Article(id=1239238141203247758, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239238136711139764, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2023-0639, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1696089600000, receivedDateStr=2023-10-01, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773386996794, onlineDateStr=2026-03-13, pubDate=1722355200000, pubDateStr=2024-07-31, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773386996794, onlineIssueDateStr=2026-03-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773386996794, creator=13701087609, updateTime=1773386996794, updator=13701087609, issue=Issue{id=1239238136711139764, tenantId=1146029695717560320, journalId=1205117023404326918, year='2024', volume='44', issue='7', pageStart='1105', pageEnd='1284', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773386995723, creator=13701087609, updateTime=1773387118529, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1239238651851370909, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239238136711139764, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1239238651851370910, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1239238136711139764, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1195, endPage=1201, ext={EN=ArticleExt(id=1239238141471683222, articleId=1239238141203247758, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Simultaneous determination of 13 components in Yankening tablets by UPLC, columnId=1239148838318043851, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Ingredient Analys, runingTitle=null, highlight=null, articleAbstract=
Objective:

To establish an UPLC quantitative analysis method for the simultaneous determination of 13 components in Yankening tablets, including phellodendrine, coptisine, baicalin, palmatine, berberine, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, chrysophanol, physcion.

Methods:

Agilent Eclipse Plus C18(100 mm×2.1 mm, 1.8 μm) column was used with acetonitrile -0.1% phosphoric acid as mobile phase, and gradient elution at a flow rate of 0.3 mL·min-1. The detection wavelengths were 210 nm and 254 nm. The column temperature was 40 ℃.

Results:

Phellodendrine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride, baicalin, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, chrysophanol, physcion showed good linear relationships within their concentration range of 0.97-48.63 μg·mL-1, 0.95-47.52 μg·mL-1, 0.86-43.15 μg·mL-1, 0.86-43.19 μg·mL-1, 0.89-44.37 μg·mL-1, 1.00-49.84 μg·mL-1, 1.02-51.01 μg·mL-1, 0.97-48.31 μg·mL-1, 0.99-49.50 μg·mL-1, 1.04-51.80 μg·mL-1, 1.00-50.04 μg·mL-1, 1.00-49.80 μg·mL-1, 1.01-50.64 μg·mL-1. The average recoveries(n=6) were 95.2%, 96.7%, 95.9%, 98.3%, 94.1%,97.6%, 99.2%, 96.6%, 95.5%, 97.2%, 97.0%, 97.8%, 98.7%, RSD values were all less than 2.0%. The contents of the 13 chemical components in 3 batches of samples were 1.367-1.488 mg·g-1(calculated as phellodendrine hydrochloride), 0.378-0.412 mg·g-1(calculated as coptisine hydrochloride), 4.611-5.505 mg·g-1, 0.324-0.407 mg·g-1(calculated as palmatine hydrochloride), 3.665-3.878 mg·g-1(calculated as berberine hydrochloride), 1.107-1.682 mg·g-1, 0.392-0.941 mg·g-1, 0.076-0.105 mg·g-1, 0.097-0.116 mg·g-1, 1.059-1.213 mg·g-1, 0.149-0.167 mg·g-1, 0.213-0.239 mg·g-1, 0.047-0.059 mg·g-1.

Conclusion:

The method is accurate, high analysis efficiency, good repeatability, it can be used to control the quality of Yankening tablets.

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目的:

采用超高效液相色谱法同时测定炎可宁片中黄柏碱、黄连碱、黄芩苷、巴马汀、小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚13个成分含量。

方法:

采用Agilent Eclipse Plus-C18(100 mm×2.1 mm,1.8 μm)色谱柱,以乙腈-0.1%磷酸水为流动相,梯度洗脱,流速0.3 mL·min-1,测定波长210、254 nm,柱温40 ℃。

结果:

