Article(id=1241779367932658080, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1241779355555266850, articleNumber=null, orderNo=null, doi=10.16155/j.0254-1793.2024.05.06, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1686758400000, receivedDateStr=2023-06-15, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773992872460, onlineDateStr=2026-03-20, pubDate=1717084800000, pubDateStr=2024-05-31, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773992872460, onlineIssueDateStr=2026-03-20, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773992872460, creator=13701087609, updateTime=1773992872460, updator=13701087609, issue=Issue{id=1241779355555266850, tenantId=1146029695717560320, journalId=1205117023404326918, year='2024', volume='44', issue='5', pageStart='737', pageEnd='920', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773992869509, creator=13701087609, updateTime=1773992925624, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1241779590985749489, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1241779355555266850, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1241779590985749490, tenantId=1146029695717560320, journalId=1205117023404326918, issueId=1241779355555266850, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=783, endPage=795, ext={EN=ArticleExt(id=1241779368255619514, articleId=1241779367932658080, tenantId=1146029695717560320, journalId=1205117023404326918, language=EN, title=Comparative study on the quality of wild and cultivated Arnebiae Radix*, columnId=1241779356482208035, journalTitle=Chinese Journal of Pharmaceutical Analysis, columnName=Column on Quality Evaluation of Arnebiae Radix, runingTitle=null, highlight=null, articleAbstract=
Objective:

To compare the quality of wild and cultivated Arnebiae Radix,using macroscopic investigation and chemometric analysis of the different components in wild and cultivated Arnebiae Radix from three different habitats.

Methods:

Wild and cultivated Arnebiae Radix were collected and their macroscopic features were compared. Using the ACQUITY UPLC BEH C18 (2.1 mm×100 mm,1.7 μm) column,with acetonitrile-0.05% formic acid water as the mobile phase,the contents of D-shikonin,acetylshikonin,β-acetoxyisovalerylshikonin,isobutyrylshikonin,β,β’-dimethylacrylalkannin and isovalerylshikonin in 48 batches of wild and cultivated Arnebiae Radix were determined. The detection wavelength was 275 nm and the flow rate was 0.2 mL·min-1. PCA and OPLS-DA were performed to reveal the differential components of wild and cultivated Arnebiae Radix.

Results:

There were great differences in macroscopic features of wild and cultivated Arnebiae Radix,and the linear relationship between the contents of six naphthoquinone components was good. The correlation coefficients were above 0.999,the average recovery rates were 93.4%-102.9%,and the RSDs were less than 3.0%. The contents of six components in different batches of wild and cultivated Arnebiae Radix were quite different,and the contents of D-shikonin and acetylshikonin in wild products were significantly higher than those in cultivated products,indicating that there were still certain differences between wild products and cultivated products. The PCA model established could distinguish wild products and cultivars,and two differentiating components in wild products and cultivars were revealed by OPLS-DA,namely isobutyryl shikonin,β,β’-dimethylacrylalkannin.

Conclusion:

By comparing the core size,cork curl degree and specific odor of wild and cultivated products,the two can be identified. The established content determination method is repeatable,specific,stable and feasible. The differential components in wild and cultivated Arnebiae Radix in three different regions are identified,which provides a basis for the quality control of Arnebiae Radix and provides ideas for expanding the source of Arnebiae Radix.

, correspAuthors=Shuang-cheng MA, Jian ZHENG, 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=Rui HUANG, Sheng-yun DAI, Dong-xue WU, Xiao-jun MA, Jie LIU, Li-nong GUO, Dao-er-jia-la, Song JING, Shuang-cheng MA, Jian ZHENG), CN=ArticleExt(id=1241779372126962248, articleId=1241779367932658080, tenantId=1146029695717560320, journalId=1205117023404326918, language=CN, title=新疆紫草野生品与栽培品的质量比较研究*, columnId=1241779358067654949, journalTitle=药物分析杂志, columnName=紫草类民族药材的品质评价研究专栏, runingTitle=null, highlight=null, articleAbstract=
目的:

对新疆紫草野生品与栽培品的质量进行比较研究,包括性状对比和结合化学计量学分析新疆紫草野生品与3个不同地区栽培品的差异性成分。

方法:

分别采集新疆紫草野生品与栽培品,对其性状进行对比;采用Waters ACQUITY UPLC BEH C18(100 mm×2.1 mm,1.7 μm)色谱柱,以乙腈-0.05%甲酸水为流动相,检测波长275 nm,流速0.2 mL·min-1,对48批野生品和栽培品中的右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁和异戊酰紫草素6个成分进行含量测定,并结合主成分分析(PCA)和正交偏最小二乘-判别分析(OPLS-DA)分析野生品和栽培品中的差异性成分。

结果:

新疆紫草野生品与栽培品在性状上存在较大差异,建立的含量测定方法线性关系良好,r>0.999,平均加样回收率93.4%~102.9%,RSD<3.0%,不同批次新疆紫草中6个成分含量差异较大,其中野生品中右旋紫草素、β-乙酰氧基异戊酰阿卡宁含量均明显高于栽培品,说明野生品与栽培品还存在一定的差异;建立的PCA模型可区分野生品和栽培品,且通过OPLS-DA确定了异丁酰紫草素和β,β’-二甲基丙烯酰阿卡宁是野生品和栽培品中的2个差异性成分。

结论:

通过对比野生品和栽培品的木心大小、栓皮卷曲程度以及特异性气味,可对二者进行初步鉴别;建立的含量测定方法重复性好,专属性强,稳定可行;确定了新疆紫草野生品与3个不同地区栽培品的差异性成分,为新疆紫草的质量控制提供依据,为扩大紫草药源提供了思路。

, correspAuthors=马双成, 郑健, authorNote=null, correspAuthorsNote=
** 马双成 Tel:(010)53852076;E-mail:;
郑健 Tel:(010)53852080;E-mail:
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黄瑞 Tel:(010)53852078;E-mail:;

戴胜云 Tel:(010)53852078;E-mail:

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Chin J Pharm Anal201636 (9):1526, articleTitle=HPLC specific chromatogram of Arnebia euchroma and determination of six naphthoquinones in Boraginaceous Herbs, refAbstract=null)], funds=[Fund(id=1241779385414517749, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, awardId=2021X4, language=CN, fundingSource=*中国食品药品检定研究院学科带头人培养基金(2021X4), fundOrder=null, country=null), Fund(id=1241779385510986745, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, awardId=null, language=CN, fundingSource=新疆维吾尔自治区药品监督管理局智力援疆创新拓展人才计划——“新疆特色民族药研究(以新疆紫草为例)团队”项目, fundOrder=null, country=null)], companyList=[AuthorCompany(id=1241779372403786334, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, xref=1., ext=[AuthorCompanyExt(id=1241779372412174943, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, companyId=1241779372403786334, language=EN, 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caption=Comparison of morphological features of wild and cultivated Arnebiae Radix, figureFileSmall=q4I708nwl+/rY4dpz6vFfw==, figureFileBig=7Rzh6Dq8IH8Vg50H2qsdbA==, tableContent=null), ArticleFig(id=1241779381555757978, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图1, caption=新疆紫草野生品与栽培品性状比较图

