Article(id=1320882349508771905, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320880138208784845, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025060203, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1750176000000, receivedDateStr=2025-06-18, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792852492930, onlineDateStr=2026-10-24, pubDate=1785513600000, pubDateStr=2026-08-01, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792852492930, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792852492930, creator=13701087609, updateTime=1792852492930, updator=13701087609, issue=Issue{id=1320880138208784845, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='15', pageStart='1', pageEnd='524', issueExtLink='null', onlineDate='null', pubDate='1785513600000', pubDateStr='2026-08-01', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792851965715, creator='13701087609', updateTime=1792851965715, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=310, endPage=317, ext={EN=ArticleExt(id=1320882349760430146, articleId=1320882349508771905, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Visualized Rapid Detection of Zearalenone and Aflatoxin Based on Aptamer Sensor and Smartphone Recognition, columnId=1261343843585241413, journalTitle=Science and Technology of Food Industry, columnName=Analysis and Determination, runingTitle=null, highlight=null, articleAbstract=

To enable rapid and convenient detection of zearalenone (ZEN) and aflatoxins (AFB1) in coarse cereals, this work developed an aptamer-based sensor integrated with image analysis software. The sensor utilized specific aptamers targeting ZEN and AFB1 as recognition elements, gold nanoparticles (AuNPs) as the colorimetric indicator, and NaCl as the chromogenic agent. Images of samples captured by a smartphone were analyzed using ImageJ software through measuring the integrated pixel density, allowing for specific and rapid detection of ZEN and AFB1 in ten different types of coarse cereals. Under optimal conditions, the aptasensor based on microplate reader absorbance measurement showed linear detection ranges of 0.05~3 ng/g for ZEN and 0.2~8 ng/g for AFB1, with calibration equations y=0.021x+0.3353 (R2=0.9983) and y=0.0207x+0.3075 (R2=0.9986), and detection limits of 0.035 ng/g and 0.1 ng/g, respectively. When using smartphone image acquisition combined with ImageJ analysis as an alternative method, secondary calibration equations were derived: y=−16.141x+189.88 (R2=0.991) for ZEN and y=−12.928x+247.08 (R2=0.9901) for AFB1, with LODs of 0.037 ng/g and 0.106 ng/g. Method validation confirmed good accuracy and precision, with spike recoveries ranging from 85.88%~101.13% (RSD 1.56%~5.34%) for ZEN and 87.76%~109.30% (RSD 0.9%~5.07%) for AFB1. The aptasensor successfully detected ZEN and AFB1 in all ten tested coarse cereal samples. By integrating smartphone image analysis, this approach achieves highly sensitive, rapid (15 min) and accurate detection of both mycotoxins in coarse cereals, making it an effective tool for on-site screening applications.

, authors=Runzhong YU1, Lifeng LU2, Xinyi LIU2, Liyuan ZHANG2, 3, 4, *, Zengwang GUO5, Zhiqiang HAO6, authorsList=Runzhong YU, Lifeng LU, Xinyi LIU, Liyuan ZHANG, Zengwang GUO, Zhiqiang HAO, authorCompany=null, correspAuthors=Liyuan ZHANG, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright © 2026 Science and Technology of Food Industry. All rights reserved., 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, fund=null), CN=ArticleExt(id=1320882353036181585, articleId=1320882349508771905, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=基于智能手机识别的适配体传感器可视化快速检测玉米赤霉烯酮和黄曲霉毒素, columnId=1261343846139572559, journalTitle=食品工业科技, columnName=分析检测, runingTitle=null, highlight=null, articleAbstract=

为了实现快速便捷检测杂粮中玉米赤霉烯酮(ZEN)和黄曲霉毒素(AFB1),本研究设计一种通过图像分析软件检测ZEN和AFB1的适配体传感器。该传感器以ZEN和AFB1适配体为识别元件,AuNPs为指示剂,NaCl为显色剂,通过智能手机采集图像,再利用ImageJ软件分析样品的图像,测量图像像元综合密度,实现对10种不同杂粮中ZEN和AFB1特异性快速检测。结果表明,在最佳条件下,基于酶标仪吸光度检测的适配体传感器对ZEN和AFB1的检测线性范围分别为ZEN 0.05~3 ng/g、AFB1 0.2~8 ng/g,线性方程为y=0.021x+0.3353(R2=0.9983)、y=0.0207x+0.3075(R2=0.9986),检出限分别为0.035 ng/g和0.1 ng/g。利用智能手机进行图像采集,通过ImageJ进行图像分析,线性方程分别为y=−16.141x+189.88(R2=0.991)、y=−12.928x+247.08(R2=0.9901),检出限分别为0.037 ng/g和0.106 ng/g,ZEN加标回收率为85.88%~101.13%,相对标准偏差(RSD)为1.56%~5.34%,AFB1加标回收率为87.76%~109.30%,相对标准偏差(RSD)为0.9%~5.07%。使用该传感器成功的检测出10种杂粮中ZEN和AFB1,结合智能手机图像分析,实现了杂粮中ZEN和AFB1的高灵敏、快速(15 min)、准确检测,为现场筛查提供了有效工具。

