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Article(id=1195814100342190965, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1195814096831561845, articleNumber=1001-2494(2024)23-2249-09, orderNo=null, doi=10.11669/cpj.2024.23.006, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1699200000000, receivedDateStr=2023-11-06, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1763033898672, onlineDateStr=2025-11-13, pubDate=1733587200000, pubDateStr=2024-12-08, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1763033898672, onlineIssueDateStr=2025-11-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1763033898672, creator=13701087609, updateTime=1763033898672, updator=13701087609, issue=Issue{id=1195814096831561845, tenantId=1146029695717560320, journalId=1190317699101192196, year='2024', volume='59', issue='23', pageStart='2199', pageEnd='2298', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1763033897836, creator=13701087609, updateTime=1763034755097, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1195817692507718044, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1195814096831561845, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1195817692507718045, tenantId=1146029695717560320, journalId=1190317699101192196, issueId=1195814096831561845, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2249, endPage=2257, ext={EN=ArticleExt(id=1195814100606432130, articleId=1195814100342190965, tenantId=1146029695717560320, journalId=1190317699101192196, language=EN, title=Chuanxiong Rhizoma and Angelicase Sinensis Radix Volatile Oil Study and Transdermal Effect Based on the Concept of Unification of Drugs and Excipients, columnId=null, journalTitle=Chinese Pharmaceutical Journal, columnName=null, runingTitle=null, highlight=null, articleAbstract=

OBJECTIVE To study the extraction method, quality, transdermal permeability, and transdermal promoting effects of Chuanxiong Rhizoma Hort. volatile oil (LVO) and Angelicase Sinensis Radix volatile oil (AVO) combined with the characteristic advantages of “unification of drugs and excipients” of traditional Chinese medicine volatile oil. METHODS The orthogonal design was used to screen and optimize extraction process of LVO and AVO. Quality control was carried out by detecting the physical parameters and indicator component content of LVO and AVO. Franz diffusion cells were used to conduct the studies of skin permeation in vitro. The contents of ligustilide and resveratrol(RSV) were determined by HPLC, and the steady-state transdermal rate(Jss) and enhancement factor (EF) compared with blank were used as evaluation indicators. And the transdermal permeability and transdermal promoting effects of different concentrations of LVO, AVO and mixed were investigated respectively. RESULTS The extraction process for LVO and AVO involves 8 times the amount of water, heating and refluxing for 8 h, collecting with ethyl acetate, dehydrating anhydrous sodium sulfate, and then volatilizing under reduced pressure. Perfect quality control methods were established for LVO and AVO, including the properties, relative density, pH, optical rotation, viscosity and ligustilide content. LVO and AVO have good permeability and promote RSV transdermal effect in vitro, and the maximum Jss values within 10 h were (48.23±6.53) and (27.84±3.03), respectively, and the maximum EF values were 4.51 and 19.34, respectively. CONCLUSION LVO and AVO have good transdermal permeability and transdermal promoting effects of RSV when used alone or in combination. The transdermal permeability of LVO was significantly higher than that of AVO, while the transdermal promoting effects of AVO was higher than that of LVO. The combination of LVO and AVO's own pharmacological effects and good transdermal absorption promoting effect plays a role of “unification of drugs and excipients”, which has a good advantage in clinical use, and lays a foundation for their development and application in transdermal drug delivery preparations of traditional Chinese medicine.

, correspAuthors=Aijun ZHANG, 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=Hongmei ZHU, Aijun ZHANG, Shuai LI), CN=ArticleExt(id=1195814168260554759, articleId=1195814100342190965, tenantId=1146029695717560320, journalId=1190317699101192196, language=CN, title=基于药辅合一理念的川芎与当归挥发油透皮作用研究, columnId=1190352405612040510, journalTitle=中国药学杂志, columnName=论著, runingTitle=null, highlight=null, articleAbstract=

目的 结合中药挥发油“药辅合一”的特性优势,研究川芎挥发油(Chuanxiong Rhizoma volatile oil,LVO)、当归挥发油(Angelicae Sinensis Radix volatile oil,AVO)提取方式、质量控制及其经皮渗透性与促透皮作用。方法 采用正交设计优选挥发油提取工艺;通过LVO、AVO物理参数与指标成分含量检测进行其质量控制;采用Franz扩散池法进行体外透皮研究,高效液相色谱(HPLC)法测定指标成分藁本内酯与白藜芦醇(resveratrol,RSV)的含量,以稳态透皮速率(Jss)和增透倍数(EF)为评价指标,分别考察不同浓度LVO、AVO及混合使用经皮渗透性及促透皮作用的影响。结果 确定LVO、AVO提取工艺为8倍量水,蒸馏时间8 h,用乙酸乙酯收集,无水硫酸钠脱水后减压挥干;对LVO、AVO建立了完善的质量控制方法,其中包括性状、相对密度、pH值、旋光度、黏度及藁本内酯含量测定;LVO、AVO均具有良好的渗透性和促RSV体外透皮作用,10 h内Jss最大分别为(48.23±6.53)(27.84±3.03)μg·cm-2·h-1,EF值最大分别为4.51、19.34。。结论 LVO、AVO单独和混合使用均具有良好的渗透性和促RSV透皮吸收作用,LVO的经皮渗透性明显高于AVO,而AVO的促渗透性则高于LVO。LVO、AVO良好的经皮渗透性与透皮吸收促进作用相结合,起到“药辅合一”的作用,具有良好的临床使用优势,为其在中药透皮给药制剂的开发和应用奠定基础。

, correspAuthors=张爱军, authorNote=null, correspAuthorsNote=
* 张爱军,女,硕士,研究员 研究方向:中药新制剂及成药性药学评价 Tel:13880898533
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朱红梅,女,硕士,助理研究员 研究方向:中药新药开发与制剂研究

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朱红梅,女,硕士,助理研究员 研究方向:中药新药开发与制剂研究

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朱红梅,女,硕士,助理研究员 研究方向:中药新药开发与制剂研究