A.野生品(wild) B~E.栽培品(cultivated) B.二年生(two years) C.三年生(three years) D.四年生(four years) E.五年生(five years)

, figureFileSmall=q4I708nwl+/rY4dpz6vFfw==, figureFileBig=7Rzh6Dq8IH8Vg50H2qsdbA==, tableContent=null), ArticleFig(id=1241779381782250402, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Fig.2, caption=UPLC chromatograms of mixed reference substance(A) and sample(B), figureFileSmall=xrPxU7Pn7RW+C7Qs+vy81A==, figureFileBig=mV1DNhGrCZ+Rs4jLZdy+wg==, tableContent=null), ArticleFig(id=1241779381933245351, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图2, caption=混合对照品(A)和样品(B)的UPLC色谱图

1.右旋紫草素(D-shikonin) 2.乙酰紫草素(acetylshikonin) 3.β-乙酰氧基异戊酰阿卡宁(β-acetoxyisovalerylshikonin) 4.异丁酰紫草素(isobutyrylshikonin) 5. β,β’-二甲基丙烯酰阿卡宁(β,β’-dimethylacrylalkannin) 6.异戊酰紫草素(isovalerylshikonin)

, figureFileSmall=xrPxU7Pn7RW+C7Qs+vy81A==, figureFileBig=mV1DNhGrCZ+Rs4jLZdy+wg==, tableContent=null), ArticleFig(id=1241779382046491563, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Fig.3, caption=Content determination results of 6 components in 48 batches of Arnebiae Radix, figureFileSmall=0DYkhekVKS9ySw+y45LhJw==, figureFileBig=lWknL0aPF5RJMZ5J6SXBjw==, tableContent=null), ArticleFig(id=1241779382147154864, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图3, caption=48批新疆紫草中6个成分含量测定结果, figureFileSmall=0DYkhekVKS9ySw+y45LhJw==, figureFileBig=lWknL0aPF5RJMZ5J6SXBjw==, tableContent=null), ArticleFig(id=1241779382298149814, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Fig.4, caption=Score plot of PCA of the wild and cultivated Arnebiae Radix, figureFileSmall=1LwtdvP+3TutUvI7xNG2CA==, figureFileBig=BbYmpaH7pweOAgPR43bTuQ==, tableContent=null), ArticleFig(id=1241779382398813116, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图4, caption=新疆紫草PCA得分图, figureFileSmall=1LwtdvP+3TutUvI7xNG2CA==, figureFileBig=BbYmpaH7pweOAgPR43bTuQ==, tableContent=null), ArticleFig(id=1241779382507865023, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Fig.5, caption=Score plot of OPLS-DA of the wild and cultivated Arnebiae Radix, figureFileSmall=dYgVDfWnqaFMCMT1uM2oHQ==, figureFileBig=L+cEUQciJWWVCXd9YuuMgg==, tableContent=null), ArticleFig(id=1241779382587556804, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图5, caption=新疆紫草OPLS-DA得分散点图, figureFileSmall=dYgVDfWnqaFMCMT1uM2oHQ==, figureFileBig=L+cEUQciJWWVCXd9YuuMgg==, tableContent=null), ArticleFig(id=1241779382650471369, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Fig.6, caption=OPLS-DA score plot of wild and cultivated Arnebiae Radix from different origins, figureFileSmall=oqmbjTu23e8TXb5HA1HwlQ==, figureFileBig=8hn8+P7i9Ch32ngsg0M2JA==, tableContent=null), ArticleFig(id=1241779382738551756, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图6, caption=新疆紫草野生品与不同产地栽培品的OPLS-DA得分图, figureFileSmall=oqmbjTu23e8TXb5HA1HwlQ==, figureFileBig=8hn8+P7i9Ch32ngsg0M2JA==, tableContent=null), ArticleFig(id=1241779382860186574, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Fig.7, caption=Content determination results of 6 components in Arnebiae Radix from Bayingbrooke cultivars of different cultivation years, figureFileSmall=QyIqPCtHUvZ1o857FbUFLA==, figureFileBig=JLO4pLnmrd7duAXYvJ/tMQ==, tableContent=null), ArticleFig(id=1241779382981821393, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=图7, caption=不同栽培年限巴音布鲁克栽培品新疆紫草中6个成分含量, figureFileSmall=QyIqPCtHUvZ1o857FbUFLA==, figureFileBig=JLO4pLnmrd7duAXYvJ/tMQ==, tableContent=null), ArticleFig(id=1241779383074096087, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Tab.1, caption=

Information of wild and cultivated Arnebiae Radix samples

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(No.)
品种
(variety)
产地
(habitat)
栽培年限
(cultivation year)
1野生品(wild)博尔塔拉州(Bortala Prefecture)-
2野生品(wild)昭苏(Zhaosu)-
3野生品(wild)博尔塔拉州(Bortala Prefecture)-
4野生品(wild)巴音布鲁克(Bayingbrooke)-
5野生品(wild)那拉提(Narat)-
6野生品(wild)那拉提(Narat)-
7野生品(wild)博尔塔拉州(Bortala Prefecture)-
8野生品(wild)伊犁哈萨克自治州(Ili Kazakh Autonomous Prefecture)-
9野生品(wild)新疆(Xinjiang)-
10野生品(wild)伊犁哈萨克自治州(Ili Kazakh Autonomous Prefecture)-
11野生品(wild)新疆(Xinjiang)-
12野生品(wild)新疆(Xinjiang)-
13野生品(wild)新疆(Xinjiang)-
14野生品(wild)霍城县(Huocheng)-
15野生品(wild)新疆(Xinjiang)-
16野生品(wild)新疆(Xinjiang)-
17野生品(wild)新疆(Xinjiang)-
18野生品(wild)新疆(Xinjiang)-
19野生品(wild)新疆(Xinjiang)-
20野生品(wild)新疆(Xinjiang)-
21野生品(wild)新疆(Xinjiang)-
22野生品(wild)新疆(Xinjiang)-
23野生品(wild)新疆(Xinjiang)-
24野生品(wild)新疆(Xinjiang)-
25野生品(wild)新疆(Xinjiang)-
26野生品(wild)新疆(Xinjiang)-
27野生品(wild)新疆(Xinjiang)-
28栽培品(cultivated)巴音布鲁克(Bayingbrooke)二年生(two years)
29栽培品(cultivated)巴音布鲁克(Bayingbrooke)二年生(two years)
30栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
31栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
32栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
33栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
34栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
35栽培品(cultivated)巴音布鲁克(Bayingbrooke)四年生(four years)
36栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
37栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
38栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
39栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
40栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
41栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
42栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
43栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
44栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
45栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
46栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
47栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
48栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
), ArticleFig(id=1241779383170565082, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=表1, caption=

