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Article(id=1151881498182431208, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250325002, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1742832000000, receivedDateStr=2025-03-25, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752559550089, onlineDateStr=2025-07-15, pubDate=1748102400000, pubDateStr=2025-05-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752559550089, onlineIssueDateStr=2025-07-15, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752559550089, creator=13701087609, updateTime=1752559550089, updator=13701087609, issue=Issue{id=1151881493552394994, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='10', pageStart='1', pageEnd='324', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752559548986, creator=13701087609, updateTime=1756202008453, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1167159075906265916, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1167159075906265917, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151881493552394994, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=28, endPage=34, ext={EN=ArticleExt(id=1151923895184536282, articleId=1151881498182431208, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Determination of dehydroepiandrosterone content in health foods by liquid chromatography, columnId=1151881695071924743, journalTitle=Journal of Food Safety & Quality, columnName=Highlight: The 9th Academic Conference Papers on Food Quality, Safety and Intelligent Testing, runingTitle=null, highlight=null, articleAbstract=

Objective To establish an analytical method for the determination of dehydroepiandrosterone (DHEA) content in variety of health food by liquid chromatography. Methods Tablets and capsules were extracted with acetonitrile:water (60:40, V:V); oral liquid was extracted with acetonitrile and added sodium chloride to salt out the targets from water; gel candy was dissolved in warm water first, and extracted with acetonitrile, after that ammonium sulfate was used to precipitate gelatin and made acetonitrile water stratification simultaneously. Finally, DHEA in health product was effectively extracted, then determined by liquid chromatography with diode array detection or liquid chromatography tandem mass spectrometry and quantified by external standard method. Results The established methods were effective in extracting DHEA from health foods, with limit of detection for solid or powder sample was 0.15 g/kg and for liquid was 1.5 mg/L and limit of quantification was 0.5 g/kg for solid or powder samples and 5 mg/L for liquid, respectively. The recovery of this method was 95.7%-104.0%, and the relative standard deviation was 1.10%-3.76%. It was found that the content of DHEA in imported dietary supplements was mostly consistent with the label, with a few samples not matching the label in real sample. Conclusion This method is sensitive, simple and convenient, and can be used not only to monitor the illegal addition of DHEA in domestic health foods, but also to evaluate the content of DHEA in cross-border dietary supplements.

, correspAuthors=Juan MENG, 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=Qiao-Zhen GUO, Jing ZHANG, Xin ZHANG, Bing SHAO, Juan MENG), CN=ArticleExt(id=1151923911533932760, articleId=1151881498182431208, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=液相色谱法测定保健食品中脱氢表雄酮含量, columnId=1151881695206142473, journalTitle=食品安全质量检测学报, columnName=专栏：第九届食品质量安全与智能检测学术会议论文, runingTitle=null, highlight=null, articleAbstract=

目的 建立液相色谱法检测多种类型保健食品中脱氢表雄酮(dehydroepiandrosterone, DHEA)含量的分析方法。方法 片剂和胶囊采用乙腈:水(60:40, V:V)提取; 口服液用乙腈提取, 加入氯化钠使乙腈水分层; 凝胶糖果用温水溶解后, 乙腈提取, 利用硫酸铵沉淀明胶且实现乙腈水分层, 最终有效提取保健食品中DHEA, 利用液相色谱二极管阵列检测或液相色谱-串联四极杆质谱法测定, 外标法定量。结果 建立的方法能有效提取检测保健食品中DHEA, 液相色谱二极管阵列检测固体或粉末样品方法检出限为0.15 g/kg, 定量限为0.5 g/kg; 液体样品方法检出限为1.5 mg/L, 定量限为5 mg/L; 方法回收率为95.7%~104.0%, 相对标准偏差为1.10%~3.76%。将该方法用于实际样品检测, 发现大多数电商平台的膳食补充剂中DHEA的含量与标签一致, 少量样品与标签不符。结论 该方法灵敏, 简单便捷, 不仅可用于监测国内功能性保健食品中DHEA的非法添加, 同时可实现对跨境膳食补充剂中DHEA的含量评价。

, correspAuthors=孟娟, authorNote=null, correspAuthorsNote=
* 孟娟(1971—), 女, 副主任技师, 主要研究方向为食品安全检测。E-mail:
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郭巧珍(1984—), 女, 博士, 副研究员, 主要研究方向为食品安全检测。E-mail:

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郭巧珍(1984—), 女, 博士, 副研究员, 主要研究方向为食品安全检测。E-mail:

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School of Public Health, Capital Medical University, Beijing 100089, China), AuthorCompanyExt(id=1167158653804098229, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, companyId=1167158653787321012, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.首都医科大学公共卫生学院, 北京 100089)])], figs=[ArticleFig(id=1167158655590871787, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Fig.1, caption=Maximum absorption of DHEA when the mobile phase is methanol and acetonitrile, figureFileSmall=9gC/Ik4v0YjrII8tPnj2mw==, figureFileBig=nJvyVMj4+m146zLdROsV2A==, tableContent=null), ArticleFig(id=1167158655741866734, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=图1, caption=流动相为甲醇和乙腈时DHEA的最大吸收, figureFileSmall=9gC/Ik4v0YjrII8tPnj2mw==, figureFileBig=nJvyVMj4+m146zLdROsV2A==, tableContent=null), ArticleFig(id=1167158655892861681, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Fig.2, caption=Effects of flow (A) and column temperature (B) on DHEA, figureFileSmall=eG9+RGWzEqSnYklNyxKH7A==, figureFileBig=KTL4atQyT9Jm5/x3E6Y7jA==, tableContent=null), ArticleFig(id=1167158656035468020, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=图2, caption=流速(A)和柱温(B)对DHEA的影响, figureFileSmall=eG9+RGWzEqSnYklNyxKH7A==, figureFileBig=KTL4atQyT9Jm5/x3E6Y7jA==, tableContent=null), ArticleFig(id=1167158656127742710, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Fig.3, caption=Chromatograms of DHEA standard solution, figureFileSmall=iOWflUYWcrGJ77GjdXd/AA==, figureFileBig=ikk7dJ36HalxqdDQWlHAqQ==, tableContent=null), ArticleFig(id=1167158656220017400, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=图3, caption=DHEA标准溶液色谱图

注: A. LC-二极管阵列检测色谱图(50 μg/mL); B. LC-MS谱图(1 ng/mL)(上下两图分别是DHEA的定量离子对和定性离子对)。

, figureFileSmall=iOWflUYWcrGJ77GjdXd/AA==, figureFileBig=ikk7dJ36HalxqdDQWlHAqQ==, tableContent=null), ArticleFig(id=1167158656282931962, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Fig.4, caption=Liquid chromatography-mass spectrometry spectrums of real sample, figureFileSmall=GPHgeVJc0IBHs2qq6JufZQ==, figureFileBig=fa1hC4u06Uv71WUUydFQXw==, tableContent=null), ArticleFig(id=1167158656396178171, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=图4, caption=实际样品的液相色谱-质谱谱图

注: 左右两图分别是定量离子对和定性离子对。

, figureFileSmall=GPHgeVJc0IBHs2qq6JufZQ==, figureFileBig=fa1hC4u06Uv71WUUydFQXw==, tableContent=null), ArticleFig(id=1167158656484258557, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Table 1, caption=

Recoveries and precisions of the method (n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品基质 本底 添加浓度
0.5 g/kg 5.0 g/kg 10.0 g/kg
回收率/% RSDs/% 回收率/% RSDs/% 回收率/% RSDs/%
硬胶囊 ND 95.7 1.86 96.7 1.17 96.9 2.14
片剂 ND 96.1 2.90 98.6 1.10 97.7 3.34
凝胶糖果 ND 99.6 3.53 103.0 2.22 104.0 2.04
口服液 ND 102.0 3.76 101.0 2.29 101.0 2.76
), ArticleFig(id=1167158656559756031, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=表1, caption=

方法的回收率和精密度(n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品基质 本底 添加浓度
0.5 g/kg 5.0 g/kg 10.0 g/kg
回收率/% RSDs/% 回收率/% RSDs/% 回收率/% RSDs/%
硬胶囊 ND 95.7 1.86 96.7 1.17 96.9 2.14
片剂 ND 96.1 2.90 98.6 1.10 97.7 3.34
凝胶糖果 ND 99.6 3.53 103.0 2.22 104.0 2.04
口服液 ND 102.0 3.76 101.0 2.29 101.0 2.76
), ArticleFig(id=1167158656652030720, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Table 2, caption=