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Hebei Med J (河北医药), 2019, 41(13):2043-2048., articleTitle=Research progress on resveratrol and resveratrol derivants, refAbstract=null), Reference(id=1196079251305841232, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, doi=null, pmid=null, pmcid=null, year=2019, volume=28, issue=3, pageStart=212, pageEnd=218, url=null, language=null, rfNumber=[20], rfOrder=19, authorNames=ZHUANG Y, HUANG H J, QUAN M M, journalName=Acta Laser Biol Sin, refType=null, unstructuredReference=ZHUANG Y, HUANG H J, QUAN M M, et al. Biological properties and its mechanism of resveratrol[J]. 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P(2020)Vol Ⅳ(中国药典 2020年版:四部)[S]. 2020., articleTitle=null, refAbstract=null), Reference(id=1196079251456836178, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, doi=null, pmid=null, pmcid=null, year=2014, volume=20, issue=14, pageStart=224, pageEnd=227, url=null, language=null, rfNumber=[22], rfOrder=21, authorNames=WU Z F, WANG S J, YANG M, journalName=Chin J Exp Tradit Med Form, refType=null, unstructuredReference=WU Z F, WANG S J, YANG M, et al. Research status and problem analysis of extraction process of volatile oil from traditional Chinese medicine[J]. 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Chin J Exp Tradit Med Form (中国实验方剂学杂志), 2016, 22(7):15-18., articleTitle=Effects of different permeation enhancer on in vitro percutaneous absorption of naringin in sanwei dieda rheumatism gel paste, refAbstract=null), Reference(id=1196079251637191252, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, doi=null, pmid=null, pmcid=null, year=null, volume=12, issue=11, pageStart=2795, pageEnd=2798, url=null, language=null, rfNumber=[24], rfOrder=23, authorNames=WANG M, DONG X R, LI J, journalName=World Chin Med(世界中医药)2017, refType=null, unstructuredReference=WANG M, DONG X R, LI J, et al. Effects of different enhancers on in vitro transdermal absorption of artemether ointment[J]. World Chin Med(世界中医药)2017, 12(11):2795-2798., articleTitle=Effects of different enhancers on in vitro transdermal absorption of artemether ointment, refAbstract=null), Reference(id=1196079251700105813, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, doi=null, pmid=null, pmcid=null, year=2006, volume=32, issue=6, pageStart=747, pageEnd=755, url=null, language=null, rfNumber=[25], rfOrder=24, authorNames=CUI F, FENG L, HU J, journalName=Drug Dev Ind Pharm, refType=null, unstructuredReference=CUI F, FENG L, HU J, et al. Factors affecting stability of z-ligustilide in the volatile oil of radix angelicae sinensis and ligusticum chuanxiong and its stability prediction[J]. Drug Dev Ind Pharm, 2006, 32(6):747-755., articleTitle=Factors affecting stability of z-ligustilide in the volatile oil of radix angelicae sinensis and ligusticum chuanxiong and its stability prediction, refAbstract=null)], funds=[Fund(id=1196079248516629051, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, awardId=2020YFS0566, language=CN, fundingSource=四川省科技计划重点研发项目(2020YFS0566), fundOrder=null, country=null), Fund(id=1196079248583737916, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, awardId=A-2020N-18, language=CN, fundingSource=四川省公益性科研院所基本科研项目资助[2020JDKY0013(A-2020N-18), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1196079243647042055, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, xref=null, ext=[AuthorCompanyExt(id=1196079243659624968, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, companyId=1196079243647042055, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Sichuan Academy of Traditional Chinese Medicine, Chengdu 610041, China), AuthorCompanyExt(id=1196079243668013577, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, companyId=1196079243647042055, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=四川省中医药科学院, 成都 610041)])], figs=[ArticleFig(id=1196079246268482083, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Fig.1, caption=The chromatomap of sample and blank of the specificity study, figureFileSmall=QY2ElMIbN2VeBbWry8A0RA==, figureFileBig=bTsefmtZlC6pIA35hIJkpw==, tableContent=null), ArticleFig(id=1196079246323008036, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=图1, caption=专属性研究中样品溶液与空白溶剂色谱图, figureFileSmall=QY2ElMIbN2VeBbWry8A0RA==, figureFileBig=bTsefmtZlC6pIA35hIJkpw==, tableContent=null), ArticleFig(id=1196079246415282725, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Fig.2, caption=The curve of Jss and EF of LVO and AVO in different concentrations, figureFileSmall=6IEwfUWvkMS0sFiT/hdQuA==, figureFileBig=gk4U2ZnyacWUp5EmawkSqg==, tableContent=null), ArticleFig(id=1196079246469808678, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=图2, caption=不同浓度 LVO、AVO 稳态透皮速率(Jss)与增透倍数(EF)曲线图, figureFileSmall=6IEwfUWvkMS0sFiT/hdQuA==, figureFileBig=gk4U2ZnyacWUp5EmawkSqg==, tableContent=null), ArticleFig(id=1196079246524334631, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.1, caption=

Qrthogonal design results with analysis of variance of volatile oil extraction process

, figureFileSmall=null, figureFileBig=null, tableContent=
Factor A/times B/h C/h D(blank) Results
1 1 1 1 1 0.204 2
2 1 2 2 2 0.304 2
3 1 3 3 3 0.291 9
4 2 1 2 3 0.444 7
5 2 2 3 1 0.411 9
6 2 3 1 2 0.240 7
7 3 1 3 2 0.541 3
8 3 2 1 3 0.383 5
9 3 3 2 1 0.442 8
K1 0.267 0.397 0.276 0.353
K2 0.366 0.367 0.397 0.362
K3 0.456 0.325 0.415 0.373
S 0.189 0.072 0.139 0.02
Factor SS f F Critical value P
A 0.054 2 54 19 1)
B 0.008 2 8 19
C 0.034 2 34 19 1)
SSE 0 2
), ArticleFig(id=1196079246599832104, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表1, caption=

挥发油提取工艺正交设计结果与方差分析

, figureFileSmall=null, figureFileBig=null, tableContent=
Factor A/times B/h C/h D(blank) Results
1 1 1 1 1 0.204 2
2 1 2 2 2 0.304 2
3 1 3 3 3 0.291 9
4 2 1 2 3 0.444 7
5 2 2 3 1 0.411 9
6 2 3 1 2 0.240 7
7 3 1 3 2 0.541 3
8 3 2 1 3 0.383 5
9 3 3 2 1 0.442 8
K1 0.267 0.397 0.276 0.353
K2 0.366 0.367 0.397 0.362
K3 0.456 0.325 0.415 0.373
S 0.189 0.072 0.139 0.02
Factor SS f F Critical value P
A 0.054 2 54 19 1)
B 0.008 2 8 19
C 0.034 2 34 19 1)
SSE 0 2
), ArticleFig(id=1196079246696301097, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.2, caption=

The results of physical constant determination of LVO and AVO

, figureFileSmall=null, figureFileBig=null, tableContent=
Parameter LVO AVO
CX210701 CX210603 CX211108 DG201101 DG210501 DG211108
d(20 ℃) 0.996 2 0.992 8 1.030 7 1.019 1 1.005 2 1.038 3
Average d(20 ℃) 1.006 6 1.020 9
pH 4.47 4.31 4.15 4.14 4.36 3.97
Average pH 4.31 4.16
[α] -6.37° -5.57° -6.39° +0.99° +0.79° +0.98°
Average [α] -6.11° +0.92°
ν 6.697 2 6.779 9 6.649 8 6.127 6 6.345 8 6.227 4
Average ν 6.708 9 6.233 6
), ArticleFig(id=1196079246788575786, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表2, caption=

川芎挥发油(LVO)和当归挥发油(AVO)物理常数测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
Parameter LVO AVO
CX210701 CX210603 CX211108 DG201101 DG210501 DG211108
d(20 ℃) 0.996 2 0.992 8 1.030 7 1.019 1 1.005 2 1.038 3
Average d(20 ℃) 1.006 6 1.020 9
pH 4.47 4.31 4.15 4.14 4.36 3.97
Average pH 4.31 4.16
[α] -6.37° -5.57° -6.39° +0.99° +0.79° +0.98°
Average [α] -6.11° +0.92°
ν 6.697 2 6.779 9 6.649 8 6.127 6 6.345 8 6.227 4
Average ν 6.708 9 6.233 6
), ArticleFig(id=1196079246855684651, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.3, caption=

