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Article(id=1308828729019822120, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308828670878376007, articleNumber=null, orderNo=null, doi=10.16438/j.0513-4870.2025-1129, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1758470400000, receivedDateStr=2025-09-22, revisedDate=1767456000000, revisedDateStr=2026-01-04, acceptedDate=null, acceptedDateStr=null, onlineDate=1789978685859, onlineDateStr=2026-09-21, pubDate=1773244800000, pubDateStr=2026-03-12, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1789978685859, onlineIssueDateStr=2026-09-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1789978685859, creator=13701087609, updateTime=1789978685859, updator=13701087609, issue=Issue{id=1308828670878376007, tenantId=1146029695717560320, journalId=1189982191388893191, year='2026', volume='61', issue='3', pageStart='677', pageEnd='1000', issueExtLink='null', onlineDate='null', pubDate='1773244800000', pubDateStr='2026-03-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1789978671998, creator='13701087609', updateTime=1789984247322, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1308852055549248371, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308828670878376007, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1308852055549248372, tenantId=1146029695717560320, journalId=1189982191388893191, issueId=1308828670878376007, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=881, endPage=886, ext={EN=ArticleExt(id=1308828729401503785, articleId=1308828729019822120, tenantId=1146029695717560320, journalId=1189982191388893191, language=EN, title=Simultaneous determination of eleven components in Jinchai Qingre Granule by QAMS and HPLC-DAD, columnId=1190335348761793317, journalTitle=Acta Pharmaceutica Sinica, columnName=Original Articles, runingTitle=null, highlight=null, articleAbstract=

The study aims to establish a method for the content determination of eleven components in Jinchai Qingre Granule by high performance liquid chromatography-diode array detector (HPLC-DAD) through quantitative analysis of multi-components by single marker (QAMS) and verify the feasibility and applicability of the method in its quality control. Agilent ZORBAX SB-C18 column (250 mm × 4.6 mm, 5 μm) was used; acetonitrile-0.1% phosphoric acid aqueous solution was used as mobile phase; gradient elution and the detection wavelength were set at 327, 278, 250 nm; flow rate was 1.0 mL·min-1; column temperature was 25 ℃. Using puerarin as the internal reference, the relative correction factors of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, daidzin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, baicalin, daidzein and arctiin was established, and the eleven components were determined. Eleventeen components showed good linear relationships within their own ranges (r ≥ 0.999 5), whose average recoveries were 98.05%-100.90% with the RSDs of 0.98%-2.35% (n = 6). The method showed good durability under different conditions such as instruments, columns, column temperature, injection volume, and flow rate. There was no significant difference between the calculated values of eleven components in ten batches of samples by the QAMS and the measured values of the external standard method (P > 0.05). The method established in the study is simple and stable, and can be used for the synchronous quality control of eleven index components in Jinchai Qingre Granule.

, authors=Jin-e NIE1, Shao-yun WANG1, Fang-fang QIAN1, Yuan-jun QU1, *, Le LI2, Jin-miao XUE2, Qiu-jing LIU2, authorsList=Jin-e NIE, Shao-yun WANG, Fang-fang QIAN, Yuan-jun QU, Le LI, Jin-miao XUE, Qiu-jing LIU, authorCompany=null, correspAuthors=Yuan-jun QU, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2026 Acta Pharmaceutica Sinica. All rights reserved., 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, fund=null), CN=ArticleExt(id=1308828730211004460, articleId=1308828729019822120, tenantId=1146029695717560320, journalId=1189982191388893191, language=CN, title=HPLC-DAD波长转换结合一测多评法测定金柴清热颗粒中的11种成分, columnId=1190335348896011050, journalTitle=药学学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