Testing results of the real samples

, figureFileSmall=null, figureFileBig=null, tableContent=
样品 剂型 重量
/(g/粒)		 标签含量/(mg/粒) 实测含量/(mg/粒) 相对偏差/%
样品1 胶囊 0.260 25 25.7 2.8
样品2 胶囊 0.199 25 27.2 8.8
样品3 片剂 0.685 100 ND ND
样品4 胶囊 0.272 100 94.3 -5.7
样品5 片剂 0.437 50 46.7 -6.6
), ArticleFig(id=1167158656731722497, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=表2, caption=

实际样品检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
样品 剂型 重量
/(g/粒)		 标签含量/(mg/粒) 实测含量/(mg/粒) 相对偏差/%
样品1 胶囊 0.260 25 25.7 2.8
样品2 胶囊 0.199 25 27.2 8.8
样品3 片剂 0.685 100 ND ND
样品4 胶囊 0.272 100 94.3 -5.7
样品5 片剂 0.437 50 46.7 -6.6
), ArticleFig(id=1167158656794637058, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Table 3, caption=

Recoveries and precisions of DHEA in different laboratories (n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品基质 实验室编号 本底值
/(g/kg)		 低浓度 中浓度 高浓度
平均回收率/% RSDs/% 平均回收率/% RSDs/% 平均回收率/% RSDs/%
硬胶囊 Lab 1 ND 104.0 1.76 97.1 0.58 96.8 0.36
Lab 2 ND 95.4 2.48 95.5 0.77 96.8 0.65
Lab 3 ND 95.2 4.94 98.3 2.98 104 1.31
Lab 4 ND 95.6 4.37 99.6 1.91 95.6 0.64
Lab 5 ND 101.0 4.98 97.9 2.05 98.3 2.18
回收率/% 95.2~104.0 95.5~99.6 95.6~104.0
RSDs/% 1.76~4.98 0.58~2.98 0.36~2.18
片剂 Lab 1 ND 102.0 1.99 100.0 1.17 97.4 1.11
Lab 2 ND 95.7 1.70 99.4 1.09 96.6 0.27
Lab 3 ND 98.8 4.99 97.3 4.56 103.0 1.24
Lab 4 ND 96.5 2.33 105.0 0.54 97.5 0.32
Lab 5 ND 103.0 2.40 99.7 2.76 100.0 0.84
回收率/% 95.7~103.0 97.3~105.0 96.6~103.0
RSDs/% 1.70~4.99 0.54~4.56 0.27~1.24
凝胶糖果 Lab 1 ND 102.0 1.91 102.0 1.93 98.8 4.91
Lab 2 ND 104.0 3.89 100.0 2.05 102.0 1.29
Lab 3 ND 105.0 4.16 104.0 3.45 97.6 4.46
Lab 4 ND 98.3 3.31 101.0 1.19 99.5 0.24
Lab 5 ND 103.0 1.70 102.0 1.70 99.5 0.59
回收率/% 98.3~105.0 100.0~104.0 97.6~102.0
RSDs/% 1.70~4.16 1.19~3.45 0.24~4.91
), ArticleFig(id=1167158656882717444, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=表3, caption=

DHEA在不同实验室的回收率和精密度(n=6)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品基质 实验室编号 本底值
/(g/kg)		 低浓度 中浓度 高浓度
平均回收率/% RSDs/% 平均回收率/% RSDs/% 平均回收率/% RSDs/%
硬胶囊 Lab 1 ND 104.0 1.76 97.1 0.58 96.8 0.36
Lab 2 ND 95.4 2.48 95.5 0.77 96.8 0.65
Lab 3 ND 95.2 4.94 98.3 2.98 104 1.31
Lab 4 ND 95.6 4.37 99.6 1.91 95.6 0.64
Lab 5 ND 101.0 4.98 97.9 2.05 98.3 2.18
回收率/% 95.2~104.0 95.5~99.6 95.6~104.0
RSDs/% 1.76~4.98 0.58~2.98 0.36~2.18
片剂 Lab 1 ND 102.0 1.99 100.0 1.17 97.4 1.11
Lab 2 ND 95.7 1.70 99.4 1.09 96.6 0.27
Lab 3 ND 98.8 4.99 97.3 4.56 103.0 1.24
Lab 4 ND 96.5 2.33 105.0 0.54 97.5 0.32
Lab 5 ND 103.0 2.40 99.7 2.76 100.0 0.84
回收率/% 95.7~103.0 97.3~105.0 96.6~103.0
RSDs/% 1.70~4.99 0.54~4.56 0.27~1.24
凝胶糖果 Lab 1 ND 102.0 1.91 102.0 1.93 98.8 4.91
Lab 2 ND 104.0 3.89 100.0 2.05 102.0 1.29
Lab 3 ND 105.0 4.16 104.0 3.45 97.6 4.46
Lab 4 ND 98.3 3.31 101.0 1.19 99.5 0.24
Lab 5 ND 103.0 1.70 102.0 1.70 99.5 0.59
回收率/% 98.3~105.0 100.0~104.0 97.6~102.0
RSDs/% 1.70~4.16 1.19~3.45 0.24~4.91
), ArticleFig(id=1167158656979186437, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=EN, label=Table 4, caption=