The results of ligustilide content determination of LVO and AVO

, figureFileSmall=null, figureFileBig=null, tableContent=
Parameter LVO AVO
CX210701 CX210603 CX20211108 DG201101 DG210501 DG20211108
Percent content 26.00 30.07 36.65 57.25 57.00 53.84
Average percent content 30.91 56.03
-30% of average percent content 21.63 39.22
), ArticleFig(id=1196079246956347948, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表3, caption=

LVO和AVO中藁本内酯含量测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
Parameter LVO AVO
CX210701 CX210603 CX20211108 DG201101 DG210501 DG20211108
Percent content 26.00 30.07 36.65 57.25 57.00 53.84
Average percent content 30.91 56.03
-30% of average percent content 21.63 39.22
), ArticleFig(id=1196079247157674541, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.4, caption=

The investigation results of receive liquid of in vitro transdermal experiment. n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Item
sampling time/h		 RSV-Qn/μg·cm-2
Physiological saline Phosphate buffer
1 0.98±0.70 1.69±0.69
2 28.03±5.82 17.14±7.14
4 83.36±13.59 67.19±27.63
6 146.73±11.88 115.03±40.05
10 218.15±21.76 168.86±36.10
Qn-t y=24.551 x-17.484 y=19.178 x-14.24
r 0.985 9 0.981 3
Jss 24.551±2.143 19.178±4.096
), ArticleFig(id=1196079247317058094, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表4, caption=

体外透皮实验接收液考察结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Item
sampling time/h		 RSV-Qn/μg·cm-2
Physiological saline Phosphate buffer
1 0.98±0.70 1.69±0.69
2 28.03±5.82 17.14±7.14
4 83.36±13.59 67.19±27.63
6 146.73±11.88 115.03±40.05
10 218.15±21.76 168.86±36.10
Qn-t y=24.551 x-17.484 y=19.178 x-14.24
r 0.985 9 0.981 3
Jss 24.551±2.143 19.178±4.096
), ArticleFig(id=1196079247400944175, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.5, caption=

The results of percutaneous permeability of LVO in different concentrations. n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 Ligustilide-Qn/μg·cm-2
0.5% LVO 1% LVO 2% LVO 3% LVO 4% LVO
1 18.58±5.85 10.48±3.08 35.68±31.06 27.97±25.19 33.16±17.94
2 43.95±13.23 31.03±5.31 77.40±45.13 83.37±62.12 93.40±32.37
4 127.95±31.49 105.98±9.80 201.38±74.40 178.37±99.14 202.63±42.10
6 202.61±33.37 174.32±14.90 300.39±76.63 261.22±118.49 298.73±49.06
8 265.72±31.03 250.82±23.06 392.06±91.64 323.37±112.32 363.25±64.45
10 293.55±29.02 300.06±20.84 455.51±87.31 388.38±118.74 433.66±55.08
Qn-t y=32.46 x-8.98 y=33.54 x-27.87 y=48.23 x-5.46 y=39.82 x+4.70 y=44.45 x+7.83
r 0.981 4 0.996 3 0.991 0 0.989 7 0.986 9
Jss 32.46±2.98 33.54±2.33 48.23±6.53 39.82±9.43 44.45±4.59
), ArticleFig(id=1196079247526773296, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表5, caption=

不同浓度LVO经皮渗透性结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 Ligustilide-Qn/μg·cm-2
0.5% LVO 1% LVO 2% LVO 3% LVO 4% LVO
1 18.58±5.85 10.48±3.08 35.68±31.06 27.97±25.19 33.16±17.94
2 43.95±13.23 31.03±5.31 77.40±45.13 83.37±62.12 93.40±32.37
4 127.95±31.49 105.98±9.80 201.38±74.40 178.37±99.14 202.63±42.10
6 202.61±33.37 174.32±14.90 300.39±76.63 261.22±118.49 298.73±49.06
8 265.72±31.03 250.82±23.06 392.06±91.64 323.37±112.32 363.25±64.45
10 293.55±29.02 300.06±20.84 455.51±87.31 388.38±118.74 433.66±55.08
Qn-t y=32.46 x-8.98 y=33.54 x-27.87 y=48.23 x-5.46 y=39.82 x+4.70 y=44.45 x+7.83
r 0.981 4 0.996 3 0.991 0 0.989 7 0.986 9
Jss 32.46±2.98 33.54±2.33 48.23±6.53 39.82±9.43 44.45±4.59
), ArticleFig(id=1196079247606465073, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.6, caption=

The results of percutaneous permeability of AVO in different concentrations.n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 Ligustilide-Qn/μg·cm-2
0.5% AVO 1% AVO 2% AVO 3% AVO 4% AVO
1 2.91±2.65 14.48±3.92 8.81±1.63 4.75±1.02 10.06±0.98
2 15.44±3.66 39.70±19.34 29.23±5.69 20.28±5.29 29.74±4.14
4 50.80±6.82 95.46±48.63 80.81±9.34 70.95±15.43 83.03±12.06
6 86.29±9.35 148.55±82.35 135.66±12.67 129.26±20.91 140.60±18.60
8 118.65±11.46 200.57±102.38 183.61±16.66 191.08±25.21 201.45±22.79
10 157.60±15.09 255.35±121.72 134.35±9.43 246.26±31.74 256.33±25.61
Qn-t y=17.25 x-17.19 y=26.78 x-12.66 y=25.35 x-18.92 y=27.49 x-31.63 y=27.84 x-23.66
r 0.998 8 0.999 9 0.999 5 0.996 2 0.998 4
Jss 17.25±1.36 26.78±13.36 25.35±1.09 27.49±3.40 27.84±3.03
), ArticleFig(id=1196079247669379634, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表6, caption=

不同浓度 AVO 经皮渗透性结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 Ligustilide-Qn/μg·cm-2
0.5% AVO 1% AVO 2% AVO 3% AVO 4% AVO
1 2.91±2.65 14.48±3.92 8.81±1.63 4.75±1.02 10.06±0.98
2 15.44±3.66 39.70±19.34 29.23±5.69 20.28±5.29 29.74±4.14
4 50.80±6.82 95.46±48.63 80.81±9.34 70.95±15.43 83.03±12.06
6 86.29±9.35 148.55±82.35 135.66±12.67 129.26±20.91 140.60±18.60
8 118.65±11.46 200.57±102.38 183.61±16.66 191.08±25.21 201.45±22.79
10 157.60±15.09 255.35±121.72 134.35±9.43 246.26±31.74 256.33±25.61
Qn-t y=17.25 x-17.19 y=26.78 x-12.66 y=25.35 x-18.92 y=27.49 x-31.63 y=27.84 x-23.66
r 0.998 8 0.999 9 0.999 5 0.996 2 0.998 4
Jss 17.25±1.36 26.78±13.36 25.35±1.09 27.49±3.40 27.84±3.03
), ArticleFig(id=1196079247732294195, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.7, caption=

