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Article(id=1153429500160364865, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241011003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1728576000000, receivedDateStr=2024-10-11, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752928622522, onlineDateStr=2025-07-19, pubDate=1741968000000, pubDateStr=2025-03-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752928622522, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752928622522, creator=13701087609, updateTime=1752928622522, updator=13701087609, issue=Issue{id=1153429493357203682, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='5', pageStart='1', pageEnd='326', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752928620900, creator=13701087609, updateTime=1758690311058, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1177595773500932351, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1177595773500932352, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=127, endPage=133, ext={EN=ArticleExt(id=1153429500663681360, articleId=1153429500160364865, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Simultaneous determination of functional components of 7 kinds of medicinal and food homologous substances by high performance liquid chromatography, columnId=1151895322591638525, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Functional Foods and Functional Components, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a method for the simultaneous determination of functional components in extracts from 7 kinds of medicinal and food homologous substances: Pueraria lobata, Sophora japonica, Lonicera japonica, Cistanche deserticola, Siraitia grosvenorii, Panax ginseng and Hovenia dulcis by high performance liquid chromatography (HPLC). Methods The analysis was conducted using an Agilent Eclipse XDB-C18 column (4.6 mm× 250 mm, 5 μm) with a mobile phase gradient elution of acetonitrile-0.1% phosphoric acid at a flow rate of 0.6 mL/min, a column temperature of 30 ℃, a detection wavelength of 210 nm, and an injection volume of 5 μL. This method was employed to quantify the content of puerarin, rutin, chlorogenic acid, echinacoside, verbascoside, mogroside V, ginsenoside Re and dihydromyricetin. Results The results demonstrated that 8 kinds of functional components exhibited excellent linear relationships within their respective concentration ranges, with correlation coefficients (r²) exceeding 0.999. The limits of detection ranged from 0.02 to 1.88 mg/L, and the limits of quantification ranged from 0.08 to 3.62 mg/L. The precision experiment results showed that the relative standard deviation (RSD) was less than 3%, the average recoveries of spiked samples ranged from 95.49% to 109.87%. Conclusion This method is simple, rapid and highly accurate, making it suitable for the qualitative and quantitative analysis of the functional components in the aforementioned 7 kinds of medicinal and food homologous substances.

, correspAuthors=Yun TANG, 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=Hui-Hui GAO, Ya-Xiong ZHAO, Zhang-Bin HAN, Hai-Tian LI, Cheng-Yan YANG, Yun TANG), CN=ArticleExt(id=1153429520418853603, articleId=1153429500160364865, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=高效液相色谱法同时测定7种药食同源物质功能成分, columnId=1151895323909124661, journalTitle=食品安全质量检测学报, columnName=本期专题：功能性食品与功能性成分, runingTitle=null, highlight=null, articleAbstract=

目的 建立高效液相色谱法(high performance liquid chromatography, HPLC)同时测定药食同源性物质葛根、槐花、金银花、肉苁蓉、罗汉果、人参、枳椇子提取物中功能成分的检测方法。方法 采用Agilent Eclipse XDB-C18色谱柱(4.6 mm×250 mm, 5 μm), 以乙腈-0.1%磷酸为流动相梯度洗脱, 流速0.6 mL/min, 柱温30 ℃, 检测波长210 nm, 进样量5 μL, 测定葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re及二氢杨梅素的含量。结果 8种功能成分在各自浓度范围内线性关系良好, 相关系数(r2)>0.999, 检出限为0.02~1.88 mg/L, 定量限为0.08~3.62 mg/L, 精密度实验结果为相对标准偏差(relative standard deviation, RSD)<3%, 平均加标回收率为95.49%~109.87%。结论 该方法简便快速, 准确度高, 可作为上述7种药食同源性物质功能成分的定性定量分析方法。

, correspAuthors=唐云, authorNote=null, correspAuthorsNote=
* 唐云(1974—), 男, 主要研究方向为酿酒生产工艺与白酒质量管控。E-mail:
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高惠惠(1998—), 女, 硕士, 主要研究方向为食品安全检测分析。E-mail:

