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Article(id=1236731779709718847, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236731777142804746, articleNumber=null, orderNo=null, doi=10.20251/j.cnki.1003-3734.2025.17.012, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=null, receivedDateStr=null, revisedDate=null, revisedDateStr=null, acceptedDate=1707321600000, acceptedDateStr=2024-02-08, onlineDate=1772789433648, onlineDateStr=2026-03-06, pubDate=1757865600000, pubDateStr=2025-09-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772789433648, onlineIssueDateStr=2026-03-06, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772789433648, creator=13701087609, updateTime=1772789433648, updator=13701087609, issue=Issue{id=1236731777142804746, tenantId=1146029695717560320, journalId=1235980733773295621, year='2025', volume='34', issue='17', pageStart='1793', pageEnd='1904', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=0, createTime=1772789433036, creator=13701087609, updateTime=1772801868692, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1236783936152596961, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236731777142804746, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1236783936152596962, tenantId=1146029695717560320, journalId=1235980733773295621, issueId=1236731777142804746, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1865, endPage=1871, ext={EN=ArticleExt(id=1236731779932016969, articleId=1236731779709718847, tenantId=1146029695717560320, journalId=1235980733773295621, language=EN, title=Quality analysis of Reyihan granules, columnId=null, journalTitle=Chinese Journal of New Drugs, columnName=null, runingTitle=null, highlight=null, articleAbstract=
Objective:

To set up the quality standards of Reyihan granules.

Methods:

The identification items for Ocimi basilici Fructus, Roses rugosae Flos, Nardostachyos radlx Et Rhizoma, and Foeniculi fructus were established using TLC. The characteristic chromatogram was established, and the contents of gallic acid and rosmarinic acid in Reyihan granules were determined using HPLC.

Results:

The TLC identification method for Ocimi basilici Fructus, Roses rugosae Flos, Nardostachyos radlx Et Rhizoma, Foeniculi fructus had clear spots, with no interference from negative controls. There were 35 characteristic peaks in the HPLC characteristic chromatogram of Reyihan granules. Through comparison with reference standards, 13 compounds were identified. Gallic acid showed a good linear relationship within the range of 1.303 2~13.032 0 μg·mL-1 (r=0.999 7), with an average recovery rate of 97.23% and an RSD of 1.20% (n=6). Rosmarinic acid showed a good linear relationship within the range of 2.462 4~24.624 0 μg·mL-1 (r=0.999 3), with an average recovery rate of 102.79% and RSD of 1.10% (n=6).

Conclusion:

This method has strong specificity, good repeatability and stability, and can achieve quality control of Reyihan granules.

, correspAuthors=Cheng-hui HE, 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=De-yong LIU, Xin-chun WANG, Payman HERMITY, Chuan-sheng HUANG, Cheng-hui HE), CN=ArticleExt(id=1236731781249028477, articleId=1236731779709718847, tenantId=1146029695717560320, journalId=1235980733773295621, language=CN, title=维药热依汗颗粒质量分析研究, columnId=1236330556003242277, journalTitle=中国新药杂志, columnName=实验研究, runingTitle=null, highlight=null, articleAbstract=
目的:

建立热依汗颗粒质量标准。

方法:

采用TLC法建立罗勒、玫瑰花、甘松、小茴香的鉴别项；采用HPLC法建立热依汗颗粒的特征图谱及没食子酸、迷迭香酸含量测定项。

结果:

罗勒、玫瑰花、甘松、小茴香TLC法斑点清晰，阴性无干扰。热依汗颗粒HPLC特征图谱中有35个特征峰。通过对照品比对，鉴别出13种化合物。没食子酸在（1.303 2～13.032 0 μg·mL-1）范围内呈良好线性关系（r=0.999 7），平均加样回收率为97.23%，RSD为1.20%（n=6）；迷迭香酸在（2.462 4～24.624 0 μg·mL-1）范围内呈良好线性关系（r=0.999 3），平均加样回收率为102.79%，RSD为1.10%（n=6）。

结论:

该方法专属性强、重复性、稳定性好，可实现对热依汗颗粒的质量控制。

, correspAuthors=何承辉, authorNote=null, correspAuthorsNote=
*何承辉，女，硕士生导师，研究员，主要从事中药民族药新制剂、新剂型及新药开发。E-mail：。
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刘德永，男，硕士研究生，主要从事中药制备新技术及质量控制研究。E-mail：。

