Article(id=1240688794928739281, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240688783964819588, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024-0180, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1710604800000, receivedDateStr=2024-03-17, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773732859594, onlineDateStr=2026-03-17, pubDate=1727625600000, pubDateStr=2024-09-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773732859594, onlineIssueDateStr=2026-03-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773732859594, creator=13701087609, updateTime=1773732859594, updator=13701087609, issue=Issue{id=1240688783964819588, tenantId=1146029695717560320, journalId=1205117023404326918, year='2024', volume='44', issue='9', pageStart='1463', pageEnd='1645', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773732856979, creator=13701087609, updateTime=1773733032821, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1240689521587712627, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240688783964819588, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1240689521587712628, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1240688783964819588, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1497, endPage=1503, ext={EN=ArticleExt(id=1240688795276866544, articleId=1240688794928739281, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Comparative study of eight components in branches and leaves of Xanthoceras sorbifolia Bunge at different growth stages by UHPLC-MS/MS method, columnId=1239256891872833779, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Ingredient Analysi, runingTitle=null, highlight=null, articleAbstract=

Objective: To establish an UHPLC-MS/MS method for determining eight primary components (catechin, epigallocatechin, rutin, quercitrin, epicatechin, (+)-dihydromyricetin, myricitrin and dihydroqurcetin) in the young branches and leaves of Xanthoceras sorbifolia Bunge, a medicinal plant from Mongolia, and to compare their contents in samples at different growth stages. Methods: A Waters CORTECS C18 (100 mm×2.1 mm, 1.6 μm) chromatographic column was adopted using the mobile phase comprised of water containing 0.1% formic acid (A) and acetonitrile (B) with gradient elution(0-1 min, 5% B; 1-10 min, 5% B→28% B; 10-11 min, 28% B→95% B; 11-14 min, 95% B; 14-15 min, 95% B→5% B) at a flow rate of 0.3 mL·min-1. The temperature of the column was set at 40 ℃. Injecting volume was 2 μL. Detection was conducted using electrospray ionization (ESI) in negative ion mode with multiple reaction monitoring (MRM). Results: The linearity of the eight chemical components was found to be excellent in the tested concentration ranges, with correlation coefficients above 0.997 6. Precision, repeatability and stability were satisfactory and the average recoveries were between 97.4% and 106.0% with RSDs≤5.0%. In six batches of leaves, contents of catechin, epigallocatechin, rutin, quercitrin, epicatechin, (+)-dihydromyricetin, myricitrin and dihydroqurcetin were in the ranges of 0.090-0.904 mg·g-1, 0.093-2.258 mg·g-1, 0.001-0.005 mg·g-1, 0.530-6.176 mg·g-1, 0.158-1.561 mg·g-1, 0.002-0.056 mg·g-1, 4.008-10.218 mg·g-1 and 1.049-16.990 mg·g-1, respectively. In six batches of young branches, the contents ranged from 0.384-1.025 mg·g-1, 0.911-2.427 mg·g-1, 0.008-0.127 mg·g-1, 0.870-2.295 mg·g-1, 0.659-1.746 mg·g-1, 0.125-1.079 mg·g-1, 2.296-4.681 mg·g-1 and 1.958-4.946 mg·g-1, respectively. The contents of eight components varied a lot in samples from different parts. The contents of myricitrin, rutin and quercitrin in the leaves exhibited noticeable changes with the growth cycle, suggesting their potential as quality control markers for leaves of Xanthoceras sorbifolia. Conclusion: The method is accurate, sensitive, stable, repeatable, and suitable for simultaneous determination of eight components in Xanthoceras sorbifolia Bunge, offering reference for quality control of its leaves and branches.

