2900.2160.1270.1840.076
RSD/%13.334.99.213.026.2
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中药蝴蝶果UPLC指纹图谱及含量测定*
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江洁怡 1 , 杨敏娟 2 , 汤瑞茵 2 , 李国卫 3 , 肖观林 1 , 张靖年 1, ** , 胥爱丽 1
药物分析杂志 | 成分分析 2024,44(2): 233-241
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药物分析杂志 | 成分分析 2024, 44(2): 233-241
中药蝴蝶果UPLC指纹图谱及含量测定*
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江洁怡1 , 杨敏娟2, 汤瑞茵2, 李国卫3, 肖观林1, 张靖年1, ** , 胥爱丽1
作者信息
  • 1.广东省中医药工程技术研究院 广东省中医药研究开发重点实验室,广州 510095
  • 2.广州中医药大学第五临床医学院,广州 510095
  • 3.广东一方制药有限公司,佛山 528244
  • Tel:(020)83482098;E-mail:

通讯作者:

**Tel:(020)83482098;E-mail:
Study on UPLC fingerprint and simultaneous determination of 5 components in Frucius Aceris Fabri*
Jie-yi JIANG1 , Min-juan YANG2, Rui-yin TANG2, Guo-wei LI3, Guan-lin XIAO1, Jing-nian ZHANG1, ** , Ai-li XU1
Affiliations
  • 1.Guangdong Province Engineering and Technology Research Institute of Traditional Chinese Medicine, Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine, Guangzhou 510095, China
  • 2. The Fifth Clinical College of Guangzhou University of Chinese Medicine, Guangzhou 510095, China
  • 3.Guangdong Yi Fang Pharmaceutical Co., Ltd., Foshan 528244, China
出版时间: 2024-02-29 doi: 10.16155/j.0254-1793.2024.02.05
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目的:

建立中药蝴蝶果超高效液相色谱(UPLC)指纹图谱及没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷5个成分的定量分析方法,为中药蝴蝶果质量控制提供依据。

方法:

采用Waters CORTECS UPLC T3色谱柱(150 mm×2.1 mm,1.6 μm),以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,柱温30 ℃,流速0.20 mL·min-1,检测波长300 nm,进样量为1 μL;采用“中药色谱指纹图谱相似度评价系统”生成对照指纹图谱并进行相似度分析,结合聚类分析、TOPSIS多元统计分析法进行评价。

结果:

建立了中药蝴蝶果UPLC指纹图谱,共标示了14个共有峰,指认了其中没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷5个成分,并建立了该5个成分的定量分析方法。10批次中药蝴蝶果的指纹图谱相似度均在0.9以上,其中来源于江西的样品S9聚为Ⅰ类,质量最佳;来源于广西和广东的样品聚为Ⅱ类,质量次之;来源于江西的样品S1、S4、S6聚为Ⅲ类,质量最差。10批样品中上述5个成分的含量分别为0.236~0.356、0.118~0.398、0.108~0.141、0.146~0.222、0.046~0.104 mg·g-1

结论:

所建立的蝴蝶果指纹图谱及含量测定分析方法精密度高,稳定性好,可用于中药蝴蝶果的质量控制及评价。

中药蝴蝶果  /  指纹图谱  /  多成分含量测定  /  聚类分析  /  TOPSIS多元统计分析
Objective:

To establish a method of UPLC fingerprint and quantitative analysis of gallic acid, protocatechuic acid, p-hydroxycinnamic acid, myricitrin and quercitrin for Frucius Aceris Fabri, and to provide a reference for the quality control of Frucius Aceris Fabri.

Methods:

The determination was performed on a Waters CORTECS UPLC T3 column (150 mm×2.1 mm, 1.6 μm), with mobile phase consisting of acetonitrile -0.1% phosphoric acid by gradually elution at a flow rate of 0.20 mL·min-1. The column temperature was 30 ℃, and detection wavelength was set at 300 nm. The quality of 10 batches of Frucius Aceris Fabri was evaluated by similarity analysis, CA and TOPSIS analysis of fingerprints.