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高惠惠(1998—), 女, 硕士, 主要研究方向为食品安全检测分析。E-mail:

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高惠惠(1998—), 女, 硕士, 主要研究方向为食品安全检测分析。E-mail:

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Wang by high-performance liquid chromatography coupled with a diode array detection method, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1177619443925528742, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, xref=null, ext=[AuthorCompanyExt(id=1177619443929723047, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, companyId=1177619443925528742, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Jinhui Liquor Co., Ltd., Longnan 742308, China), AuthorCompanyExt(id=1177619443938111656, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, companyId=1177619443925528742, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=金徽酒股份有限公司, 陇南 742308)])], figs=[ArticleFig(id=1177619446588911828, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Fig.1, caption=Spectral scanning images of 8 kinds of functional components, figureFileSmall=q69U9GiTdd+1fgXL5+kd4Q==, figureFileBig=J9EY8eUv/RKmwSCisTPFDw==, tableContent=null), ArticleFig(id=1177619446668603605, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=图1, caption=8种功能成分的光谱扫描图

注: A. 绿原酸; B. 二氢杨梅素; C. 人参皂苷Re; D. 芦丁; E. 松果菊苷; F. 葛根素; G. 毛蕊花糖苷; H. 罗汉果甜苷V。

, figureFileSmall=q69U9GiTdd+1fgXL5+kd4Q==, figureFileBig=J9EY8eUv/RKmwSCisTPFDw==, tableContent=null), ArticleFig(id=1177619446807015638, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Fig.2, caption=HPLC chromatograms of 4 kinds of gradient elution, figureFileSmall=hL/nLG4qKJlo9TY2DLXuGA==, figureFileBig=D2pyTCLiBvu10EJkGB8PNQ==, tableContent=null), ArticleFig(id=1177619446890901719, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=图2, caption=4种梯度洗脱的HPLC色谱图

注: A. 梯度洗脱一; B. 梯度洗脱二; C. 梯度洗脱三; D. 梯度洗脱四。

, figureFileSmall=hL/nLG4qKJlo9TY2DLXuGA==, figureFileBig=D2pyTCLiBvu10EJkGB8PNQ==, tableContent=null), ArticleFig(id=1177619447016730840, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Fig.3, caption=HPLC chromatograms of 4 kinds of mobile phase standard solutions, figureFileSmall=fjH/u0ZMggoGOYiyaIVomg==, figureFileBig=xk86xMRP2YFrArcQTakB0g==, tableContent=null), ArticleFig(id=1177619447092228313, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=图3, caption=4种流动相标准溶液HPLC色谱图

注: A. 甲醇-水溶液; B. 甲醇-0.1%磷酸水溶液; C. 乙腈-水; D. 乙腈-0.1%磷酸水溶液。

, figureFileSmall=fjH/u0ZMggoGOYiyaIVomg==, figureFileBig=xk86xMRP2YFrArcQTakB0g==, tableContent=null), ArticleFig(id=1177619447243223258, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Table 1, caption=

Gradient elution procedure

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 乙腈/% 0.1%磷酸水溶液/%
0 5 95
10.0 15 85
30.0 30 70
30.1 5 95
35.0 5 95
), ArticleFig(id=1177619447490687195, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=表1, caption=

梯度洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 乙腈/% 0.1%磷酸水溶液/%
0 5 95
10.0 15 85
30.0 30 70
30.1 5 95
35.0 5 95
), ArticleFig(id=1177619447616516316, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Table 2, caption=

Optimization of gradient elution program

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 流动相A/% 流动相B/% 时间/min 流动相A/% 流动相B/%
梯度洗脱 一 0 5 95 梯度洗脱 二 0 5 95
10 30 70 10 20 80
30 35 5 30 25 75
35 40 60 35 30 70
40 5 95 40 5 95
45 5 95 45 5 95
梯度洗脱 三 0 5 95 梯度洗脱 四 0 5 95
10 15 85 10.0 15 85
30 20 80 30.0 30 70
35 25 75 30.1 5 95
40 5 95 35.0 5 95
45 5 95
), ArticleFig(id=1177619447725568221, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=表2, caption=