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目的:建立UHPLC-MS/MS测定方法测定蒙药文冠木枝和叶中儿茶素、表没食子儿茶素、芦丁、槲皮苷、表儿茶素、二氢杨梅素、杨梅苷和二氢槲皮素的不同周期含量对比研究。方法:采用WATER-SCORTECS 反相C18(100 mm×2.1 mm;1.6 μm)色谱柱,以0.1%甲酸水溶液为流动相A,乙腈为流动相B,梯度洗脱(0~1 min,5% B;1~10 min,5% B→28% B;10~11 min,28% B→95% B;11~14 min,95% B;14~15 min,95% B→5% B),柱温40 ℃,流速0.3 mL·min-1,进样体积2 μL,多反应监测(MRM)方法中选择负离子进行检测。结果:在测定的浓度范围内8个化学成分线性关系良好(r>0.997 6),精密度、重复性和稳定性良好,平均加样回收率为97.4%~106.0%,RSD≤5.0%。6批叶样品中8个成分测定结果分别为0.090~0.904 mg·g-1、0.093~2.258 mg·g-1、0.001~0.005 mg·g-1、0.530~6.176 mg·g-1、0.158~1.561 mg·g-1、0.002~0.056 mg·g-1、4.008~10.218 mg·g-1和1.049~16.990 mg·g-1,枝中8个成分测定结果分别为0.384~1.025 mg·g-1、0.911~2.427 mg·g-1、0.008~0.127 mg·g-1、0.870~2.295 mg·g-1、0.659~1.746 mg·g-1、0.125~1.079 mg·g-1、2.296~4.681 mg·g-1和1.958~4.946 mg·g-1。含量测定结果显示8个成分在文冠木不同部位中的含量存在较大差异,叶中杨梅苷、芦丁、槲皮苷随生长周期变化明显,可作为文冠木叶质量控制的主要指标。结论:该方法准确度和灵敏度高,且稳定性和重复性好,适用于蒙药文冠木药材中8个成分的同时检测,为枝和叶的质量控制提供实验基础。

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1.表没食子儿茶素(epigallocatechin) 2.儿茶素(catechin) 3.表儿茶素(epicatechin) 4.二氢杨梅素[(+)-dihydromyricetin] 5.杨梅苷(myricitrin) 6.芦丁(rutin) 7.二氢槲皮素(dihydroqurcetin) 8.槲皮苷(quercitrin)

, figureFileSmall=ai5Ep+x9wNAsSLvn3DwIxg==, figureFileBig=4TJvHaoI1A63Av6rPkoUgA==, tableContent=null), ArticleFig(id=1240704454564049008, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=EN, label=Fig.2, caption=Changes of 8 components in leaves and braches at different periods, figureFileSmall=SxJFNVnFmJFSF9n+FkF/gg==, figureFileBig=FIv97NzpwbcTuaYj0WreYA==, tableContent=null), ArticleFig(id=1240704454652129400, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=CN, label=图2, caption=叶、枝中8个成分在不同周期的变化图, figureFileSmall=SxJFNVnFmJFSF9n+FkF/gg==, figureFileBig=FIv97NzpwbcTuaYj0WreYA==, tableContent=null), ArticleFig(id=1240704454710849661, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=EN, label=Tab.1, caption=

Mass spectrometric data for 8 chemical components in Xanthoceras sorbifolia Bunge

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
相对分子质量
(relative molecular mass)
母离子
(precursor ion)m/z
子离子
(product ion)m/z
去簇电压
(declustering potential)/V
碰撞能量
(collision energy)/V
表没食子儿茶素(epigallocatechin)306.27305.2125.1-108-28
儿茶素(catechins)290.27289.0245.3-130-21
表儿茶素(epicatechin)290.27288.9202.9-120-26
二氢杨梅素(dihydromyricetin)320.25319.0193.2-110-15
二氢槲皮素(dihydroqurcetin)464.38463.1316.1-241-38
杨梅苷(myricetin)610.52609.1271.1-200-69
芦丁(rutin)304.25302.9285.1-120-16
槲皮苷(queritrin)448.38447.3300.0-140-40
), ArticleFig(id=1240704454811512962, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=CN, label=表1, caption=