Results:

The UPLC fingerprint of Frucius Aceris Fabri was established, and 14 peaks were selected as the characteristic fingerprint peaks. Five chemical components were identified, which were gallic acid, protocatechuic acid, p-hydroxycinnamic acid, myricitrin and quercitrin. And a quantitative method for the determination of the 5 chemical components was established. Good similarities were found in the established fingerprint through similarity analysis. CA and TOPSIS analysis showed that 10 batches of Frucius Aceris Fabri samples could be clustered into 3 groups. The sample S9 from Jiangxi was classified as class Ⅰ with best quality. The samples from Guangxi and Guangdong were classified as Class Ⅱ with medium quality. The samples S1, S4 and S6 from Jiangxi were classified as Ⅲ with worst quality. The linear relationship of the 5 chemical components was good, and the r values were all above 0.999. The contents of the 5 chemical components in 10 batches of samples were 0.236-0.356 mg·g-1, 0.118-0.398 mg·g-1, 0.108-0.141 mg·g-1, 0.146-0.222 mg·g-1 and 0.046-0.104 mg·g-1, respectively.

Conclusion:

The established UPLC fingerprint and quantitative analysis methods of Frucius Aceris Fabri are precise and stable, which can be used for evaluating and controlling the quality of Frucius Aceris Fabri.