建立高效液相色谱–二极管阵列检测法(high performance liquid chromatography-diode array detector, HPLC-DAD) 波长转换结合一测多评法(quantitative analysis of multi-components by single marker, QAMS) 同时测定金柴清热颗粒中11种成分含量的方法, 为其质量控制提供科学依据及质量保障。采用Agilent ZORBAX SB-C18色谱柱(250 mm × 4.6 mm, 5 μm), 以乙腈-0.1%磷酸水溶液为流动相, 梯度洗脱, 检测波长327、278、250 nm, 流速1.0 mL·min-1, 柱温25 ℃, 以葛根素为内参物, 建立葛根素与新绿原酸、绿原酸、隐绿原酸、大豆苷、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷的相对校正因子, 计算含量; 11种成分在各自浓度范围内线性关系良好(r ≥ 0.999 5), 平均加样回收率为98.05%~100.90%, RSD为0.98%~2.35% (n = 6); 不同设备、色谱柱、柱温、进样体积、流速等色谱条件下耐用性良好, 与外标法(external standard method, ESM) 相比较, 以葛根素为内参物的QAMS法对其余10种成分的测定结果无显著性差异。结果表明, 该方法简便、稳定, 可实现金柴清热颗粒中11种成分的质量控制, 为其质量标准化提供技术支撑。

, authors=聂金娥1, 王少芸1, 钱芳芳1, 曲远均1, *, 李乐2, 薛金苗2, 刘秋静2, authorsList=聂金娥, 王少芸, 钱芳芳, 曲远均, 李乐, 薛金苗, 刘秋静, authorCompany=null, correspAuthors=曲远均, authorNote=null, correspAuthorsNote=
曲远均, E-mail:
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AnalyteRegressive equationsr2Linear range/μg·mL-1
Neochlorogenic acidy = 10.351x + 0.836 60.999 73.19-31.90
Chlorogenic acidy = 15.185x - 2.2000.999 84.238-42.380
Cryptochlorogenic acidy = 11.003x - 1.256 10.999 93.68-36.80
Puerariny = 42.596x + 31.9510.999 919.25-192.54
Daidziny = 48.213x - 1.902 40.999 83.53-35.32
Isochlorogenic acid By = 16.957x - 0.634 10.999 92.53-25.34
Isochlorogenic acid Ay = 22.639x + 0.282 90.999 91.10-11.96
Isochlorogenic acid Cy = 20.941x - 1.890 20.999 82.05-20.46
Baicaliny = 36.111x - 8.1780.999 916.05-160.50
Daidzeiny = 23.526x + 0.390 20.999 91.07-10.66
Arctiiny = 6.173 1x + 1.107 30.999 88.59-85.86
), ArticleFig(id=1308834471701475367, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1308828729019822120, language=CN, label=Tab.1, caption=

Regressive equations, correlation coefficients and linear ranges

, figureFileSmall=null, figureFileBig=null, tableContent=
AnalyteRegressive equationsr2Linear range/μg·mL-1
Neochlorogenic acidy = 10.351x + 0.836 60.999 73.19-31.90
Chlorogenic acidy = 15.185x - 2.2000.999 84.238-42.380
Cryptochlorogenic acidy = 11.003x - 1.256 10.999 93.68-36.80
Puerariny = 42.596x + 31.9510.999 919.25-192.54
Daidziny = 48.213x - 1.902 40.999 83.53-35.32
Isochlorogenic acid By = 16.957x - 0.634 10.999 92.53-25.34
Isochlorogenic acid Ay = 22.639x + 0.282 90.999 91.10-11.96
Isochlorogenic acid Cy = 20.941x - 1.890 20.999 82.05-20.46
Baicaliny = 36.111x - 8.1780.999 916.05-160.50
Daidzeiny = 23.526x + 0.390 20.999 91.07-10.66
Arctiiny = 6.173 1x + 1.107 30.999 88.59-85.86
), ArticleFig(id=1308834471781167144, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1308828729019822120, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
Sample No.C4C1C2C3C5C6C7C8C9C10C11
ESMESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMS
M116.403.903.903.383.354.414.392.912.902.022.000.700.702.022.0017.9017.910.440.418.788.76
M216.443.883.873.383.364.484.352.872.852.022.010.710.682.022.0117.9317.950.450.438.818.82
M316.393.873.873.333.334.464.372.872.842.021.990.710.632.022.0017.8917.920.440.428.688.69
M416.373.803.793.363.354.694.492.862.861.961.940.690.701.971.9917.7317.740.430.448.768.77
M516.363.923.913.353.344.714.522.862.842.022.000.710.672.011.9817.8617.800.440.438.538.55
M616.464.014.003.343.354.664.632.872.882.032.010.720.712.022.0017.9317.900.430.458.698.68
M716.253.793.803.303.314.414.422.822.801.951.940.650.661.951.9817.6917.740.420.428.558.56
M816.293.823.823.313.304.454.402.882.851.981.970.670.681.931.9517.7517.730.410.438.568.53
M916.303.913.903.333.304.594.552.832.841.981.990.690.701.991.9717.7517.690.430.448.638.65
M1016.453.994.003.403.384.754.702.882.872.052.070.750.782.052.0417.9917.940.480.498.828.80
), ArticleFig(id=1308834471865053225, tenantId=1146029695717560320, journalId=1189982191388893191, articleId=1308828729019822120, language=CN, label=Tab.2, caption=