梯度洗脱程序的优化

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 流动相A/% 流动相B/% 时间/min 流动相A/% 流动相B/%
梯度洗脱 一 0 5 95 梯度洗脱 二 0 5 95
10 30 70 10 20 80
30 35 5 30 25 75
35 40 60 35 30 70
40 5 95 40 5 95
45 5 95 45 5 95
梯度洗脱 三 0 5 95 梯度洗脱 四 0 5 95
10 15 85 10.0 15 85
30 20 80 30.0 30 70
35 25 75 30.1 5 95
40 5 95 35.0 5 95
45 5 95
), ArticleFig(id=1177619447914311902, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Table 3, caption=

Linear ranges, linear equations, correlation coefficients, limits of detection and limits of quantification of 8 kinds of functional components

, figureFileSmall=null, figureFileBig=null, tableContent=
功能成分 线性范围/(mg/L) 线性方程 相关系数(r2) 检出限/(mg/L) 定量限/(mg/L)
绿原酸 4.815~96.30 Y=18.0171X-18.2149 0.9996 0.04 0.10
葛根素 19.36~290.40 Y=29.1162X+0.7483 0.9998 0.03 0.14
松果菊苷 18.36~91.80 Y=11.8842X-1.8745 0.9999 0.19 0.64
二氢杨梅素 19.60~98.00 Y=71.8638X+8.0685 0.9997 0.02 0.08
芦丁 18.32~366.40 Y=31.8854X-33.5051 0.9995 0.03 0.10
毛蕊花糖苷 4.88~97.60 Y=15.7228X-7.8008 0.9998 0.08 0.23
罗汉果甜苷V 19.70~197.00 Y=1.2788X-0.8140 0.9999 0.73 2.39
人参皂苷Re 19.38~96.90 Y=1.2285X-1.6321 0.9992 1.88 3.62
), ArticleFig(id=1177619447985615071, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=表3, caption=

8种功能成分的线性范围、线性方程、相关系数、检出限及定量限

, figureFileSmall=null, figureFileBig=null, tableContent=
功能成分 线性范围/(mg/L) 线性方程 相关系数(r2) 检出限/(mg/L) 定量限/(mg/L)
绿原酸 4.815~96.30 Y=18.0171X-18.2149 0.9996 0.04 0.10
葛根素 19.36~290.40 Y=29.1162X+0.7483 0.9998 0.03 0.14
松果菊苷 18.36~91.80 Y=11.8842X-1.8745 0.9999 0.19 0.64
二氢杨梅素 19.60~98.00 Y=71.8638X+8.0685 0.9997 0.02 0.08
芦丁 18.32~366.40 Y=31.8854X-33.5051 0.9995 0.03 0.10
毛蕊花糖苷 4.88~97.60 Y=15.7228X-7.8008 0.9998 0.08 0.23
罗汉果甜苷V 19.70~197.00 Y=1.2788X-0.8140 0.9999 0.73 2.39
人参皂苷Re 19.38~96.90 Y=1.2285X-1.6321 0.9992 1.88 3.62
), ArticleFig(id=1177619448115638496, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Table 4, caption=

Precision test results of analysis of 8 kinds of functional components

, figureFileSmall=null, figureFileBig=null, tableContent=
功能成分 RSD/%
绿原酸 0.10
葛根素 0.11
松果菊苷 0.41
二氢杨梅素 0.18
芦丁 0.14
毛蕊花糖苷 0.16
罗汉果甜苷V 0.21
人参皂苷Re 0.11
), ArticleFig(id=1177619448174358753, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=表4, caption=

8种功能成分分析精密度实验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
功能成分 RSD/%
绿原酸 0.10
葛根素 0.11
松果菊苷 0.41
二氢杨梅素 0.18
芦丁 0.14
毛蕊花糖苷 0.16
罗汉果甜苷V 0.21
人参皂苷Re 0.11
), ArticleFig(id=1177619448270827746, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=EN, label=Table 5, caption=