文冠木中8个化学成分的质谱数据

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
相对分子质量
(relative molecular mass)
母离子
(precursor ion)m/z
子离子
(product ion)m/z
去簇电压
(declustering potential)/V
碰撞能量
(collision energy)/V
表没食子儿茶素(epigallocatechin)306.27305.2125.1-108-28
儿茶素(catechins)290.27289.0245.3-130-21
表儿茶素(epicatechin)290.27288.9202.9-120-26
二氢杨梅素(dihydromyricetin)320.25319.0193.2-110-15
二氢槲皮素(dihydroqurcetin)464.38463.1316.1-241-38
杨梅苷(myricetin)610.52609.1271.1-200-69
芦丁(rutin)304.25302.9285.1-120-16
槲皮苷(queritrin)448.38447.3300.0-140-40
), ArticleFig(id=1240704454907981961, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=EN, label=Tab.2, caption=

Regression equation,LOD and LOQ

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
线性方程
(linear equation)
r线性范围
(linear range)/(ng·mL-1)
LOD/(ng·mL-1)LOQ/(ng·mL-1)
表没食子儿茶素(epigallocatechin)Y=3 895X-148.60.999 60.133~1330.053 20.106 4
儿茶素(catechins)Y=3 617X-19.440.999 90.580~1160.046 40.928
表儿茶素(epicatechin)Y=1 365X-225.41.0000.960~1920.307 21.536
二氢杨梅素(dihydromyricetin)Y=10 191X-143.50.999 90.147~1470.023 50.117 6
二氢槲皮素(dihydroqurcetin)Y=14 822X+269.10.999 70.124~62.00.049 60.198 4
杨梅苷(myricetin)Y=1 202X-712.20.999 10.630~6300.299 21.496
芦丁(rutin)Y=5 177X+234.01.0000.187~9350.010 10.029 9
槲皮苷(quercitrin)Y=19 671X-1 7370.999 90.118~5900.011 00.026 5
), ArticleFig(id=1240704455012839566, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=CN, label=表2, caption=

回归方程、LOD和LOQ

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
线性方程
(linear equation)
r线性范围
(linear range)/(ng·mL-1)
LOD/(ng·mL-1)LOQ/(ng·mL-1)
表没食子儿茶素(epigallocatechin)Y=3 895X-148.60.999 60.133~1330.053 20.106 4
儿茶素(catechins)Y=3 617X-19.440.999 90.580~1160.046 40.928
表儿茶素(epicatechin)Y=1 365X-225.41.0000.960~1920.307 21.536
二氢杨梅素(dihydromyricetin)Y=10 191X-143.50.999 90.147~1470.023 50.117 6
二氢槲皮素(dihydroqurcetin)Y=14 822X+269.10.999 70.124~62.00.049 60.198 4
杨梅苷(myricetin)Y=1 202X-712.20.999 10.630~6300.299 21.496
芦丁(rutin)Y=5 177X+234.01.0000.187~9350.010 10.029 9
槲皮苷(quercitrin)Y=19 671X-1 7370.999 90.118~5900.011 00.026 5
), ArticleFig(id=1240704455113502865, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=EN, label=Tab.3, caption=

Determination of 8 chemical components in 6 batches of Xanthoceras sorbifolia leaves and branches