Comparison of content determination results between ESM and QAMS (mg·g-1). C stands for components; C1: Neochlorogenic acid; C2: Chlorogenic acid; C3: Cryptochlorogenic acid; C4: Puerarin; C5: Daidzin; C6: Isochlorogenic acid B; C7: Isochlorogenic acid A; C8: Isochlorogenic acid C; C9: Baicalin; C10: Daidzein; C11: Arctiin; M1: 24070501; M2: 24070502; M3: 24071101; M4: 24071102; M5: 24071801; M6: 24071802; M7: 24072201; M8: 24072202; M9: 24072901; M10: 24072902. ESM: External standard method; QAMS: Quantitative analysis of multi-components by single marker

, figureFileSmall=null, figureFileBig=null, tableContent=
Sample No.C4C1C2C3C5C6C7C8C9C10C11
ESMESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMSESMQAMS
M116.403.903.903.383.354.414.392.912.902.022.000.700.702.022.0017.9017.910.440.418.788.76
M216.443.883.873.383.364.484.352.872.852.022.010.710.682.022.0117.9317.950.450.438.818.82
M316.393.873.873.333.334.464.372.872.842.021.990.710.632.022.0017.8917.920.440.428.688.69
M416.373.803.793.363.354.694.492.862.861.961.940.690.701.971.9917.7317.740.430.448.768.77
M516.363.923.913.353.344.714.522.862.842.022.000.710.672.011.9817.8617.800.440.438.538.55
M616.464.014.003.343.354.664.632.872.882.032.010.720.712.022.0017.9317.900.430.458.698.68
M716.253.793.803.303.314.414.422.822.801.951.940.650.661.951.9817.6917.740.420.428.558.56
M816.293.823.823.313.304.454.402.882.851.981.970.670.681.931.9517.7517.730.410.438.568.53
M916.303.913.903.333.304.594.552.832.841.981.990.690.701.991.9717.7517.690.430.448.638.65
M1016.453.994.003.403.384.754.702.882.872.052.070.750.782.052.0417.9917.940.480.498.828.80
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HPLC-DAD波长转换结合一测多评法测定金柴清热颗粒中的11种成分
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聂金娥 1 , 王少芸 1 , 钱芳芳 1 , 曲远均 1, * , 李乐 2 , 薛金苗 2 , 刘秋静 2
药学学报 | 研究论文 2026,61(3): 881-886
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药学学报 |研究论文 2026 , 61 (3) : 881 -886
HPLC-DAD波长转换结合一测多评法测定金柴清热颗粒中的11种成分
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聂金娥1, 王少芸1, 钱芳芳1, 曲远均1, * , 李乐2, 薛金苗2, 刘秋静2
作者信息
  • 1山东中医药大学附属医院, 山东 济南 250000
  • 2山东中医药大学, 山东 济南 250000
通讯作者:
曲远均, E-mail:
Simultaneous determination of eleven components in Jinchai Qingre Granule by QAMS and HPLC-DAD
Jin-e NIE1, Shao-yun WANG1, Fang-fang QIAN1, Yuan-jun QU1, * , Le LI2, Jin-miao XUE2, Qiu-jing LIU2
Affiliations
  • 1Affiliated Hospital of Shandong University of TCM, Jinan 250000, China
  • 2Shandong University of Traditional Chinese Medicine, Jinan 250000, China
出版时间: 2026-03-12 doi: 10.16438/j.0513-4870.2025-1129
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摘要
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建立高效液相色谱–二极管阵列检测法(high performance liquid chromatography-diode array detector, HPLC-DAD) 波长转换结合一测多评法(quantitative analysis of multi-components by single marker, QAMS) 同时测定金柴清热颗粒中11种成分含量的方法, 为其质量控制提供科学依据及质量保障。采用Agilent ZORBAX SB-C18色谱柱(250 mm × 4.6 mm, 5 μm), 以乙腈-0.1%磷酸水溶液为流动相, 梯度洗脱, 检测波长327、278、250 nm, 流速1.0 mL·min-1, 柱温25 ℃, 以葛根素为内参物, 建立葛根素与新绿原酸、绿原酸、隐绿原酸、大豆苷、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷的相对校正因子, 计算含量; 11种成分在各自浓度范围内线性关系良好(r ≥ 0.999 5), 平均加样回收率为98.05%~100.90%, RSD为0.98%~2.35% (n = 6); 不同设备、色谱柱、柱温、进样体积、流速等色谱条件下耐用性良好, 与外标法(external standard method, ESM) 相比较, 以葛根素为内参物的QAMS法对其余10种成分的测定结果无显著性差异。结果表明, 该方法简便、稳定, 可实现金柴清热颗粒中11种成分的质量控制, 为其质量标准化提供技术支撑。