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刘德永，男，硕士研究生，主要从事中药制备新技术及质量控制研究。E-mail：。

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刘德永，男，硕士研究生，主要从事中药制备新技术及质量控制研究。E-mail：。

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1:石吊兰素对照品;2～4:供试品（批号：20231105）;5:缺罗勒阴性对照

, figureFileSmall=qAFsL61/qZLb4qLAuIPrag==, figureFileBig=fuuCZydpYyZXZBR2pBTlWA==, tableContent=null), ArticleFig(id=1236792933840646951, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=EN, label=null, caption=null, figureFileSmall=aD8GunoRARBUF1sWt07Ljw==, figureFileBig=qNPqI8erKP+F0JScoIIQWA==, tableContent=null), ArticleFig(id=1236792933911950123, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=图2, caption=玫瑰花TLC鉴别图

A:紫外光灯（254 nm）;B:日光灯;1:没食子酸对照品;2:玫瑰花对照药材;3～4:供试品（批号：20231101）;5～6:缺玫瑰花阴性对照

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1:甘松对照药材;2～4:供试品（批号：20231102）;5:缺甘松阴性对照

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1:小茴香对照药材;2～3:供试品（批号：20231105）;4:缺小茴香阴性对照

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A:供试品与混合对照品;B:供试品与阴性样品;C:供试品与单味药材样品;2:没食子酸;3:新绿原酸;6:绿原酸;7:咖啡酸;15:芦丁;16:金丝桃苷;17:异槲皮苷;23:迷迭香酸;27:木犀草素;29:芹菜素;30:山柰酚;34:石吊兰素;35:甘松新酮

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编号药味批号产地来源
1罗勒Q30048109新疆新疆麦迪森药业有限公司饮片分公司
2牛舌草C30010406海南
3薰衣草M30048022新疆
4甘松G30031508四川
5玫瑰花H30016415新疆
6菟丝草C30059708新疆
7小茴香22010602甘肃新疆仁誉中药饮片有限公司
), ArticleFig(id=1236792936055239499, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=表1, caption=

热依汗颗粒药材来源信息

, figureFileSmall=null, figureFileBig=null, tableContent=
编号药味批号产地来源
1罗勒Q30048109新疆新疆麦迪森药业有限公司饮片分公司
2牛舌草C30010406海南
3薰衣草M30048022新疆
4甘松G30031508四川
5玫瑰花H30016415新疆
6菟丝草C30059708新疆
7小茴香22010602甘肃新疆仁誉中药饮片有限公司
), ArticleFig(id=1236792936168485711, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min流动相A流动相B
0397
10397
581783
601486
681486
1103070
1123268
1304555
1384555
145397
150397
), ArticleFig(id=1236792936252371793, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=表2, caption=

特征图谱测定的梯度洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min流动相A流动相B
0397
10397
581783
601486
681486
1103070
1123268
1304555
1384555
145397
150397
), ArticleFig(id=1236792936323674964, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min流动相A流动相B
0298
15298
201882
621882
656535
776535
80298
85298
), ArticleFig(id=1236792936424338264, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=表3, caption=

没食子酸、迷迭香酸含量测定的梯度洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min流动相A流动相B
0298
15298
201882
621882
656535
776535
80298
85298
), ArticleFig(id=1236792936520807260, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
成分色谱柱型号含量/mg·g-1RSD/%
没食子酸Agilent250 mm×4.6 mm，5 μm0.950 92.30
SHIMADZU0.918 8
Waters0.910 6
迷迭香酸Agilent250 mm×4.6 mm，5 μm2.262 32.95
SHIMADZU2.284 5
Waters2.160 9
), ArticleFig(id=1236792936604693343, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=表4, caption=