, authors=于润众1, 卢利丰2, 刘鑫怡2, 张丽媛2, 3, 4, *, 郭增旺5, 郝志强6, authorsList=于润众, 卢利丰, 刘鑫怡, 张丽媛, 郭增旺, 郝志强, authorCompany=null, correspAuthors=张丽媛, authorNote=

于润众(1981−),男,硕士,副教授,研究方向:数据挖掘与分析,E-mail:

, correspAuthorsNote=
张丽媛(1981−),女,博士,教授,研究方向:食品安全与质量溯源,E-mail:。
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companyName=null, departmentName=null, remark=6黑龙江谱尼测试科技有限公司,黑龙江哈尔滨 151000)])], figs=[ArticleFig(id=1320882357578612880, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.1, caption=Flowchart of digital image detection, figureFileSmall=1XXO+vdHg2KkYBjXmcgWBA==, figureFileBig=CwIldthXwHp+WjeWyKUM7Q==, tableContent=null), ArticleFig(id=1320882357637333137, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图1, caption=数字化图像检测流程图

注:a为ZEN和AFB1检测原理;b为真菌毒素智能检测流程;c为ImageJ 图像分析过程。

, figureFileSmall=1XXO+vdHg2KkYBjXmcgWBA==, figureFileBig=CwIldthXwHp+WjeWyKUM7Q==, tableContent=null), ArticleFig(id=1320882357830271122, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.2, caption=Linear relationship between different concentrations of ZEN and A650 nm in mixed standards, figureFileSmall=fo0cNIIcgVjf1diB9Oik6w==, figureFileBig=xZCyUfgv0s25zlpqLz2OBA==, tableContent=null), ArticleFig(id=1320882357905768595, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图2, caption=混标中不同浓度ZEN与A650 nm的线性关系

注:a表示混标溶液中0~10 ng/g ZEN的添加量与吸光度的曲线;b表示0.05~3 ng/g ZEN的添加量与吸光度的标准曲线。

, figureFileSmall=fo0cNIIcgVjf1diB9Oik6w==, figureFileBig=xZCyUfgv0s25zlpqLz2OBA==, tableContent=null), ArticleFig(id=1320882357972877460, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.3, caption=Linear relationship between different concentrations of AFB1 and A650 nm in mixed standards, figureFileSmall=lmkzxnznwTWKGkj8mCuSHQ==, figureFileBig=jmHWGMhl0vxDFAn7ZK55lA==, tableContent=null), ArticleFig(id=1320882358035792021, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图3, caption=混标中不同浓度AFB1与A650 nm的线性关系

注:a表示混标溶液中0~10 ng/g AFB1的添加量与吸光度的曲线;b表示0.2~8.0 ng/g AFB1的添加量与吸光度的标准曲线。

, figureFileSmall=lmkzxnznwTWKGkj8mCuSHQ==, figureFileBig=jmHWGMhl0vxDFAn7ZK55lA==, tableContent=null), ArticleFig(id=1320882358102900886, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.4, caption=3D calibration plot of image pixel integrated density and ZEN concentration, figureFileSmall=iLqLSVmmiXv5/3D5Oghfog==, figureFileBig=IlJXbtE6BCz6yjpxbYUBHg==, tableContent=null), ArticleFig(id=1320882358174204055, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图4, caption=图像像元综合密度与ZEN浓度的3D校准图

注:插图为不同ZEN浓度下溶液颜色。

, figureFileSmall=iLqLSVmmiXv5/3D5Oghfog==, figureFileBig=IlJXbtE6BCz6yjpxbYUBHg==, tableContent=null), ArticleFig(id=1320882358241312920, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.5, caption=Linear relationship diagram between integrated density and ZEN concentration, figureFileSmall=Q3S7fqvFOhekiH+s3C/UeA==, figureFileBig=H/zHjclHKloZoYJs60vSFg==, tableContent=null), ArticleFig(id=1320882358325199001, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图5, caption=综合密度与ZEN浓度的线性关系图, figureFileSmall=Q3S7fqvFOhekiH+s3C/UeA==, figureFileBig=H/zHjclHKloZoYJs60vSFg==, tableContent=null), ArticleFig(id=1320882358388113562, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.6, caption=3D calibration plot of image pixel integrated density and AFB1 concentration, figureFileSmall=dtkW2jlTiQLYBYK39SaZaA==, figureFileBig=wP67cCupySNlSjk9P7GmNg==, tableContent=null), ArticleFig(id=1320882358467805339, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图6, caption=图像像元综合密度与AFB1浓度的3D校准图