The results of percutaneous permeability of LVO and AVO mixing in different concentrations. n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 Ligustilide-Qn/μg·cm-2
2% AVO+0.5% LVO 2% AVO+1% LVO 2% LVO+0.5% AVO 2% LVO+1% AVO
1 6.29±4.76 2.85±0.70 13.37±5.50 12.39±7.12
2 30.80±13.18 16.66±1.41 46.52±15.14 41.28±19.10
4 99.04±27.56 60.45±7.16 118.08±29.18 108.22±33.00
6 171.63±40.87 115.23±1.27 187.71±35.12 173.10±38.08
8 250.59±53.23 171.00±3.43 250.53±40.86 237.92±43.43
10 314.82±58.96 219.88±1.67 309.24±45.14 297.92±46.91
Qn-t y=35.10 x-35.84 y=24.79 x-30.40 y=33.21 x-17.33 y=32.04 x-20.43
r 0.998 2 0.995 4 0.998 7 0.999 7
Jss 35.10±6.18 24.79±0.57 33.21±4.25 32.04±4.16
), ArticleFig(id=1196079247799403060, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表7, caption=

不同浓度 LVO+AVO 混合经皮渗透性结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 Ligustilide-Qn/μg·cm-2
2% AVO+0.5% LVO 2% AVO+1% LVO 2% LVO+0.5% AVO 2% LVO+1% AVO
1 6.29±4.76 2.85±0.70 13.37±5.50 12.39±7.12
2 30.80±13.18 16.66±1.41 46.52±15.14 41.28±19.10
4 99.04±27.56 60.45±7.16 118.08±29.18 108.22±33.00
6 171.63±40.87 115.23±1.27 187.71±35.12 173.10±38.08
8 250.59±53.23 171.00±3.43 250.53±40.86 237.92±43.43
10 314.82±58.96 219.88±1.67 309.24±45.14 297.92±46.91
Qn-t y=35.10 x-35.84 y=24.79 x-30.40 y=33.21 x-17.33 y=32.04 x-20.43
r 0.998 2 0.995 4 0.998 7 0.999 7
Jss 35.10±6.18 24.79±0.57 33.21±4.25 32.04±4.16
), ArticleFig(id=1196079247874900533, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.8, caption=

The results of the transdermal effect on promoting RSV of LVO in different concentrations.n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 RSV-Qn/μg·cm-2
Blank 0.5% LVO 1% LVO 2% LVO 3% LVO 4% LVO
1 1.83±2.47 6.99±8.73 4.00±1.10 37.42±38.54 21.80±18.73 28.19±11.97
2 7.45±9.55 21.47±20.64 17.31±2.76 86.61±60.65 73.31±45.23 80.54±23.43
4 20.00±21.42 65.91±46.50 64.88±7.20 195.95±77.16 159.71±68.80 160.76±23.19
6 36.15±35.67 115.25±61.66 112.59±8.56 272.58±85.91 220.85±59.12 228.87±24.76
8 54.36±54.42 158.50±62.83 152.10±11.79 321.34±108.32 274.85±57.44 268.71±25.44
10 73.00±66.17 185.51±52.51 178.07±9.70 351.92±115.61 310.73±53.28 297.90±22.11
Qn-t y=7.93 x-8.84 y=20.84 x-15.40 y=20.36 x-17.01 y=35.83 x+25.84 y=32.14 x+10.81 y=30.11 x+21.91
r 0.992 4 0.993 6 0.991 8 0.956 5 0.974 1 0.960 5
Jss 7.93±7.20 20.84±5.36 20.35±1.07 35.83±11.92 32.14±3.13 30.11±1.19
EF - 2.63 2.57 4.51 4.05 3.80
), ArticleFig(id=1196079248000729654, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表8, caption=

不同浓度LVO促白藜芦醇(RSV)体外透皮作用结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 RSV-Qn/μg·cm-2
Blank 0.5% LVO 1% LVO 2% LVO 3% LVO 4% LVO
1 1.83±2.47 6.99±8.73 4.00±1.10 37.42±38.54 21.80±18.73 28.19±11.97
2 7.45±9.55 21.47±20.64 17.31±2.76 86.61±60.65 73.31±45.23 80.54±23.43
4 20.00±21.42 65.91±46.50 64.88±7.20 195.95±77.16 159.71±68.80 160.76±23.19
6 36.15±35.67 115.25±61.66 112.59±8.56 272.58±85.91 220.85±59.12 228.87±24.76
8 54.36±54.42 158.50±62.83 152.10±11.79 321.34±108.32 274.85±57.44 268.71±25.44
10 73.00±66.17 185.51±52.51 178.07±9.70 351.92±115.61 310.73±53.28 297.90±22.11
Qn-t y=7.93 x-8.84 y=20.84 x-15.40 y=20.36 x-17.01 y=35.83 x+25.84 y=32.14 x+10.81 y=30.11 x+21.91
r 0.992 4 0.993 6 0.991 8 0.956 5 0.974 1 0.960 5
Jss 7.93±7.20 20.84±5.36 20.35±1.07 35.83±11.92 32.14±3.13 30.11±1.19
EF - 2.63 2.57 4.51 4.05 3.80
), ArticleFig(id=1196079248076227127, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.9, caption=

The results of the transdermal effect on promoting RSV of AVO in different concentrations.n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 RSV-Qn/μg·cm-2
Blank 0.5% AVO 1% AVO 2% AVO 3% AVO 4% AVO
4 0±0 0±0 0.50±0.54 0.69±0.32 0.98±0.53 2.13±1.89
6 0.11±0.20 0±0 1.27±1.17 4.49±5.87 4.16±0.61 3.74±3.32
8 0.75±0.69 1.25±0.11 7.30±7.18 8.82±3.59 9.17±4.50 14.92±17.20
10 1.38±0.84 4.37±0.66 27.36±26.85 29.58±5.19 28.18±10.66 29.11±18.12
Qn-t y=0.24 x-1.11 y=0.718 x-3.62 y=4.33 x-21.21 y=4.55 x-20.96 y=4.33 x-19.70 y=4.61 x-19.765
r 0.933 5 0.807 9 0.794 5 0.830 4 0.842 6 0.910 5
Jss 0.24±0.15 0.72±0.10 4.33±4.25 4.55±0.62 4.33±1.71 4.61±3.14
EF - 3.02 18.68 19.11 18.19 19.34
), ArticleFig(id=1196079248168501816, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表9, caption=

不同浓度AVO促RSV体外透皮作用结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 RSV-Qn/μg·cm-2
Blank 0.5% AVO 1% AVO 2% AVO 3% AVO 4% AVO
4 0±0 0±0 0.50±0.54 0.69±0.32 0.98±0.53 2.13±1.89
6 0.11±0.20 0±0 1.27±1.17 4.49±5.87 4.16±0.61 3.74±3.32
8 0.75±0.69 1.25±0.11 7.30±7.18 8.82±3.59 9.17±4.50 14.92±17.20
10 1.38±0.84 4.37±0.66 27.36±26.85 29.58±5.19 28.18±10.66 29.11±18.12
Qn-t y=0.24 x-1.11 y=0.718 x-3.62 y=4.33 x-21.21 y=4.55 x-20.96 y=4.33 x-19.70 y=4.61 x-19.765
r 0.933 5 0.807 9 0.794 5 0.830 4 0.842 6 0.910 5
Jss 0.24±0.15 0.72±0.10 4.33±4.25 4.55±0.62 4.33±1.71 4.61±3.14
EF - 3.02 18.68 19.11 18.19 19.34
), ArticleFig(id=1196079248256582201, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=EN, label=Tab.10, caption=