新疆紫草野生品与栽培品的来源信息

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(No.)
品种
(variety)
产地
(habitat)
栽培年限
(cultivation year)
1野生品(wild)博尔塔拉州(Bortala Prefecture)-
2野生品(wild)昭苏(Zhaosu)-
3野生品(wild)博尔塔拉州(Bortala Prefecture)-
4野生品(wild)巴音布鲁克(Bayingbrooke)-
5野生品(wild)那拉提(Narat)-
6野生品(wild)那拉提(Narat)-
7野生品(wild)博尔塔拉州(Bortala Prefecture)-
8野生品(wild)伊犁哈萨克自治州(Ili Kazakh Autonomous Prefecture)-
9野生品(wild)新疆(Xinjiang)-
10野生品(wild)伊犁哈萨克自治州(Ili Kazakh Autonomous Prefecture)-
11野生品(wild)新疆(Xinjiang)-
12野生品(wild)新疆(Xinjiang)-
13野生品(wild)新疆(Xinjiang)-
14野生品(wild)霍城县(Huocheng)-
15野生品(wild)新疆(Xinjiang)-
16野生品(wild)新疆(Xinjiang)-
17野生品(wild)新疆(Xinjiang)-
18野生品(wild)新疆(Xinjiang)-
19野生品(wild)新疆(Xinjiang)-
20野生品(wild)新疆(Xinjiang)-
21野生品(wild)新疆(Xinjiang)-
22野生品(wild)新疆(Xinjiang)-
23野生品(wild)新疆(Xinjiang)-
24野生品(wild)新疆(Xinjiang)-
25野生品(wild)新疆(Xinjiang)-
26野生品(wild)新疆(Xinjiang)-
27野生品(wild)新疆(Xinjiang)-
28栽培品(cultivated)巴音布鲁克(Bayingbrooke)二年生(two years)
29栽培品(cultivated)巴音布鲁克(Bayingbrooke)二年生(two years)
30栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
31栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
32栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
33栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
34栽培品(cultivated)巴音布鲁克(Bayingbrooke)三年生(three years)
35栽培品(cultivated)巴音布鲁克(Bayingbrooke)四年生(four years)
36栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
37栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
38栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
39栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
40栽培品(cultivated)巴音布鲁克(Bayingbrooke)五年生(five years)
41栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
42栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
43栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
44栽培品(cultivated)乌鲁木齐南山(Nanshan,Urumqi)四年生(four years)
45栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
46栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
47栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
48栽培品(cultivated)昭苏(Zhaosu)三年生(three years)
), ArticleFig(id=1241779383271228381, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Tab.2, caption=

Investigation results of regression equation and linear range of 6 components

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
线性范围
(linear range)/(μg·mL-1
r
右旋紫草素(D-shikonin)Y=1.060×104X+1.775×1040.495~4950.999 8
乙酰紫草素(acetylshikonin)Y=1.350×104X+2.058×1040.513~5130.999 9
β-乙酰氧基异戊酰阿卡宁(β-acetoxyisovalerylshikonin)Y=1.076×104X-1.256×1030.507~5070.999 9
异丁酰紫草素(isobutyrylshikonin)Y=1.163×104X-4.095×1030.465~4650.999 9
β,β’-二甲基丙烯酰阿卡宁(β,β’-dimethylacrylalkannin)Y=1.255×104X-1.733×1040.501~5010.999 9
异戊酰紫草素(isovalerylshikonin)Y=9.023×103X-8.259×1030.480~4800.999 9
), ArticleFig(id=1241779383380280290, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=表2, caption=

6个成分的回归方程和线性范围考察结果

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
回归方程
(regression equation)
线性范围
(linear range)/(μg·mL-1
r
右旋紫草素(D-shikonin)Y=1.060×104X+1.775×1040.495~4950.999 8
乙酰紫草素(acetylshikonin)Y=1.350×104X+2.058×1040.513~5130.999 9
β-乙酰氧基异戊酰阿卡宁(β-acetoxyisovalerylshikonin)Y=1.076×104X-1.256×1030.507~5070.999 9
异丁酰紫草素(isobutyrylshikonin)Y=1.163×104X-4.095×1030.465~4650.999 9
β,β’-二甲基丙烯酰阿卡宁(β,β’-dimethylacrylalkannin)Y=1.255×104X-1.733×1040.501~5010.999 9
异戊酰紫草素(isovalerylshikonin)Y=9.023×103X-8.259×1030.480~4800.999 9
), ArticleFig(id=1241779383485137897, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Tab.3, caption=

The recoveries of six hydroxyl naphthoquinones

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
称样量
(weight of sample)/g
样品中的量
(content)/mg
加入量
(added)/mg
测得量
(measured)/mg
回收率
(recovery)/%
平均回收率
(average recovery)/%
RSD/%
右旋紫草素(D-shikonin)0.257 40.183 60.196 60.389 5104.7102.82.5
0.249 00.177 60.196 60.376 3101.1
0.243 40.173 60.196 60.369 099.4
0.255 60.182 30.196 60.381 6101.4
0.267 20.190 60.196 60.399 1106.0
0.258 70.184 50.196 60.389 8104.4
乙酰紫草素(acetylshikonin)0.250 23.283 23.2506.678 8104.5101.12.3
0.245 23.217 53.2506.451 699.5
0.250 13.282 73.2506.544 7100.4
0.250 73.290 53.2506.639 0103.0
0.250 43.286 63.2506.569 4101.0
0.249 43.273 43.2506.458 098.0
β-乙酰氧基异戊酰阿卡宁(β-acetoxyisovalerylshikonin)0.252 30.247 20.251 00.485 194.893.42.0
0.253 30.248 20.251 00.485 594.6
0.251 30.246 20.251 03.514 492.1
0.254 20.249 10.251 00.476 690.6
0.253 30.248 20.251 00.487 695.4
0.250 40.245 40.251 00.479 193.1
异丁酰紫草素(isobutyrylshikonin)0.251 80.452 80.251 00.941 199.697.13.0
0.251 10.451 60.490 10.942 1100.1
0.251 00.451 40.490 10.936 899.0
0.245 00.440 60.490 10.908 095.4
0.251 70.452 80.490 10.920 595.4
0.248 90.447 60.490 10.903 493.0
β,β’-二甲基丙烯酰阿卡宁(β,β’-dimethylacrylalkannin)0.250 51.375 51.4862.79795.697.63.0
0.257 71.415 21.4862.83095.2
0.258 31.418 61.4862.941102.4
0.256 31.407 61.4862.84096.4
0.258 51.419 81.4862.90499.9
0.253 51.392 11.4862.81595.7
异戊酰紫草素(isovalerylshikonin)0.252 72.427 82.4304.78196.995.11.4
0.253 22.432 62.4304.76395.9
0.253 12.431 72.4304.70293.4
0.253 32.433 12.4304.74495.1
0.253 52.435 52.4304.71493.8
0.247 12.374 12.4304.69495.5
), ArticleFig(id=1241779383569023978, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=表3, caption=