Determination results of spiked recovery rates for 8 kinds of functional components

, figureFileSmall=null, figureFileBig=null, tableContent=
功能成分 加标量/(mg/L) 平均回收率/%
绿原酸 3.8720 102.78
8.1730
11.1370
二氢杨梅素 13.6340 105.63
27.2680
59.9260
松果菊苷 6.7120 107.00
22.5000
42.4050
毛蕊花糖苷 4.9020 109.87
8.8520
11.3630
葛根素 37.8104 102.05
79.6132
160.9379
芦丁 51.5597 98.87
114.6000
224.1990
罗汉果甜苷V 22.3079 104.43
45.8516
96.8513
人参皂苷Re 9.1690 95.49
18.1460
36.3500
), ArticleFig(id=1177619448388268259, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429500160364865, language=CN, label=表5, caption=

8种功能成分加标回收率测定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
功能成分 加标量/(mg/L) 平均回收率/%
绿原酸 3.8720 102.78
8.1730
11.1370
二氢杨梅素 13.6340 105.63
27.2680
59.9260
松果菊苷 6.7120 107.00
22.5000
42.4050
毛蕊花糖苷 4.9020 109.87
8.8520
11.3630
葛根素 37.8104 102.05
79.6132
160.9379
芦丁 51.5597 98.87
114.6000
224.1990
罗汉果甜苷V 22.3079 104.43
45.8516
96.8513
人参皂苷Re 9.1690 95.49
18.1460
36.3500
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高效液相色谱法同时测定7种药食同源物质功能成分
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高惠惠 , 赵亚雄 , 韩张斌 , 李海恬 , 杨成彦 , 唐云 *
食品安全质量检测学报 | 本期专题：功能性食品与功能性成分 2025,16(5): 127-133
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食品安全质量检测学报 | 本期专题：功能性食品与功能性成分 2025, 16(5): 127-133
高效液相色谱法同时测定7种药食同源物质功能成分
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高惠惠 , 赵亚雄, 韩张斌, 李海恬, 杨成彦, 唐云*
作者信息
  • 金徽酒股份有限公司, 陇南 742308

    • 高惠惠(1998—), 女, 硕士, 主要研究方向为食品安全检测分析。E-mail:

通讯作者:

* 唐云(1974—), 男, 主要研究方向为酿酒生产工艺与白酒质量管控。E-mail:
Simultaneous determination of functional components of 7 kinds of medicinal and food homologous substances by high performance liquid chromatography
Hui-Hui GAO , Ya-Xiong ZHAO, Zhang-Bin HAN, Hai-Tian LI, Cheng-Yan YANG, Yun TANG*
Affiliations
  • Jinhui Liquor Co., Ltd., Longnan 742308, China
出版时间: 2025-03-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241011003
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摘要
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目的 建立高效液相色谱法(high performance liquid chromatography, HPLC)同时测定药食同源性物质葛根、槐花、金银花、肉苁蓉、罗汉果、人参、枳椇子提取物中功能成分的检测方法。方法 采用Agilent Eclipse XDB-C18色谱柱(4.6 mm×250 mm, 5 μm), 以乙腈-0.1%磷酸为流动相梯度洗脱, 流速0.6 mL/min, 柱温30 ℃, 检测波长210 nm, 进样量5 μL, 测定葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re及二氢杨梅素的含量。结果 8种功能成分在各自浓度范围内线性关系良好, 相关系数(r2)>0.999, 检出限为0.02~1.88 mg/L, 定量限为0.08~3.62 mg/L, 精密度实验结果为相对标准偏差(relative standard deviation, RSD)<3%, 平均加标回收率为95.49%~109.87%。结论 该方法简便快速, 准确度高, 可作为上述7种药食同源性物质功能成分的定性定量分析方法。