Frucius Aceris Fabri  /  fingerprint  /  multi-component determination  /  cluster analysis  /  TOPSIS multivariate statistical analysis
江洁怡, 杨敏娟, 汤瑞茵, 李国卫, 肖观林, 张靖年, 胥爱丽. 中药蝴蝶果UPLC指纹图谱及含量测定*. 药物分析杂志, 2024 , 44 (2) : 233 -241 . DOI: 10.16155/j.0254-1793.2024.02.05
Jie-yi JIANG, Min-juan YANG, Rui-yin TANG, Guo-wei LI, Guan-lin XIAO, Jing-nian ZHANG, Ai-li XU. Study on UPLC fingerprint and simultaneous determination of 5 components in Frucius Aceris Fabri*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (2) : 233 -241 . DOI: 10.16155/j.0254-1793.2024.02.05
中药蝴蝶果又名红蝴蝶,为槭树科植物罗浮槭Acer fabri Hance及桂林槭A. kweilinense Fang et Fang f.的果实,味甘、微苦,性凉,具有清热解毒的功效,主要用于治疗声音嘶哑、咽喉炎、扁桃体炎、肝炎、肺结核等症[1-2]。蝴蝶果广泛分布于我国江西、湖北、湖南、广东、广西、贵州、海南、四川等地,资源丰富,具有很好的开发应用前景。研究表明,槭树科植物富含黄酮、单宁、苯丙素、萜、甾、生物碱及二芳基庚烷衍生物等类化学成分,具有降低胆固醇、预防脑血管疾病、减少冠心病、高血压和延缓神经衰老等多种药效活性[3-5],然而现阶段针对罗浮槭及桂林槭的研究较少,主要集中在其作为观赏植物的引种、栽培等方面[6-7],其作为中药材的药理、化学成分研究却鲜有报道;质量标准方面,也仅见《广西中药材标准》(1990年版)对其性状鉴别作了相关规定[8],缺乏必要的质控指标及方法,严重限制了其进一步开发利用。
中药指纹图谱可整体、全面地反映中药的化学成分信息,特别是超高效液相色谱法(UPLC)指纹图谱,具有整体性好,特征性强,分析效率高等优点,被广泛应用于中药材、饮片及其复方制剂的质量控制中[9-12]。本研究拟建立蝴蝶果UPLC指纹图谱,同时测定其中没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷5个成分的含量,为蝴蝶果的质量控制提供科学依据,为其进一步的开发利用提供研究基础。
Waters ACQUITY H-Class色谱仪(Agilent公司),XS205DU十万分之一电子分析天平(梅特勒-托利多公司),Milli-Q Advantage型A10自动型纯水机(Millipore公司),KQ-700DE型数控超声波清洗机(昆山市超声仪器有限公司)。
对照品没食子酸[批号(下同)110831-201906,含量(下同)91.5%]、原儿茶酸(110809-201906,97.7%)、杨梅苷(111860-201703,95.2%)、槲皮苷(111538-202007,93.5%)均购自中国食品药品检定研究院,对羟基肉桂酸(21101401,99.94%)购自成都普菲德生物科技有限公司;10批蝴蝶果药材由广东一方制药有限公司提供,经广东省中医药工程技术研究院刘法锦教授鉴定为槭树科植物罗浮槭Acer fabri Hance及桂林槭Acer kweilinense Fang et Fang f.的果实,产地信息见表1
乙腈、磷酸等液相用试剂为色谱纯(Merck公司),甲醇等其余试剂均为分析纯(广州化学试剂厂),水为超纯水(Advantage A10超纯水系统)。
Waters CORTECS UPLC T3色谱柱(150 mm×2.1 mm,1.6 μm),以乙腈(A)-0.1%磷酸(B)为流动相,梯度洗脱(0~5 min,5%A→10%A;5~7 min,10%%A→15%A;7~15 min,15%%A→20%A;15~25 min,20%%A→25%A),流速0.20 mL·min-1,检测波长300 nm,进样量1 μL,柱温30 ℃。
取蝴蝶果药材粉末约1.0 g,精密称量,置具塞锥形瓶中,精密加入70%甲醇水溶液15 mL,称量,加热回流1 h,取出,放冷,再次称量,用70%甲醇水溶液补足减失的量,摇匀,过0.22 μm的微孔滤膜,取续滤液,即得。
分别取没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷的对照品适量,精密称定,加甲醇制成质量浓度分别为330.113、348.984、410.154、165.088、660.110 μg·mL-1的对照品储备液。取上述各对照品储备液适量,制成系列浓度的混合对照品溶液。