耐用性考察结果

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成分色谱柱型号含量/mg·g-1RSD/%
没食子酸Agilent250 mm×4.6 mm，5 μm0.950 92.30
SHIMADZU0.918 8
Waters0.910 6
迷迭香酸Agilent250 mm×4.6 mm，5 μm2.262 32.95
SHIMADZU2.284 5
Waters2.160 9
), ArticleFig(id=1236792936722133856, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
成分已知量/mg加入量/mg测得量/mg加样回收率/%平均加样回收率/%RSD/%
没食子酸0.230 20.244 80.464 795.7997.231.20
0.230 70.244 80.473 799.26
0.230 50.244 80.468 397.11
0.230 90.244 80.467 696.69
0.231 00.244 80.468 396.92
0.230 70.244 80.469 797.64
迷迭香酸0.489 80.503 31.010 1103.38102.791.10
0.490 90.503 31.010 9103.30
0.490 60.503 30.997 3100.69
0.491 30.503 31.006 2102.31
0.491 50.503 31.012 9103.58
0.490 90.503 31.011 7103.47
), ArticleFig(id=1236792936806019940, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=表5, caption=

加样回收率考察结果

, figureFileSmall=null, figureFileBig=null, tableContent=
成分已知量/mg加入量/mg测得量/mg加样回收率/%平均加样回收率/%RSD/%
没食子酸0.230 20.244 80.464 795.7997.231.20
0.230 70.244 80.473 799.26
0.230 50.244 80.468 397.11
0.230 90.244 80.467 696.69
0.231 00.244 80.468 396.92
0.230 70.244 80.469 797.64
迷迭香酸0.489 80.503 31.010 1103.38102.791.10
0.490 90.503 31.010 9103.30
0.490 60.503 30.997 3100.69
0.491 30.503 31.006 2102.31
0.491 50.503 31.012 9103.58
0.490 90.503 31.011 7103.47
), ArticleFig(id=1236792936910877545, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=EN, label=null, caption=null, figureFileSmall=null, figureFileBig=null, tableContent=
样品没食子酸迷迭香酸
202311010.840 11.589 4
202311020.887 61.783 9
202311030.926 51.656 0
202311040.928 71.847 2
202311050.922 31.999 1
202311060.949 52.051 8
202311070.948 02.153 0
), ArticleFig(id=1236792937007346541, tenantId=1146029695717560320, journalId=1235980733773295621, articleId=1236731779709718847, language=CN, label=表6, caption=

没食子酸、迷迭香酸含量测定结果

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样品没食子酸迷迭香酸
202311010.840 11.589 4
202311020.887 61.783 9
202311030.926 51.656 0
202311040.928 71.847 2
202311050.922 31.999 1
202311060.949 52.051 8
202311070.948 02.153 0
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刘德永 1 , 王新春 1, 3 , 帕依曼·亥米提 2 , 黄川生 1, 3 , 何承辉 2, *
中国新药杂志 | 实验研究 2025,34(17): 1865-1871
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中国新药杂志 | 实验研究 2025, 34(17): 1865-1871
维药热依汗颗粒质量分析研究
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刘德永1 , 王新春1, 3, 帕依曼·亥米提2, 黄川生1, 3, 何承辉2, *
作者信息
  • 1石河子大学药学院，石河子　832003
  • 2新疆维吾尔自治区药物研究所，乌鲁木齐　830004
  • 3石河子大学医学院第一附属医院药剂科，石河子　832008

    • 刘德永，男，硕士研究生，主要从事中药制备新技术及质量控制研究。E-mail：。

通讯作者:

*何承辉，女，硕士生导师，研究员，主要从事中药民族药新制剂、新剂型及新药开发。E-mail：。
Quality analysis of Reyihan granules
De-yong LIU1 , Xin-chun WANG1, 3, Payman HERMITY2, Chuan-sheng HUANG1, 3, Cheng-hui HE2, *
Affiliations
  • 1College of Pharmacy, Shihezi University, Shihezi 832003, China
  • 2Xinjiang Uygur Autonomous Region Institute of Meteria Medica, Urumqi 830004, China
  • 3Department of Pharmacy The First Affiliated Hospital of the Medical College, Shihezi University, Shihezi 832008, China
出版时间: 2025-09-15 doi: 10.20251/j.cnki.1003-3734.2025.17.012
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摘要
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目的:

建立热依汗颗粒质量标准。

方法:

采用TLC法建立罗勒、玫瑰花、甘松、小茴香的鉴别项；采用HPLC法建立热依汗颗粒的特征图谱及没食子酸、迷迭香酸含量测定项。

结果:

罗勒、玫瑰花、甘松、小茴香TLC法斑点清晰，阴性无干扰。热依汗颗粒HPLC特征图谱中有35个特征峰。通过对照品比对，鉴别出13种化合物。没食子酸在（1.303 2～13.032 0 μg·mL-1）范围内呈良好线性关系（r=0.999 7），平均加样回收率为97.23%，RSD为1.20%（n=6）；迷迭香酸在（2.462 4～24.624 0 μg·mL-1）范围内呈良好线性关系（r=0.999 3），平均加样回收率为102.79%，RSD为1.10%（n=6）。

结论:

该方法专属性强、重复性、稳定性好，可实现对热依汗颗粒的质量控制。

热依汗颗粒  /  TLC法  /  特征图谱  /  HPLC法  /  质量标准
Objective:

To set up the quality standards of Reyihan granules.