高效液相色谱法  /  同时测定  /  药食同源  /  功能成分

Objective To establish a method for the simultaneous determination of functional components in extracts from 7 kinds of medicinal and food homologous substances: Pueraria lobata, Sophora japonica, Lonicera japonica, Cistanche deserticola, Siraitia grosvenorii, Panax ginseng and Hovenia dulcis by high performance liquid chromatography (HPLC). Methods The analysis was conducted using an Agilent Eclipse XDB-C18 column (4.6 mm× 250 mm, 5 μm) with a mobile phase gradient elution of acetonitrile-0.1% phosphoric acid at a flow rate of 0.6 mL/min, a column temperature of 30 ℃, a detection wavelength of 210 nm, and an injection volume of 5 μL. This method was employed to quantify the content of puerarin, rutin, chlorogenic acid, echinacoside, verbascoside, mogroside V, ginsenoside Re and dihydromyricetin. Results The results demonstrated that 8 kinds of functional components exhibited excellent linear relationships within their respective concentration ranges, with correlation coefficients (r²) exceeding 0.999. The limits of detection ranged from 0.02 to 1.88 mg/L, and the limits of quantification ranged from 0.08 to 3.62 mg/L. The precision experiment results showed that the relative standard deviation (RSD) was less than 3%, the average recoveries of spiked samples ranged from 95.49% to 109.87%. Conclusion This method is simple, rapid and highly accurate, making it suitable for the qualitative and quantitative analysis of the functional components in the aforementioned 7 kinds of medicinal and food homologous substances.