注:插图为不同AFB1浓度下溶液颜色。

, figureFileSmall=dtkW2jlTiQLYBYK39SaZaA==, figureFileBig=wP67cCupySNlSjk9P7GmNg==, tableContent=null), ArticleFig(id=1320882358543302812, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Fig.7, caption=Linear relationship diagram between integrated density and AFB1 concentration, figureFileSmall=i3y8M0P4iQraHsBT9rm3OQ==, figureFileBig=ZU/gmmNUDBR2v4poFCIG5Q==, tableContent=null), ArticleFig(id=1320882358614605981, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=图7, caption=综合密度与AFB1浓度的线性关系图, figureFileSmall=i3y8M0P4iQraHsBT9rm3OQ==, figureFileBig=ZU/gmmNUDBR2v4poFCIG5Q==, tableContent=null), ArticleFig(id=1320882358685909150, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Table 1, caption=

Spike recoveries of ZEN and AFB1 in actual samples

, figureFileSmall=null, figureFileBig=null, tableContent=
样品名称ZEN添加量
(ng/g)
AFB1添加量
(ng/g)
ZEN的加标回收率
(%)
AFB1的加标回收率
(%)
ZEN相对标准偏差
(%)
AFB1相对标准偏差
(%)
小米10010087.46±2.7587.76±2.783.143.18
20020094.52±2.3895.4±4.832.525.07
300400100.58±1.84102.05±1.211.831.18
小麦10010089.05±4.7692.59±2.795.343.01
20020093.73±3.6497.02±3.683.873.80
30040097.94±1.59101.25±1.841.621.83
高粱10010085.88±2.3889.37±3.943.24.41
20020098.49±2.7592.99±1.972.802.12
300400101.13±5.8109.3±1.001.560.90
), ArticleFig(id=1320882358744629407, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=表1, caption=

实际样品中ZEN和AFB1的加标回收率

, figureFileSmall=null, figureFileBig=null, tableContent=
样品名称ZEN添加量
(ng/g)
AFB1添加量
(ng/g)
ZEN的加标回收率
(%)
AFB1的加标回收率
(%)
ZEN相对标准偏差
(%)
AFB1相对标准偏差
(%)
小米10010087.46±2.7587.76±2.783.143.18
20020094.52±2.3895.4±4.832.525.07
300400100.58±1.84102.05±1.211.831.18
小麦10010089.05±4.7692.59±2.795.343.01
20020093.73±3.6497.02±3.683.873.80
30040097.94±1.59101.25±1.841.621.83
高粱10010085.88±2.3889.37±3.943.24.41
20020098.49±2.7592.99±1.972.802.12
300400101.13±5.8109.3±1.001.560.90
), ArticleFig(id=1320882358815932576, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=EN, label=Table 2, caption=

Detection results of ZEN and AFB1 in real samples

, figureFileSmall=null, figureFileBig=null, tableContent=
样品名称ZEN浓度(ng/g)AFB1浓度(ng/g)ZEN相对标准偏差(%)AFB1相对标准偏差(%)
大米0.99±0.051.75±0.14.805.68
黄豆2.81±0.143.66±0.15.072.71
小麦1.93±0.082.82±0.44.221.4
玉米2.83±0.080.99±0.042.873.98
红芸豆1.34±0.051.39±0.064.054.32
高粱(北旗)2.32±0.091.43±0.083.745.53
甜荞2.39±0.082.17±0.063.612.78
苦荞1.52±0.063.94±0.144.083.61
小米1.24±0.031.94±0.12.095.12
高粱(龙杂13)2.61±0.023.16±0.078.132.16
), ArticleFig(id=1320882358895624353, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320882349508771905, language=CN, label=表2, caption=

实际样品中ZEN和AFB1的检测结果

, figureFileSmall=null, figureFileBig=null, tableContent=
样品名称ZEN浓度(ng/g)AFB1浓度(ng/g)ZEN相对标准偏差(%)AFB1相对标准偏差(%)
大米0.99±0.051.75±0.14.805.68
黄豆2.81±0.143.66±0.15.072.71
小麦1.93±0.082.82±0.44.221.4
玉米2.83±0.080.99±0.042.873.98
红芸豆1.34±0.051.39±0.064.054.32
高粱(北旗)2.32±0.091.43±0.083.745.53
甜荞2.39±0.082.17±0.063.612.78
苦荞1.52±0.063.94±0.144.083.61
小米1.24±0.031.94±0.12.095.12
高粱(龙杂13)2.61±0.023.16±0.078.132.16
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基于智能手机识别的适配体传感器可视化快速检测玉米赤霉烯酮和黄曲霉毒素
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于润众 1 , 卢利丰 2 , 刘鑫怡 2 , 张丽媛 2, 3, 4, * , 郭增旺 5 , 郝志强 6
食品工业科技 | 分析检测 2026,47(15): 310-317
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食品工业科技 |分析检测 2026 , 47 (15) : 310 -317
基于智能手机识别的适配体传感器可视化快速检测玉米赤霉烯酮和黄曲霉毒素
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于润众1 , 卢利丰2, 刘鑫怡2, 张丽媛2, 3, 4, * , 郭增旺5, 郝志强6
作者信息
  • 1黑龙江八一农垦大学电气与信息学院,黑龙江大庆 163319
  • 2黑龙江八一农垦大学食品学院,黑龙江大庆 163319
  • 3国家杂粮工程技术中心,黑龙江大庆 163319
  • 4黑龙江农产品加工与质量安全重点实验室,黑龙江大庆 163319
  • 5东北农业大学食品学院,黑龙江哈尔滨 151000
  • 6黑龙江谱尼测试科技有限公司,黑龙江哈尔滨 151000
通讯作者:
张丽媛(1981−),女,博士,教授,研究方向:食品安全与质量溯源,E-mail:。
作者简介:

于润众(1981−),男,硕士,副教授,研究方向:数据挖掘与分析,E-mail:

Visualized Rapid Detection of Zearalenone and Aflatoxin Based on Aptamer Sensor and Smartphone Recognition
Runzhong YU1 , Lifeng LU2, Xinyi LIU2, Liyuan ZHANG2, 3, 4, * , Zengwang GUO5, Zhiqiang HAO6
Affiliations
  • 1College of Electrical and Information Engineering, Heilongjiang Bayi Agricultural University, Daqing 163319, China
  • 2College of Food Science, Heilongjiang Bayi Agricultural University, Daqing 163319, China
  • 3National Engineering Technology Center for Coarse Cereals, Daqing 163319, China
  • 4Heilongjiang Key Laboratory of Agricultural Products Processing and Quality Safety, Daqing 163319, China
  • 5Food Science College, Northeast Agricultural University, Harbin 151000, China
  • 6Heilongjiang Pony Testing Technical Co., Ltd., Harbin 151000, China
出版时间: 2026-08-01 doi: 10.13386/j.issn1002-0306.2025060203
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为了实现快速便捷检测杂粮中玉米赤霉烯酮(ZEN)和黄曲霉毒素(AFB1),本研究设计一种通过图像分析软件检测ZEN和AFB1的适配体传感器。该传感器以ZEN和AFB1适配体为识别元件,AuNPs为指示剂,NaCl为显色剂,通过智能手机采集图像,再利用ImageJ软件分析样品的图像,测量图像像元综合密度,实现对10种不同杂粮中ZEN和AFB1特异性快速检测。结果表明,在最佳条件下,基于酶标仪吸光度检测的适配体传感器对ZEN和AFB1的检测线性范围分别为ZEN 0.05~3 ng/g、AFB1 0.2~8 ng/g,线性方程为y=0.021x+0.3353(R2=0.9983)、y=0.0207x+0.3075(R2=0.9986),检出限分别为0.035 ng/g和0.1 ng/g。利用智能手机进行图像采集,通过ImageJ进行图像分析,线性方程分别为y=−16.141x+189.88(R2=0.991)、y=−12.928x+247.08(R2=0.9901),检出限分别为0.037 ng/g和0.106 ng/g,ZEN加标回收率为85.88%~101.13%,相对标准偏差(RSD)为1.56%~5.34%,AFB1加标回收率为87.76%~109.30%,相对标准偏差(RSD)为0.9%~5.07%。使用该传感器成功的检测出10种杂粮中ZEN和AFB1,结合智能手机图像分析,实现了杂粮中ZEN和AFB1的高灵敏、快速(15 min)、准确检测,为现场筛查提供了有效工具。

适配体  /  金纳米粒子  /  玉米赤霉烯酮  /  黄曲霉毒素B1  /  比色适配体传感器  /  图像分析法

To enable rapid and convenient detection of zearalenone (ZEN) and aflatoxins (AFB1) in coarse cereals, this work developed an aptamer-based sensor integrated with image analysis software. The sensor utilized specific aptamers targeting ZEN and AFB1 as recognition elements, gold nanoparticles (AuNPs) as the colorimetric indicator, and NaCl as the chromogenic agent. Images of samples captured by a smartphone were analyzed using ImageJ software through measuring the integrated pixel density, allowing for specific and rapid detection of ZEN and AFB1 in ten different types of coarse cereals. Under optimal conditions, the aptasensor based on microplate reader absorbance measurement showed linear detection ranges of 0.05~3 ng/g for ZEN and 0.2~8 ng/g for AFB1, with calibration equations y=0.021x+0.3353 (R2=0.9983) and y=0.0207x+0.3075 (R2=0.9986), and detection limits of 0.035 ng/g and 0.1 ng/g, respectively. When using smartphone image acquisition combined with ImageJ analysis as an alternative method, secondary calibration equations were derived: y=−16.141x+189.88 (R2=0.991) for ZEN and y=−12.928x+247.08 (R2=0.9901) for AFB1, with LODs of 0.037 ng/g and 0.106 ng/g. Method validation confirmed good accuracy and precision, with spike recoveries ranging from 85.88%~101.13% (RSD 1.56%~5.34%) for ZEN and 87.76%~109.30% (RSD 0.9%~5.07%) for AFB1. The aptasensor successfully detected ZEN and AFB1 in all ten tested coarse cereal samples. By integrating smartphone image analysis, this approach achieves highly sensitive, rapid (15 min) and accurate detection of both mycotoxins in coarse cereals, making it an effective tool for on-site screening applications.