加样回收率试验(n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
成分
(component)
称样量
(weight of sample)/g
样品中的量
(content)/mg
加入量
(added)/mg
测得量
(measured)/mg
回收率
(recovery)/%
平均回收率
(average recovery)/%
RSD/%
右旋紫草素(D-shikonin)0.257 40.183 60.196 60.389 5104.7102.82.5
0.249 00.177 60.196 60.376 3101.1
0.243 40.173 60.196 60.369 099.4
0.255 60.182 30.196 60.381 6101.4
0.267 20.190 60.196 60.399 1106.0
0.258 70.184 50.196 60.389 8104.4
乙酰紫草素(acetylshikonin)0.250 23.283 23.2506.678 8104.5101.12.3
0.245 23.217 53.2506.451 699.5
0.250 13.282 73.2506.544 7100.4
0.250 73.290 53.2506.639 0103.0
0.250 43.286 63.2506.569 4101.0
0.249 43.273 43.2506.458 098.0
β-乙酰氧基异戊酰阿卡宁(β-acetoxyisovalerylshikonin)0.252 30.247 20.251 00.485 194.893.42.0
0.253 30.248 20.251 00.485 594.6
0.251 30.246 20.251 03.514 492.1
0.254 20.249 10.251 00.476 690.6
0.253 30.248 20.251 00.487 695.4
0.250 40.245 40.251 00.479 193.1
异丁酰紫草素(isobutyrylshikonin)0.251 80.452 80.251 00.941 199.697.13.0
0.251 10.451 60.490 10.942 1100.1
0.251 00.451 40.490 10.936 899.0
0.245 00.440 60.490 10.908 095.4
0.251 70.452 80.490 10.920 595.4
0.248 90.447 60.490 10.903 493.0
β,β’-二甲基丙烯酰阿卡宁(β,β’-dimethylacrylalkannin)0.250 51.375 51.4862.79795.697.63.0
0.257 71.415 21.4862.83095.2
0.258 31.418 61.4862.941102.4
0.256 31.407 61.4862.84096.4
0.258 51.419 81.4862.90499.9
0.253 51.392 11.4862.81595.7
异戊酰紫草素(isovalerylshikonin)0.252 72.427 82.4304.78196.995.11.4
0.253 22.432 62.4304.76395.9
0.253 12.431 72.4304.70293.4
0.253 32.433 12.4304.74495.1
0.253 52.435 52.4304.71493.8
0.247 12.374 12.4304.69495.5
), ArticleFig(id=1241779385150276590, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=EN, label=Tab.4, caption=

Contents of six components in 48 batches of wild and cultivated Arnebiae Radix samples

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(No.)
含量(content)/%总量
(total)/%
右旋紫草素
D-shikonin)
乙酰紫草素
(acetyl-shikonin)
β-乙酰氧基异戊酰阿卡宁
(β-acetoxy-isovalerylshikonin)
异丁酰紫草素
(isobutyryl-shikonin)
β,β’-二甲基丙烯酰阿卡宁
(β,β’-dimethylac-rylalkannin)
异戊酰紫草素
(isovalerylshikonin)
10.100.760.360.370.311.002.90
20.180.350.170.320.610.812.44
30.150.540.330.510.951.293.77
40.050.440.070.110.610.711.99
50.040.590.190.470.590.902.78
60.121.500.180.380.741.694.61
70.061.040.090.140.671.133.13
80.121.330.170.280.521.884.30
90.030.100.010.050.270.530.99
100.080.540.740.550.431.343.68
110.152.060.510.510.933.227.38
120.010.050.270.080.810.611.83
130.190.690.430.540.551.213.61
140.151.100.310.370.872.235.03
150.110.840.100.590.751.263.65
160.040.270.120.160.420.691.70
170.080.490.180.240.491.012.49
180.070.440.210.320.500.762.30
190.080.650.230.550.571.163.24
200.110.450.230.450.670.992.90
210.050.370.290.180.420.802.11
220.030.190.040.110.330.611.31
230.060.180.200.240.560.601.84
240.030.170.220.160.490.601.67
250.030.240.290.211.461.073.30
260.030.170.150.170.570.591.68
270.020.210.070.110.460.671.54
280.141.670.100.600.881.625.01
290.090.710.150.110.390.992.44
300.081.410.110.190.621.163.57
310.141.620.180.290.582.084.89
320.172.350.270.400.812.486.48
330.091.750.210.320.692.185.24
340.142.190.330.470.802.416.34
350.142.060.150.260.592.145.34
360.232.140.230.300.912.756.56
370.121.750.150.260.741.604.62
380.091.840.150.390.752.225.44
390.142.050.310.420.822.346.08
400.172.700.220.450.802.737.07
410.090.470.120.100.320.501.60
420.100.100.070.040.150.190.65
430.130.410.090.080.330.531.57
440.050.160.050.040.150.270.72
45-0.020.040.010.060.050.18
46-0.020.070.010.050.060.21
47--0.03-0.030.030.09
480.01-0.110.030.140.150.44
), ArticleFig(id=1241779385280300020, tenantId=1146029695717560320, journalId=1205117023404326918, articleId=1241779367932658080, language=CN, label=表4, caption=

48批新疆紫草野生品与栽培品中6个成分含量测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
编号
(No.)
含量(content)/%总量
(total)/%
右旋紫草素
D-shikonin)
乙酰紫草素
(acetyl-shikonin)
β-乙酰氧基异戊酰阿卡宁
(β-acetoxy-isovalerylshikonin)
异丁酰紫草素
(isobutyryl-shikonin)
β,β’-二甲基丙烯酰阿卡宁
(β,β’-dimethylac-rylalkannin)
异戊酰紫草素
(isovalerylshikonin)
10.100.760.360.370.311.002.90
20.180.350.170.320.610.812.44
30.150.540.330.510.951.293.77
40.050.440.070.110.610.711.99
50.040.590.190.470.590.902.78
60.121.500.180.380.741.694.61
70.061.040.090.140.671.133.13
80.121.330.170.280.521.884.30
90.030.100.010.050.270.530.99
100.080.540.740.550.431.343.68
110.152.060.510.510.933.227.38
120.010.050.270.080.810.611.83
130.190.690.430.540.551.213.61
140.151.100.310.370.872.235.03
150.110.840.100.590.751.263.65
160.040.270.120.160.420.691.70
170.080.490.180.240.491.012.49
180.070.440.210.320.500.762.30
190.080.650.230.550.571.163.24
200.110.450.230.450.670.992.90
210.050.370.290.180.420.802.11
220.030.190.040.110.330.611.31
230.060.180.200.240.560.601.84
240.030.170.220.160.490.601.67
250.030.240.290.211.461.073.30
260.030.170.150.170.570.591.68
270.020.210.070.110.460.671.54
280.141.670.100.600.881.625.01
290.090.710.150.110.390.992.44
300.081.410.110.190.621.163.57
310.141.620.180.290.582.084.89
320.172.350.270.400.812.486.48
330.091.750.210.320.692.185.24
340.142.190.330.470.802.416.34
350.142.060.150.260.592.145.34
360.232.140.230.300.912.756.56
370.121.750.150.260.741.604.62
380.091.840.150.390.752.225.44
390.142.050.310.420.822.346.08
400.172.700.220.450.802.737.07
410.090.470.120.100.320.501.60
420.100.100.070.040.150.190.65
430.130.410.090.080.330.531.57
440.050.160.050.040.150.270.72
45-0.020.040.010.060.050.18
46-0.020.070.010.050.060.21
47--0.03-0.030.030.09
480.01-0.110.030.140.150.44
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新疆紫草野生品与栽培品的质量比较研究*
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黄瑞 1, 2 , 戴胜云 1 , 吴东雪 1 , 马晓军 3 , 刘杰 1 , 过立农 1 , 道尔加拉 4 , 敬松 5 , 马双成 1, ** , 郑健 1, **
药物分析杂志 | 紫草类民族药材的品质评价研究专栏 2024,44(5): 783-795
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药物分析杂志 | 紫草类民族药材的品质评价研究专栏 2024, 44(5): 783-795
新疆紫草野生品与栽培品的质量比较研究*
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黄瑞1, 2 , 戴胜云1 , 吴东雪1, 马晓军3, 刘杰1, 过立农1, 道尔加拉4, 敬松5, 马双成1, ** , 郑健1, **
作者信息
  • 1.中国食品药品检定研究院,北京 102629
  • 2.中国药科大学,南京 211198
  • 3.伊犁哈萨克自治州检验检测认证研究院,伊犁835000
  • 4.新疆德立凯农牧科技发展有限公司,乌鲁木齐 830000
  • 5.新疆欣嘉农业科技开发有限公司,伊犁 835400
  • 黄瑞 Tel:(010)53852078;E-mail:;