Results of inter laboratory comparison of real samples

, figureFileSmall=null, figureFileBig=null, tableContent=
编号 DHEA含量/(g/kg)
Lab 1 Lab 2 Lab 3 Lab 4 Lab 5 Lab 6* RSDs/%
样品1
硬胶囊DHEA-01S		 97.7 102.0 106.0 104.0 98.7 103.0 3.43
样品2
硬胶囊DHEA-02S-2		 386.0 374.0 405.0 375.0 371.0 387.0 3.65
样品3
片剂DHEA-03S		 78.5 73.6 78.3 76.0 71.7 76.6 3.92
), ArticleFig(id=1167158657071461126, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151881498182431208, language=CN, label=表4, caption=

实际样品实验室间比对结果

, figureFileSmall=null, figureFileBig=null, tableContent=
编号 DHEA含量/(g/kg)
Lab 1 Lab 2 Lab 3 Lab 4 Lab 5 Lab 6* RSDs/%
样品1
硬胶囊DHEA-01S		 97.7 102.0 106.0 104.0 98.7 103.0 3.43
样品2
硬胶囊DHEA-02S-2		 386.0 374.0 405.0 375.0 371.0 387.0 3.65
样品3
片剂DHEA-03S		 78.5 73.6 78.3 76.0 71.7 76.6 3.92
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郭巧珍 1, 2 , 张晶 1, 2 , 张昕 1 , 邵兵 1, 2 , 孟娟 1, *
食品安全质量检测学报 | 专栏：第九届食品质量安全与智能检测学术会议论文 2025,16(10): 28-34
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食品安全质量检测学报 | 专栏：第九届食品质量安全与智能检测学术会议论文 2025, 16(10): 28-34
液相色谱法测定保健食品中脱氢表雄酮含量
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郭巧珍1, 2 , 张晶1, 2, 张昕1, 邵兵1, 2, 孟娟1, *
作者信息
  • 1.北京市疾病预防控制中心, 食物中毒诊断溯源技术北京重点实验室, 北京 100013
  • 2.首都医科大学公共卫生学院, 北京 100089

    • 郭巧珍(1984—), 女, 博士, 副研究员, 主要研究方向为食品安全检测。E-mail:

通讯作者:

* 孟娟(1971—), 女, 副主任技师, 主要研究方向为食品安全检测。E-mail:
Determination of dehydroepiandrosterone content in health foods by liquid chromatography
Qiao-Zhen GUO1, 2 , Jing ZHANG1, 2, Xin ZHANG1, Bing SHAO1, 2, Juan MENG1, *
Affiliations
  • 1. Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Control and Prevention, Beijing 100013, China
  • 2. School of Public Health, Capital Medical University, Beijing 100089, China
出版时间: 2025-05-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250325002
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摘要
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目的 建立液相色谱法检测多种类型保健食品中脱氢表雄酮(dehydroepiandrosterone, DHEA)含量的分析方法。方法 片剂和胶囊采用乙腈:水(60:40, V:V)提取; 口服液用乙腈提取, 加入氯化钠使乙腈水分层; 凝胶糖果用温水溶解后, 乙腈提取, 利用硫酸铵沉淀明胶且实现乙腈水分层, 最终有效提取保健食品中DHEA, 利用液相色谱二极管阵列检测或液相色谱-串联四极杆质谱法测定, 外标法定量。结果 建立的方法能有效提取检测保健食品中DHEA, 液相色谱二极管阵列检测固体或粉末样品方法检出限为0.15 g/kg, 定量限为0.5 g/kg; 液体样品方法检出限为1.5 mg/L, 定量限为5 mg/L; 方法回收率为95.7%~104.0%, 相对标准偏差为1.10%~3.76%。将该方法用于实际样品检测, 发现大多数电商平台的膳食补充剂中DHEA的含量与标签一致, 少量样品与标签不符。结论 该方法灵敏, 简单便捷, 不仅可用于监测国内功能性保健食品中DHEA的非法添加, 同时可实现对跨境膳食补充剂中DHEA的含量评价。

