Article(id=1217779719177683649, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217779717386715826, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250403001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1743609600000, receivedDateStr=2025-04-03, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768270910305, onlineDateStr=2026-01-13, pubDate=1750780800000, pubDateStr=2025-06-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768270910305, onlineIssueDateStr=2026-01-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768270910305, creator=13701087609, updateTime=1768270910305, updator=13701087609, issue=Issue{id=1217779717386715826, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='12', pageStart='1', pageEnd='320', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1768270909877, creator=13701087609, updateTime=1768299620707, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1217900139386163208, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217779717386715826, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1217900139386163209, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217779717386715826, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=145, endPage=150, ext={EN=ArticleExt(id=1217779719609696975, articleId=1217779719177683649, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Rapid determination of multiple pesticide residues in fruits and vegetables on bioluminescence inhibition of enzyme receptor, columnId=1217529311867883548, journalTitle=Journal of Food Safety & Quality, columnName=Highlight: Analysis and Monitoring of Toxic and Harmful Substances in Food, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a rapid analytical protocol for the detection of multiple pesticide residues in fruits and vegetables on bioluminescence inhibition of enzyme receptor. Methods Regarding sensitivity as the index of investigating, the effects of the extraction solvent and reaction time were separately investigated. The pretreatment protocol and detection conditions parameters were optimized. Then, the sensitivity, false positive rate, false negative rate and cross-reactivity of the method were evaluated. Results The findings demonstrated that the enzyme receptor bioluminescence technique was capable of concurrently identifying several pesticide residues within fruits and vegetables, with a sensitivity that complies with the GB 2763—2021 National food safety standards-Maximum residue limits of pesticides in food. The false negative rate was 0%, the false positive rate was less than 5%, and the cross-reactivity rate was less than or equal to 0.10%. Comparative analysis utilizing both the bioluminescence inhibition of enzyme receptor and the conventional national standard method was conducted on fruit and vegetable samples sourced from the market, yielding congruent results. Conclusion The bioluminescence inhibition of enzyme receptor established in the present study is deemed suitable for the rapid assessment of pesticide residues in fruits and vegetables. It is appropriated for oversight by local regulatory bodies and assists companies in improving their internal quality control, which is highly important for boosting the quick detection ability of pesticide residues.

, correspAuthors=Yun LING, 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=Shi-Ge XING, Gui-Hong YAO, Mei-Yi YAO, Long-Fei LIU, Yun LING), CN=ArticleExt(id=1217779720154956522, articleId=1217779719177683649, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=酶受体生物荧光法快速检测水果和蔬菜中多农药残留, columnId=1217529312056627244, journalTitle=食品安全质量检测学报, columnName=本期重点:食品中有毒有害物质分析与监测, runingTitle=null, highlight=null, articleAbstract=

目的 建立酶受体生物荧光法快速检测水果和蔬菜中多农药残留的方法。方法 以灵敏度为指标, 分别考察提取溶剂、反应时间对结果的影响, 确定最佳前处理方法及检测条件; 对方法的灵敏度、假阳性率、假阴性率和交叉反应性进行评价。结果 该方法同时检测水果蔬菜中的多种农药残留, 灵敏度满足GB 2763—2021《食品安全国家标准 食品中农药最大残留限量》的限量要求, 假阴性率为0%, 假阳性率<5%, 交叉反应率≤0.10%。采用酶受体生物荧光法和国家标准方法对市售水果和蔬菜样品进行检测, 结果一致。结论 本研究建立的酶受体生物荧光法可用于水果和蔬菜中农药残留快速测定, 适用于基层监管部门进行监管, 也有助于企业内部更好地进行质量控制, 对于提升农药残留的快速检测能力具有重要意义。

, correspAuthors=凌云, authorNote=null, correspAuthorsNote=
*凌云(1979—), 女, 博士, 研究员, 主要研究方向为营养与食品卫生学。E-mail:
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邢仕歌(1980—), 女, 博士, 研究员, 主要研究方向为食品安全快检技术。E-mail:

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Rapid qualitative determination of organophosphorus and n-methylcarbamate pesticide residues in grain based on bioluminescence inhibition of enzyme[J]. Journal of the Chinese Cereals and Oils, 2022, 37(2): 167-174., articleTitle=Rapid qualitative determination of organophosphorus and n-methylcarbamate pesticide residues in grain based on bioluminescence inhibition of enzyme, refAbstract=null), Reference(id=1217833938425397617, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, doi=null, pmid=null, pmcid=null, year=2023, volume=12, issue=15, pageStart=2828, pageEnd=null, url=null, language=null, rfNumber=[28], rfOrder=34, authorNames=JIANG Z, ZHUANG Y, GUO S, journalName=Journal of Agricultural and Food Chemistry, refType=null, unstructuredReference=JIANG Z, ZHUANG Y, GUO S, et al. Advances in microfluidics techniques for rapid detection of pesticide residues in food[J]. Journal of Agricultural and Food Chemistry, 2023, 12(15): 2828., articleTitle=Advances in microfluidics techniques for rapid detection of pesticide residues in food, refAbstract=null), Reference(id=1217833938513478004, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, doi=null, pmid=null, pmcid=null, year=2025, volume=73, issue=9, pageStart=4982, pageEnd=4997, url=null, language=null, rfNumber=[29], rfOrder=35, authorNames=LI C, ZHANG W, XU X, journalName=Journal of Agricultural and Food Chemistry, refType=null, unstructuredReference=LI C, ZHANG W, XU X, et al. Applications and challenges of fluorescent probes for the detection of pesticide residues in food[J]. Journal of Agricultural and Food Chemistry, 2025, 73(9): 4982-4997., articleTitle=Applications and challenges of fluorescent probes for the detection of pesticide residues in food, refAbstract=null), Reference(id=1217833938609947000, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, doi=null, pmid=null, pmcid=null, year=2022, volume=70, issue=41, pageStart=13093, pageEnd=13117, url=null, language=null, rfNumber=[30], rfOrder=36, authorNames=XU L, ABD-EL-ATY AM, EUN JB, journalName=Foods, refType=null, unstructuredReference=XU L, ABD-EL-ATY AM, EUN JB. Recent advances in rapid detection techniques for pesticide residue: A review[J]. Foods, 2022, 70(41): 13093-13117., articleTitle=Recent advances in rapid detection techniques for pesticide residue: A review, refAbstract=null)], funds=[Fund(id=1217833932603703372, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, awardId=2024JK029, language=CN, fundingSource=中国质量检验检测科学研究院基本科研业务费项目(2024JK029), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1217833924605166141, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, xref=1, ext=[AuthorCompanyExt(id=1217833924613554751, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, companyId=1217833924605166141, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 Institute of Food Safety, Chinese Academy of Quality and Inspection & Testing, Beijing 100176, China), AuthorCompanyExt(id=1217833924772938323, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, companyId=1217833924605166141, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 中国质量检验检测科学研究院食品安全研究所, 北京 100176)]), AuthorCompany(id=1217833924865213021, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, xref=2, ext=[AuthorCompanyExt(id=1217833924877795934, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, companyId=1217833924865213021, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 Beijing Biotai Co., Ltd., Beijing 100176, China), AuthorCompanyExt(id=1217833924886184543, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, companyId=1217833924865213021, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 北京中检葆泰生物技术有限公司, 北京 100176)])], figs=[ArticleFig(id=1217833929940321250, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=EN, label=Fig.1, caption=Extraction effects of different solvent, figureFileSmall=4ZxKIpcSHTxMbO/ArmIevw==, figureFileBig=IMXigFN7jq7Ud3424Bck5w==, tableContent=null), ArticleFig(id=1217833930036790253, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=CN, label=图1, caption=不同溶剂提取效果, figureFileSmall=4ZxKIpcSHTxMbO/ArmIevw==, figureFileBig=IMXigFN7jq7Ud3424Bck5w==, tableContent=null), ArticleFig(id=1217833930162619384, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=EN, label=Table 1, caption=

Maximum residue limits of 23 kinds of pesticides in vegetables and fruits

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编号 农药种类 水果/(mg/kg) 蔬菜/(mg/kg)
1 苯线磷 0.02 0.02
2 地虫硫磷 0.01 0.01
3 毒虫畏 0.01 0.01
4 对硫磷 0.01 0.01
5 庚烯磷 0.01 0.01
6 甲拌磷 0.01 0.01
7 甲基硫环磷 0.03 0.03
8 久效磷 0.03 0.03
9 磷胺 0.05 0.05
10 氯唑磷 0.01 0.01
11 灭线磷 0.02 0.02
12 杀虫畏 0.01 0.01
13 杀扑磷 0.05 0.05
14 速灭磷 0.01 0.01
15 特丁硫磷 0.01 0.01
16 氧乐果 0.02 0.02
17 乙酰甲胺磷 0.02 0.02
18 蝇毒磷 0.05 0.05
19 治螟磷 0.01 0.01
20 内吸磷 0.02 0.02
21 丁硫克百威 0.01 0.01
22 克百威 0.02 0.02
23 灭多威 0.20 0.20
), ArticleFig(id=1217833930254893058, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=CN, label=表1, caption=