, figureFileSmall=null, figureFileBig=null, tableContent=
来源
(source)
批号
(lot No.)
含量(content)/(mg·g-1
表没食子儿茶素
(epigallocatechin)
儿茶素
(catechins)
表儿茶素
(epicatechin)
二氢杨梅素
(dihydromyricetin)
二氢槲皮素
(dihydroqurcetin)
杨梅苷
(myricetin)
芦丁
(rutin)
槲皮苷
(quercitrin)
文冠木叶
Xanthoceras sorbifolia leaves)
202305070.5700.0940.1740.01116.99010.2180.0016.176
202305141.3820.3120.5300.05612.0977.7370.0015.016
202305212.2580.9041.5610.0267.6087.6180.0013.938
202306040.4800.2700.4700.0279.8385.9600.0033.944
202306131.1150.5700.9770.0127.7625.0950.0014.388
202307160.2710.3450.5770.0025.3904.0080.0053.880
文冠木枝
Xanthoceras sorbifolia branches)
202305071.1660.8411.4310.1921.9583.1830.0290.870
202305142.4270.9041.5441.0793.2653.6210.0741.132
202305210.9110.3840.6590.1251.9462.9590.0081.882
202306041.3921.1211.1210.1704.7164.6810.0262.295
202306131.3491.0251.7460.8802.6412.2960.1271.341
202307161.3760.7391.2780.2163.3633.3460.0341.954
), ArticleFig(id=1240704455193194644, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1240688794928739281, language=CN, label=表3, caption=

6批文冠木叶和枝中8个化学成分的含量测定结果(n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
来源
(source)
批号
(lot No.)
含量(content)/(mg·g-1
表没食子儿茶素
(epigallocatechin)
儿茶素
(catechins)
表儿茶素
(epicatechin)
二氢杨梅素
(dihydromyricetin)
二氢槲皮素
(dihydroqurcetin)
杨梅苷
(myricetin)
芦丁
(rutin)
槲皮苷
(quercitrin)
文冠木叶
Xanthoceras sorbifolia leaves)
202305070.5700.0940.1740.01116.99010.2180.0016.176
202305141.3820.3120.5300.05612.0977.7370.0015.016
202305212.2580.9041.5610.0267.6087.6180.0013.938
202306040.4800.2700.4700.0279.8385.9600.0033.944
202306131.1150.5700.9770.0127.7625.0950.0014.388
202307160.2710.3450.5770.0025.3904.0080.0053.880
文冠木枝
Xanthoceras sorbifolia branches)
202305071.1660.8411.4310.1921.9583.1830.0290.870
202305142.4270.9041.5441.0793.2653.6210.0741.132
202305210.9110.3840.6590.1251.9462.9590.0081.882
202306041.3921.1211.1210.1704.7164.6810.0262.295
202306131.3491.0251.7460.8802.6412.2960.1271.341
202307161.3760.7391.2780.2163.3633.3460.0341.954
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UHPLC-MS/MS法测定不同周期蒙药文冠木枝和叶中8个成分的对比研究
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布仁满达 1, 2 , 吴杰斯 2 , 敖敦格日乐 1 , 黄格尔柱拉 1 , 胡阿荣 1 , 其日格尔 1 , 拉喜那木吉拉 1, *
药物分析杂志 | 成分分析 2024,44(9): 1497-1503
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药物分析杂志 | 成分分析 2024, 44(9): 1497-1503
UHPLC-MS/MS法测定不同周期蒙药文冠木枝和叶中8个成分的对比研究
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布仁满达1, 2 , 吴杰斯2, 敖敦格日乐1, 黄格尔柱拉1, 胡阿荣1, 其日格尔1, 拉喜那木吉拉1, *
作者信息
  • 1.内蒙古民族大学国家药监局中药(蒙药)质量控制重点实验室,通辽 028000
  • 2.内蒙古民族大学附属医院,通辽 028000
  • Tel: 14747502383; E-mail:

通讯作者:

* Tel: 13789710541; E-mail:
Comparative study of eight components in branches and leaves of Xanthoceras sorbifolia Bunge at different growth stages by UHPLC-MS/MS method
Bu-ren-man-da1, 2 , Jie-si WU2, Ao-dun-ge-ri-le1, Huang-ge-er-zhu-la1, A-rong HU1, Qi-ri-ge-er1, La-xi-na-mu-ji-la1, *
Affiliations
  • 1.Key Laboratory of Quality Control of Traditional Mongolian Medicine, State Administration of Traditional Chinese Medicine, Inner Mongolia Minzu University, Tongliao 028000, China
  • 2.Affiliated Hospital of Inner Mongolia Minzu University, Tongliao 028000, China
出版时间: 2024-09-30 doi: 10.16155/j.0254-1793.2024-0180
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目的:建立UHPLC-MS/MS测定方法测定蒙药文冠木枝和叶中儿茶素、表没食子儿茶素、芦丁、槲皮苷、表儿茶素、二氢杨梅素、杨梅苷和二氢槲皮素的不同周期含量对比研究。方法:采用WATER-SCORTECS 反相C18(100 mm×2.1 mm;1.6 μm)色谱柱,以0.1%甲酸水溶液为流动相A,乙腈为流动相B,梯度洗脱(0~1 min,5% B;1~10 min,5% B→28% B;10~11 min,28% B→95% B;11~14 min,95% B;14~15 min,95% B→5% B),柱温40 ℃,流速0.3 mL·min-1,进样体积2 μL,多反应监测(MRM)方法中选择负离子进行检测。结果:在测定的浓度范围内8个化学成分线性关系良好(r>0.997 6),精密度、重复性和稳定性良好,平均加样回收率为97.4%~106.0%,RSD≤5.0%。6批叶样品中8个成分测定结果分别为0.090~0.904 mg·g-1、0.093~2.258 mg·g-1、0.001~0.005 mg·g-1、0.530~6.176 mg·g-1、0.158~1.561 mg·g-1、0.002~0.056 mg·g-1、4.008~10.218 mg·g-1和1.049~16.990 mg·g-1,枝中8个成分测定结果分别为0.384~1.025 mg·g-1、0.911~2.427 mg·g-1、0.008~0.127 mg·g-1、0.870~2.295 mg·g-1、0.659~1.746 mg·g-1、0.125~1.079 mg·g-1、2.296~4.681 mg·g-1和1.958~4.946 mg·g-1。含量测定结果显示8个成分在文冠木不同部位中的含量存在较大差异,叶中杨梅苷、芦丁、槲皮苷随生长周期变化明显,可作为文冠木叶质量控制的主要指标。结论:该方法准确度和灵敏度高,且稳定性和重复性好,适用于蒙药文冠木药材中8个成分的同时检测,为枝和叶的质量控制提供实验基础。

文冠木  /  枝和叶  /  化学成分  /  定量分析  /  超高效液相串联质谱  /  多反应监测

Objective: To establish an UHPLC-MS/MS method for determining eight primary components (catechin, epigallocatechin, rutin, quercitrin, epicatechin, (+)-dihydromyricetin, myricitrin and dihydroqurcetin) in the young branches and leaves of Xanthoceras sorbifolia Bunge, a medicinal plant from Mongolia, and to compare their contents in samples at different growth stages. Methods: A Waters CORTECS C18 (100 mm×2.1 mm, 1.6 μm) chromatographic column was adopted using the mobile phase comprised of water containing 0.1% formic acid (A) and acetonitrile (B) with gradient elution(0-1 min, 5% B; 1-10 min, 5% B→28% B; 10-11 min, 28% B→95% B; 11-14 min, 95% B; 14-15 min, 95% B→5% B) at a flow rate of 0.3 mL·min-1. The temperature of the column was set at 40 ℃. Injecting volume was 2 μL. Detection was conducted using electrospray ionization (ESI) in negative ion mode with multiple reaction monitoring (MRM). Results: The linearity of the eight chemical components was found to be excellent in the tested concentration ranges, with correlation coefficients above 0.997 6. Precision, repeatability and stability were satisfactory and the average recoveries were between 97.4% and 106.0% with RSDs≤5.0%. In six batches of leaves, contents of catechin, epigallocatechin, rutin, quercitrin, epicatechin, (+)-dihydromyricetin, myricitrin and dihydroqurcetin were in the ranges of 0.090-0.904 mg·g-1, 0.093-2.258 mg·g-1, 0.001-0.005 mg·g-1, 0.530-6.176 mg·g-1, 0.158-1.561 mg·g-1, 0.002-0.056 mg·g-1, 4.008-10.218 mg·g-1 and 1.049-16.990 mg·g-1, respectively. In six batches of young branches, the contents ranged from 0.384-1.025 mg·g-1, 0.911-2.427 mg·g-1, 0.008-0.127 mg·g-1, 0.870-2.295 mg·g-1, 0.659-1.746 mg·g-1, 0.125-1.079 mg·g-1, 2.296-4.681 mg·g-1 and 1.958-4.946 mg·g-1, respectively. The contents of eight components varied a lot in samples from different parts. The contents of myricitrin, rutin and quercitrin in the leaves exhibited noticeable changes with the growth cycle, suggesting their potential as quality control markers for leaves of Xanthoceras sorbifolia. Conclusion: The method is accurate, sensitive, stable, repeatable, and suitable for simultaneous determination of eight components in Xanthoceras sorbifolia Bunge, offering reference for quality control of its leaves and branches.