盐酸黄柏碱、盐酸黄连碱、黄芩苷、盐酸巴马汀、盐酸小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚的的线性范围分别为0.97~48.63、0.95~47.52、0.86~43.15、0.86~43.19、0.89~44.37、1.00~49.84、1.02~51.01、0.97~48.31、0.99~49.50、1.04~51.80、1.00~50.04、1.00~49.80、1.01~50.64 μg·mL-1;平均回收率(n=6)分别为95.2%、96.7%、98.3%、94.1%、95.9%、97.6%、99.2%、96.6%、95.5%、97.2%、97.0%、97.8%、98.7%,RSD均<2.0%。3批样品中上述13个成分的含量分别为1.367~1.488(以盐酸黄柏碱计)、0.378~0.412(以盐酸黄连碱计)、4.611~5.505、0.324~0.407(以盐酸巴马汀计)、3.665~3.878(以盐酸小檗碱计)、1.107~1.682、0.392~0.941、0.076~0.105、0.097~0.116、1.059~1.213、0.149~0.167、0.213~0.239、0.047~0.059 mg·g-1

结论:

该方法准确,分析效率高,重复性好,适用于炎可宁片的质量控制。

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articleId=1239238141203247758, language=CN, orderNo=9, keyword=汉黄芩苷), Keyword(id=1239238148115461097, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=10, keyword=黄芩素), Keyword(id=1239238148220318703, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=11, keyword=芦荟大黄素), Keyword(id=1239238148325176311, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=12, keyword=大黄酸), Keyword(id=1239238148442616831, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=13, keyword=汉黄芩素), Keyword(id=1239238148560056321, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=14, keyword=大黄素), Keyword(id=1239238148669108230, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=15, keyword=大黄酚), Keyword(id=1239238150208417807, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=16, keyword=大黄素甲醚), Keyword(id=1239238150304886803, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=17, keyword=含量测定), Keyword(id=1239238150418133019, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, orderNo=18, keyword=质量控制)], refs=[Reference(id=1239238151227633750, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, doi=null, pmid=null, pmcid=null, year=1993, volume=null, issue=null, pageStart=104, pageEnd=null, url=null, language=null, rfNumber=[1], rfOrder=0, authorNames=null, journalName=null, refType=null, unstructuredReference=卫生部药品标准. 中药成方制剂. 第七册[S]. 1993: 104, articleTitle=null, refAbstract=null), 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1.黄柏碱(phellodendrine) 2.酸黄连碱(coptisine) 3黄芩苷(baicalin) 4.酸巴马汀(palmatine) 5.小檗碱(berberine) 6.汉黄芩苷(wogonoside) 7.黄芩素(baicalein) 8.芦荟大黄素(aloe-emodin) 9.大黄酸(rhein) 10.汉黄芩素(wogonin) 11.大黄素(emodin) 12.大黄酚(chrysophanol) 13.大黄素甲醚(physcion)

, figureFileSmall=IEFd85BqOtkG8X52O5udBg==, figureFileBig=pO1vWPf1IgKUcfMyh5OnMg==, tableContent=null), ArticleFig(id=1239238150795620411, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=EN, label=Tab.1, caption=

Linear relationship

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物
(compound)
回归方程
(regression equation)
r线性范围
(linear range)/(μg·mL-1)
盐酸黄柏碱(phellodendrine hydrochloride)Y=16 489.08X-25.550.999 90.97~48.63
盐酸黄连碱(coptisine hydrochloride)Y=10 216.06X-19.670.999 90.95~47.52
黄芩苷(baicalin)Y=42 197.37X-63.430.999 80.86~43.15
盐酸巴马汀(palmatine hydrochloride)Y=11 825.12X-52.050.999 90.86~43.19
盐酸小檗碱(berberine hydrochloride)Y=43 662.12X+29.190.999 90.89~44.37
汉黄芩苷(wogonoside)Y=10 447.04X-16.360.999 91.00~49.84
黄芩素(baicalein)Y=17 251.04X+54.310.999 91.02~51.01
芦荟大黄素(aloe-emodin)Y=12 365.51X-31.460.999 90.97~48.31
大黄酸(rhein)Y=13 474.14X-33.150.999 90.99~49.50
汉黄芩素(wogonin)Y=11 104.17X-14.780.999 81.04~51.80
大黄素(emodin)Y=12 028.78X-50.070.999 91.00~50.04
大黄酚(chrysophanol)Y=12 353.72X+22.710.999 81.00~49.80
大黄素甲醚(physcion)Y=12 434.09X-23.480.999 91.01~50.64
), ArticleFig(id=1239238150875312192, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, label=表1, caption=