金柴清热颗粒  /  高效液相色谱法  /  一测多评法  /  相对校正因子  /  葛根素

The study aims to establish a method for the content determination of eleven components in Jinchai Qingre Granule by high performance liquid chromatography-diode array detector (HPLC-DAD) through quantitative analysis of multi-components by single marker (QAMS) and verify the feasibility and applicability of the method in its quality control. Agilent ZORBAX SB-C18 column (250 mm × 4.6 mm, 5 μm) was used; acetonitrile-0.1% phosphoric acid aqueous solution was used as mobile phase; gradient elution and the detection wavelength were set at 327, 278, 250 nm; flow rate was 1.0 mL·min-1; column temperature was 25 ℃. Using puerarin as the internal reference, the relative correction factors of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, daidzin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, baicalin, daidzein and arctiin was established, and the eleven components were determined. Eleventeen components showed good linear relationships within their own ranges (r ≥ 0.999 5), whose average recoveries were 98.05%-100.90% with the RSDs of 0.98%-2.35% (n = 6). The method showed good durability under different conditions such as instruments, columns, column temperature, injection volume, and flow rate. There was no significant difference between the calculated values of eleven components in ten batches of samples by the QAMS and the measured values of the external standard method (P > 0.05). The method established in the study is simple and stable, and can be used for the synchronous quality control of eleven index components in Jinchai Qingre Granule.