Methods:

The identification items for Ocimi basilici Fructus, Roses rugosae Flos, Nardostachyos radlx Et Rhizoma, and Foeniculi fructus were established using TLC. The characteristic chromatogram was established, and the contents of gallic acid and rosmarinic acid in Reyihan granules were determined using HPLC.

Results:

The TLC identification method for Ocimi basilici Fructus, Roses rugosae Flos, Nardostachyos radlx Et Rhizoma, Foeniculi fructus had clear spots, with no interference from negative controls. There were 35 characteristic peaks in the HPLC characteristic chromatogram of Reyihan granules. Through comparison with reference standards, 13 compounds were identified. Gallic acid showed a good linear relationship within the range of 1.303 2~13.032 0 μg·mL-1 (r=0.999 7), with an average recovery rate of 97.23% and an RSD of 1.20% (n=6). Rosmarinic acid showed a good linear relationship within the range of 2.462 4~24.624 0 μg·mL-1 (r=0.999 3), with an average recovery rate of 102.79% and RSD of 1.10% (n=6).

Conclusion:

This method has strong specificity, good repeatability and stability, and can achieve quality control of Reyihan granules.

Reyihan granules  /  TLC  /  characteristic chromatogram  /  HPLC  /  quality standard
刘德永, 王新春, 帕依曼·亥米提, 黄川生, 何承辉. 维药热依汗颗粒质量分析研究. 中国新药杂志, 2025 , 34 (17) : 1865 -1871 . DOI: 10.20251/j.cnki.1003-3734.2025.17.012
De-yong LIU, Xin-chun WANG, Payman HERMITY, Chuan-sheng HUANG, Cheng-hui HE. Quality analysis of Reyihan granules[J]. Chinese Journal of New Drugs, 2025 , 34 (17) : 1865 -1871 . DOI: 10.20251/j.cnki.1003-3734.2025.17.012
正文