保健食品  /  脱氢表雄酮  /  液相色谱法

Objective To establish an analytical method for the determination of dehydroepiandrosterone (DHEA) content in variety of health food by liquid chromatography. Methods Tablets and capsules were extracted with acetonitrile:water (60:40, V:V); oral liquid was extracted with acetonitrile and added sodium chloride to salt out the targets from water; gel candy was dissolved in warm water first, and extracted with acetonitrile, after that ammonium sulfate was used to precipitate gelatin and made acetonitrile water stratification simultaneously. Finally, DHEA in health product was effectively extracted, then determined by liquid chromatography with diode array detection or liquid chromatography tandem mass spectrometry and quantified by external standard method. Results The established methods were effective in extracting DHEA from health foods, with limit of detection for solid or powder sample was 0.15 g/kg and for liquid was 1.5 mg/L and limit of quantification was 0.5 g/kg for solid or powder samples and 5 mg/L for liquid, respectively. The recovery of this method was 95.7%-104.0%, and the relative standard deviation was 1.10%-3.76%. It was found that the content of DHEA in imported dietary supplements was mostly consistent with the label, with a few samples not matching the label in real sample. Conclusion This method is sensitive, simple and convenient, and can be used not only to monitor the illegal addition of DHEA in domestic health foods, but also to evaluate the content of DHEA in cross-border dietary supplements.