Jinchai Qingre Granule  /  high-performance liquid chromatography  /  quantitative analysis of multi-components with single marker  /  relative correction factor  /  puerarin
聂金娥, 王少芸, 钱芳芳, 曲远均, 李乐, 薛金苗, 刘秋静. HPLC-DAD波长转换结合一测多评法测定金柴清热颗粒中的11种成分. 药学学报, 2026 , 61 (3) : 881 -886 . DOI: 10.16438/j.0513-4870.2025-1129
Jin-e NIE, Shao-yun WANG, Fang-fang QIAN, Yuan-jun QU, Le LI, Jin-miao XUE, Qiu-jing LIU. Simultaneous determination of eleven components in Jinchai Qingre Granule by QAMS and HPLC-DAD[J]. Acta Pharmaceutica Sinica, 2026 , 61 (3) : 881 -886 . DOI: 10.16438/j.0513-4870.2025-1129
正文
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金柴清热颗粒是山东中医药大学附属医院急症科多年经方研发而来, 由金银花、柴胡、葛根、牛蒡子、黄芩等十味药组成, 具有疏风解表、清热利咽之功效, 主要用于普通感冒、流行性感冒属感冒风热证者。近年来, 我国陆续发布了支持中药制剂发展的利好政策, 鼓励医疗机构制剂的研发与应用, 同时, 也提出提升中药质量促进中医药产业高质量发展的高要求, 以更好增进人民健康福祉和服务中国式现代化[1-3]。目前, 针对本制剂的研究仅局限于柴胡和葛根的薄层色谱鉴别、浸出物测定及颗粒剂通则检查, 在质量标准中尚未开展成分定量测定工作[4], 也未有同时定量分析多成分的研究报道。金柴清热颗粒作为传统经方与现代技术相结合的结晶, 在我院临床使用广泛, 临床效果显著, 为确保该制剂临床安全性和有效性, 其质量控制显得尤为关键。
金柴清热颗粒组方及化学成分复杂, 多成分含量测定能够更好地控制该制剂的质量以保证临床有效性。一测多评(quantitative analysis of multi-components by single marker, QAMS) 是通过测定中药中某一关键成分(内参物) 的含量, 结合其他成分与关键成分之间的相对校正因子, 推算出其他成分的含量[5-7]。这种方法能够简化分析流程, 降低分析成本, 同时提高分析效率。本研究采用HPLC-DAD波长转换扫描结合QAMS测定金柴清热颗粒中的11种成分, 该方法不仅符合当前中药质量控制的发展趋势, 也为金柴清热颗粒的临床应用提供了更为可靠的质量保障。
材料与方法
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样品与试剂   金柴清热颗粒(山东中医药大学附属医院, 批号: 24070501、24070502、24071101、24071102、24071801、24071802、24072201、24072202、24072901、24072902); 葛根素(批号: 110752-202217, 质量浓度为96.8%), 绿原酸(批号: 110753-202119, 质量浓度为96.3%), 隐绿原酸(批号111111-202401, 质量浓度为96.7%), 黄芩苷(批号: 110715-202223, 质量浓度为97.2%), 牛蒡苷(批号: 110819-202414, 质量浓度为96.6%), 大豆苷(批号: 111738-202305, 质量浓度为96.3%), 大豆苷元(批号: 111502-202304, 质量浓度为99.3%), 均购于中国食品药品检定研究院; 新绿原酸(批号: B21396, 质量浓度≥ 98%), 异绿原酸A (批号: B21539, 质量浓度≥ 98%), 异绿原酸B (批号: B21540, 质量浓度≥ 98%), 异绿原酸C (批号: B21541, 质量浓度≥ 98%) 均购于上海源叶生物科技有限公司; 乙腈、甲醇(色谱纯, 安徽天地生命科技有限公司)、磷酸(色谱纯, 天津市科密欧化学试剂有限公司)、水(娃哈哈水, 杭州娃哈哈集团有限公司); 其他试剂均为分析纯。
仪器   Agilent高效液相色谱仪(Agilent 1260 Infinity Ⅱ; 美国安捷伦科技有限公司); Waters高效液相色谱仪(Waters 2695; 美国沃特世科技有限公司); 数控超声波清洗器(KQ-250B型; 昆山市超声仪器有限公司); 电子天平(XPE205; 瑞士梅特勒-托利多仪器公司); 恒温水浴锅(KHW-S-4; 北京永光明有限公司); 鼓风干燥箱(101-1EBS型; 北京永光明有限公司); 分析天平(ME204E/02; 瑞士梅特勒-托利多仪器公司) 等。