线性关系

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物
(compound)
回归方程
(regression equation)
r线性范围
(linear range)/(μg·mL-1)
盐酸黄柏碱(phellodendrine hydrochloride)Y=16 489.08X-25.550.999 90.97~48.63
盐酸黄连碱(coptisine hydrochloride)Y=10 216.06X-19.670.999 90.95~47.52
黄芩苷(baicalin)Y=42 197.37X-63.430.999 80.86~43.15
盐酸巴马汀(palmatine hydrochloride)Y=11 825.12X-52.050.999 90.86~43.19
盐酸小檗碱(berberine hydrochloride)Y=43 662.12X+29.190.999 90.89~44.37
汉黄芩苷(wogonoside)Y=10 447.04X-16.360.999 91.00~49.84
黄芩素(baicalein)Y=17 251.04X+54.310.999 91.02~51.01
芦荟大黄素(aloe-emodin)Y=12 365.51X-31.460.999 90.97~48.31
大黄酸(rhein)Y=13 474.14X-33.150.999 90.99~49.50
汉黄芩素(wogonin)Y=11 104.17X-14.780.999 81.04~51.80
大黄素(emodin)Y=12 028.78X-50.070.999 91.00~50.04
大黄酚(chrysophanol)Y=12 353.72X+22.710.999 81.00~49.80
大黄素甲醚(physcion)Y=12 434.09X-23.480.999 91.01~50.64
), ArticleFig(id=1239238150963392581, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=EN, label=Tab.2, caption=

Content determination results of 3 batches of samples

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物
(compound)
含量(content)/mg per tablet
lot No. 180306lot No. 190502lot No. 201005
盐酸黄柏碱(phellodendrine hydrochloride)1.4881.3671.411
盐酸黄连碱(coptisine hydrochloride)0.3930.3780.412
黄芩苷(baicalin)5.5054.7354.611
盐酸巴马汀(palmatine hydrochloride)0.3240.4070.368
盐酸小檗碱(berberine hydrochloride)3.6653.8783.823
汉黄芩苷(wogonoside)1.2351.6821.107
黄芩素(baicalein)0.3920.9410.608
芦荟大黄素(aloe-emodin)0.0880.1050.076
大黄酸(rhein)0.0970.1160.117
汉黄芩素(wogonin)1.0591.2131.147
大黄素(emodin)0.1540.1670.149
大黄酚(chrysophanol)0.2130.2280.239
大黄素甲醚(physcion)0.0550.0590.047
), ArticleFig(id=1239238151064055884, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1239238141203247758, language=CN, label=表2, caption=

3批样品含量测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
化合物
(compound)
含量(content)/mg per tablet
lot No. 180306lot No. 190502lot No. 201005
盐酸黄柏碱(phellodendrine hydrochloride)1.4881.3671.411
盐酸黄连碱(coptisine hydrochloride)0.3930.3780.412
黄芩苷(baicalin)5.5054.7354.611
盐酸巴马汀(palmatine hydrochloride)0.3240.4070.368
盐酸小檗碱(berberine hydrochloride)3.6653.8783.823
汉黄芩苷(wogonoside)1.2351.6821.107
黄芩素(baicalein)0.3920.9410.608
芦荟大黄素(aloe-emodin)0.0880.1050.076
大黄酸(rhein)0.0970.1160.117
汉黄芩素(wogonin)1.0591.2131.147
大黄素(emodin)0.1540.1670.149
大黄酚(chrysophanol)0.2130.2280.239
大黄素甲醚(physcion)0.0550.0590.047
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UPLC法同时测定炎可宁片中13个成分含量
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董晓茜 1 , 鄢长余 2 , 马进 1, *
药物分析杂志 | 成分分析 2024,44(7): 1195-1201
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药物分析杂志 | 成分分析 2024, 44(7): 1195-1201
UPLC法同时测定炎可宁片中13个成分含量
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董晓茜1 , 鄢长余2, 马进1, *
作者信息
  • 1.辽宁省中医药研究院(辽宁中医药大学附属第二医院),沈阳 110034
  • 2.大连市检验检测认证技术服务中心,大连 116021
  • Tel:(024)86803333-8734;E-mail:

通讯作者:

* Tel:(024)86803333-8213;E-mail:
Simultaneous determination of 13 components in Yankening tablets by UPLC
Xiao-qian DONG1 , Chang-yu YAN2, Jin MA1, *
Affiliations
  • 1.Liaoning Institute of Traditional Chinese Medicine (The Second Hospital Affiliated to Liaoning University of Traditional Chinese Medicine), Shenyang 110034, China
  • 2.Dalian Inspection and Certification Technical Service Center, Dalian 116021, China
出版时间: 2024-07-31 doi: 10.16155/j.0254-1793.2023-0639
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目的:

采用超高效液相色谱法同时测定炎可宁片中黄柏碱、黄连碱、黄芩苷、巴马汀、小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚13个成分含量。

方法:

采用Agilent Eclipse Plus-C18(100 mm×2.1 mm,1.8 μm)色谱柱,以乙腈-0.1%磷酸水为流动相,梯度洗脱,流速0.3 mL·min-1,测定波长210、254 nm,柱温40 ℃。

结果:

盐酸黄柏碱、盐酸黄连碱、黄芩苷、盐酸巴马汀、盐酸小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚的的线性范围分别为0.97~48.63、0.95~47.52、0.86~43.15、0.86~43.19、0.89~44.37、1.00~49.84、1.02~51.01、0.97~48.31、0.99~49.50、1.04~51.80、1.00~50.04、1.00~49.80、1.01~50.64 μg·mL-1;平均回收率(n=6)分别为95.2%、96.7%、98.3%、94.1%、95.9%、97.6%、99.2%、96.6%、95.5%、97.2%、97.0%、97.8%、98.7%,RSD均<2.0%。3批样品中上述13个成分的含量分别为1.367~1.488(以盐酸黄柏碱计)、0.378~0.412(以盐酸黄连碱计)、4.611~5.505、0.324~0.407(以盐酸巴马汀计)、3.665~3.878(以盐酸小檗碱计)、1.107~1.682、0.392~0.941、0.076~0.105、0.097~0.116、1.059~1.213、0.149~0.167、0.213~0.239、0.047~0.059 mg·g-1

结论:

该方法准确,分析效率高,重复性好,适用于炎可宁片的质量控制。

炎可宁片  /  超高效液相色谱法  /  消炎止痢  /  黄柏碱  /  黄连碱  /  黄芩苷  /  巴马汀  /  小檗碱  /  汉黄芩苷  /  黄芩素  /  芦荟大黄素  /  大黄酸  /  汉黄芩素  /  大黄素  /  大黄酚  /  大黄素甲醚  /  含量测定  /  质量控制
Objective:

To establish an UPLC quantitative analysis method for the simultaneous determination of 13 components in Yankening tablets, including phellodendrine, coptisine, baicalin, palmatine, berberine, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, chrysophanol, physcion.

Methods:

Agilent Eclipse Plus C18(100 mm×2.1 mm, 1.8 μm) column was used with acetonitrile -0.1% phosphoric acid as mobile phase, and gradient elution at a flow rate of 0.3 mL·min-1. The detection wavelengths were 210 nm and 254 nm. The column temperature was 40 ℃.

Results:

Phellodendrine hydrochloride, coptisine hydrochloride, palmatine hydrochloride, berberine hydrochloride, baicalin, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, chrysophanol, physcion showed good linear relationships within their concentration range of 0.97-48.63 μg·mL-1, 0.95-47.52 μg·mL-1, 0.86-43.15 μg·mL-1, 0.86-43.19 μg·mL-1, 0.89-44.37 μg·mL-1, 1.00-49.84 μg·mL-1, 1.02-51.01 μg·mL-1, 0.97-48.31 μg·mL-1, 0.99-49.50 μg·mL-1, 1.04-51.80 μg·mL-1, 1.00-50.04 μg·mL-1, 1.00-49.80 μg·mL-1, 1.01-50.64 μg·mL-1. The average recoveries(n=6) were 95.2%, 96.7%, 95.9%, 98.3%, 94.1%,97.6%, 99.2%, 96.6%, 95.5%, 97.2%, 97.0%, 97.8%, 98.7%, RSD values were all less than 2.0%. The contents of the 13 chemical components in 3 batches of samples were 1.367-1.488 mg·g-1(calculated as phellodendrine hydrochloride), 0.378-0.412 mg·g-1(calculated as coptisine hydrochloride), 4.611-5.505 mg·g-1, 0.324-0.407 mg·g-1(calculated as palmatine hydrochloride), 3.665-3.878 mg·g-1(calculated as berberine hydrochloride), 1.107-1.682 mg·g-1, 0.392-0.941 mg·g-1, 0.076-0.105 mg·g-1, 0.097-0.116 mg·g-1, 1.059-1.213 mg·g-1, 0.149-0.167 mg·g-1, 0.213-0.239 mg·g-1, 0.047-0.059 mg·g-1.

Conclusion:

The method is accurate, high analysis efficiency, good repeatability, it can be used to control the quality of Yankening tablets.