    戴胜云 Tel:(010)53852078;E-mail:

通讯作者:

** 马双成 Tel:(010)53852076;E-mail:;
郑健 Tel:(010)53852080;E-mail:
Comparative study on the quality of wild and cultivated Arnebiae Radix*
Rui HUANG1, 2 , Sheng-yun DAI1 , Dong-xue WU1, Xiao-jun MA3, Jie LIU1, Li-nong GUO1, Dao-er-jia-la4, Song JING5, Shuang-cheng MA1, ** , Jian ZHENG1, **
Affiliations
  • 1.National Institutes for Food and Drug Control, Beijing 102629, China
  • 2.China Pharmaceutical University, Nanjing 211198, China
  • 3.Yili Institute of Inspection, Testing and Certification, Yili 835000, China
  • 4.Xinjiang Delikai Agriculture and Animal Husbandry Science and Technology Development Co., Ltd., Urumqi 830000, China
  • 5.Xinjiang Xinjia Agricultural Science and Technology Development Co., Ltd., Ili 835400, China
出版时间: 2024-05-31 doi: 10.16155/j.0254-1793.2024.05.06
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目的:

对新疆紫草野生品与栽培品的质量进行比较研究,包括性状对比和结合化学计量学分析新疆紫草野生品与3个不同地区栽培品的差异性成分。

方法:

分别采集新疆紫草野生品与栽培品,对其性状进行对比;采用Waters ACQUITY UPLC BEH C18(100 mm×2.1 mm,1.7 μm)色谱柱,以乙腈-0.05%甲酸水为流动相,检测波长275 nm,流速0.2 mL·min-1,对48批野生品和栽培品中的右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁和异戊酰紫草素6个成分进行含量测定,并结合主成分分析(PCA)和正交偏最小二乘-判别分析(OPLS-DA)分析野生品和栽培品中的差异性成分。

结果:

新疆紫草野生品与栽培品在性状上存在较大差异,建立的含量测定方法线性关系良好,r>0.999,平均加样回收率93.4%~102.9%,RSD<3.0%,不同批次新疆紫草中6个成分含量差异较大,其中野生品中右旋紫草素、β-乙酰氧基异戊酰阿卡宁含量均明显高于栽培品,说明野生品与栽培品还存在一定的差异;建立的PCA模型可区分野生品和栽培品,且通过OPLS-DA确定了异丁酰紫草素和β,β’-二甲基丙烯酰阿卡宁是野生品和栽培品中的2个差异性成分。

结论:

通过对比野生品和栽培品的木心大小、栓皮卷曲程度以及特异性气味,可对二者进行初步鉴别;建立的含量测定方法重复性好,专属性强,稳定可行;确定了新疆紫草野生品与3个不同地区栽培品的差异性成分,为新疆紫草的质量控制提供依据,为扩大紫草药源提供了思路。

新疆紫草  /  栽培品  /  野生品  /  差异性分析  /  β,β’-二甲基丙烯酰阿卡宁  /  β-乙酰氧基异戊酰阿卡宁  /  异丁酰紫草素
Objective:

To compare the quality of wild and cultivated Arnebiae Radix,using macroscopic investigation and chemometric analysis of the different components in wild and cultivated Arnebiae Radix from three different habitats.

Methods:

Wild and cultivated Arnebiae Radix were collected and their macroscopic features were compared. Using the ACQUITY UPLC BEH C18 (2.1 mm×100 mm,1.7 μm) column,with acetonitrile-0.05% formic acid water as the mobile phase,the contents of D-shikonin,acetylshikonin,β-acetoxyisovalerylshikonin,isobutyrylshikonin,β,β’-dimethylacrylalkannin and isovalerylshikonin in 48 batches of wild and cultivated Arnebiae Radix were determined. The detection wavelength was 275 nm and the flow rate was 0.2 mL·min-1. PCA and OPLS-DA were performed to reveal the differential components of wild and cultivated Arnebiae Radix.

Results:

There were great differences in macroscopic features of wild and cultivated Arnebiae Radix,and the linear relationship between the contents of six naphthoquinone components was good. The correlation coefficients were above 0.999,the average recovery rates were 93.4%-102.9%,and the RSDs were less than 3.0%. The contents of six components in different batches of wild and cultivated Arnebiae Radix were quite different,and the contents of D-shikonin and acetylshikonin in wild products were significantly higher than those in cultivated products,indicating that there were still certain differences between wild products and cultivated products. The PCA model established could distinguish wild products and cultivars,and two differentiating components in wild products and cultivars were revealed by OPLS-DA,namely isobutyryl shikonin,β,β’-dimethylacrylalkannin.

Conclusion:

By comparing the core size,cork curl degree and specific odor of wild and cultivated products,the two can be identified. The established content determination method is repeatable,specific,stable and feasible. The differential components in wild and cultivated Arnebiae Radix in three different regions are identified,which provides a basis for the quality control of Arnebiae Radix and provides ideas for expanding the source of Arnebiae Radix.