混合对照品溶液制备    精密称取新绿原酸、绿原酸、隐绿原酸、葛根素、大豆苷、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷对照品适量, 置于25 mL量瓶中, 加甲醇溶解并稀释至刻度, 摇匀, 即得新绿原酸、绿原酸、隐绿原酸、葛根素、大豆苷、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷质量浓度分别为136.8、211.9、184.0、962.7、176.6、116.7、59.8、102.3、802.5、53.3、429.3 μg·mL-1的混合对照品贮备液; 精密量取混合对照品贮备液1 mL, 置于10 mL量瓶中, 加甲醇稀释并定容至刻度, 摇匀, 即得(各成分质量浓度分别为13.68、21.19、18.4、96.27、17.66、11.67、5.98、10.23、80.25、5.33、42.93 μg·mL-1)。
供试品溶液制备   取金柴清热颗粒适量, 研细, 取细粉0.3 g, 精密称定, 置于具塞锥形瓶中, 精密加入70%甲醇50 mL, 称定重量, 超声(电压220 V, 频率50 kHz) 提取30 min, 冷却, 用70%甲醇补足减失的重量, 摇匀, 滤过, 取续滤液, 即得。
色谱条件   色谱柱: Agilent ZORBAX-C18色谱柱(250 mm × 4.6 mm, 5 μm, 美国安捷伦科技有限公司); 流动相: 乙腈(A)-0.1%磷酸溶液(B), 梯度洗脱: 0~10 min, 7%→10% A; 10~15 min, 10% A; 15~40 min, 10%→20% A; 40~65 min, 20%→40% A; 记录时间为65 min; 流速1.0 mL·min-1; 检测波长: 0~20 min, 327 nm; 20~40 min, 250 nm; 40~50 min, 327 nm; 51~53.9 min, 250 nm; 54~65 min, 278 nm; 柱温25 ℃; 进样量10 μL。
方法学考察
专属性  按处方比例分别制备缺金银花阴性样品、缺葛根阴性样品、缺黄芩阴性样品、缺牛蒡子的阴性样品, 照“供试品溶液制备”方法, 分别制备缺金银花阴性样品溶液、缺葛根阴性样品溶液、缺黄芩阴性样品溶液、缺牛蒡子阴性样品溶液。取混合对照品溶液、供试品溶液及各阴性对照溶液, 照“色谱条件”下进行测定。
线性关系  分别精密吸取混合对照品溶液2、5、10、15、20 μL注入高效液相色谱仪, 以峰面积为纵坐标(y), 对照品进样量(μg·mL-1) 为横坐标(x), 绘制标准曲线。
精密度  取金柴清热颗粒(批号: 24070501), 按“供试品溶液制备”方法制备供试品溶液, 连续进样6次, 测定11种成分的峰面积, 计算相对标准偏差(RSD) 值。
重复性  取金柴清热颗粒(批号: 24070501) 样品, 按“供试品溶液制备”方法制备供试品溶液6份, 照“色谱条件”下进行测定, 计算11种成分的含量, 计算RSD值。
稳定性  取金柴清热颗粒(批号: 24070501) 制备供试品溶液, 按“色谱条件”项下分别于0、2、4、6、8、12、24 h进样测定, 记录11种成分的峰面积并计算RSD值。
加样回收率  取已知含量的金柴清热颗粒(批号: 24070501), 研细, 取粉末约0.15 g, 精密称定, 平行6份, 置于具塞锥形瓶中, 分别按已知含量的100%精密加入含新绿原酸、绿原酸、隐绿原酸、葛根素、大豆苷、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷的混合对照品溶液25 mL, 再按照“供试品溶液制备”方法制备溶液, 照“色谱条件”进样测定, 记录11种成分的峰面积并计算各成分的加样回收率和RSD。
相对校正因子的确定[8-10]  取“混合对照品溶液”, 按“色谱条件”进样测定。以葛根素为内参物, 计算相对校正因子。fs/i = fs/fi = (As/Cs) / (Ai/Ci), 式中As为内参物对照品峰面积, Cs为内参物对照品浓度, Ai为待测成分对照品峰面积, Ci为待测成分对照品浓度。
耐用性考察[11-13]  取“混合对照品溶液”, 分别考察不同进样体积(2、5、10、15、20 μL)、不同柱温(25、30、35 ℃)、不同流速(0.8、1.0、1.2 mL·min-1)、不同仪器及色谱柱对相对校正因子的影响, 计算平均值和RSD值。
待测色谱峰定位[14-16]  取“混合对照品溶液”, 分别采用2种不同色谱仪和3种色谱柱测定, 计算各待测成分对葛根素的相对保留时间(r ＝ tRi/tRs) 及RSD。
一测多评法与外标法测定结果比较   取金柴清热颗粒10批, 研细, 取细粉0.3 g, 精密称定, 按“供试品溶液制备”方法制备, 按“色谱条件”进样测定, 分别用ESM和QAMS测定各成分含量[10, 17, 18], 两者结果进行比较。
结果
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1　方法学考察
1.1　专属性