23种农药在蔬菜水果中的最大残留限量

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编号 农药种类 水果/(mg/kg) 蔬菜/(mg/kg)
1 苯线磷 0.02 0.02
2 地虫硫磷 0.01 0.01
3 毒虫畏 0.01 0.01
4 对硫磷 0.01 0.01
5 庚烯磷 0.01 0.01
6 甲拌磷 0.01 0.01
7 甲基硫环磷 0.03 0.03
8 久效磷 0.03 0.03
9 磷胺 0.05 0.05
10 氯唑磷 0.01 0.01
11 灭线磷 0.02 0.02
12 杀虫畏 0.01 0.01
13 杀扑磷 0.05 0.05
14 速灭磷 0.01 0.01
15 特丁硫磷 0.01 0.01
16 氧乐果 0.02 0.02
17 乙酰甲胺磷 0.02 0.02
18 蝇毒磷 0.05 0.05
19 治螟磷 0.01 0.01
20 内吸磷 0.02 0.02
21 丁硫克百威 0.01 0.01
22 克百威 0.02 0.02
23 灭多威 0.20 0.20
), ArticleFig(id=1217833930397499404, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=EN, label=Table 2, caption=

Design of orthogonal factor horizontal

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水平 因素
孵育温度/℃ 时间1+时间2/min
1 30 8+3
2 35 8+5
3 40 10+5
), ArticleFig(id=1217833931693539345, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=CN, label=表2, caption=

正交因素水平设计

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水平 因素
孵育温度/℃ 时间1+时间2/min
1 30 8+3
2 35 8+5
3 40 10+5
), ArticleFig(id=1217833931823562783, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=EN, label=Table 3, caption=

Effects of measurement conditions on the sensitivity results

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组别 30 ℃ 35 ℃ 40 ℃
8 min+3 min 8 min+5 min 10 min+5 min 8 min+3 min 8 min+5 min 10 min+5 min 8 min+3 min 8 min+5 min 10 min+5 min
砂糖橘 21.7 41.3 65.2 54.3 78.3 100.0 43.5 69.6 80.4
苹果 19.6 43.5 67.4 56.5 80.4 100.0 41.3 71.7 78.3
桃子 21.7 43.5 63.0 50.0 78.3 100.0 47.8 65.2 82.6
葡萄 23.9 37.0 65.2 54.3 78.3 100.0 39.1 69.6 78.3
香蕉 21.7 41.3 65.2 54.3 76.1 100.0 43.5 69.6 80.4
哈密瓜 26.1 37.0 65.2 54.3 78.3 100.0 43.5 69.6 80.4
洋葱 17.4 43.5 65.2 56.5 78.3 100.0 45.7 67.4 80.4
花椰菜 21.7 41.3 65.2 54.3 78.3 100.0 43.5 69.6 80.4
大白菜 10.9 32.6 54.3 54.3 78.3 100.0 43.5 69.6 80.4
茄子 21.7 41.3 65.2 58.7 84.8 100.0 43.5 69.6 80.4
黄瓜 21.7 41.3 65.2 54.3 78.3 100.0 45.7 69.6 80.4
菜豆 21.7 39.1 65.2 54.3 78.3 100.0 43.5 69.6 80.4
莴苣 21.7 37.0 65.2 54.3 80.4 100.0 43.5 69.6 80.4
萝卜 21.7 41.3 65.2 56.5 78.3 100.0 43.5 69.6 80.4
莲藕 21.7 45.7 65.2 54.3 73.9 100.0 43.5 69.6 80.4
绿豆芽 21.7 43.5 63 54.3 78.3 100.0 43.5 69.6 80.4
), ArticleFig(id=1217833931970363435, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=CN, label=表3, caption=