The results of the transdermal effect on promoting RSV of LVO and AVO mixing in different concentrations. n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 RSV-Qn/μg·cm-2
Blank 2% AVO+0.5% LVO 2% AVO+1% LVO 2% LVO+0.5% AVO 2% LVO+1% AVO
1 0.85±0.76 5.89±5.04 1.66±0.26 11.95±5.76 11.20±5.98
2 4.41±2.57 29.91±18.50 10.94±1.10 44.85±14.46 41.97±14.85
4 11.69±5.41 91.30±40.92 44.13±8.36 110.82±24.20 104.90±23.13
6 20.16±6.94 154.96±59.51 78.52±0.78 175.03±28.25 162.34±29.14
8 27.59±6.46 220.08±63.88 112.95±1.13 229.35±28.72 213.48±24.30
10 36.46±7.66 264.37±63.38 140.64±0.92 267.57±26.66 252.05±23.10
Qn-t y=3.95 x-3.53 y=29.67 x-25.50 y=15.98 x-17.76 y=29.06 x-10.22 y=27.23 x-9.69
r 0.999 0 0.997 4 0.997 6 0.992 0 0.993 5
Jss 3.95±0.71 29.67±6.73 15.98±0.35 29.06±2.23 27.23±1.57
EF - 7.52 4.05 7.36 6.90
), ArticleFig(id=1196079248374022714, tenantId=1146029695717560320, journalId=1190317699101192196, articleId=1195814100342190965, language=CN, label=表10, caption=

不同浓度 LVO+AVO 混合促 RSV 体外透皮作用结果。n=3,$\bar{x}±s$

, figureFileSmall=null, figureFileBig=null, tableContent=
Sampling
time/h		 RSV-Qn/μg·cm-2
Blank 2% AVO+0.5% LVO 2% AVO+1% LVO 2% LVO+0.5% AVO 2% LVO+1% AVO
1 0.85±0.76 5.89±5.04 1.66±0.26 11.95±5.76 11.20±5.98
2 4.41±2.57 29.91±18.50 10.94±1.10 44.85±14.46 41.97±14.85
4 11.69±5.41 91.30±40.92 44.13±8.36 110.82±24.20 104.90±23.13
6 20.16±6.94 154.96±59.51 78.52±0.78 175.03±28.25 162.34±29.14
8 27.59±6.46 220.08±63.88 112.95±1.13 229.35±28.72 213.48±24.30
10 36.46±7.66 264.37±63.38 140.64±0.92 267.57±26.66 252.05±23.10
Qn-t y=3.95 x-3.53 y=29.67 x-25.50 y=15.98 x-17.76 y=29.06 x-10.22 y=27.23 x-9.69
r 0.999 0 0.997 4 0.997 6 0.992 0 0.993 5
Jss 3.95±0.71 29.67±6.73 15.98±0.35 29.06±2.23 27.23±1.57
EF - 7.52 4.05 7.36 6.90
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基于药辅合一理念的川芎与当归挥发油透皮作用研究
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朱红梅 , 张爱军 * , 李帅
中国药学杂志 | 论著 2024,59(23): 2249-2257
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中国药学杂志 | 论著 2024, 59(23): 2249-2257
基于药辅合一理念的川芎与当归挥发油透皮作用研究
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朱红梅, 张爱军*, 李帅
作者信息
  • 四川省中医药科学院, 成都 610041

    • 朱红梅,女,硕士,助理研究员 研究方向:中药新药开发与制剂研究

通讯作者:

* 张爱军,女,硕士,研究员 研究方向:中药新制剂及成药性药学评价 Tel:13880898533
Chuanxiong Rhizoma and Angelicase Sinensis Radix Volatile Oil Study and Transdermal Effect Based on the Concept of Unification of Drugs and Excipients
Hongmei ZHU, Aijun ZHANG*, Shuai LI
Affiliations
  • Sichuan Academy of Traditional Chinese Medicine, Chengdu 610041, China
出版时间: 2024-12-08 doi: 10.11669/cpj.2024.23.006
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摘要
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目的 结合中药挥发油“药辅合一”的特性优势,研究川芎挥发油(Chuanxiong Rhizoma volatile oil,LVO)、当归挥发油(Angelicae Sinensis Radix volatile oil,AVO)提取方式、质量控制及其经皮渗透性与促透皮作用。方法 采用正交设计优选挥发油提取工艺;通过LVO、AVO物理参数与指标成分含量检测进行其质量控制;采用Franz扩散池法进行体外透皮研究,高效液相色谱(HPLC)法测定指标成分藁本内酯与白藜芦醇(resveratrol,RSV)的含量,以稳态透皮速率(Jss)和增透倍数(EF)为评价指标,分别考察不同浓度LVO、AVO及混合使用经皮渗透性及促透皮作用的影响。结果 确定LVO、AVO提取工艺为8倍量水,蒸馏时间8 h,用乙酸乙酯收集,无水硫酸钠脱水后减压挥干;对LVO、AVO建立了完善的质量控制方法,其中包括性状、相对密度、pH值、旋光度、黏度及藁本内酯含量测定;LVO、AVO均具有良好的渗透性和促RSV体外透皮作用,10 h内Jss最大分别为(48.23±6.53)(27.84±3.03)μg·cm-2·h-1,EF值最大分别为4.51、19.34。。结论 LVO、AVO单独和混合使用均具有良好的渗透性和促RSV透皮吸收作用,LVO的经皮渗透性明显高于AVO,而AVO的促渗透性则高于LVO。LVO、AVO良好的经皮渗透性与透皮吸收促进作用相结合,起到“药辅合一”的作用,具有良好的临床使用优势,为其在中药透皮给药制剂的开发和应用奠定基础。

川芎  /  当归  /  挥发油  /  质量控制  /  经皮渗透

OBJECTIVE To study the extraction method, quality, transdermal permeability, and transdermal promoting effects of Chuanxiong Rhizoma Hort. volatile oil (LVO) and Angelicase Sinensis Radix volatile oil (AVO) combined with the characteristic advantages of “unification of drugs and excipients” of traditional Chinese medicine volatile oil. METHODS The orthogonal design was used to screen and optimize extraction process of LVO and AVO. Quality control was carried out by detecting the physical parameters and indicator component content of LVO and AVO. Franz diffusion cells were used to conduct the studies of skin permeation in vitro. The contents of ligustilide and resveratrol(RSV) were determined by HPLC, and the steady-state transdermal rate(Jss) and enhancement factor (EF) compared with blank were used as evaluation indicators. And the transdermal permeability and transdermal promoting effects of different concentrations of LVO, AVO and mixed were investigated respectively. RESULTS The extraction process for LVO and AVO involves 8 times the amount of water, heating and refluxing for 8 h, collecting with ethyl acetate, dehydrating anhydrous sodium sulfate, and then volatilizing under reduced pressure. Perfect quality control methods were established for LVO and AVO, including the properties, relative density, pH, optical rotation, viscosity and ligustilide content. LVO and AVO have good permeability and promote RSV transdermal effect in vitro, and the maximum Jss values within 10 h were (48.23±6.53) and (27.84±3.03), respectively, and the maximum EF values were 4.51 and 19.34, respectively. CONCLUSION LVO and AVO have good transdermal permeability and transdermal promoting effects of RSV when used alone or in combination. The transdermal permeability of LVO was significantly higher than that of AVO, while the transdermal promoting effects of AVO was higher than that of LVO. The combination of LVO and AVO's own pharmacological effects and good transdermal absorption promoting effect plays a role of “unification of drugs and excipients”, which has a good advantage in clinical use, and lays a foundation for their development and application in transdermal drug delivery preparations of traditional Chinese medicine.