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热依汗颗粒是由罗勒、玫瑰花、甘松、小茴香等7味药材组成的维药复方制剂，为新疆墨玉县维吾尔医医院的院内制剂（批准单位：新疆维吾尔自治区食品药品监督管理局；批件号：MZJ-W-0292-2009）。其中罗勒收载于《新疆维吾尔自治区中药维吾尔药饮片炮制规范》2020年版[1]，牛舌草、薰衣草、菟丝草收载于《中华人民共和国卫生部药品标准》（维药分册）[2]，其他药材收载于《中华人民共和国药典》2020年版一部[3]。功能主治为活血通阻，消食，补血，养心补脑，增强心脑功能。用于神经衰弱引起的头疼、失眠、健忘、多梦，心功能不全，心律不齐，贫血，胃炎，胆囊炎，也可用于白癜风的辅助治疗。此院内制剂具有长久的临床应用历史，其疗效确切，且安全性好。但原制剂质量标准为院内制剂内部标准，只有玫瑰花TLC定性鉴别，存在标准较低、质量稳定性差等问题。在原有制剂的研究基础上应用TLC及HPLC分析技术，对热依汗颗粒进行定性和定量分析研究。本文建立了罗勒、甘松、小茴香、玫瑰花TLC鉴别项，没食子酸、迷迭香酸HPLC含量测定项及HPLC特征图谱项，保证了热依汗颗粒的有效性和可控性。在此研究基础上建立一套规范化的质量标准，可为该制剂中药新药开发奠定基础。
材料
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1　药物与试剂
没食子酸（批号：110831-201906，纯度：91.5%）、咖啡酸（批号：110885-201703）、芦丁（批号：100080-202012）、金丝桃苷（批号：111521-201507）、异槲皮苷（批号：111809-202205）、迷迭香酸（批号：111871-202007，纯度：98.1%）、芹菜素（批号：111901-202205）、山柰酚（批号：110861-201310）、石吊兰素（批号：111555-200602）、甘松新酮（批号：111832-201603）、甘松对照药材（批号：121402-201805）、小茴香对照药材（批号：121198-202304）均购自中国食品药品检定研究院；新绿原酸（批号：MUST-23040213）、绿原酸（批号：MUST-22111711）均购自成都曼思特生物科技有限公司；木犀草素（批号：111520-200504，中国药品生物制品检定所）；乙腈（色谱纯，赛默飞世尔科技有限公司）；甲醇（分析纯，天津市致远化学试剂有限公司）；其余试剂均为分析纯；水为超纯水。
处方饮片来源信息见表1，方中各味药材由新疆维吾尔自治区药物研究所何承辉研究员鉴定为正品。罗勒为唇形科植物罗勒（Ocimum basilicum L.）的干燥地上部分。牛舌草为紫草科植物意大利牛舌草（Anchusa italica Retz.）的干燥全草。薰衣草为唇形科植物狭叶薰衣草（Lavandula angustifolia Mill.）的干燥地上部分。甘松为败酱科植物甘松（Nardostachys jatamansi DC.）的干燥根及根茎。玫瑰花为蔷薇科植物玫瑰（Rosa rugosa Thunb.）的干燥花蕾。菟丝草为旋花科植物菟丝子（Cuscuta chinensis Lam.）的干燥地上部分。小茴香为伞形科植物茴香（Foeniculum vulgare Mill.）的干燥成熟果实。
2　仪器
LC—2030 Plus型HPLC仪（日本岛津公司）；BS110S型电子天平（北京赛多利斯天平有限公司）；AB135—S型电子天平（梅特勒-托利多公司）；WP-UP-WF—20型超纯水机（四川沃特尔水处理设备有限公司）；KQ5200B型超声波清洗器（昆山市超声仪器有限公司）。
方法与结果
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1　TLC鉴别
1.1　罗勒的TLC鉴别[4]
取热依汗颗粒（批号：20231105）10 g，研磨成细粉，样品中加入70%甲醇100 mL并进行超声提取，时间为20 min，过滤除去杂质并将滤液蒸干，加50 mL水溶解蒸干后的残渣并用水饱和的正丁醇提取2次，每次提取量为50 mL，将2次正丁醇提取液合并混匀后再次蒸干，将得到的残渣加1.5 mL乙醇溶解，即得供试品溶液。取石吊兰素对照品，加甲醇制成每1 mL含0.7 mg的对照品溶液。同供试品溶液制备方法制得缺罗勒阴性对照溶液。展开条件为二氯甲烷-乙酸乙酯-甲酸（5∶5∶1）、硅胶G薄层板，点样量3 μL，展开至边缘1 cm处取出晾干，喷以5%三氯化铁乙醇溶液，105 ℃加热至斑点显色清晰。TLC图见图1。供试品色谱中，与对照品色谱相应的位置上，显相同颜色的斑点，没有检测到干扰的阴性结果。
1.2　玫瑰花的TLC鉴别[5-7]