Xanthoceras sorbifolia Bunge  /  leaves and branches  /  chemical components  /  quantitative analysis  /  UHPLC-MS/MS  /  multiple reaction monitoring (MRM)
布仁满达, 吴杰斯, 敖敦格日乐, 黄格尔柱拉, 胡阿荣, 其日格尔, 拉喜那木吉拉. UHPLC-MS/MS法测定不同周期蒙药文冠木枝和叶中8个成分的对比研究. 药物分析杂志, 2024 , 44 (9) : 1497 -1503 . DOI: 10.16155/j.0254-1793.2024-0180
Bu-ren-man-da, Jie-si WU, Ao-dun-ge-ri-le, Huang-ge-er-zhu-la, A-rong HU, Qi-ri-ge-er, La-xi-na-mu-ji-la. Comparative study of eight components in branches and leaves of Xanthoceras sorbifolia Bunge at different growth stages by UHPLC-MS/MS method[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (9) : 1497 -1503 . DOI: 10.16155/j.0254-1793.2024-0180
蒙药文冠木是蒙藏医常用的药材,为无患子科植物文冠果(Xanthoceras sorbifolia Bunge)经过干燥处理的枝茎,森登为蒙、藏医通用名称。森登记载于《蒙医月帝》、《中华本草》、《四部医典》、《蒙古学百科全书·医学卷》和《无误蒙药鉴》等蒙医藏医经典著作中[1-5],其性味为甘、涩,微苦,性凉,具有燥血、协日乌素、清热、消肿、止痛的功效[6]。蒙药文冠木作为蒙医药中常用的燥协日乌素药材[7-8],临床应用广泛,被纳入多种方剂[9],对治疗风湿性关节炎和关节水肿疾病[10-11]显示出显著疗效。目前有关蒙药文冠木的报道主要集中在临床应用[12-13]方面,其主要化合物包括黄酮类等成分[14-17]。在对文冠木的化学成分进行研究时,发现主要包含黄酮、香豆素、萜、甾体、蒽醌等类成分[18-20],本研究采用UHPLC-MS/MS分析技术[21-22]对采集于种植基地的6批文冠木枝、叶中的8个成分进行对比研究。
LC-30A超高液相色谱仪(岛津公司),5500QTRAP三重四极杆线性离子阱质谱仪(AB SCIEX公司),数据采集使用Analyst TF 1.7.2软件(公司),数据处理软件版本为SCIEX OS 2.1.6,HA225SM型十万分之一电子分析天平(普利赛思公司)以及KH-500DE型数子电控超声波清洗器(功率500 W,频率40 kHz,昆山禾创超声仪器有限公司)。
对照品表没食子儿茶素(批号PY19009,含量98%)、儿茶素(批号BC12106,含量98%)、表儿茶素(批号FA40877,含量98%)、二氢杨梅素(批号JU09416,含量98%)、二氢槲皮素(批号HN10375,含量98%)、芦丁(批号BL40842,含量98%)购于河南标准物质研究中心,杨梅苷(批号DSTDY010401,含量98%)购自德思特生物,槲皮苷(批号J12HB186308,含量98%)购自上海源叶生物科技有限公司。乙腈(色谱纯,赛默飞公司),水选择屈臣氏蒸馏水,甲醇(色谱纯,赛默飞公司),甲酸(色谱纯,Sigma公司),其他试剂均为分析纯。0.22 μm微孔滤膜(天津市富诚达科技有限公司)。文冠木采摘于内蒙古自治区通辽左翼中旗花吐古拉镇文冠木种植基地,均经内蒙古民族大学吴香杰教授鉴定为无患子科植物文冠果(Xanthoceras sorbifolia Bunge)的干燥枝茎及叶。批号按采集日期命名(20230504、20230514、20230521、20230528、20230604、20230613、20230716),材料采集后阴凉处晒干。