Chuanxiong Rhizoma Hort.  /  Angelicase Sinensis Radix  /  volatile oil  /  quality  /  transdermal permeability
朱红梅, 张爱军, 李帅. 基于药辅合一理念的川芎与当归挥发油透皮作用研究. 中国药学杂志, 2024 , 59 (23) : 2249 -2257 . DOI: 10.11669/cpj.2024.23.006
Hongmei ZHU, Aijun ZHANG, Shuai LI. Chuanxiong Rhizoma and Angelicase Sinensis Radix Volatile Oil Study and Transdermal Effect Based on the Concept of Unification of Drugs and Excipients[J]. Chinese Pharmaceutical Journal, 2024 , 59 (23) : 2249 -2257 . DOI: 10.11669/cpj.2024.23.006
正文
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中药挥发油的“药辅合一”是指其在发挥自身药理作用的同时,还可利用其形、色、气、味等理化特性和助悬、分散、吸附、助磨、增稠、促渗等功能特性来作为辅料以辅助制剂的成型与稳定[1-4]。国内外学者对中药挥发油的物质基础和药理作用等做了一定的研究,但基于“药辅合一”理论,将其既作为辅料又作为药效物质用于中药制剂中的研究还相对较少[5-9]。
近年来,中药挥发油作为经皮给药促透剂的研究已成热点。传统中医理论认为,中药中的挥发油或芳香成分具有芳香走窜的性质,可快速发挥疗效;现代药物研究表明,中药挥发油相对分子质量小,脂溶性强,能促进药物的吸收与跨膜转运,兼有促进渗透与治疗疾病的双重特点,可作为透皮促进剂。与现有化学合成的透皮吸收促进剂相比,中药挥发油属于天然的透皮吸收促进剂,无致敏性,对皮肤刺激性小,不良反应少,而且还具有一定的治疗效果,能够与透皮吸收的药物产生协同作用[10-12],正日益受到广泛关注。
川芎和当归作为传统中药,组成的药对是应用中常见的配伍形式,具有理气活血,化瘀止痛的功效[13-15],两者的挥发油成分含量较大亦具有一定的药理作用。川芎挥发油(Chuanxiong Rhizoma volatile oil,LVO)具有解热、镇痛镇静、改善血流变、保护神经细胞、抗炎、降压等多种药效作用[16]。当归挥发油(Angelicae Sinensis Radix volatile oil,AVO)具有活血化瘀、行气止痛、降血压、降血脂、保护心肌细胞、抑菌、抗氧化、抑制消化道平滑肌、抑制气管平滑肌、抗肿瘤等作用[17-18]。并且LVO、AVO作为中药挥发油,无皮肤刺激性,所含的成分多为小分子脂溶性化合物,自身具有较好的透皮吸收特性和特殊的理化性质,既能作为“药”又能作为“辅”用于中药制剂。白藜芦醇(resveratrol,RSV)是由多种植物产生的非黄酮类多酚化合物,被认为是天然植物抗毒素,具有抗肿瘤、抗氧化、抗炎、抗辐射、抗衰老、抗自由基、心血管保护和免疫调节等生物学功能,已广泛应用于食品、美容、保健品和化工等领域[19-20]。
本实验对LVO、AVO的提取工艺、质量控制方法、经皮渗透性进行研究,并以RSV为透皮吸收对象,研究LVO、AVO单独使用与混合使用的促透皮作用。以期阐明中药挥发油在透皮给药的优势,促进LVO、AVO在中药透皮给药制剂开发中的应用。
1 仪器与材料
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1.1 仪器与设备
Agilent1290型超高效液相色谱仪(美国Agilent公司);AUW220D型电子天平(日本岛津公司);TK-12D型透皮扩散试验仪(上海锴凯科技贸易有限公司);ZNCL-TS智能磁力搅拌器(上海标和仪器有限公司);B-100Bath旋转蒸发仪(瑞士BUCHI公司);DZKW-4型恒温水浴锅(北京中兴伟业仪器有限公司);KQ-300B型超声波清洗器(昆山市超声仪器有限公司);JY5002型电子天平(上海良平仪器有限公司);DB207型电热恒温鼓风干燥箱(成都电烘箱厂);雷磁PHS-25型pH计(上海仪电科学仪器股份有限公司);WZZ-2S型数显自动旋光仪(上海索光光电技术有限公司);挥发油测定器、比重瓶、平式毛细管黏度计(成都科隆化学品有限责任公司)。
1.2 药品与试剂
川芎(批号:20210624、201603、210701,四川,经鉴定为伞形科植物川芎(Ligusticum chuangxiong Hort.)的干燥根茎,检验均合格);当归[批号:20211012、210501、201101,甘肃,经鉴定为伞形科植物当归[Angelica sinensis (Oliv.) Diels]的干燥根,检验均合格];川芎挥发油(批号:CX21108、CX201603、CX210701,实验室自制);当归挥发油(批号:DG211108、DG210501、DG201101,实验室自制);藁本内酯(批号:RFS-G01001909024,纯度:98.443%,成都瑞芬思生物科技有限公司);白藜芦醇(批号:R19J12S138196,纯度:99.6%,上海源叶生物科技有限公司)。
乙酸乙酯、无水硫酸钠、磷酸氢二钾、磷酸二氢钾、无水乙醇、磷酸均为分析纯;体积分数95%乙醇为食用级;氯化钠注射液(B20093704,四川科伦药业股份有限公司);甲醇、乙腈为色谱级。
1.3 实验动物
健康雄性 SPF 级昆明种小鼠,体质量(20±2)g,由四川省中医药科学院动物中心提供,实验动物许可证号为:SYXK(川)2018-100,饲养于四川省中医药科学院动物中心SPF屏障系统,温度20~22 ℃,相对湿度50%左右,每日定时喂养,12 h/12 h明暗交替,实验方案由四川省中医药科学院动物伦理委员会批准。
2 方法与结果
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2.1 提取工艺研究
中药挥发油传统的提取方法技术成熟,但有提取率低、杂质多、加热时间长、热敏性成分易破坏、选择性差等不足。并且LVO、AVO的相对密度接近水,采收期和干燥方式也会影响挥发油的相对密度,在蒸馏后油水不易分离,制备得到的挥发油纯度较低,且药材的粉碎程度对其挥发油一定的影响。
LVO、AVO均采用水蒸汽蒸馏,乙酸乙酯收集,无水硫酸钠脱水后减压挥干,以得油率和藁本内酯含量综合评分为考察指标,正交实验进行工艺优化:加水量A(4、6、8倍),浸泡时间B(0、1、2 h),蒸馏时间C(4、6、8 h)确定挥发油最佳提取工艺,实验设计与方差分析结果见表1。结果显示,加水量和蒸馏时间对LVO、AVO的提取有极显著性影响,浸泡时间均无显著性影响。结合生产实际与成本效益确定LVO、AVO的提取为8倍量水,蒸馏时间8 h,乙酸乙酯收集,无水硫酸钠脱水后减压挥干,于4 ℃下冷藏避光保存备用。