high performance liquid chromatography  /  simultaneous determination  /  medicinal and food homologous  /  functional components
高惠惠, 赵亚雄, 韩张斌, 李海恬, 杨成彦, 唐云. 高效液相色谱法同时测定7种药食同源物质功能成分. 食品安全质量检测学报, 2025 , 16 (5) : 127 -133 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241011003
Hui-Hui GAO, Ya-Xiong ZHAO, Zhang-Bin HAN, Hai-Tian LI, Cheng-Yan YANG, Yun TANG. Simultaneous determination of functional components of 7 kinds of medicinal and food homologous substances by high performance liquid chromatography[J]. Journal of Food Safety & Quality, 2025 , 16 (5) : 127 -133 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241011003
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0 引言
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如今人们对健康的关注度日益提升, 保健食品领域也随之蓬勃发展。药食同源物质因其功效丰富、来源广泛, 在该领域中占据着重要地位[1]。众多白酒企业敏锐地捕捉到这一趋势, 将“药食同源”的中药材及其提取物作为主要生产原料, 用于露酒、保健酒等功能性白酒的研发[2]。随着消费者保健意识的增强, 单一型白酒功能产品已难以满足市场需要, 复合型多功能白酒逐步上市。为了使产品功能达到最优化, 药食同源药材常常交互使用, 葛根、槐花、金银花、肉苁蓉、罗汉果、人参、枳椇子等作为露酒及保健酒研发的常用原料, 备受关注并得到了广泛的应用研究[3]。
葛根具透疹、升阳止泻、通经活络之功效, 含有黄酮、皂苷和三萜等成分[4-5]。葛根素是从葛根中提取的黄酮类化合物, 具有保护心脏、降低炎症[6]、保护肝脏[7]、改善心脑血管疾病[8]的药理作用。槐花具有清肝、泻火之功效, 芦丁是其发挥作用的重要成分, 具有抗炎、降血脂的作用[9]。金银花具有抗炎、解热之功效, 含有绿原酸、木樨素、挥发油等多种药理成分, 绿原酸是其重要的抗菌、抗氧化的有效成分[10-11]。肉苁蓉具有“沙漠人参”之称, 有补肾、增免之功效[12], 主要功能成分包括松果菊苷、毛蕊花糖苷和粗多糖等[13]。罗汉果有清热润肺、滑肠通便之功效, 对其多集中于甜苷类活性成分的研究[14-15]。人参有滋元、补气、生津之功效[16], 皂苷是其发挥功效的主要活性成分[17], 功能成分人参皂苷Re在神经炎症、免疫调节中有重要作用[18]。枳椇子是典型的解酒保肝中药材, 含黄酮、生物碱等化合物[19], 二氢杨梅素作为一种重要活性成分, 具有降压、抗菌等功效[20]。
目前葛根素[21-22]、芦丁[23-24]、绿原酸[25-26]、松果菊苷和毛蕊花糖苷[27-28]、罗汉果甜苷V[15]、人参皂苷Re[17]、二氢杨梅素[29-31]的检测方法优化、开发的文献较多, 但多是基于单一成分测定方法的建立。随着复合型多功能产品的上市, 需要一种可以快速检测常用多种药食同源原料的方法。现有的单一成分测定方法虽然可以用于特定成分的分析, 但其适用范围相对较窄, 无法满足复合型产品多成分分析的需求。《中国药典》及其他相关标准[GB/T 22251—2008《保健食品中葛根素的测定》、T/CNHFA 111.178—2024《保健食品用原料 槐米》、GB/T 22250—2008《保健食品中绿原酸的测定》、GB 1886.77—2016《食品安全国家标准 食品添加剂罗汉果甜苷》、NY/T 1842—2010《人参皂苷的测定》]规定了葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re是作为特定药材的重要的质量评价控制指标, 明确了其检测方法(枳椇子中二氢杨梅素的测定方法尚未明确), 但仍缺乏一种能够同时检测多种成分的标准方法用以对复合型保健产品中多种药食同源物质功能成分的含量测定分析。若仅依赖《中国药典》和现行标准方法对复合型产品进行测定, 可能会出现检测成本高昂、耗时耗力, 效率低等问题。
基于此, 本研究旨在建立高效液相色谱法(high performance liquid chromatography, HPLC)实现对葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re及二氢杨梅素这8种功能成分的同时测定, 为中药材及其提取物的质量控制和品质评价提供更为便捷的技术手段。