WATER-SCORTECS 色谱柱(100 mm×2.1 mm,1.6 μm),流动相为0.1%甲酸水溶液(A)-乙腈(B),梯度洗脱(0~1 min,5% B;1~10 min,5% B→28% B;10~11 min,28% B→95% B;11~14 min,95% B;14~15 min,95% B→5% B),柱温40 ℃,流速0.3 mL·min-1,进样体积2 μL。
采用ESI源,多反应监测(MRM)的负离子扫描模式;离子化电压为-4 500 V,气源选择高纯度氮气,吹扫离子源以500 ℃进行烘烤(TEM),气帘气(CUR,N2)为170 kPa碰撞气(CAD)压力为选择medium,每个离子对在四极杆中停留采集数据时间为80 ms,去簇电压(DP)和碰撞能电压(CE)见表1
精密称取表没食子儿茶素1.33 mg、儿茶素1.16 mg、表儿茶素1.92 mg、二氢杨梅素1.47 mg、二氢槲皮素1.24 mg、杨梅苷1.26 mg、芦丁1.87 mg、槲皮苷1.18 mg,置10 mL量瓶中,用甲醇定容。取各溶液1 mL,分别置于10 mL量瓶中,用甲醇定容,得上述各成分质量浓度分别为13.3、11.6、19.2、14.7、12.4、12.6、18.7、11.8 μg·mL-1的单一对照品溶液,精密量取各单一对照品溶液1 mL至10 mL量瓶中,用甲醇定容,即得。各对照品色谱图见图1
精确称取药材粗粉1.0 g,置于50 mL带塞锥形瓶中,加入90%甲醇溶液20 mL,密封瓶口,称量,超声(500 W,100 kHz)处理30 min。再称量,并用90%甲醇溶液补足损失的量。摇匀,滤过,取滤液即得。
取“2.3.1”项混合对照品溶液适量,用甲醇逐级稀释,以对照品质量浓度X为横坐标,峰面积Y为纵坐标,计算回归方程,见表2
混合对照品溶液用甲醇稀释,进样测定得信噪比S/N=3为检测限,及信噪比S/N=10为定量限(LOQ),结果见表2
取“2.3.1”项混合对照品溶液,连续进样6次,测定峰面积。结果表没食子儿茶素、儿茶素、表儿茶素、二氢杨梅素、二氢槲皮素、杨梅苷、芦丁、槲皮苷峰面积的RSD(n=6)分别为2.5%、0.87%、4.4%、3.0%、4.6%、3.1%、4.5%、2.2%,表明仪器精密度良好。
按“2.3.2”项方法制备供试品溶液,分别在0、5、10、12、24 h进样测定峰面积,计算表没食子儿茶素、儿茶素、表儿茶素、二氢杨梅素、二氢槲皮素、杨梅苷、芦丁、槲皮苷峰面积的RSD(n=6)分别为2.9%、2.6%、5.2%、2.1%、2.3%、2.4%、2.2%、6.2%,表明24 h内供试品溶液的稳定性良好。
取样品(批号20230514),按“2.3.2”项方法制备6份供试品溶液,进样检测。计算得供试品溶液的表没食子儿茶素、儿茶素、表儿茶素、二氢杨梅素、二氢槲皮素、杨梅苷、芦丁、槲皮苷平均含量分别为1.383、0.312、0.530、0.056、12.097、7.737、0.001、5.016 mg·g-1,RSD分别为1.8%、4.1%、3.1%、2.9%、2.3%、2.1%、1.4%、8.0%。该方法重复性良好。