Arnebiae Radix  /  cultivated species  /  wild species  /  assay  /  β,β’-dimethylacrylalkannin  /  β-acetoxyisovalerylshikonin  /  isobutyrylshikonin
黄瑞, 戴胜云, 吴东雪, 马晓军, 刘杰, 过立农, 道尔加拉, 敬松, 马双成, 郑健. 新疆紫草野生品与栽培品的质量比较研究*. 药物分析杂志, 2024 , 44 (5) : 783 -795 . DOI: 10.16155/j.0254-1793.2024.05.06
Rui HUANG, Sheng-yun DAI, Dong-xue WU, Xiao-jun MA, Jie LIU, Li-nong GUO, Dao-er-jia-la, Song JING, Shuang-cheng MA, Jian ZHENG. Comparative study on the quality of wild and cultivated Arnebiae Radix*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44 (5) : 783 -795 . DOI: 10.16155/j.0254-1793.2024.05.06
紫草是常见的传统药材,始载于《神农本草经》[1],在我国有着非常悠久的药用历史。紫草在古代最初作为染料使用,《本草纲目》中记载“此草花紫根紫,可以染紫,故名紫草”[2],作为染料,紫草有重要的经济价值,《齐民要术》中记载“种蓝十亩,敌谷田一顷。能自染青者,其利又倍矣”[3]。宋代以后,其药用价值逐渐被人们熟悉且重用,由此紫草作为药材在临床的使用日渐成熟。新疆紫草Arnebia euchroma(Royle)Johnst.来源于软紫草属,具有清热凉血、活血解毒、透疹消斑的功效,可用于治疗血热毒盛、斑疹紫黑、麻疹不透、疮疡、湿疹、水火烫伤等[4],主要分布在我国新疆南北部高寒草原以及西藏西部高海拔地区,多长于路边、田野、荒山、山地草丛中和干燥的灌木林下[5],生长环境严苛,对自然环境要求较高,具有耐寒、喜湿的特性,生长缓慢,种群更新能力弱[6]。近年来,随着医疗需求的增加与相关产业链蓬勃发展、气候环境的恶化以及野生资源的大肆开发与过度采挖,使得野生新疆紫草资源面临资源枯竭的状态。针对这个原因以及对紫草资源的合理开发,人工栽培品逐步推广应用,在传统栽培经验基础上结合人工种植经验,制定了相对完整的新疆紫草栽培技术规程[7]。受地理环境和气候条件的影响,新疆不同地区的栽培品质量不明确,在市场上暂未见样品流通。近年来有关新疆紫草的文献报道主要集中在来源产地、化学成分、药理活性等[8-13],而未见有对野生品与栽培品的外观性状、化学成分研究的文献报道。
基于上述现状,本研究将对新疆紫草野生品与栽培品的性状进行比较,并对右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁和异戊酰紫草素等6个萘醌类成分进行含量测定,进一步结合主成分分析(PCA)和正交偏最小二乘法-判别分析(OPLS-DA)2种化学模式识别方法,筛选出可区分新疆紫草野生品与栽培品的差异性成分,以期为新疆紫草栽培品的质量控制奠定基础,为扩大紫草药源提供科学依据,实现野生资源的保护和可持续开发。
Acquity UPLC H-Class超高效液相色谱仪,四元溶剂管理器,PDA检测器,工作站Empower 3.0版本(Waters公司);XS105DU型万分之一电子分析天平(Mettler Toledo公司);XPE105型万分之一电子天平(Mettler Toledo公司);Milli-Q Advantage A10超纯水器(Millipore公司);KQ-500DE超声波清洗器(昆山市超声仪器有限公司);Labofuge 400离心机(Centrifuge公司)。
乙腈:色谱纯,霍尼韦尔贸易上海有限公司;甲酸:色谱纯,国药集团化学试剂有限公司;石油醚:60~90 ℃,国药集团化学试剂有限公司;水:纯净水,娃哈哈有限公司。
对照品:右旋紫草素(批号ST78490120,纯度98.0%)、乙酰紫草素(批号ST78820120,纯度98.0%)、β-乙酰氧基异戊酰阿卡宁(批号ST17190120,纯度98.0%)、异戊酰紫草素(批号STC9540105,纯度98.0%),上海诗丹德标准技术服务有限公司;异丁酰紫草素(批号DST220318-341,纯度98%),乐美天医药德思特生物公司;β,β’-二甲基丙烯酰阿卡宁(批号111689-201805,纯度98.0%),中国食品药品检定研究院。
药材样品来自药材市场或产地采集,经中国食品药品检定研究院郑健研究员鉴定为紫草科软紫草属新疆紫草Arnebia euchroma(Royle)Johnst.的干燥根。新疆紫草样品48批,包括27批野生品和3个不同地区的21批栽培品。样品来源信息见表1
选取2批自行采集的新疆紫草野生品与4批同一栽培基地不同年限的栽培品,进行性状比较分析。根据2020年版《中华人民共和国药典》(简称《中国药典》)、《中药鉴定学》及相关参考文献资料,从形状、大小、色泽、断面、质地及气味等方面对比野生品与栽培品的特点。
新疆紫草野生品与栽培品在性状上存在较大差异,具体描述如下:(1)野生品:呈不规则的长圆柱形,多扭曲,长7~20 cm,直径1~2.5 cm;表面紫红色或紫褐色,皮部疏松,颜色为紫红色,呈条形片状,常10余层重叠,易剥落;顶端有的可见分歧的茎残基,被灰白色短硬毛[6];质松,体轻,易折断,折断面不整齐,木部较小,黄白色或黄色;气特异,味微苦、涩。如图1-A所示。
(2)栽培品:呈不规则的长圆柱形,略弯曲,长6~23 cm,直径0.7~2.1 cm。表面紫褐色或灰棕色,栓皮较少,颜色为紫褐色或灰褐色,呈不规则长条状,少见重叠,随着栽培年限的增长增多且卷曲,可见灰白色短硬毛。质硬,体轻,易折断,折断面不平坦,木部较大,木心大小随栽培年限增长而缩小,淡黄白色或灰褐色;气微特异,味微苦、涩。如图1-B~E所示。
采用Waters Acquity UPLC BEH C18(100 mm×2.1 mm,1.7 μm)色谱柱,柱温35 ℃,流速0.2 mL·min-1,以乙腈-0.05%甲酸水溶液(70:30 )为流动相,检测波长275 nm,进样量2 μL。色谱图见图2
取右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁和异戊酰紫草素的对照品各约10 mg,精密称定,分别置10 mL量瓶中,用乙腈溶解并定容至刻度,即得各单一成分对照品储备液。分别取各上述储备液适量,置同一10 mL量瓶中,加乙腈配制成上述6个成分质量浓度分别为50.11、100.05、50.43、48.45、100.02、49.90 μg·mL-1的混合溶液,即得。