2.2 质量研究
LVO和AVO的质量控制主要集中在物理常数、挥发油中藁本内酯含量测定及相关性状分析。物理常数的测定包括相对密度、pH值、相对黏度、旋光度等;主要成分藁本内酯含量测定主要采用高效液相色谱法。
2.2.1 性状
LVO和AVO均为浅黄色或深黄色的澄清液体,气香,味甘、辛、微苦。在甲醇、乙醇、乙酸乙酯中易溶,几乎不溶于水。
2.2.2 物理常数
相对密度(d):参考《中国药典》2020年版四部[21]通则0601相对密度测定法,采用比重瓶法,于20 ℃下测定3批LVO、AVO的相对密度。
pH值:参考《中国药典》2020年版四部[21]通则0631 pH值测定法,3批LVO、AVO分别用无水乙醇稀释10倍后测定。
比旋度[α] :参考《中国药典》2020年版四部[21]通则0621旋光度测定法,3批LVO、AVO加乙醇制成每1 mL中含50 mg的溶液,分别测定旋光度。
运动黏度(ν):参考《中国药典》2020年版四部[21]通则0633黏度测定法,采用平式毛细管黏度计,分别测定3批LVO、AVO的运动黏度。
LVO、AVO物理常数测定结果见表2,两种挥发油的相对密度、pH值、旋光度及黏度均有一定差异。
2.2.3 藁本内酯含量测定
色谱条件: 采用Agilent Eclipse XDB-C18(4.6 mm×150 mm,5 μm)色谱柱;流动相:乙腈-水(75∶25);流速:1 mL·min-1;检测波长:350 nm;柱温:35 ℃;进样量:10 μL。
对照品溶液的制备: 取藁本内酯适量,精密称定,置棕色量瓶中,加甲醇制成每1 mL含50 μg的溶液,即得。
供试品溶液的制备: 取挥发油约10 mg,精密称定,加甲醇溶解至5 mL量瓶中,摇匀,即得。
线性关系考察: 精密配制质量浓度分别为5.38、26.92、67.30、134.59、201.89、269.18、403.77、672.96 μg·mL-1的藁本内酯对照品溶液,按上述色谱条件检测,以藁本内酯对照品浓度为横坐标(X),峰面积为纵坐标(Y),得回归方程为Y=18.877X+54.884,r=0.999 9,结果表明藁本内酯在5.383 6~672.956 μg·mL-1线性关系良好。
精密度考察:取同一供试品溶液,按上述色谱条件检测,重复进样6次,记录色谱峰面积并计算其RSD值。RSD值为0.27%,表明仪器精密度良好。
重复性试验:取同一批挥发油平行制备6份供试品溶液,按上述色谱条件检测,记录峰面积并计算藁本内酯含量的RSD值,RSD为0.78%,表明方法重复性良好。
稳定性试验:取一供试品溶液按上述色谱条件,分别在制备后的0、1、2、4、8、12、24 h进样,按“2.2.3”项下色谱条件检测,记录峰面积,并计算峰面积RSD值。结果,RSD值为0.27%,表明供试品溶液在24 h内稳定性良好。
加样回收率试验:分别按样品中藁本内酯量与加入量1∶0.5、1∶1、1∶1.5配制供试品溶液,每个浓度平行3份,按上述色谱条件检测,记录峰面积并计算平均加样回收率。结果平均加样回收率为99.77%,RSD值为0.85%,表明该方法准确性良好。
含量测定:根据上述方法分别测定3批LVO、AVO中藁本内酯的含量,结果见表3,并根据平均百分含量-30%为含量最低值设定限度。对比分析,AVO中藁本内酯的含量明显高于LVO。
2.3 经皮渗透性与促透皮作用研究
2.3.1 样品中藁本内酯、RSV含量测定
色谱条件:色谱柱 phenomenex C18(4.6 mm×150 mm,5 μm);C18保护柱(4.6 mm×10 mm);流速:1 mL·min-1,进样量:10 μL;藁本内酯流动相:甲醇-水60∶40;柱温:35 ℃;检测波长:350 nm;RSV流动相:0.1%磷酸(A)-乙腈(B)梯度洗脱,0~25 min,20%→95%(B);柱温:30 ℃;检测波长:306 nm。
方法专属性 取不同时间点接收液,分别测定藁本内酯和RSV含量,并随空白溶剂对照,结果见图1,空白溶剂对检测指标均无干扰。
2.3.2 体外透皮试验方法
离体皮肤制备 健康雄性小鼠,剃去腹部毛,脱颈处死,取其剑突以下腹部去毛皮肤。除去皮下黏膜和脂肪组织,用生理盐水冲洗至皮肤洗液无白色浑浊,滤纸吸干皮肤表面生理盐水,冷冻备用。
供试液的配制 首先配制一定浓度的RSV溶液,用该溶液分别配制成含不同浓度挥发油的川芎挥发油乳剂(Emulsion of LVO,LVOE)和当归挥发油乳剂(Emulsion of AVO,AVOE)。
体外透皮试验方法与计算公式 将制备好的离体皮肤分别固定在Franz扩散池的供药池和接收池之间,供药池中分别加入1 mL相应乳剂,接收池中加入接收液,排空气泡,池外水浴循环加热(37±0.5)℃,300 r·min-1磁力搅拌,并以一定浓度的RSV溶液作为空白对照。分别于1、2、4、6、8、10 h取样,每次取样后补足等体积的生理盐水并排空气泡。取样液体过0.45 μm针头滤器后按照“2.3.1”方法测定藁本内酯、RSV的含量。
按公式1计算接收液中指标成分的单位面积累计释放量(Qn),并以单位面积累计释放量为纵坐标,时间为横坐标,进行回归,所得的直线斜率为稳态透皮速率Jss(μg·cm-2·h-1),增透倍数按公式2计算:
Qn= ρ n V + ∑ i = 1 n - 1 ρ i V i A
EF=Jss测定组/Jss对照组
式中,ρn为第n个取样点测得的药物质量浓度(μg·mL-1);V为接收液的体积;ρi为第i个取样点测得的药物质量浓度(μg·mL-1);Vi为取样体积;A为渗透面积。
接收液的筛选:分别考察磷酸盐缓冲液(pH 7.8~8.0)、生理盐水作为接收液时RSV的稳态渗透速率,确定接受液,结果见表4。以生理盐水为接受液时RSV的稳态渗透速率高于磷酸盐缓冲液(pH 7.8~8.0),且线性关系良好(r=0.985 9),故选用生理盐水作为接收液。
2.3.3 经皮渗透
采用体外透皮试验方法,对不同浓度LVO、AVO及两者混合使用的经皮渗透情况进行研究,每个浓度相应乳剂平行3次试验,指标成分藁本内酯不同时间单位面积累计透过量Qn、Qn-t、r值、Jss等结果见表5~7。
LVO、AVO及LVO+AVO中指标性成分藁本内酯的经皮渗透均呈零级动力学过程,累计透过量与时间线性关系良好,r值均大于0.9,10 h内稳态透皮速率Jss(μg·cm-2·h-1)比较,2%>4%>3%>1%>0.5% LVO,4%>3%>1%>2%>0.5% AVO,2% AVO+0.5% LVO>2% AVO+1% LVO;2% LVO+0.5% AVO与2% LVO+1% AVO无明显差别。不同浓度的LVO、AVO均具有良好的经皮渗透性,且川芎和当归在临床上经常是对药的方式联合使用,在为达到两种中药自身药效的基础上,将2种挥发油混合使用同时也具备了经皮渗透的优势。