health food  /  dehydroepiandrosterone  /  liquid chromatography
郭巧珍, 张晶, 张昕, 邵兵, 孟娟. 液相色谱法测定保健食品中脱氢表雄酮含量. 食品安全质量检测学报, 2025 , 16 (10) : 28 -34 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250325002
Qiao-Zhen GUO, Jing ZHANG, Xin ZHANG, Bing SHAO, Juan MENG. Determination of dehydroepiandrosterone content in health foods by liquid chromatography[J]. Journal of Food Safety & Quality, 2025 , 16 (10) : 28 -34 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250325002
正文
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0 引言
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脱氢表雄酮(dehydroepiandrosterone, DHEA)是人体血液循环中最为丰富的甾体物质[1-2], 是肾上腺与性腺分泌的一种C19类固醇素, 是睾酮等多种激素的前体, 被称为“荷尔蒙之母”, 起到保持荷尔蒙平衡、支持整体健康的作用。DHEA被认为具有抗衰老[3]、抗癌[4]、改善骨质疏松[5]、减轻焦虑和抑郁[6-9], 以及调节血脂[10]、调节炎性细胞因子[11-12]、改善糖尿病[13-14]等功能, 并且被用于治疗妇女卵巢储备减退[15]、更年期症状[16-18], 也应用于生殖领域[19]。生理剂量的DHEA未见明显的毒副作用, 但大剂量长期使用DHEA仍然存在安全问题[20], 摄入过多DHEA不但影响人体激素的平衡, 而且会导致脸部皮肤出油、多毛、长青春痘等, 还有致癌风险[21-22], 使用不当, 会造成不孕。因此, 在美国, DHEA被列为处方药并禁止其作为保健品销售, 但由于监管标准的差异和市场需求驱动, 中国市场对于DHEA的需求持续增长, 部分消费者希望通过使用DHEA来增加生育机会、提高抵抗力等。《辅助生殖促排卵药物治疗专家共识》指出[23], DHEA可以应用于卵巢早衰患者, 通常的推荐用量为25 mg/次。消费者应关注其潜在的风险和副作用, 合理使用保健品。
数据显示2023年全球DHEA胶囊市场销售额达到上亿美元, 且中国市场规模也达到上百万美元[24]。尽管目前我国还没有批准此类保健品, 但国外多款含DHEA的膳食补充剂在跨境电商平台上热销, 目前进口产品剂型主要有片剂和胶囊, 含量约25~100 mg/片(粒)。为应对消费市场剂型的多样化和对产品质量的保障, 有必要建立检测适用于多种剂型保健食品中DHEA的分析方法。另一方面, DHEA被归类为合成代谢雄性类固醇, 为世界反兴奋剂机构(World Anti-Doping Agency, WADA)禁用名单上的药物, 在体育赛事运动员[25]的饮食中不得检出。由于DHEA具有多种功效[2], 建立不同剂型保健品中DHEA的分析方法是防止功能性保健食品中违法添加检测的需要[26]。DHEA的检测方法主要有气相色谱法和液相色谱法。张丽霞等[27]建立了功能性保健食品(减肥类)中DHEA的气相色谱-质谱检测方法, 以监督保健食品中DHEA的非法添加。临床血液[28-30]和动物源性食品[31]中DHEA的检测主要采用液相色谱-质谱联用技术。GB/T 5009.193—2003《保健食品中脱氢表雄甾酮(DHEA)的测定》采用甲醇-水为流动相, 液相色谱紫外检测[32], 基质覆盖片剂和胶囊。
本研究一方面针对跨境电商膳食补充剂DHEA产品中DHEA的含量采用液相色谱法进行评价, 并利用液相色谱-串联四极杆质谱法(ultra performance liquid chromatography- tandem mass spectrometry, UPLC-MS/MS)对结果进行确证; 另一方面对于功能性保健食品采用UPLC-MS/MS检测是否存在DHEA的非法添加。以期为保健食品中DHEA的监测提供依据。
1 材料与方法
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1.1 试剂与材料
DHEA标准溶液(配制在甲醇中, 质量浓度100 μg/mL, 纯度>99.9%)(天津阿尔塔科技有限公司); 甲醇、乙腈(色谱纯, 美国Honeywell公司); 甲酸(色谱纯, 美国Sigma公司); 氯化钠、硫酸铵(分析纯, 国药集团化学试剂有限公司); 超低元素水(Mili-Q Water System纯化制备); 0.22 μm有机滤膜(天津津腾实验设备有限公司)。
1.2 设备与仪器
UPLC-PDA-TQXS MS超高效液相色谱-二极管阵列-串联四极杆质谱仪、ACQUITY UPLC BEH C18色谱柱(100 mm× 2.1 mm, 1.7 μm)(美国Waters公司); AL204DU-1C电子天平(感量为0.1 mg, 德国梅特勒-托利多公司); JJ200 G&G电子天平(感量为0.01 g, 常熟双杰测试仪器厂); G560E Vortex-gene2旋涡振荡器(美国Scientific Industries公司); Allegra X-30R冷冻离心机(转速≥6000 r/min, 美国Beckman公司); SB25-12DTD超声波清洗器(宁波新芝生物科技股份有限公司); Milli-Q去离子水发生器(美国Millipore公司)。