取样品(批号20230514)9份,取每份0.5 g,已知含量的样品中按0.5、1、1.5倍量加入混合好的对照品,按“2.3.2”项方法制备溶液,进样测定并计算回收率。表没食子儿茶素、儿茶素、表儿茶素、二氢杨梅素、二氢槲皮素、杨梅苷、芦丁、槲皮苷的平均回收率(n=9)分别为102.9%、103.1%、102.6%、101.8%、104.1%、100.4%、98.5%和99.6%,RSD为2.8%、3.1%、1.9%、0.67%、5.3%、2.1%、1.1%和1.9%。表明该方法准确度良好。
取6批样品,按“2.3.2”项方法制备供试品溶液,分别精密吸取2 μL进样测定,得到色谱峰面积,计算样品中8个化学成分的含量。结果见表3
本试验以100 kHz的超声条件为基础,首先考察了不同甲醇浓度(50%、60%、70%、80%、90%、100%)对提取率的影响,结果显示90%甲醇水提取率最佳。再次考察不同液料比 (g/mL)1∶10、1∶15、1∶20、1∶30对提取效果的影响,发现1∶20为最佳比例。在提取时间方面(20、30、40、60,30 min)的提取时间效果最佳。对待测成分进行质谱条件优化时发现选择的8个化学成分均在负离子模式下响应良好,故采取MRM的扫描方式选择负离子扫描模式。
采集不同时间段的枝和叶研究发现,枝中儿茶素、表儿茶素、二氢杨梅素、槲皮苷含量显著高于叶;且枝中杨梅苷、芦丁、槲皮苷的含量明显低于叶。枝中的8个成分的变化规律不明显,但叶中的杨梅苷、芦丁、槲皮苷含量随着叶的生长呈现负相关性(见图2)。
经分析,杨梅苷、芦丁、槲皮苷可作为叶的质量控制主要指标。批号20230507的叶样品正处于嫩芽期,此时叶中杨梅苷、芦丁、槲皮苷含量最高,表明嫩芽期是文冠木叶中活性成分积累的关键时期。 研究结果表明,生长周期对文冠木叶子中杨梅苷、芦丁、槲皮苷成分的含量有显著影响,对确定文冠木叶的采集时间具有重要意义。
本文建立了UHPLC-MS/MS法同时测定蒙药文冠木枝和叶中儿茶素和表没食子儿茶素、芦丁及槲皮苷、表儿茶素、二氢杨梅素、杨梅苷和二氢槲皮素的含量,该法前处理方法简便(仅超声处理),色谱分析时间短,灵敏度高,专属性强,准确度高,为药材及叶子的质量控制提供实验基础,杨梅苷、芦丁、槲皮苷可用于叶的质量控制指标。
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2024年第44卷第9期
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doi: 10.16155/j.0254-1793.2024-0180
  • 接收时间:2024-03-17
  • 首发时间:2026-03-17
  • 出版时间:2024-09-30
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  • 收稿日期:2024-03-17
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    1.内蒙古民族大学国家药监局中药(蒙药)质量控制重点实验室,通辽 028000
    2.内蒙古民族大学附属医院,通辽 028000

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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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