取同一蝴蝶果药材供试品溶液(S3),按“2.1”项色谱条件,连续进样6次,记录色谱图,以3号峰(原儿茶酸)为参照,计算得到各共有峰的相对保留时间(relative retention time,RRT)及相对峰面积(relative common peak area,RPA)的RSD均<3.0%,表明所用仪器精密度良好。
分别取同一批次蝴蝶果药材(S3),照“2.2”项方法平行制备6份供试品溶液,按“2.1”项色谱条件进样并记录色谱图,以3号峰(原儿茶酸)为参照,计算得到各共有峰的RRT及RPA的RSD均<3.0%,表明所建立方法重复性良好。
取同一蝴蝶果药材供试品溶液(S3),按“2.1”项色谱条件,分别在0、2、4、8、16、24 h进样,记录色谱图,以3号峰(原儿茶酸)为参照,计算得到各共有峰的RRT及RPA的RSD均<3.0%,表明供试品溶液24 h内稳定性良好。
取10批蝴蝶果药材,照“2.2”项方法分别制备供试品溶液,精密吸取各供试品溶液及混合对照品溶液1 μL,按“2.1”项色谱条件依次进样测定,记录色谱图,将数据导入“中药色谱指纹图谱相似度评价系统(2012版)”,以S1号样品色谱图为参照图谱,采用中位数法,时间窗宽度为0.1 min,全谱自动匹配,生成对照指纹图谱,10批样品叠加图谱见图1。根据匹配数据,共标定14个共有峰,通过与对照品保留时间及紫外全波长扫描图比对,指认了其中5个共有峰,分别为没食子酸(峰1)、原儿茶酸(峰3)、对羟基肉桂酸(峰7)、杨梅苷(峰11)、槲皮苷(峰14),见图2。以3号峰原儿茶酸为参照(S),计算其他各共有峰的RRT和RPA,结果见表23
将10批样品数据导入“中药色谱指纹图谱相似度评价系统(2012版)”,计算10批蝴蝶果药材供试品指纹图谱与所生成对照指纹图谱的相似度分别为0.988、0.959、0.996、0.979、0.996、0.989、0.974、0.978、0.988、0.993,均>0.9,不同产地间蝴蝶果药材化学成分基本一致、质量稳定。
聚类分析亦称为群分析,其能把相似的对象通过静态分类的方法分成不同的组别或者更多的子集,是研究样品或指标分类问题的多元统计方法。以各批样品的14个共有峰的相对峰面积为原始数据,经归一化处理后,导入SPSS 26.0软件,采用组间联接聚类法,选取平方Euclidean距离作为度量标准进行聚类分析。结果表明,当类间距>10时,10批蝴蝶果药材可聚为3类,S1、S4、S6聚为一类,均来源于江西;S9聚为一类,来源于江西;其余批次聚为一类,来源于广西和广东,见图3
TOPSIS法是常用的样品优劣综合评价方法,能充分利用原始数据的信息,精确地反映各评价目标之间的差距。其原理是通过归一化后的数据规范化矩阵,找出多个目标中的最优目标(理想解)和最劣目标(反理想解),分别计算各评价目标与理想解、反理想解的距离,获得各目标与理想解的贴近度并进行排序,以此作为评价目标优劣的依据。以各批样品的14个共有峰的RPA为原始数据,导入在线SPSS分析软件,对数据进行归一化处理后,运行TOPSIS分析,比较不同批次蝴蝶果药材的质量差异。结果表明,S9的质量最佳,S1、S4、S6的质量最次,见表4
精密吸取没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷的对照品储备液适量,加甲醇稀释得不同浓度梯度的系列混合对照品溶液,精密吸取上述溶液各1 μL,按“2.1”项色谱条件测定,记录各色谱峰峰面积,以峰面积(Y)为纵坐标,对照品溶液浓度(X)为横坐标进行回归,得到线性回归方程,各成分在相应的含量范围内线性关系良好,见表5
取同一蝴蝶果药材供试品溶液(S3),按“2.1”项色谱条件,连续测定6次,记录各成分峰面积,计算得到没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷峰面积的RSD分别为0.73%、0.64%、0.60%、0.68%、0.48%,表明所用仪器精密度良好。
取同一蝴蝶果药材供试品溶液(S3),按“2.1”项色谱条件,分别在0、2、4、8、16、24 h进样,记录各成分峰面积,计算得到没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷峰面积的RSD分别为1.1%、0.82%、1.6%、0.89%、1.2%,表明供试品溶液24 h内稳定性良好。
分别取同一批次蝴蝶果药材(S3),照“2.2”项的方法平行制备6份供试品溶液,并按“2.1”项色谱条件测定,记录各成分峰面积并计算含量,计算得到没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷的平均含量分别为0.246、0.222、0.134、0.205、0.064 mg·g-1,RSD分别为1.9%、1.1%、2.7%、1.7%、1.5%,表明该方法重复性较好。
取已知含量的蝴蝶果药材样品(S3),分别精密加入各对照品适量,照“2.2”项方法平行制备6份供试溶液,并按“2.1”项色谱条件测定,记录各成分峰面积并计算含量,计算得到没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷的平均加样回收率分别为93.9%、99.3%、94.0%、100.0%、98.8%,RSD分别为1.6%、1.1%、1.5%、1.6%、2.5%,符合2020年版《中国药典》规定。