1 材料与方法
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1.1 材料、试剂及仪器
葛根提取物葛根素、槐花提取物芦丁、金银花提取物绿原酸、肉苁蓉提取物松果菊苷和毛蕊花糖苷、罗汉果提取物罗汉果甜苷V、人参提取物人参皂苷Re及枳椇子提取物二氢杨梅素均购自陕西华泽生物科技有限公司。
葛根素(含量96.8%)、芦丁(含量91.6%)、绿原酸(含量96.3%)、松果菊苷(含量91.8%)、毛蕊花糖苷(含量97.6%)、罗汉果皂苷V(含量98.5%)、人参皂苷Re(含量96.9%)(中国食品药品检定研究院); 二氢杨梅素(含量98%, 北京索莱宝科技有限公司); 甲醇、乙腈(色谱纯, 德国默克股份股份两合公司); 磷酸(分析纯, 国药集团化学试剂有限公司)。
1260 lnfinity II型高效液相色谱仪、Agilent Eclipse XDB-C18 (4.6 mm×250 mm, 5 μm)[安捷伦科技(中国)有限公司]; TU-1810紫外可见分光光度计(北京普析通用仪器有限责任公司); Milli-Q型超纯水仪[默克化工技术(上海)有限公司]; SQP十万分之一电子天平[赛多利斯科学仪器(北京)有限公司]; KQ5200 V型超声波清洗器(昆山市超声仪器有限公司)。
1.2 方法
1.2.1 标准溶液的配制
单个标准储备液: 分别称取葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re及二氢杨梅素标准品10.00 mg置于10 mL容量瓶中, 加甲醇超声溶解, 定容至刻度线, 制备成1000 mg/L的标准储备液。4 ℃冰箱保存, 有效期1个月。
混合标准使用液: 分别吸取8种标准储备液适量, 依次稀释, 配制成5个不同质量浓度的混合标准系列使用液。4 ℃冰箱保存, 有效期7 d。
1.2.2 样品处理
称取金银花提取物绿原酸、枳椇子提取物二氢杨梅素、肉苁蓉提取物松果菊苷和毛蕊花糖苷各5.00 mg, 加入15 mL甲醇超声溶解30 min, 待冷却后用甲醇定容至20 mL容量瓶中, 摇匀; 称取葛根提取物葛根素、槐花提取物芦丁、人参提取物人参皂苷Re、罗汉果提取物罗汉果甜苷V各5.00 mg, 加入20 mL甲醇超声溶解30 min, 待冷却后用甲醇定容至25 mL容量瓶中, 摇匀。试样经0.22 μm微孔滤膜过滤, 上机待测。
1.2.3 色谱条件
色谱柱: Agilent Eclipse XDB-C18 (4.6 mm×250 mm, 5 μm); 紫外检测波长: 210 nm; 柱温: 30 ℃; 流速: 0.6 mL/min; 进样量: 5 μL; 梯度洗脱程序详见表1。
1.2.4 样品含量测定
在规定色谱条件下, 以混合标准品溶液在色谱图上的保留时间对8种功能成分进行定性分析, 以峰面积定量, 根据标准曲线得到各样品含量。
1.3 数据处理
使用Excel 2016软件对数据进行处理, Origin 2022软件作图。
2 结果与分析
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2.1 检测波长的选择
采用紫外检测器, 分别扫描上述8种标准品溶液在190~400 nm波长范围内的光谱图, 结果见图1。实验发现葛根素和罗汉果甜苷V在200 nm处有最大吸收值, 响应最强; 毛蕊花糖苷、松果菊苷和人参皂苷Re在205 nm处有最大吸收值; 二氢杨梅素的最大吸收波长是210 nm; 绿原酸和芦丁的最大吸收波长是220 nm。综合选择在波长210 nm下同时测定葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re及二氢杨梅素。
2.2 洗脱梯度的选择
本研究中进行了14次梯度优化, 考察了不同洗脱程序下各成分的分离度、响应值、峰型、检测周期, 以确定最佳洗脱梯度参数。选择了实验中4个具有代表性的洗脱梯度, 用以说明优化效果。梯度洗脱程序见表2, 洗脱效果见图2。
结果表明(图2), 采用梯度一(图2A)进行洗脱时, 观察到各成分峰形相互折叠, 不能有效分离, 应调整流动相比例。采用梯度二(图2B)洗脱时, 罗汉果甜苷V和人参皂苷Re未出峰未被洗脱出, 没有出现峰图。其余6种成分被洗脱出来, 且各成分之间有了一定程度的分离, 但绿原酸、葛根素、松果菊苷和毛蕊花糖苷洗脱效果不理想, 未被完全分离, 出现双峰, 无法进行定性定量, 仍需进一步优化洗脱程序。采用梯度三(图2C)洗脱时, 8种成分均被洗脱, 葛根素和松果菊苷峰型得到改善, 但毛蕊花糖苷仍为双峰, 绿原酸峰型较差。最后采用梯度四(图2D)洗脱时, 所有成分完全分离, 且分离度高、峰型佳、对称性好, 满足定性定量分析要求, 检测时间缩短, 为最优洗脱梯度。
2.3 流动相的选择
本研究探究了甲醇-水、甲醇-0.1%磷酸水溶液、乙腈-水和乙腈-0.1%的磷酸水溶液4种流动相体系对上述8种成分的洗脱效果(图3)。