1.3 方法
1.3.1 样品前处理
(1)硬胶囊和片剂等固体试样
硬胶囊取不少于20粒或不低于5 g样品, 剪开取其内容物, 研细(必要时), 混匀; 片剂取不少于20片或不低于10 g试样, 经高速粉碎机或研钵磨成粉状, 混匀。
准确称取0.2 g试样(准确至0.01 g)于50 mL离心管中, 加入20 mL流动相, 涡旋1 min, 至分散, 超声提取5 min后高速离心5 min (转速10000 r/min), 上清液经0.22 μm滤膜过滤后上机测定。
(2)凝胶糖果固体试样
凝胶糖果取不低于10粒(对于不同色泽或风味混装的试样, 则按色泽或种类均匀取样), 剪碎。准确称取5.0 g (准确至0.01 g)剪碎的凝胶糖果, 用500 mL温水溶解, 放置常温后, 取5 mL溶解后的液体样品加入5 mL乙腈提取, 再加入硫酸铵至有固体析出。涡旋30 s, 超声提取5 min后, 高速离心5 min(转速10000 r/min), 取上层(有机层)经0.22 μm滤膜过滤后上机测定。
(3)口服液等液体试样
取5支独立包装或不少于20 mL样品充分混匀。准确量取5.0 mL试样(精确至0.01 mL)于50 mL离心管中, 加入5 mL乙腈提取, 再加入氯化钠至有固体析出。涡旋30 s, 超声提取5 min后, 高速离心5 min(转速10000 r/min), 取上层(有机层)经0.22 μm滤膜过滤后上机测定。
1.3.2 色谱质谱条件
色谱柱: ACQUITY UPLC BEH C18柱(2.1 mm×100 mm, 1.7 μm)。柱温: 35 ℃; 流动相: 乙腈-0.1%甲酸水(60:40, V:V); 流速: 0.3 mL/min; 进样量: 5 μL; 检测波长: 200 nm。质谱条件: 电离模式: 电喷雾离子源正离子(electro spray ionization, ESI+); 毛细管电压: 3 kV; 锥孔电压: 15 V; 源温: 150 ℃; 脱溶剂气温度: 450 ℃; 脱溶剂气流量: 800 L/Hr (N2), 锥孔气流量: 150 L/Hr (N2), 碰撞气流量: 0.13 mL/min (Ar)。质谱扫描方式: 多反应监测(multi reaction monitorin, MRM); 离子对信息: 289.2>253.2(定量), 碰撞能量: 11 eV; 289.2>271.2, 碰撞能量: 7 eV。
1.4 数据处理
数据通过Masslynx 4.2采集并定量分析。本研究数据处理及统计图表绘制采用Microsoft Office Excel 2007软件。
2 结果与分析
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2.1 色谱及质谱条件的优化
参考GB/T 5009.193—2003和文献[31,33], 选择C18柱作为分离DHEA的色谱柱。二极管阵列检测器(photo-diode array, PDA)扫描测定DHEA的最大吸收波长为195~200 nm, 见图1, 乙腈和甲醇的截止波长分别为190 nm和210 nm, 乙腈中DHEA响应高于甲醇, 同时为减少干扰, 选择乙腈为有机相。实验发现在水相中加入0.1%甲酸能有效地提高DHEA灵敏度(特别是对UPLC-MS/MS体系)。色谱柱温度和流速的改变可以影响目标化合物的出峰时间和响应强度, 实验通过优化柱温和流速(结果见图2), 发现流速越快, 柱温越高, 保留时间越靠前, 受基质影响较大, 响应越低, 通过优化条件, 最终采用乙腈-甲酸水(0.1%)的体积比为60:40, 等度洗脱, 流速为0.3 mL/min, 柱温为35 ℃, 其色谱图见图3A。采用DHEA标准溶液优化得到质谱参数, 如母离子、子离子和最优碰撞能量(见1.3.2), 标准溶液的LC-MS/MS谱图见图3B。
2.2 样品前处理的优化
常见保健品配料有: 片剂的基质成分主要是麦芽糊精、大豆分离蛋白、乳糖等; 硬胶囊的基质成分主要是麦芽糊精、微晶纤维素、羟甲基淀粉钠、羟丙基甲基纤维素、二氧化硅、硬脂酸镁; 这些配料易溶于水, 兼顾DHEA易溶于正己烷、甲醇等有机相的特点以及减少UPLC-MS/MS分离中的溶剂效应, 片剂和胶囊用流动相提取稀释、高速离心, 结果得到片剂和胶囊中DHEA的回收率在95.7%~98.6%之间(见表1)。口服液通常为水溶液, 利用DHEA溶于有机相的特点, 用乙腈提取, 加入氯化钠使有机相和水层实现分层, 高速离心后检测上层, 得到目标物回收率为101.0%~102.0%(见表1)。
对于凝胶糖果, 首先用温水(70 ℃)将其溶解后放置至常温, 基于前期研究发现饱和硫酸铵可以沉淀明胶[34], 同时可实现乙腈与水的分层, 因此实验采用乙腈提取后, 加入硫酸铵至饱和, 高速离心后, 测定有机相, DHEA的回收率达99.6%~104.0%(见表1)。
2.3 方法学考察
2.3.1 基质效应