分别取10批蝴蝶果药材,照“2.2”项方法制备供试品溶液,每批次平行处理2份,按“2.1”项色谱条件测定,记录各成分峰面积并计算含量,结果(图4表6)表明,10批蝴蝶果药材中没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷的平均含量分别为0.290、0.216、0.127、0.184、0.076。
为了更全面、充分地展现蝴蝶果药材中所含化学成分,采用紫外210~400 nm全波长扫描对检测波长进行筛选,结果显示在300 nm各色谱峰数目、丰度、分离度均为最优,故选择300 nm为检测波长,并先后考察了不同色谱柱(Waters CORTECS UPLC T3、Waters ACQUITY UPLC BEH Shield RP C18、Waters ACQUITY UPLC BEH C18)、流动相体系(乙腈-水、乙腈-0.1%甲酸水溶液、乙腈-0.1%磷酸水溶液)、柱温(25、30、35 ℃)以及流速(0.17、0.20、0.23 mL·min-1)对分离效果的影响,最终确定了“2.1”项色谱条件;考察了不同提取溶剂(50%甲醇水、70%甲醇水、纯甲醇)、提取方式(超声、加热回流)、提取时间(0.5、1、2 h)对提取效果的影响,最终确定了“2.2”项供试品处理方法。
本研究建立了蝴蝶果药材的指纹图谱,对10批不同产地样品进行测定,并采用相似度评价、CA、TOPSIS等多元统计分析法对其进行分析评价。相似度评价结果显示各批次样品的相似度均在0.9以上,表明蝴蝶果药材总体质量较好、较稳定。CA分析能较好在将不同产地的蝴蝶果药材进行分类,其中广西和广东的蝴蝶果样品聚为一类,可能与两广地区地理位置比邻且气候特点相近有关系;而来源于江西的样品则分为两类,结合TOPSIS分析法得到的不同批次蝴蝶果药材的质量优劣排序结果,S9的质量最佳,广西和广东的样品次之,S1、S4、S6的质量最差,由此可知,江西样品分为两类,与个别样品的质量差异有关,这可能与样品的采收季节等因素有关。由于本研究只代表性地选取了3个产地10批样品进行分析,且样品存在个体差异,统计结果仅为蝴蝶果样品的质量评价提供参考,尚未能充分地反映我国蝴蝶果药材的整体情况,而后续将继续收集更多批次药材,采集更多数据,丰富统计结果。
课题组前期对蝴蝶果药材中化学成分进行了初步摸索,发现其主要含有黄酮、多酚、萜等类化学成分。本研究建立了蝴蝶果药材的UPLC指纹图谱,经与对照品比对,指认了其中5个共有峰分别为没食子酸(峰1)、原儿茶酸(峰3)、对羟基肉桂酸(峰7)、杨梅苷(峰11)、槲皮苷(峰14),其中没食子酸、原儿茶酸和对羟基肉桂酸是自然界广泛存在的天然多酚类化合物,均具有抗炎、抗氧化、自由基清除、抗菌、抗肿瘤等活性[13-20];杨梅苷、槲皮苷为多酚羟基黄酮苷类化合物,均具有抗氧化、改善心血管功能、护肝、抗炎、降脂等药理作用[21-24]。本研究在指纹图谱研究的基础上同步建立了蝴蝶果药材中没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷等5种成分的含量测定方法,并对10批蝴蝶果药材进行测定,结果10批样品中没食子酸、原儿茶酸、对羟基肉桂酸、杨梅苷、槲皮苷平均含量分别为0.290、0.216、0.127、0.184、0.076 mg·g-1。若对各成分的含量以平均含量的70%设限,10批样品中,存在1批样品(S8)原儿茶酸含量低于标准,2批样品(S1、S4)槲皮苷含量低于标准,其余含量均在限度内。另外对10批样品的含测数据分析可发现,蝴蝶果药材中槲皮苷的含量较低,大多在万分之一以下,为了保证含量测定的准确性及质量控制的合理性,建议质量标准中可不对其进行控制。
  • *2021年省属科研机构稳定性支持项目(粤财科教[2021]113号)
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doi: 10.16155/j.0254-1793.2024.02.05
  • 接收时间:2023-01-12
  • 首发时间:2026-03-13
  • 出版时间:2024-02-29
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*2021年省属科研机构稳定性支持项目(粤财科教[2021]113号)
作者信息
    1.广东省中医药工程技术研究院 广东省中医药研究开发重点实验室,广州 510095
    2.广州中医药大学第五临床医学院,广州 510095
    3.广东一方制药有限公司,佛山 528244

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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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