aptamer  /  gold nanoparticles  /  zearalenone  /  aflatoxins B1  /  color matching aptamer sensor  /  image analysis method
于润众, 卢利丰, 刘鑫怡, 张丽媛, 郭增旺, 郝志强. 基于智能手机识别的适配体传感器可视化快速检测玉米赤霉烯酮和黄曲霉毒素. 食品工业科技, 2026 , 47 (15) : 310 -317 . DOI: 10.13386/j.issn1002-0306.2025060203
Runzhong YU, Lifeng LU, Xinyi LIU, Liyuan ZHANG, Zengwang GUO, Zhiqiang HAO. Visualized Rapid Detection of Zearalenone and Aflatoxin Based on Aptamer Sensor and Smartphone Recognition[J]. Science and Technology of Food Industry, 2026 , 47 (15) : 310 -317 . DOI: 10.13386/j.issn1002-0306.2025060203
玉米赤霉烯酮(Zearalenone,ZEN)又称F-2毒素,是一种由多种镰刀菌通过聚酮途径生物合成的非甾体雌激素类真菌毒素,广泛存在于玉米、小麦、燕麦、大麦、大豆等粮食中[1−3],ZEN可以通过食物链进行富集,饲料中ZEN污染会对畜禽健康构成严重威胁。并且ZEN残留物可能存在于由受污染的原材料制成的食品中[4]。此外,被ZEN污染的植物和动物源性食品在摄入后会在体内积聚,从而影响人类健康[5]。黄曲霉毒素(Aflatoxin,AFB1)为黄曲霉和寄生曲霉菌产生的多肽复合物,是影响人类健康的毒性最严重的真菌毒素之一[6]。目前,已分离出20余种黄曲霉毒素及其衍生物,其中AFB1的毒性最强(约20 mg即可致成人死亡),且在自然界中产量最高、稳定性最强[7−8]。目前针对ZEN和AFB1的检测主要包括高效液相色谱法(HPLC)[9]、高效液相色谱-串联质谱(HPLC-MS)[10]、薄层色谱分析法(TLC)[11]、酶联免疫吸附法(ELISA)[12],尽管这些方法灵敏度高、选择性好,但仍存在操作流程复杂、耗时较长、成本高昂、需专业人员实施等局限,难以满足快速及现场检测需求。
适配体是通过指数富集适配体系统进化(SELEX)在体外选择过程获得的一段单链寡核苷酸(ssDNA或ssRNA)分子,折叠的适配体可形成茎环、发夹等特异性二级或三级结构,与目标分子结合,具有较高的亲和力和特异性。适配体仅由四个碱基(DNA 或 RNA 的 A、G、C、T 或U)组成[13]。适配体虽具备与抗体相当的高特异性和高亲和力,但其靶标识别范围远广于抗体。主要包括离子[14]、小分子[15]、蛋白质[16]、病毒[17]、细菌[18]、寄生虫[19],甚至整个活细胞[20]。此外,适配体具有亲和力高、特异性强、分子量小、无批量变异、易修饰合成、稳定性好、易运输储存、无免疫原性等优点[21],被认为是最具潜力的抗体替代品。适配体已广泛应用于分析化学[22]、生物学[23]、食品学[24]和医学[25–26]等多个领域。适配体技术已深度赋能真菌毒素检测,尤其在电化学/光电化学传感器中展现出超高灵敏度和现场适用性[27]。通过标准化适配体筛选、开发多毒素联检平台及智能终端集成,有望成为替代传统方法的主流快检工具,为食品安全监管提供技术支撑[28]。
基于此,本研究提出一种利用图像分析软件对ZEN与AFB1进行可视化检测的比色适配体传感器方案。ZEN与AFB1适配体能够有效地保护AuNPs,从而极大地提高AuNPs在高浓度盐溶液中的稳定性。利用ImageJ软件对检测样品进行图像分析,可以通过测量图像颜色的综合密度与ZEN、AFB1浓度联系起来,进而可以通过监测颜色的变化来确定ZEN和AFB1的浓度,实现对ZEN和AFB1的定量检测。最后,使用该传感器进行加标回收实验以及对10种不同粮食样品的检测,来验证该传感器的实用性。本研究旨在为粮食中真菌毒素的快速、现场检测提供一种有效的技术手段。
核酸适配体(ZEN Apt5'-GAT GGG GAA AGG GTC CCC CTG GGT TGG AGC ATC GGA CA-3'、AFB1 Apt5'-GTT GGG CAC GTG TTG TCT CTC TGT GTC TCG TGC CCT TCG CTA GGC CCA CA-3') 本研究中所使用的寡核苷酸序列均由上海生工生物技术有限公司合成和纯化;氯金酸(HAuCl4) 国药集团化学试剂有限公司;玉米赤霉烯酮标准品、黄曲霉毒素B1标准品 青岛普瑞帮生物工程有限公司;柠檬酸钠、氯化钠、无水硫酸钠、无水硫酸镁、硫酸铵 上海麦克林生物科技有限公司;C18E 深圳安普生物医药有限公司;重铬酸钾、乙腈、甲酸、丙酮 上海麦克林生物科技有限公司;小米、小麦、黄豆、高粱(北旗)、高粱(龙杂13)、苦荞麦、甜荞麦、玉米、红芸豆、大米 市售。