结果表明以甲醇-水为流动相时, 基线漂移, 仅有二氢杨梅素出峰; 以甲醇-0.1%磷酸水溶液为流动相时, 二氢杨梅素和绿原酸被洗脱出来, 其他成分均未出峰, 且绿原酸色谱峰型差, 无法进行定量分析; 以乙腈-水为流动相时, 除绿原酸外的7种成分均出峰, 说明相较于甲醇, 乙腈的洗脱能力更强。以乙腈-0.1%磷酸水溶液为流动相时, 葛根素、芦丁、绿原酸、松果菊苷、毛蕊花糖苷、罗汉果甜苷V、人参皂苷Re及二氢杨梅素这8种功能成分均被洗脱出来, 基线平稳, 峰型较好, 且能完全分离, 分离度大于1.5。因此选用乙腈-0.1%的磷酸水溶液作为流动相。
2.4 方法学考察
2.4.1 标准曲线、检出限、定量限和线性范围
按照“1.2.3”项下色谱条件, 将配制好的混合标准使用液依次进样, 以质量浓度为横坐标(X, mg/L), 峰面积为纵坐标(Y)绘制标准曲线, 得出线性回到方程。将8种功能成分标准品溶液进行适当稀释得到不同浓度溶液, 按照3倍信噪比(S/N)作为检出限, 10倍信噪比作为定量限。线性范围、线性方程、方法检出限和定量限结果见表3。结果表明8种功能成分在各自质量浓度范围内线性关系良好, 相关系数(r2)均大于0.999, 检出限为0.02~1.88 mg/L, 定量限为0.08~3.62 mg/L, 满足检测要求。
2.4.2 精密度实验
按照“1.2.3”项下色谱条件, 取同一浓度的混合标准溶液连续进样6次, 记录峰面积, 测定结果见表4。绿原酸、葛根素、松果菊苷、二氢杨梅素、芦丁、毛蕊花糖苷、罗汉果甜苷V及人参皂苷Re峰面积测定结果的RSD分别是0.10%、0.11%、0.41%、0.18%、0.14%、0.16%、0.21%、0.11%, 均小于3%, 得出数据较为一致, 离散程度低, 表明仪器精密度良好。
2.5 样品含量测定及加标回收实验
按照“1.2.2”样品处理方法和“1.2.3”色谱条件, 测得金银花提取物绿原酸含量为5.7570 mg/L、枳椇子提取物二氢杨梅素含量为32.6869 mg/L、肉苁蓉提取物松果菊苷和毛蕊花糖苷含量分别为23.8608 mg/L和6.7236 mg/L、葛根提取物葛根素含量为79.1286 mg/L、槐花提取物芦丁含量为92.5780 mg/L、罗汉果提取物罗汉果甜苷V含量为50.6113 mg/L、人参提取物人参皂苷Re含量为21.3980 mg/L。取8种制备的提取物试液, 按照提取物中各成分含量的低、中、高3个浓度水平, 按照体积比为1:1依次添加标准品溶液, 计算各功能成分的加标回收率, 结果见表5。由表5可知, 各个功能成分的平均加标回收率分别是绿原酸102.78%、二氢杨梅素105.63%、松果菊苷107.00%、毛蕊花糖苷109.87%、葛根素102.05%、芦丁98.87%、罗汉果甜苷V 104.43%、人参皂苷Re 95.49%, 表明该测定方法具有可行性。
3 结论
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本研究建立了HPLC同时测定药食同源性物质葛根提取物葛根素、槐花提取物芦丁、金银花提取物绿原酸、肉苁蓉提取物松果菊苷和毛蕊花糖苷、罗汉果提取物罗汉果甜苷V、人参提取物人参皂苷Re、枳椇子提取物二氢杨梅素的检测方法, 这8种功能成分在各自浓度范围内线性关系良好, 相关系数(r2)>0.999, 检出限为0.02~1.88 mg/L, 定量限为0.08~3.62 mg/L, 精密度实验结果为RSD<3%, 平均回收率为95.49%~109.87%。该方法高效快速, 可在35 min内完成上述8种功能成分的同时检测, 解决了一些常用药食同源物质多个成分含量测定分析时耗时耗力、试剂消耗的问题, 降低了检测成本, 提高了检测效率, 具有较佳的实用价值, 可为药食同源性物质及其提取物质量控制和品质评价提供更为便捷的检测技术手段。
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2025年第16卷第5期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241011003
  • 接收时间：2024-10-11
  • 首发时间：2025-07-19
  • 出版时间：2025-03-15
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  • 收稿日期：2024-10-11
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    金徽酒股份有限公司, 陇南 742308

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* 唐云(1974—), 男, 主要研究方向为酿酒生产工艺与白酒质量管控。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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