测定条件对灵敏度结果的影响

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组别 30 ℃ 35 ℃ 40 ℃
8 min+3 min 8 min+5 min 10 min+5 min 8 min+3 min 8 min+5 min 10 min+5 min 8 min+3 min 8 min+5 min 10 min+5 min
砂糖橘 21.7 41.3 65.2 54.3 78.3 100.0 43.5 69.6 80.4
苹果 19.6 43.5 67.4 56.5 80.4 100.0 41.3 71.7 78.3
桃子 21.7 43.5 63.0 50.0 78.3 100.0 47.8 65.2 82.6
葡萄 23.9 37.0 65.2 54.3 78.3 100.0 39.1 69.6 78.3
香蕉 21.7 41.3 65.2 54.3 76.1 100.0 43.5 69.6 80.4
哈密瓜 26.1 37.0 65.2 54.3 78.3 100.0 43.5 69.6 80.4
洋葱 17.4 43.5 65.2 56.5 78.3 100.0 45.7 67.4 80.4
花椰菜 21.7 41.3 65.2 54.3 78.3 100.0 43.5 69.6 80.4
大白菜 10.9 32.6 54.3 54.3 78.3 100.0 43.5 69.6 80.4
茄子 21.7 41.3 65.2 58.7 84.8 100.0 43.5 69.6 80.4
黄瓜 21.7 41.3 65.2 54.3 78.3 100.0 45.7 69.6 80.4
菜豆 21.7 39.1 65.2 54.3 78.3 100.0 43.5 69.6 80.4
莴苣 21.7 37.0 65.2 54.3 80.4 100.0 43.5 69.6 80.4
萝卜 21.7 41.3 65.2 56.5 78.3 100.0 43.5 69.6 80.4
莲藕 21.7 45.7 65.2 54.3 73.9 100.0 43.5 69.6 80.4
绿豆芽 21.7 43.5 63 54.3 78.3 100.0 43.5 69.6 80.4
), ArticleFig(id=1217833932150718517, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779719177683649, language=EN, label=Table 4, caption=

Consistency of bioluminescence inhibition of enzyme receptor and national standard method

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编号 样品 酶受体生物荧光法 国家标准方法 编号 样品 酶受体生物荧光法 国家标准方法
1 砂糖橘 - ND 5 圆白菜 - ND
2 苹果 - ND 6 韭菜 - ND
3 葡萄 - ND 7 黄瓜 - ND
4 花椰菜 - ND 8 豇豆 + 0.03 mg/kg(氧乐果)
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酶受体生物荧光法与国家标准方法一致性

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编号 样品 酶受体生物荧光法 国家标准方法 编号 样品 酶受体生物荧光法 国家标准方法
1 砂糖橘 - ND 5 圆白菜 - ND
2 苹果 - ND 6 韭菜 - ND
3 葡萄 - ND 7 黄瓜 - ND
4 花椰菜 - ND 8 豇豆 + 0.03 mg/kg(氧乐果)
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酶受体生物荧光法快速检测水果和蔬菜中多农药残留
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邢仕歌 1 , 姚桂红 1 , 姚美伊 1 , 刘龙飞 2 , 凌云 1, *
食品安全质量检测学报 | 本期重点:食品中有毒有害物质分析与监测 2025,16(12): 145-150
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食品安全质量检测学报 | 本期重点:食品中有毒有害物质分析与监测 2025, 16(12): 145-150
酶受体生物荧光法快速检测水果和蔬菜中多农药残留
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邢仕歌1 , 姚桂红1, 姚美伊1, 刘龙飞2, 凌云1, *
作者信息
  • 1 中国质量检验检测科学研究院食品安全研究所, 北京 100176
  • 2 北京中检葆泰生物技术有限公司, 北京 100176
  • 邢仕歌(1980—), 女, 博士, 研究员, 主要研究方向为食品安全快检技术。E-mail:

通讯作者:

*凌云(1979—), 女, 博士, 研究员, 主要研究方向为营养与食品卫生学。E-mail:
Rapid determination of multiple pesticide residues in fruits and vegetables on bioluminescence inhibition of enzyme receptor
Shi-Ge XING1 , Gui-Hong YAO1, Mei-Yi YAO1, Long-Fei LIU2, Yun LING1, *
Affiliations
  • 1 Institute of Food Safety, Chinese Academy of Quality and Inspection & Testing, Beijing 100176, China
  • 2 Beijing Biotai Co., Ltd., Beijing 100176, China
出版时间: 2025-06-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250403001
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目的 建立酶受体生物荧光法快速检测水果和蔬菜中多农药残留的方法。方法 以灵敏度为指标, 分别考察提取溶剂、反应时间对结果的影响, 确定最佳前处理方法及检测条件; 对方法的灵敏度、假阳性率、假阴性率和交叉反应性进行评价。结果 该方法同时检测水果蔬菜中的多种农药残留, 灵敏度满足GB 2763—2021《食品安全国家标准 食品中农药最大残留限量》的限量要求, 假阴性率为0%, 假阳性率<5%, 交叉反应率≤0.10%。采用酶受体生物荧光法和国家标准方法对市售水果和蔬菜样品进行检测, 结果一致。结论 本研究建立的酶受体生物荧光法可用于水果和蔬菜中农药残留快速测定, 适用于基层监管部门进行监管, 也有助于企业内部更好地进行质量控制, 对于提升农药残留的快速检测能力具有重要意义。