VICTOR Nivo 酶标仪 珀金埃尔默企业管理(上海)有限公司;UV-1500PC紫外分光光度计 翰艺仪器(上海)有限公司;JEM-2100F透射电镜 日本电子株式会社;KQ-500DE超声波清洗器 昆山市超声仪器有限公司;Vortex-Genie 2漩涡振荡器 美国Scientific Industries公司;DFY-500高速粉碎机 浙江省永康市金穗机械制造厂;TH2-82T水浴恒温振荡器 常州荣华仪器有限公司;H2-16KR台式高速离心机 湖南可成仪器设备有限公司;CYNS-12水浴氮吹仪 上海力辰仪器科技有限公司;SMART-RO-15纯水仪 上海和泰仪器有限公司;ZEN3700激光粒度仪 上海百吉仪器有限公司。
取100 μL 100 μg/mL的ZEN与AFB1单标溶液各一份,置于1 mL试管中混合。加入800 μL乙腈,涡旋振荡混匀,配制得10 μg/mL混合标准储备液。取该储备液,用乙腈梯度稀释,配制浓度系列为5、10、20、50、100、150、200、300、400、600、800、1000 ng/mL的混合标准工作溶液。
用研磨机将高粱样品磨成粉末,并通过80目筛。将AFB1、ZEN混合标准溶液(5、10、20、50、100、150、200、300、400、600、800、1000 ng/mL)加入样品中,制成含有混合标准溶液的加标样品。每个加标水平做3组平行实验。共获得30个样品和10个加标水平(加标实验仅选取了上述12组浓度中的10个浓度,每个浓度3个平行,因此为30个样品;所列12组浓度为配制标准曲线所用)。为了确保混合标准溶液的良好分布,在每个样品中加入适量的丙酮以湿润样品粉末,并进行充分搅拌,然后在通风环境下风干24 h后再进行萃取。加标的样品保存在密封的瓶子里。
对现有的QuEChERS[27−30]样品预处理方法适当改进,用电子天平称取5.00 g(精确到0.01 g)样品粉末,置于50 mL的离心管内,并向离心管内加入5 mL的蒸馏水,用力摇匀使样品完全润湿,然后向离心管内加入10 mL 1%(甲酸酸化过)的乙腈溶液,涡旋振荡3 min后,超声萃取10 min。再向离心管内加入1 g NaCl和3 g (NH4)2SO4,涡旋振荡2 min,利用冷冻离心机在4 ℃下以11000 r/min离心10 min。用移液枪吸取5 mL上清液(有机相),置于含有0.2 g C18的10 mL离心管中,涡旋振荡2 min,并利用冷冻离心机在4 °C下以11000 r/min离心10 min。吸取4 mL上清液用移液枪转移到10 mL氮吹瓶中,在水温为40 °C下利用氮吹仪对溶液进行氮气吹干浓缩,接着将浓缩后的样品用1 mL乙腈溶液定容,并将定容后的溶液用0.22 µm的PTEE滤膜过滤到2 mL样品瓶中备用。
通过柠檬酸钠还原法制备AuNPs溶液的具体操作步骤:首先将100 mL超纯水和1 mL 1%(HAuCl4)溶液加入预先用铬酸洗液浸泡过的250 mL烧杯中,对含有氯金酸(HAuCl4)的水溶液进行加热并搅拌直至沸腾。向沸腾的溶液中加入2.5 mL 1%柠檬酸钠溶液,迅速搅拌10 min,可以看到溶液的颜色由浅黄色转变为黑色,然后转变为紫红色,最终转变为酒红色且透明。当溶液的颜色不再发生变化,应停止加热,接着搅拌10 min。将AuNPs溶液冷却至室温并用10 mL试管进行分装,将其放入冰箱中4 ℃保存备用。
向微孔板孔中加入100 µL金纳米粒子(AuNPs),再分别加入10 µL浓度为5 µmol/L的ZEN适配体溶液和10 µL浓度为0.7 µmol/L的AFB1适配体溶液,室温孵育5 min。接着,分别加入10 µL经过QuEChERS法处理的不同浓度粮食提取液(0、5、10、20、50、100、150、200、300、400、600、800、1000 ng/mL),在室温下孵育5 min。最后,将10 µL浓度为2 mol/L的NaCl溶液分别加入到溶液中,在室温下反应5 min。用微孔板阅读器测定溶液的吸光度值(A650 nm),以吸光度值为纵坐标,ZEN和AFB1浓度为横坐标,分别建立工作曲线。
首先使用智能手机将样品图像拍摄下来,然后通过智能手机上的ImageJ软件对图像进行分析。分析流程为:打开软件、选择8位图像类型、点击流程、减去背景、点击“椭圆选择”、利用矩形工具,在离心管上选一个固定正方形区为分析区、点击“分析测量”,得出RGB综合密度,然后移至下一个样品图像进行成像分析。最后,对ZEN和AFB1浓度(X轴)与颜色综合密度(Y轴)进行线性回归分析,得到校准曲线图。
为探讨该方法的可行性和实用性,用本文设计的传感器对小米、小麦、高粱进行ZEN和AFB1的加标回收率检测,加入混合标准溶液浓度为(ZEN为100、200、300 ng/mL,AFB1为100、200、400 ng/mL)。按照实验方法进行实际样品的检测,每个加标水平做3组平行实验。计算加标回收率和RSD值。