Yankening  /  UPLC  /  anti-dysentery and anti-inflammatory  /  phellodendrine  /  coptisine  /  baicalin  /  palmatine  /  berberine  /  wogonoside  /  baicalein  /  aloe-emodin  /  rhein  /  wogonin  /  emodin  /  chrysophanol  /  physcion  /  determination  /  quality control
董晓茜, 鄢长余, 马进. UPLC法同时测定炎可宁片中13个成分含量. 药物分析杂志, 2024 , 44 (7) : 1195 -1201 . DOI: 10.16155/j.0254-1793.2023-0639
Xiao-qian DONG, Chang-yu YAN, Jin MA. Simultaneous determination of 13 components in Yankening tablets by UPLC[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (7) : 1195 -1201 . DOI: 10.16155/j.0254-1793.2023-0639
炎可宁片为临床常用的清热泻火及消炎止痢药物,用于治疗急性扁桃腺炎、细菌性肺炎、急性结膜炎、中耳炎等症,其处方由黄柏、大黄、黄芩、黄连、板蓝根5味药材组成。炎可宁片处方标准收载于卫生部部颁标准第七册[1],包含性状、显微鉴别和化学反应。文献报道中对于炎可宁片各药味成分含量多为单成分测定,无法准确控制药品的质量,因此完善提高标准十分必要。前期报道,许学丽等[2]采用HPLC法测定可宁片中9个成分含量,高洪琳等[3]采用HPLC法同时测定炎可宁片中6个成分含量,张丹等[4]采用UPLC-MS/MS法测定炎可宁片中5个成分含量,尹永芹等[5]采用HPLC法测定炎可宁片中5个蒽醌类成分含量,黄燕萍[6]测定了炎可宁片中盐酸小檗碱、黄芩苷和汉黄芩素的含量,曹莺[7]测定炎可宁片中小檗碱和巴马汀的含量,古炳明等[8]采用HPLC法测定了炎可宁胶囊中6个成分含量,李亚等[9]对炎可宁片中各个成分的含量测定研究进展进行了总结。表明多成分含量测定逐渐成为一种能更好地评价和控制复方制剂中药材质量、投料比例的检测方法。因此本文结合前期报道中的各类化学成分,并参考测定黄芩、黄连、黄柏3种药材的相关文献中的梯度洗脱条件[10-14],建立了超高效液相色谱方法,在相同的色谱条件下测定炎可宁片中黄柏碱、黄连碱、黄芩苷、巴马汀、小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚13个成分含量,可以进一步提高炎可宁片的质量控制标准。
Waters 2998 ACQUITY UPLC系统(配PDA检测器)(沃特世公司);Agilent Eclipse Plus C18(100 mm×2.1 mm,1.8 μm)色谱柱;Sartorius BP221S(0.000 1 g)、Sartorius BP221D(0.000 01 g)电子天平(赛多利斯公司)。
对照品盐酸黄柏碱(批号111895-201805,纯度94.9%)、盐酸黄连碱(批号112026-201802,纯度94.0%)、黄芩苷(批号110715-201318,纯度93.3%)、盐酸巴马汀(批号110732-201913,纯度85.7%)、盐酸小檗碱(批号110713-202015,纯度85.9%)、汉黄芩苷(批号112002-201501,纯度98.8%)、黄芩素(批号111595-201808,纯度97.9%)、芦荟大黄素(批号110795-201308,纯度97.8%)、大黄酸(批号110757-201607,纯度99.3%)、汉黄芩素(批号111514-201706,纯度100.0%)、大黄素(批号110756-201512,纯度98.7%)、大黄酚(批号110796-201621,纯度99.2%)、大黄素甲醚(批号110758-201415,纯度99.1%)均购买自中国食品药品检定研究院;甲醇、乙腈、磷酸均为色谱纯。收集炎可宁片3批(吉林玉仁制药股份有限公司,批号190502、180306、201005)。
精密称取各对照品适量,加甲醇制成每1 mL含盐酸黄柏碱5 μg、盐酸黄连碱10 μg、盐酸巴马汀10 μg、盐酸小檗碱30 μg、黄芩苷20 μg、汉黄芩苷15 μg、黄芩素5 μg、芦荟大黄素5 μg、大黄酸5 μg、汉黄芩素10 μg、大黄素10 μg、大黄酚20 μg、大黄素甲醚5 μg的混合溶液,即得。
取去除包衣层的样品20片,研细混匀后,称取细粉约2 g,精密称定,置具塞锥形瓶中,加70%甲醇水50 mL后称量,85 ℃水浴连续加热回流1 h,室温下放冷,用70%甲醇水补足减失的量,滤过后即得。
根据炎可宁片处方配伍比例,配制3种阴性样品(依次为缺黄柏、黄连阴性样品;缺黄芩阴性样品;缺大黄阴性样品)。再分别按照“2.1.2”项下方法操作,制备阴性样品溶液。
采用Agilent Eclipse Plus C18 (100 mm×2.1 mm,1.8 μm)色谱柱,以乙腈(A)-0.1%磷酸水溶液为流动相,进行梯度洗脱(0~3 min,10%A→15%A;3~7 min,15%A→35%A;7~10 min,35%A;10~15 min,35%A→45%A;15~18 min,45%A;18~20 min,45%A→75%A;20~25 min,75%A→90%A),流速0.3 mL·min-1,双波长切换(0~6 min,黄柏碱,210 nm;6~25 min,其他12个成分,254 nm)。柱温40 ℃,进样量1 μL。结果表明,13个成分分离度符合要求,且无阴性样品干扰(图1)。
精密吸取“2.1.1”项下混合对照品溶液适量,用甲醇逐级稀释成5个质量浓度的溶液,按“2.2”项下条件进样测定,记录色谱图,以对照品质量浓度(X)为横坐标,峰面积(Y)为纵坐标进行线性回归。结果见表1r均>0.999,表明各成分在相应范围内线性关系良好。