2.3.4 促RSV透皮作用
采用体外透皮试验方法,对不同浓度LVO、AVO及LVO+AVO两者混合使用促RSV体外透皮作用进行研究,每个浓度相应乳剂平行3次试验,指标成分RSV不同时间单位面积累计透过量Qn、Qn-t、r值、Jss、EF等结果见表8~10。
不同浓度LVO促RSV体外透皮作用研究发现,RSV的经皮扩散呈零级动力学过程,累计透过量与时间线性关系良好,r值均大于0.9,且不同浓度的LVO对RSV体外透皮作用均有良好的促进作用,增透倍数EF值表明,2%>3%>4%>1%>0.5% LVO。不同浓度AVO促RSV体外透皮作用研究发现,RSV的经皮扩散累计透过量与时间线性关系稍差,r值范围为0.794 5~0.910 5,但不同浓度的AVO对RSV体外透皮作用均有良好的促进作用,增透倍数EF值表明,4%>2%>1%>3%>0.5% AVO。LVO与AVO促透作用均与浓度有一定的依赖性。
LVO+AVO混合使用对RSV体外透皮作用也有良好的促进作用,RSV的经皮扩散呈零级动力学过程,累计透过量与时间线性关系良好,均大于0.9。LVO+AVO混合使用增透倍数EF值与LVO、AVO分别单独使用比较,2% AVO>2% AVO+0.5% LVO>2% AVO+1% LVO,2% LVO+0.5% AVO>2% LVO+1% AVO>2% LVO。
2.3.5 渗透性与促透性分析
根据不同浓度LVO、AVO的经皮渗透性和促RSV透皮作用研究,挥发油浓度与稳态透皮速率与增透倍数曲线图见图2。LVO的经皮渗透性明显高于AVO,而AVO的促渗透性则高于LVO。不同浓度挥发油的经皮渗透性与增透倍数均有一定的差异,但是对同一种挥发油而言,经皮渗透性与增透倍数随浓度变化趋势相似。
3 讨论
收起
LVO、AVO的相对密度接近水的密度,现有的提取工艺在蒸馏后油水不易分离,制备得到的挥发油纯度较低、杂质多,且加热时间长、热敏性成分易破坏等,故本研究在提取工艺中采用乙酸乙酯收集挥发油,并依次进行脱水和减压蒸馏,得到的LVO、AVO纯度高且得率和指标成分质量稳定。随着新技术的不断发展,提取方法也越来越倾向于多种方法的联合应用。通过不断优化提取工艺和提取设备,以期广泛地用于工业化生产,进一步增进对传统中医药的研究与探索。挥发油的提取不仅致力于提高挥发油的得率和质量,更应重视挥发油的化学成分和生物药效活性的变化,经过一系列生产工艺,尽量避免挥发油成分的变化和损失,更好地服务于临床用药和保证用药安全[22]。
“药辅合一”是川芎、当归挥发油在中药制剂研究与应用开发中的优势所在。LVO、AVO的物理常数的检测与之作为“药辅合一”中“辅”尤为重要,除理化性质外还表现出其功能特性,有助于药物的溶解或者分散、改善药物稳定性、提高药物生物利用度等;则川芎-当归挥发油的定性和定量控制与“药辅合一”中“药”密切相关。因此LVO、AVO的质量研究是其开发应用的关键。现有的市售挥发油多数以性状和部分物理常数作为质量控制指标,本研究对LVO、AVO进行了性状、相对密度、pH值、黏度、旋光度等物理常数和主要成分藁本内酯的研究,在一定程度上控制了LVO、AVO的质量,为后期开发和应用提供了保障。
本研究表明LVO、AVO均具有良好的渗透性和促RSV体外透皮作用,10 h内稳态透皮速率Jss最大分别为(48.23±6.53)(27.84±3.03) μg·cm-2·h-1,增透倍数EF值最大分别为4.51、19.34。且在为达到两种中药自身药效的基础上,将两种挥发油混合使用同时也具备了经皮渗透和促透皮作用的优势,研究中两种挥发油联合使用10 h内稳态透皮速率Jss和增透倍数EF值最大分别为(35.10±6.18)、7.52。不同浓度AVO促RSV体外透皮作用研究时,1、2 h均未能检测出指标成分RSV,故采用4 h后的结果进行数分析。在LVO、AVO促RSV体外透皮作用研究时发现,空白对照的渗透曲线均有一定差异,可能的原因是不同试验研究所用的老鼠皮肤批次不同,因此试验采取每次研究所用皮肤一致,且每个试验平行3次,从而减小误差。
研究分析发现,LVO的经皮渗透性明显高于AVO,而AVO的促渗透性则高于LVO。目前,透皮吸收促进剂已成为了增加药物透皮吸收的首选方法。常用的化学透皮吸收促进剂氮酮、丙二醇等在体外透皮吸收研究中发现透增倍数能达到2~10倍[23-24],与之相比本研究的LVO、AVO亦具有良好的透皮吸收作用,而且作为天然的透皮吸收促进剂,LVO、AVO具有低皮肤刺激性与低毒性,且其抑菌抗炎等生理活性使其非常适合于皮肤局部给药。自身药理效应与良好的透皮吸收促进作用相结合,起到“药辅合一”的作用,具有良好的临床使用优势,是LVO、AVO在中药制剂研究与应用开发中的优势所在。
但LVO、AVO的研究也存在一定的难点。LVO、AVO在水中溶解性较低,在后期处方考察和制剂研究中需考虑溶剂的影响。LVO、AVO的主要化学成分为内酯类成分,是一不稳定物质,易发生异构化反应,在室温及低温下均不稳定,水蒸气蒸馏过程中的开放体系及高温过程可能加速这一过程,长时间储存,成分的含量和种类均会发生改变[16]。Cui等[25]发现光降解、水解、氧化是影响挥发油成分发生变化的主要因素,运用合适的共溶剂、添加适量的抗氧化剂、维持pH值并防止光照可大幅延长藁本内酯的储存时间。因此,LVO、AVO的溶解性和稳定性在其制剂开发研究均值得思考。
基金
收起
  • 四川省科技计划重点研发项目(2020YFS0566)
  • 四川省公益性科研院所基本科研项目资助[2020JDKY0013(A-2020N-18)
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doi: 10.11669/cpj.2024.23.006
  • 接收时间：2023-11-06
  • 首发时间：2025-11-13
  • 出版时间：2024-12-08
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  • 收稿日期：2023-11-06
基金
四川省科技计划重点研发项目(2020YFS0566)
四川省公益性科研院所基本科研项目资助[2020JDKY0013(A-2020N-18)
作者信息
    四川省中医药科学院, 成都 610041

通讯作者:

* 张爱军,女,硕士,研究员 研究方向:中药新制剂及成药性药学评价 Tel:13880898533
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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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