新绿原酸、绿原酸、隐绿原酸、葛根素、大豆苷、异绿原酸B、异绿原酸A、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷色谱峰分离度均大于1.5, 混合对照品溶液、供试品溶液分别在其相同的保留时间处有同一色谱峰, 阴性样品溶液中无相应的色谱峰, 表明本试验专属性良好。结果如图1所示。
1.2　线性关系
11种成分在相应浓度范围内, 线性关系良好(r ≥ 0.999 5), 结果见表1。
1.3　精密度
11种成分峰面积的RSD分别为0.74%、0.84%、0.58%、1.42%、0.50%、0.55%、0.38%、0.64%、0.39%、0.47%、0.47%, 均小于2.0%, 表明该方法精密度良好, 满足测定要求。
1.4　重复性
11种成分的平均含量分别为3.90、3.36、4.57、16.40、2.87、2.01、0.71、2.01、17.87、0.44、8.71 mg·g-1, RSD分别为1.78%、0.53%、2.85%、0.24%、0.60%、1.35%、1.39%、1.06%、0.41%、1.54%、1.17%, 均小于3.0%, 表明本方法的重复性良好。
1.5　稳定性
11种成分的峰面积RSD分别为2.05%、0.60%、1.64%、0.62%、0.57%、1.55%、2.31%、1.08%、0.73%、1.68%、0.83%。结果显示供试品溶液在24 h内稳定性良好。
1.6　加样回收率
11种成分的平均加样回收率分别为100.74%、98.60%、100.90%、99.71%、98.05%、98.62%、98.83%、98.81%、99.29%、98.89%、99.24%, RSD均小于5.0%, 表明该方法回收率良好, 满足分析要求。
1.7　相对校正因子的测定
以葛根素为内参物, 新绿原酸、绿原酸、隐绿原酸、大豆苷、异绿原酸A、异绿原酸B、异绿原酸C、黄芩苷、大豆苷元、牛蒡苷的相对校正因子分别为4.162 5、2.875 6、3.952 2、0.895 6、2.561 3、1.899 0、2.080 8、1.798 3、1.822 8、7.022 4, RSD均小于3.0%。
1.8　耐用性
不同进样体积(2、5、10、15、20 μL)、不同柱温(25、30、35 ℃)、不同流速(0.8、1.0、1.2 mL·min-1)、不同仪器及色谱柱各成分相对校正因子的RSD均小于1.0%, 表明各成分在本色谱条件下耐用性良好。
1.9　待测色谱峰定位
各待测成分相对保留时间(r ＝ tRi/tRs) 的RSD均小于0.5%, 表明利用相对保留时间进行色谱峰的定位是可行的。
2　一测多评法与外标法测定结果比较
金柴清热颗粒10批, EMS实测值与QAMS计算值比较, 各成分含量具有高度一致性, 测定结果无显著性差异, 表明建立的方法具有较好的准确度与可信度, 结果见表2。
讨论
收起
本研究运用HPLC-DAD对各待测成分进行全波长扫描, 确定新绿原酸、绿原酸、隐绿原酸、异绿原酸A~C的检测波长为327 nm, 葛根素、大豆苷、大豆苷元检测波长为250 nm, 牛蒡苷、黄芩苷检测波长为278 nm。金柴清热颗粒样品标志物的选择以君臣佐使为原则, 选取了君药中金银花中的新绿原酸、绿原酸、隐绿原酸、异绿原酸A~C, 臣药葛根中的葛根素、大豆苷及大豆苷元, 佐使药牛蒡子的主要成分牛蒡苷及黄芩中黄芩苷为定量指标成分[19-23], 通过试验验证该方法目标峰处无干扰峰, 方法专属性强。
本研究采用HPLC-DAD波长转换扫描结合QAMS建立了金柴清热颗粒中11种化学成分含量的QAMS分析方法, 通过方法学考察、色谱峰定位及与ESM测定值进行比较, 结果表明, QAMS计算值与ESM实测值无显著性差异, 具有较高的重现性和稳定性, 可实现金柴清热颗粒中11种成分的质量控制, 对保障金柴清热颗粒的质量均一性、临床有效性及安全性具有重要意义, 为全面控制金柴清热颗粒的质量提供参考。

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2026年第61卷第3期
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doi: 10.16438/j.0513-4870.2025-1129
  • 接收时间：2025-09-22
  • 首发时间：2026-09-21
  • 出版时间：2026-03-12
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  • 收稿日期：2025-09-22
  • 修回日期：2026-01-04
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    1山东中医药大学附属医院, 山东 济南 250000
    2山东中医药大学, 山东 济南 250000

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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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