取热依汗颗粒5 g（批号：20231101），研磨成细粉，加40 mL水，进行超声提取，时间为20 min，过滤除去杂质并将滤液蒸干，加5 mL甲醇溶解残渣，即得供试品液。取没食子酸对照品，加甲醇制成每1 mL含0.5 mg的溶液，作为对照品溶液。取玫瑰花对照药材0.3 g，加10 mL甲醇，同法制得对照药材溶液。同法制得缺玫瑰花阴性对照溶液。展开条件为三氯甲烷-乙酸乙酯-甲酸（5∶5∶1）、硅胶GF254薄层板，点样量5 μL，展开至边缘1 cm处取出薄层板并晾干，置紫外光灯（254 nm）下检视，再喷以5%三氯化铁乙醇溶液置日光灯下检视。TLC图见图2。供试品色谱中，在与对照品、对照药材色谱相应的位置上，显相同颜色的斑点，没有检测到干扰的阴性结果。
1.3　甘松的TLC鉴别[3]
取热依汗颗粒13 g（批号：20231102），研磨成细粉，加130 mL石油醚（60～90 ℃），超声提取30 min，过滤，取滤液浓缩至干，将得到的残渣加5 mL石油醚（60～90 ℃）溶解，即得供试品溶液。取甘松对照药材0.5 g，加40 mL石油醚（60～90 ℃），其他条件同供试品溶液，制得对照药材溶液。同法制得缺甘松阴性对照溶液。展开条件为石油醚（60～90 ℃）-乙酸乙酯（4∶1）、硅胶GF254薄层板，点样量10 μL，展开至边缘1 cm处取出薄层板并晾干，置紫外光灯（254 nm）下检视。TLC图见图3。供试品色谱中，与对照药材色谱相应的位置上，显相同颜色的斑点，没有检测到干扰的阴性结果。
1.4　小茴香的TLC鉴别[7]
取热依汗颗粒（批号：20231105）5 g，研磨成细粉，加50 mL水溶解后用石油醚（60～90 ℃）-乙醚（1∶1）混合液振摇提取2次，每次提取量为50 mL，合并2次的石油醚-乙醚混合液并置水浴上蒸干混合液，将蒸干后得到的残渣加1.5 mL乙醇溶解，即得供试品溶液。取小茴香对照药材0.5 g，其他条件同供试品溶液制得对照药材溶液。同供试品溶液制备条件制得缺小茴香阴性对照溶液。展开条件为正己烷-乙酸乙酯-甲酸（18∶3∶0.1）、硅胶G薄层板，点样量为8 μL，展开至边缘1 cm处取出薄层板并晾干，喷以10%硫酸乙醇溶液，置紫外光灯（365 nm）下检视。TLC图见图4。供试品色谱中，与对照药材色谱相应的位置上，显相同颜色的斑点，没有检测到干扰的阴性结果。
2　特征图谱建立[8-11]
2.1　色谱条件
采用岛津Shim-pack VP-ODS色谱柱（250 mm×4.6 mm, 5 μm）；体积流量为1.0 mL·min-1；柱温30 ℃；检测波长330 nm；以乙腈（A）-0.1％磷酸溶液（B）为流动相，梯度洗脱见表2；进样量10 μL。
2.2　混合对照品溶液的制备
分别取对照品适量，加甲醇制成每1 mL含没食子酸、新绿原酸、绿原酸、咖啡酸、芦丁、金丝桃苷、异槲皮苷、迷迭香酸、木犀草素、槲皮素、熊果酸、芹菜素、山柰酚、齐墩果酸、石吊兰素、甘松新酮分别为335.6，9.397，6.748，1.592，27.84，9.015，14.64，6.278，9.88，17.125，17.582，6.84，13.272，32.16，7.534，386.4 μg的混合对照品溶液。
2.3　热依汗颗粒、阴性颗粒、单味药材颗粒的制备
按处方比例称取7味药材饮片，加水煎煮3次，每次1 h，过滤，合并3次滤液，减压浓缩（-0.08 MPa，70～75 ℃）至相对密度为1.15～1.25的清膏，真空干燥（-0.08 MPa，60～65 ℃）得浸膏，粉碎成细粉。加入辅料制粒，即得热依汗颗粒。按处方比例分别称取缺单味药材的饮片，同法制备阴性颗粒。称取单味药材饮片，同法制备单味药材颗粒。
2.4　供试品、阴性、单味药材样品溶液的制备
取热依汗颗粒适量，研磨至细粉，精密称取0.5 g，置具塞锥形瓶中，精密加入70%甲醇100 mL，称定重量，超声处理1 h，放冷，再称定重量，用70%甲醇补足缺失的重量，混合均匀之后，滤过，取续滤液，即得供试品溶液。同法制备阴性、单味药材样品液。
2.5　特征图谱专属性试验与色谱峰指认
取“2.2”“2.4”项下各溶液，按“2.1”项下方法进样检测，特征图谱见图5。综合供试品中各峰分离程度进行目标峰取舍，使所选特征峰最大程度反映制剂特点，共标定35个特征峰，通过对照品比对指认出13个峰，2号峰为没食子酸、3号峰为新绿原酸、6号峰为绿原酸、7号峰为咖啡酸、15号峰为芦丁、16号峰为金丝桃苷、17号峰为异槲皮苷、23号峰为迷迭香酸、27号峰为木犀草素、29号峰为芹菜素、30号峰为山柰酚、34号峰为石吊兰素、35号峰为甘松新酮。再通过阴性、单味药材溶液与供试品溶液色谱比较，对色谱峰进行归属，7，23，32号峰归属于牛舌草，13号峰归属于罗勒、薰衣草、玫瑰花，16号峰归属于罗勒、薰衣草、玫瑰花，21号峰归属于罗勒、小茴香、菟丝草，22，25，26，31号峰归属于罗勒、薰衣草、菟丝草，32号峰归属于罗勒、甘松，34号峰归属于罗勒，35号峰归属于甘松，其他色谱峰属于多种药材共有峰，表明处方中药材均有特征峰。