并选取了来自不同产地的10种不同的粮食进行了实际样品的检测,来证明该传感器的实用性,每种粮食样品做3组平行实验。按照实验方法进行实际样品的检测。
Origin 2021用于图表制作,IBM SPSS Statistics 25用于数据分析,Thermos Scientific的Velox软件用于TEM图像采集,ImageJ用于图像数字化处理。
本实验原理如图1所示,实验利用ZEN和AFB1适配体在靶标物质ZEN和AFB1存在下特异性结合形成结合物,由此构建了检测ZEN和AFB1比色传感器[29]。在没有ZEN和AFB1的情况下,ZEN和AFB1适配体与AuNPs之间通过静电吸附作用包裹在AuNPs表面。因此,ZEN和AFB1适配体可以保护AuNPs免受NaCl诱导的聚集。然而,ZEN与AFB1适配体一旦特异性的与相应的真菌毒素结合后,适配体对于AuNPs的保护作用就会消失,由此导致AuNPs在高盐浓度溶液中的聚集,溶液颜色从酒红色转变为蓝色。因此,添加不同浓度的ZEN和AFB1会产生不同程度的AuNPs聚集。溶液的吸光度值与靶标ZEN和AFB1的浓度有关,通过ImageJ软件分析样本的图像发现通过测量图像像元综合密度,可以将其与目标浓度联系起来。实现对ZEN和AFB1的定性和定量检测[30]。
将一系列不同浓度的ZEN和AFB1混合标准溶液加入粉碎的高粱样品中,使样品中ZEN的终浓度分别为0、0.05、0.1、0.2、0.5、1、1.5、2、3、4、6、8、10 ng/g。采用QuEChERS方法对样品进行处理,所绘制的工作曲线如图2a所示。随着ZEN浓度的增加,AuNPs溶液在A650 nm处的吸光度值逐渐上升;但当ZEN浓度超过3 ng/g后,吸光度值转而下降。其原因在于,在玉米赤霉烯酮(ZEN)的检测过程中,高浓度ZEN会与适配体充分结合并引发其构象变化,使适配体从AuNPs表面脱离,导致其对AuNPs的保护作用丧失,进而引起AuNPs聚集状态发生变化,共同导致吸光度下降。如图2b所示,ZEN在0.05~3 ng/g浓度范围内,A650 nm的吸光度值与ZEN浓度呈良好的线性关系,线性方程为y=0.021x+0.3353,决定系数为R2=0.9983,线性范围为0.05~3.0 ng/g。在空白溶液的优化条件下,检测限为0.035 ng/g(S/N=3),定量限为0.05 ng/g(S/N=10)。
在优化的实验条件下,将一系列不同浓度的ZEN和AFB1混合标准溶液加入粉碎的高粱样品中,使样品中ZEN的终浓度分别为0、0.05、0.1、0.2、0.5、1、1.5、2、3、4、6、8、10 ng/g。用QuEChERS方法对样品进行处理,绘制工作曲线。如图3a所示,AuNPs溶液在A650 nm处的吸光度值随AFB1浓度的增加而升高,但在AFB1浓度增至8 ng/g的阈值后,吸光度值呈现下降趋势。该现象是由于当AFB1浓度超过阈值时,过量AFB1会与适配体结合,引发其构象变化并脱离AuNPs表面,这种保护作用的丧失最终导致了AuNPs的聚集。如图3b所示,AFB1在0.2~8 ng/g浓度范围内,A650 nm的吸光度值与AFB1浓度呈良好的线性关系,线性方程为y=0.0207x+0.3075,决定系数为R2=0.9986,线性范围为0.2~8 ng/g,在空白溶液的优化条件下,检测限为0.1 ng/g(S/N=3),定量限为0.2 ng/g(S/N=10)。
利用图像分析软件对ZEN的检测结果如图4和图5所示,当混标溶液中ZEN的浓度由0.05 ng/g增加到3 ng/g时,溶液图像像元综合密度相应下降,在浓度为0.05~3 ng/g的范围内图像像元综合密度与ZEN浓度呈良好的线性关系,线性方程为y=−16.141x+189.88(R2=0.991),计算检测限为0.037 ng/g,检测结果与酶标仪分析结果一致,表明本传感器适用于现场检测ZEN。
利用图像分析软件对AFB1的检测结果如图6和图7所示,当混标溶液中AFB1的浓度由0.2 ng/g增加至8 ng/g时,溶液图像像元综合密度相应下降,在浓度为0.2~8 ng/g的范围内图像像元综合密度与AFB1浓度呈良好的线性关系,线性方程为y=−12.928x+247.08(R2=0.9901)。计算检测限为0.106 ng/g,检测结果与酶标仪分析结果一致,表明该传感器适用于现场检测AFB1。
如图4~图7所示,通过图像分析软件对ZEN和AFB1的检测结果表明:在各自浓度范围内(ZEN:0.05~3 ng/g;AFB1:0.2~8 ng/g),溶液图像像元综合密度均随毒素浓度增加而下降,且呈现良好线性关系(ZEN:y=−16.141x+189.88,R2=0.991;AFB1:y=−12.928x+247.08,R2=0.9901),检测限分别为0.037 ng/g和0.106 ng/g。两种毒素的检测结果均与酶标仪分析结果一致,证明该传感器适用于ZEN和AFB1的现场检测。智能手机图像分析作为快速筛查工具,以接近实验室的准确性、极致的便捷性和低成本的优势更适合现场筛查,为粮食安全现场监管提供高效解决方案。