取样品粉末0.5 g,精密称定,置于50 mL具塞锥形瓶中,精密加入石油醚(60~90 ℃) 25 mL,称定质量,超声(40 kHz,200 W)处理30 min,放冷,再称定质量,用石油醚(60~90 ℃)补足减失的质量,转移至50 mL离心管,以转速4 000 r·min-1离心5 min,取上清液10 mL至蒸发皿,蒸干,残渣加乙腈溶解,转移至10 mL量瓶中,加乙腈至刻度,摇匀,0.22 μm微孔滤膜滤过,即得。
精密吸取混合对照品溶液0.1、0.2、0.5、1.0 mL分别置于4个10 mL量瓶中,加乙腈定容至刻度,摇匀,即得1~4号对照品混合溶液,精密量取“2.2.2”项下各对照品储备液1 mL至2 mL量瓶中,加乙腈定容至刻度,摇匀,即得5号对照品混合溶液。分别精密吸取1~5号混合对照品溶液及单个对照品溶液各2 μL,注入液相色谱仪,记录峰面积,以各对照品的浓度( X,μg·mL-1 )为横坐标,以峰面积(Y)为纵坐标,绘制标准曲线,得到回归方程。结果显示,新疆紫草中6个成分在测定的范围内相关系数r均≥0.999,在相应的浓度范围内线性关系良好,结果见表2
精密吸取“2.2.2”项混合对照品溶液2 μL,连续进样6次,记录峰面积并计算RSD。结果显示右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁、异戊酰紫草素峰面积的RSD(n=6)分别为1.3%、1.3%、0.72%、1.9%、1.6%、1.4%,表明仪器精密度良好。
取样品粉末(样品5号)6份,分别按“2.2.3”项方法制备成供试品溶液,按“2.2.1”项方法分别注入色谱仪,计算6个成分含量及RSD。结果显示右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁、异戊酰紫草素的含量分别为0.12%、1.31%、0.17%、0.37%、0.76%、1.65%,RSD分别为2.8%、1.6%、2.6%、0.49%、2.6%、1.9%,表明方法重复性良好。
取同一批次新疆紫草(样品5号),按“2.2.3”项下方法制备供试品溶液,分别于制备后0、2、6、12、24、48 h,按“2.2.1”项下色谱条件分别注入色谱仪,记录6个成分峰面积,并计算RSD。结果显示右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁、异戊酰紫草素峰面积的RSD分别为1.3%、0.50%、1.9%、0.76%、1.8%、2.2%,表明仪器稳定性良好。
分别取右旋紫草素、乙酰紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素、β,β’-二甲基丙烯酰阿卡宁、异戊酰紫草素的对照品适量,精密称定,置量瓶中,加乙腈溶解配制成质量浓度分别为0.196 6、1.213、0.151 4、0.490 1、1.485、1.498 μg·mL-1的6个对照品溶液。取已知含量的样品粉末(样品5号),每份各约0.25 g,精密称定,分别置具塞锥形瓶中,精密加入上述6个对照品溶液各1 mL,再精密加入石油醚(60~90 ℃) 25 mL,按照“2.2.3”项下方法平行制备供试溶液,进样分析,计算回收率,结果见表3
取48批次新疆紫草野生品和栽培品粉末0.5 g,精密称定,按“2.2.3”项下方法制备供试品溶液,按“2.2.1”项下色谱条件测定6个成分的峰面积,采用外标法计算各成分含量,测定结果见表4。结果显示,不同批次新疆紫草野生品和栽培品中6个成分含量差异较大,其含量从大到小的排列顺序依次为异戊酰紫草素、乙酰紫草素、β,β’-二甲基丙烯酰阿卡宁、异丁酰紫草素、β-乙酰氧基异戊酰阿卡宁、右旋紫草素,含量平均值依次为1.21%、0.86%、0.57%、0.27%、0.19%、0.09%。48批新疆紫草中6个成分总量结果如图3所示。
新疆紫草野生品和栽培品中均含有萘醌类成分,且含量上存在差异,不同产地的栽培品也存在较大差异。产自巴音布鲁克地区的栽培品萘醌类成分总量最高,平均值为5.31%,说明该产地的栽培品质量优良,其次是野生品,平均值为2.90%。而乌鲁木齐南山地区和昭苏园区的栽培品6个萘醌成分总含量较低,平均值分别为1.14%和0.23%。
主成分分析(PCA)是一种无监督的多元处理技术,可对数据处理中变异数据进行总结,它将会产生一个视觉散点图,可定性分析多元数据中的异同。为了更加清晰的区分48批紫草样品,将6个化合物的数据进行了PCA。将48批样品6个化合物的数据导入Chempattern软件,采用PCA,样本变异量88.5(PC1 77.58%,PC2 10.92%),累积R2X=0.902,R2Y=0.415,Q2=0.344,说明所建立的模型具有良好的稳定性及预测能力。结果如图4所示,野生品与乌鲁木齐南山、昭苏园区栽培品完全分离;巴音布鲁克栽培品与乌鲁木齐南山、昭苏园区栽培品完全分离;而野生品与巴音布鲁克栽培品有部分重合,巴音布鲁克栽培品组中包含3个野生品。
PCA反映出野生品与三地区栽培品在6个萘醌成分含量上有差异,为了进一步探究他们之间的差异性,使用正交偏最小二乘-判别分析(OPLS-DA),分析如图5所示,野生品与乌鲁木齐南山、昭苏园区栽培品完全分离;巴音布鲁克栽培品与乌鲁木齐南山、昭苏园区栽培品完全分离;野生品与巴音布鲁克栽培品虽相对于PCA而言聚集性增强,但二者均未分开。提取OPLS-DA模型中6个化合物含量的变量投影重要性(VIP)值,其中VIP值>1的成分有2个,分别为异丁酰紫草素和β,β’-二甲基丙烯酰阿卡宁,且β,β’-二甲基丙烯酰阿卡宁的贡献度最大,表明这2个化学成分是区别新疆紫草野生品和3个不同地区栽培品的差异标志物,在6个萘醌类化学成分中,乙酰紫草素的含量虽较高,但其在区分差异性方面贡献率较低。
使用OPLS-DA,分析野生品与不同产地栽培品化学成分的差异,如图6所示,野生品与巴音布鲁克栽培品部分重合,与乌鲁木齐南山、昭苏园区栽培品完全分离,通过VIP对野生品和巴音布鲁克栽培品的差异标志物进行分析,发现VIP值>1的化学成分为乙酰紫草素和异戊酰紫草素,得到引起野生品、巴音布鲁克栽培品差异的主要化合物为乙酰紫草素和异戊酰紫草素;引起野生品、乌鲁木齐南山栽培品差异的主要化合物为异丁酰紫草素和β,β’-二甲基丙烯酰阿卡宁;引起野生品、昭苏园区栽培品差异的主要化合物为β-乙酰氧基异戊酰阿卡宁,可以看出不同产地的新疆紫草质量存在差异。
巴音布鲁克地区的栽培品在不同的栽培年限上存在含量差异,计算两年生、三年生、四年生以及五年生样品的含量平均值,结果如图7所示。五年生栽培品萘醌类成分总含量最高,平均值为5.95%,其次是三年生栽培品,平均值为5.31%,最后是四年生栽培品和两年生栽培品,萘醌成分总含量分别为5.30%和3.71%,提示五年生的栽培品质量最佳。以上6个成分存在2种变化趋势,右旋紫草素、β-乙酰氧基异戊酰阿卡宁、异丁酰紫草素和β,β’-二甲基丙烯酰阿卡宁这4个成分在不同年限的栽培品中变化不明显,其含量随着栽培年限的增长缓慢增加。乙酰紫草素和异戊酰紫草素的变化明显,三年生栽培品含量比起两年生栽培品含量出现明显上升,后持续平稳上升,分别由1.19%和1.31%增加到2.10%和2.33%,说明这2个成分受栽培年限的影响程度很大。