2.6　精密度试验
取“2.2”项下同一份混合对照品溶液，按“2.1”项色谱条件测定6次，选定峰面积最大、特征最明显的23号峰（迷迭香酸）为参照峰（S），计算各特征峰相对保留时间、相对峰面积的RSD值。结果显示，RSD值为0.01%～0.08%（n=6）和0.46%～3.98%（n=6），精密度符合要求。
2.7　重复性试验
取热依汗颗粒按“2.1”“2.4”项制备供试品溶液6份、进样测定，以23号峰（迷迭香酸）为S，计算各特征峰相对保留时间、相对峰面积的RSD值。结果显示，RSD值为0.01%～0.32%（n=6）和0.96%～24.12%（n=6），可以满足重复性的要求。
2.8　稳定性试验
取热依汗颗粒按“2.1”“2.4”项制备供试品溶液，设置不同的进样时间测定，分别为0，3，6，12，18，24 h，以23号峰（迷迭香酸）为S，计算各特征峰相对保留时间、相对峰面积的RSD值。结果显示，RSD值为0.02%～1.18%（n=6）和0.87%～32.12%（n=6），表明在24 h中有较高的稳定性。
3　含量测定
3.1　色谱条件[12-13]
梯度洗脱条件见表3，采用双波长模式，没食子酸检测波长为274 nm，迷迭香酸检测波长为330 nm。其他色谱条件同“2.1”项。
3.2　对照品溶液的制备
分别取对照品适量，加甲醇制成没食子酸、迷迭香酸分别为534, 513 μg·mL-1的对照品贮备液。
3.3　供试品溶液的制备
同“2.4”项。
3.4　线性关系考察
取“3.2”项下没食子酸对照品贮备液，加甲醇稀释配制1.303 2，2.606 4，5.212 8，7.819 2，13.032 0 μg·mL-1的系列对照品浓度，取迷迭香酸对照品贮备液同法配制成2.462 4，4.924 8，9.849 6，14.774 4，24.624 0 μg·mL-1的系列对照品浓度，按“3.1”项色谱条件对峰面积进行测定，并绘制对应的曲线，横纵坐标分别为溶液浓度、色谱峰峰面积，最后通过线性回归得到标准曲线。没食子酸线性方程为：Y=34 564X+3 208.5，r=0.999 7，表明没食子酸在1.303 2～13.032 0 μg·mL-1范围内线性关系良好。迷迭香酸线性方程为：Y=32 703X+14 677，r=0.999 3，表明迷迭香酸在2.462 4～24.624 0 μg·mL-1范围内线性关系良好。
3.5　精密度试验
取同一份没食子酸、迷迭香酸对照品溶液，按“3.1”项色谱条件重复进样6次，结果显示没食子酸、迷迭香酸峰面积的RSD值分别为1.56%，1.35%（n=6），精密度符合要求。
3.6　稳定性试验
根据“3.3”中的方法对供试品溶液进行制备，设置不同的进样时间，分别为0，2，4，6，8，12，24 h，按“3.1”项色谱条件对各成分的含量进行测定，并计算RSD值，最终得到的结果是2.55%，0.83%（n=7），根据上述结果可知，在24 h中保持了较高的稳定性。
3.7　日间精密度试验
根据“3.3”项下方法分别于接下来的5 d内制备供试品溶液，按“3.1”项色谱条件连续5 d进样测定。计算没食子酸和迷迭香酸的含量和RSD值，结果显示2种成分的RSD分别为1.99%，1.81%（n=5），日间精密度符合要求。
3.8　重复性试验
根据“3.1”“3.3”项制备供试品溶液6份、测定2种成分的含量。最终得到的结果显示，2种成分含量的RSD依次为0.98%，0.90%（n=6），可以满足重复性的要求。
3.9　耐用性试验
根据“3.1”“3.3”项制备供试品溶液、用3个不同品牌的色谱柱分别进样测定。计算没食子酸和迷迭香酸的含量和RSD值，实验结果见表4，结果显示2种成分含量的RSD分别为2.30%，2.95%（n=3），表明该方法耐用性良好。
3.10　加样回收率试验