基质效应(matrix effect, ME)是化学分析中因样品基质干扰导致检测结果偏差的现象, 常见于色谱质谱检测中。ME=(基质加标响应值/纯溶剂响应值)×100%, 若ME小于80%则为基质抑制, 大于120%则为基质增强, 此时需要降低ME对目标物检测的影响。本研究中DHEA的ME在80%~120%之间, ME影响可忽略, 因此本研究采用纯溶剂配制的标准曲线定量。
2.3.2 线性关系、定量限及检出限
采用液相色谱二极管阵列检测: 精密量取DHEA标准品溶液, 用流动相配制成质量浓度为5~200 μg/mL的系列溶液, 以DHEA的峰面积(Y)为纵坐标, 质量浓度(X, μg/mL)为横坐标, 绘制得到标准曲线, 线性关系系数(r2)大于0.999。采用信噪比估计法, 向空白样品基质添加DHEA, 信噪比为10或3时的添加浓度作为估算方法的定量限和检出限。当片剂、硬胶囊和凝胶糖果试样称样量为0.2 g时, 提取液体积为20 mL时, 方法的检出限为0.15 g/kg, 定量限为0.5 g/kg; 当口服液剂型试样取样量为5 mL时, 提取液体积为5 mL时, 方法的检出限为1.5 mg/L, 定量限为5 mg/L。
采用UPLC-MS/MS检测: 标准曲线线性范围为1~100 ng/mL, 线性关系(r2)大于0.998。片剂、硬胶囊和凝胶糖果基质中方法的检出限为0.5 mg/kg, 定量限为1 mg/kg; 口服液的方法检出限为0.3 μg/L, 定量限为1 μg/L。
2.3.3 回收率及精密度
回收率和精密度的考察均以液相色谱二极管阵列检测结果计算。片剂、硬胶囊、凝胶糖果以0.5、5.0和10.0 g/kg进行加标回收率实验, 口服液以5、50、100 mg/L进行6次平行独立实验, 计算每个样品中DHEA的浓度, 每个浓度的平均回收率及6次重复性实验的RSD。结果显示4种基质中DHEA的回收率在95.7%~104.0%之间, RSDs在1.10%~3.76%之间, 详见表1。
2.4 实际样品检测
根据剂型、含量以及不同品牌, 本研究采购含DHEA的电商产品共5份(表2), 采用本研究建立的方法测定, 其中4份样品与标签一致, 1份样品中未检出DHEA, 与标签不符, 该结果通过了UPLC-MS/MS确证, 见图4。购买的20余份保健品(包括提高免疫力、抗衰老、缓解焦虑等功能), 均未检出DHEA。
2.5 实验室间验证
液相色谱二极管阵列检测法通过了另外5家检测机构的实验室间协同验证, 准确度和精密度见表3, 其范围均可达到5~200 μg/mL, 回收率、精密度、检出限和定量限与2.3所列结果相同; 进一步通过实际样品检测进行评价, 5家检测机构与本实验室对随机抽取的实际样品测定结果见表4, 实验室间的RSDs均小于5%, 结果符合GB 5009.295—2023《食品安全国家标准 化学分析方法验证通则》中目标分析物浓度在10~100 g/kg (RSD≤10%), 以及>100 g/kg (RSD≤5%)限值规定。
3 结论
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本研究建立了不同剂型保健食品中DHEA的检测方法。根据不同剂型选择不同的提取净化方式, 且根据检测目的不同选择不同的检测方法, 本研究建立的方法回收率在95.7%~104.0%之间, RSDs小于5%, 进行了实验室内和实验室间的方法学评价, 各实验室间的RSDs均小于5%。此方法应用于跨境电商平台购买的膳食补充剂中DHEA含量分析, 结果显示大部分DHEA膳食补充剂产品检测含量与标签一致, 仍有部分产品含量与标签不一致。该方法操作简单, 灵敏度和准确性高, 适用于国内功能性保健食品中DHEA的非法添加检测和跨境电商DHEA膳食补充剂的质量评价。
基金
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  • 高层次公共卫生技术人才建设项目(学科骨干-03-28)
文献
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参考文献 引证文献
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2025年第16卷第10期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250325002
  • 接收时间：2025-03-25
  • 首发时间：2025-07-15
  • 出版时间：2025-05-25
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  • 收稿日期：2025-03-25
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高层次公共卫生技术人才建设项目(学科骨干-03-28)
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    1.北京市疾病预防控制中心, 食物中毒诊断溯源技术北京重点实验室, 北京 100013
    2.首都医科大学公共卫生学院, 北京 100089

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* 孟娟(1971—), 女, 副主任技师, 主要研究方向为食品安全检测。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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