选取三种不同粮食样品进行加标回收率分析。结果显示(表1),ZEN的加标回收率在85.88%~101.13%之间,相对标准偏差(RSD)为1.56%~5.34%;AFB1的加标回收率在87.76%~109.3%之间,RSD为0.9%~5.07%。结果表明,该方法具有良好的准确度和重复性,适用于复杂基质中ZEN与AFB1的定量检测。
进一步对10种不同粮食样品中ZEN与AFB1的含量进行检测。结果(表2)表明,ZEN含量范围为0.99~2.83 ng/g,RSD为2.09%~8.13%;AFB1含量范围为0.99~3.94 ng/g,RSD为1.4%~5.68%。所有检测结果均远低于国家标准GB 2761-2017《食品中真菌毒素限量》规定的AFB1限量(5 μg/kg),说明本方法在实际样品检测中具备良好的适用性与可靠性。
综上所述,本研究建立的检测方法操作简便、结果准确,可用于粮食样品中ZEN与AFB1的快速筛查与定量分析。
本研究采用QuEChERS法做为样品的前处理方法,使用ZEN、AFB1适配体和AuNPs构建了基于图像分析的比色适配体传感器,检测粮食中的ZEN和AFB1。通过观察图像像元综合密度与ZEN和AFB1浓度的3D校准图,并与谢建国[31]的研究进行比较,溶液的颜色变化与图像像元综合密度的变化规律基本相似,但是所得到的图像像元综合密度值高于文献所报的,分析造成的原因可能有几个方面。首先,所制备的AuNPs粒径不同,导致在高浓度下NaCl所诱导的AuNPs聚集程度不同,显示的颜色的强弱也不同,所以在利用软件分析时所显示的像元综合密度值不同。其次,是被检测物质的差异造成,不同的检测物质与适配体结合后形成不同结构的结合物,进而影响了AuNPs的聚集,所呈现的颜色强弱也就不同,则造成像元综合密度值的差异。最后,除了NaCl还有一些阳离子作为显色剂会引起AuNPs的聚集,但聚集的程度可能不同,显示的颜色也有所差异,经过图像分析软件分析后,所显示的图像像元综合密度也就不同。将本方法的检出限与国家标准GB 2761-2017《食品中真菌毒素限量》中规定的限量值进行比较:ZEN 的限量(如玉米中为60 μg/kg)远高于本方法的检出限(0.037 ng/g=0.037 μg/kg);AFB1 的限量(其他谷物中为5 μg/kg)也远高于本方法的检出限(0.106 ng/g=0.106 μg/kg)。这表明本方法的检测灵敏度完全满足国家标准对真菌毒素的筛查要求。同时,本方法在成本、速度和操作简便性等方面展现出显著优势,适用于现场快速检测。
本研究成功构建了一种基于图像分析的比色适配体传感器,用于粮食中ZEN和AFB1的快速可视化检测。该传感器对ZEN和AFB1的检测限分别达到0.037 ng/g和0.106 ng/g,线性范围分别为0.05~3 ng/g和0.2~8 ng/g。基于AuNPs的颜色变化,结合ImageJ软件分析,该方法可在无需精密仪器的情况下实现毒素的定性与定量分析。实验证明,该方法操作简便、快速且成本低,为现场快速筛查提供了有效工具。

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2026年第47卷第15期
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doi: 10.13386/j.issn1002-0306.2025060203
  • 接收时间:2025-06-18
  • 首发时间:2026-10-24
  • 出版时间:2026-08-01
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  • 收稿日期:2025-06-18
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    1黑龙江八一农垦大学电气与信息学院,黑龙江大庆 163319
    2黑龙江八一农垦大学食品学院,黑龙江大庆 163319
    3国家杂粮工程技术中心,黑龙江大庆 163319
    4黑龙江农产品加工与质量安全重点实验室,黑龙江大庆 163319
    5东北农业大学食品学院,黑龙江哈尔滨 151000
    6黑龙江谱尼测试科技有限公司,黑龙江哈尔滨 151000

通讯作者:

张丽媛(1981−),女,博士,教授,研究方向:食品安全与质量溯源,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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