取炎可宁片样品(批号180306),按“2.1.2”项下方法制备成供试品溶液,连续进样6次,结果黄柏碱、黄连碱、黄芩苷、巴马汀、小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚峰面积的RSD分别为0.52%、0.81%、0.37%、0.83%、0.35%、0.93%、0.83%、1.0%、0.86%、0.68%、0.63%、0.86%、1.1%,表明仪器精密度良好。
取炎可宁片样品(批号180306),按“2.1.2”项下方法平行制备供试品溶液6份,按“2.2”项下条件进样测定,结果黄柏碱(以盐酸黄柏碱计)、黄连碱(以盐酸黄连碱计)、黄芩苷、巴马汀(以盐酸巴马汀计)、小檗碱(以盐酸小檗碱计)、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚的平均含量分别为1.452、0.381、5.523、0.331、3.602、1.252、0.403、0.094、0.104、1.082、0.154、0.223、0.062 mg·片-1,RSD分别为0.75%、1.2%、0.58%、1.1%、0.98%、0.93%、1.2%、1.0%、1.1%、0.96%、1.0%、0.78%、1.0%,表明本方法重复性良好。
取炎可宁片样品(批号180306),按“2.1.2”项下方法制备供试品溶液,分别在溶液制备后0、4、8、12、24 h进样测定,结果黄柏碱、黄连碱、黄芩苷、巴马汀、小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚、大黄素甲醚峰面积的RSD分别为0.32%、0.74%、0.25%、0.67%、0.32%、0.48%、0.83%、1.0%、0.95%、0.79%、1.0%、1.0%、1.3%,表明13个成分在24 h内稳定性良好。
精密称取各对照品,分别加甲醇制成每1 mL含盐酸黄柏碱0.477 3 mg、盐酸黄连碱0.096 16 mg、黄芩苷1.887 0 mg、盐酸巴马汀0.858 7 mg、盐酸小檗碱1.345 2 mg、汉黄芩苷0.497 0 mg、黄芩素0.099 50 mg、芦荟大黄素0.051 59 mg、大黄酸0.050 30 mg、汉黄芩素0.250 2 mg、大黄素0.050 58 mg、大黄酚0.100 6 mg、大黄素甲醚0.050 59 mg的混合对照品溶液。
取已知含量的炎可宁片样品(批号180306)20片(去除包衣),研细,取约1 g,精密称定。精密加入上述混合对照品溶液10 mL,水浴挥干溶剂,按“2.1.2”项下方法制备供试溶液,按“2.2”项下条件测定并计算回收率。结果盐酸黄柏碱、盐酸黄连碱、黄芩苷、盐酸巴马汀、盐酸小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚和大黄素甲醚的平均加样回收率(n=6)分别为95.2%、96.7%、95.9%、98.3%、94.1%、97.6%、99.2%、96.6%、95.5%、97.2%、97.0%、97.8%、98.7%,RSD分别为0.88%、0.59%、0.58%、1.2%、1.2%、0.46%、0.93%、1.0%、0.82%、1.2%、0.87%、0.94%、1.1%,符合方法学要求。
取3批炎可宁片,每批称取平行样品2份,按“2.1.2”项下方法制备供试品溶液,按“2.2”项下条件,进样2次进行分析,并按外标法计算含量(黄柏碱、黄连碱、巴马汀、小檗碱的含量分别以盐酸黄柏碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱计)。结果见表2
采用PDA检测器在200~400 nm范围内对选定的黄柏碱、黄连碱、黄芩苷、巴马汀、小檗碱、汉黄芩苷、黄芩素、芦荟大黄素、大黄酸、汉黄芩素、大黄素、大黄酚和大黄素甲醚13个指标性成分进行光谱扫描,结果显示,黄柏碱在其最大吸收波长284 nm处的紫外吸收过低,供试品溶液中黄柏碱峰面积小,无法满足定量的要求,为了保证定量计算的准确性,故选择210 nm作为其测定波长[9],其他12个指标性成分在254 nm处均有较好的紫外吸收,因此最终确定以210 nm和254 nm作为测定波长。
本试验对提取溶剂和提取方法均进行了方法学考察。比较了50%甲醇水、70%甲醇水、90%甲醇水、甲醇4种溶剂的提取效率,通过对指标成分的含量、峰形、分离度、杂质等因素的综合考虑,确定用70%甲醇水作为提取溶剂。供试品溶液制备时,提取方法分别考察了超声处理和加热回流,结果表明采用加热回流提取有效成分,其效率高于超声处理;用70%甲醇分别加热回流提取0.5、1和1.5 h,结果提取1、1.5 h高于0.5 h的含量,且提取1.5 h比1 h的含量并无明显差异,故选择加热回流提取1 h作为提取方法。
本方法采用UPLC法同时测定了黄柏、黄芩、大黄、黄连药材中的13个指标性成分,提取方法简单,可操作性强,在分离度良好的情况下,25 min内完成13个指标性成分的分离,分析用时较短,与常规液相色谱分析相比,分析效率提高约3倍[2-3],且色谱峰峰形好,阴性无干扰,本方法可以更有效地控制炎可宁片中4种中药材的质量。
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doi: 10.16155/j.0254-1793.2023-0639
  • 接收时间:2023-10-01
  • 首发时间:2026-03-13
  • 出版时间:2024-07-31
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    1.辽宁省中医药研究院(辽宁中医药大学附属第二医院),沈阳 110034
    2.大连市检验检测认证技术服务中心,大连 116021

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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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