分别取对照品适量，加甲醇制成没食子酸、迷迭香酸分别为0.244 8，0.503 3 mg·mL-1的对照品溶液。精密称取已知含量的热依汗颗粒6份，每份0.25 g，分别加入上述对照品溶液1 mL，分别按照“3.3”项制备供试品溶液，测定2种成分的含量。根据最终计算的结果可知2种成分的回收率为95.79%~103.58%，RSD为1.10%~1.20%（n=6）。结合实验结果可知，此方法可以达到较高的回收率，具体如表5中所示。
3.11　样品含量测定
取7批热依汗颗粒，按“3.1”“3.3”项制备供试品溶液，进样测定并计算含量。结果见表6。在7批热依汗颗粒含量测定数据的基础上，制定指标成分含量不得低于上述结果的70%，即没食子酸含量不得低于0.588 1 mg·g-1，迷迭香酸含量不得低于1.112 6 mg·g-1。
讨论
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在TLC鉴别中，本实验基于在特征图谱中鉴别出的成分对处方中7味药材均进行了TLC色谱条件摸索，对牛舌草中咖啡酸[14]、菟丝草中金丝桃苷[15]以及采用薰衣草自身药材对薰衣草进行TLC鉴别时发现它们均有阴性干扰，通过与本实验HPLC特征图谱比对也可发现牛舌草、薰衣草、菟丝草中鉴别出的主要化合物均有阴性干扰，故不纳入质量标准。
确立的罗勒、玫瑰花、小茴香、甘松共4味药材的TLC鉴别方法，无阴性干扰，方法简便，可有效控制热依汗颗粒的质量。罗勒中含有的黄酮类化合物石吊兰素为该药材专属性特征成分，因此在罗勒TLC鉴别中选择了石吊兰素对照品。玫瑰花中含有水溶性没食子酸，张旭等[16]的研究证明没食子酸能显著降低血脂水平，且没食子酸为玫瑰花主要成分之一，因此在玫瑰花TLC鉴别中选择了没食子酸对照品。没食子酸和迷迭香酸在特征图谱中分离度良好、峰面积较大，符合HPLC可测性原则，没食子酸、迷迭香酸具有抗炎、抗氧化、抗血栓、抗血小板凝聚等功效，有很好的心血管保护作用[17-20]，故选择这2种成分进行含量测定。将没食子酸和迷迭香酸对照品在200~800 nm波长范围内进行紫外扫描，结果没食子酸在274 nm处吸收最强，迷迭香酸在330 nm处吸收最强，故采用双波长模式在274和330 nm处测定2种化合物。
本研究通过增加TLC鉴别项、HPLC特征图谱项、HPLC含量测定项，提升了维药热依汗颗粒质量标准。在原院内制剂标准基础上，修订了玫瑰花的TLC法，新增了罗勒的TLC法，弥补了热依汗颗粒中君药的鉴别项；特征图谱建立的35个特征峰及识别出的13种化合物可实现对该制剂的定性控制；增加的没食子酸、迷迭香酸含量测定项可实现对该制剂的定量控制。
基金
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  • 乌鲁木齐市重点科技开发课题资助项目(B251010001)
  • 兵团重点领域科技公关资助项目(2024AB07)
  • 自治区重大科技专项资助项目(2022A03017-1)
文献
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2025年第34卷第17期
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doi: 10.20251/j.cnki.1003-3734.2025.17.012
  • 首发时间：2026-03-06
  • 出版时间：2025-09-15
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  • 录用日期：2024-02-08
基金
乌鲁木齐市重点科技开发课题资助项目(B251010001)
兵团重点领域科技公关资助项目(2024AB07)
自治区重大科技专项资助项目(2022A03017-1)
作者信息
    1石河子大学药学院，石河子　832003
    2新疆维吾尔自治区药物研究所，乌鲁木齐　830004
    3石河子大学医学院第一附属医院药剂科，石河子　832008

通讯作者:

*何承辉，女，硕士生导师，研究员，主要从事中药民族药新制剂、新剂型及新药开发。E-mail：。
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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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