《中国药典》规定,紫草药材中指标性成分β,β’-二甲基丙烯酰阿卡宁不得少于0.30%,巴音布鲁克各个年限的栽培品均远高于上述要求,在含量方面符合规定,达到药用要求。
因巴音布鲁克产区的新疆紫草栽培品质量最优,故本研究对新疆紫草野生品与巴音布鲁克产区的栽培品进行了性状的对比,找出了能区分二者的性状特征,通过对二者进行性状对比之后得知二者均含有紫色物质,从外观形态来看,二者有明显的性状差异。野生品栓皮重叠且卷曲,木心较小,有明显的特异性气味。而巴音布鲁克产区栽培品质地较硬,栓皮较少且弯曲不明显,木心较大,特异性气味不明显。故野生品与巴音布鲁克产区栽培品可以通过性状特征来进行区分。而来源于其他产区的栽培品因样本数量过小,无代表性特征等原因,未有单独对其性状与野生品进行对比研究,后续应针对不同产地、不同年限栽培品进行性状鉴定。
产地对药材质量会有一定的影响,本研究中的27批野生品虽来源于不同的产地,但均来源于我国新疆省,在对48批新疆紫草样品进行含量测定后,将其结果进行了聚类分析,结果显示除了4个样品外,各个产地的野生品都能很好地聚为一类并与栽培品分离,故将新疆紫草野生品归为一个大组,便于后续的结果分析。
本文参考《中国药典》一部含量测定项下色谱条件选择和溶液制备方法,在其基础上进行优化,建立测定6个成分的UPLC分析方法。测定的6个萘醌类成分极性较小,故采用石油醚提取,乙腈溶解残渣。课题组前期筛选了检测波长,结果显示当波长为275 nm时,均能检测到上述6个成分,且各峰的分离度良好,因此确定检测波长为275 nm[14]。含量测定结果显示,新疆紫草野生品与栽培品在成分含量上有一定的区别,但未呈现一定的规律性,且本文只对巴音布鲁克、乌鲁木齐南山、昭苏园区3个栽培产区进行分析,不能代表所有栽培品的质量,故后续需进一步研究。此外,本文只对巴音布鲁克产区不同年限的栽培品进行成分含量对比,故后续应对其他产区不同年限的栽培品进行分析。
产于乌鲁木齐南山地区(海拔1 200 m左右)的四年生栽培品,其β,β’-二甲基丙烯酰阿卡宁平均值为0.24%,同样生长年限的巴音布鲁克(海拔2 500 m左右)栽培品,其β,β’-二甲基丙烯酰阿卡宁平均值为0.59%,产于昭苏园区(海拔800 m左右)的三年生栽培品,β,β’-二甲基丙烯酰阿卡宁平均值为0.07%,远低于同年生的巴音布鲁克栽培品0.70%的含量,不符合2020年版《中国药典》规定,推测可能受到产地海拔的影响,栽培年限过短,造成其质量不佳。延长其栽培年限是否能够增加昭苏园区栽培品的含量,后续还需要进一步研究。
乌鲁木齐南山地区栽培品含量略高于昭苏园区,二者栽培品质量都远低于巴音布鲁克栽培品,进一步证实了海拔对紫草质量的影响,海拔越高,环境越严苛,越利于其成分的累积。来自于巴音布鲁克地区不同生长年限的栽培品均达到《中国药典》的要求,但其性状与野生品相比,还有较大的差异,后续需跟进观察七年生、八年生、九年生等更多生长年限的栽培品,将其性状与野生品进行对比研究。
课题组前期对新疆紫草野生品与栽培品的DNA进行了ITS2序列测序,结果显示新疆紫草野生品与栽培品ITS2序列相同,是同一物种,但栽培品的性状与《中国药典》中规定的性状差异较大。本研究发现,生长年限与生长地区海拔均会影响其成分含量,生长年限越长、海拔越高、生长环境越苛刻,越会长出质量更优的紫草,这也是造成紫草野生资源短缺的重要原因。紫草市场资源的匮乏,导致了很多的掺伪品,后续需要更多深入的研究。
本文对新疆紫草野生品与栽培品的性状进行对比研究,可根据栓皮是否卷曲,木心的大小和有无明显的特异性气味区分野生品与巴音布鲁克产区的栽培品。本文对新疆紫草野生品与栽培品中6个萘醌类成分进行含量测定,根据含量测定结果,产于巴音布鲁克的栽培品6个萘醌类成分总含量最高,而产于昭苏园区的所有批次药材栽培品中指标性成分β,β’-二甲基丙烯酰阿卡宁均不符合《中国药典》规定,说明栽培品质量层次不齐,栽培品的质量控制尚需要进一步研究。
本研究采用UPLC建立了在同一色谱条件下,同时测定新疆紫草野生品与栽培品中6个萘醌类成分的含量测定方法,该方法的精密度、重复性、稳定性均符合规定,方法稳定可行,6个萘醌类成分可以作为控制新疆紫草质量的指标性成分;同时采用PCA和OPLS-DA 2种化学模式识别技术确定了异丁酰紫草素和β,β’-二甲基丙烯酰阿卡宁可有效区分新疆紫草野生品与栽培品。巴音布鲁克地区栽培品含量已达到《中国药典》的标准,但其性状与野生品差异较大,不符合《中国药典》标准,后续将继续开展新疆紫草栽培品的研究。本研究发现,海拔越高,环境越严苛,越利于新疆紫草的生长。正因为生长环境的严峻,才导致新疆紫草野生资源的短缺,建议加大栽培品的研究力度和种植范围,扩大规模,跟踪其生长年限,栽培出符合《中国药典》规定的药材,以解决新疆紫草资源匮乏的问题。
  • *中国食品药品检定研究院学科带头人培养基金(2021X4)
  • 新疆维吾尔自治区药品监督管理局智力援疆创新拓展人才计划——“新疆特色民族药研究(以新疆紫草为例)团队”项目
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doi: 10.16155/j.0254-1793.2024.05.06
  • 接收时间:2023-06-15
  • 首发时间:2026-03-20
  • 出版时间:2024-05-31
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  • 收稿日期:2023-06-15
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*中国食品药品检定研究院学科带头人培养基金(2021X4)
新疆维吾尔自治区药品监督管理局智力援疆创新拓展人才计划——“新疆特色民族药研究(以新疆紫草为例)团队”项目
作者信息
    1.中国食品药品检定研究院,北京 102629
    2.中国药科大学,南京 211198
    3.伊犁哈萨克自治州检验检测认证研究院,伊犁835000
    4.新疆德立凯农牧科技发展有限公司,乌鲁木齐 830000
    5.新疆欣嘉农业科技开发有限公司,伊犁 835400

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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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