酶受体生物荧光法  /  农药残留  /  快速检测  /  水果  /  蔬菜

Objective To establish a rapid analytical protocol for the detection of multiple pesticide residues in fruits and vegetables on bioluminescence inhibition of enzyme receptor. Methods Regarding sensitivity as the index of investigating, the effects of the extraction solvent and reaction time were separately investigated. The pretreatment protocol and detection conditions parameters were optimized. Then, the sensitivity, false positive rate, false negative rate and cross-reactivity of the method were evaluated. Results The findings demonstrated that the enzyme receptor bioluminescence technique was capable of concurrently identifying several pesticide residues within fruits and vegetables, with a sensitivity that complies with the GB 2763—2021 National food safety standards-Maximum residue limits of pesticides in food. The false negative rate was 0%, the false positive rate was less than 5%, and the cross-reactivity rate was less than or equal to 0.10%. Comparative analysis utilizing both the bioluminescence inhibition of enzyme receptor and the conventional national standard method was conducted on fruit and vegetable samples sourced from the market, yielding congruent results. Conclusion The bioluminescence inhibition of enzyme receptor established in the present study is deemed suitable for the rapid assessment of pesticide residues in fruits and vegetables. It is appropriated for oversight by local regulatory bodies and assists companies in improving their internal quality control, which is highly important for boosting the quick detection ability of pesticide residues.

bioluminescence inhibition of enzyme receptor  /  pesticide residues  /  rapid detection  /  fruits  /  vegetables
邢仕歌, 姚桂红, 姚美伊, 刘龙飞, 凌云. 酶受体生物荧光法快速检测水果和蔬菜中多农药残留. 食品安全质量检测学报, 2025 , 16 (12) : 145 -150 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250403001
Shi-Ge XING, Gui-Hong YAO, Mei-Yi YAO, Long-Fei LIU, Yun LING. Rapid determination of multiple pesticide residues in fruits and vegetables on bioluminescence inhibition of enzyme receptor[J]. Journal of Food Safety & Quality, 2025 , 16 (12) : 145 -150 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250403001
由于农药的过量使用和滥用, 农药残留在食物中引发中毒的情况较为突出。过量残留的农药可经消化道、呼吸道及完整的皮肤和黏膜进入人体, 在人体内积累, 诱发心脑血管病、糖尿病、癌症等慢性疾病; 农药残留还会影响胎儿正常发育, 甚至引起致癌、致畸和致突变[1-2]。有机磷类和氨基甲酸酯类农药是目前最常用的农药之一, 由此引发的中毒事件也最多[3-4]。我国对农药残留的监控十分重视, GB 2763—2021《食品安全国家标准 食品中农药最大残留限量》和GB 2763.1—2022《食品安全国家标准 食品中2,4-滴丁酸钠盐等112种农药最大残留限量》中蔬菜、水果等居民日常消费的重点农产品的限量标准数量增长明显, 涉及到有机磷和氨基甲酸酯类农药的有20余项。
农药残检测方法主要有色谱法和快速检测方法两类[5-6]。色谱法具有灵敏度高、检出限低等特点[7-9], 但对检测人员的技术要求高, 样品的前处理复杂, 检测流程耗时, 设备维护成本高, 不适宜基层部门使用[10-14]。快速检测方法操作简便, 检测时间短, 成本低, 无需大型仪器, 因而适合于现场的快速筛查。快速检测技术[15-17]则有生物传感器[18-22]、免疫层析法[23-24]和酶抑制法[25-26]等, 目前应用较为广泛的为后两种。但是胶体金法多针对单一农药快速检测, 无法实现多农药残留检测。酶抑制法基于有机磷和氨基甲酸酯类农药对乙酰胆碱酯酶活性的抑制效应, 间接快速检测这些农药残留, 但是目前普遍使用的胆碱酯酶易失活, 对环境要求较高, 因而方法不稳定, 部分目标物的灵敏度无法满足新的国家限量要求。鉴于我国农药残留检测的现状和需求, 建立适用于现场应用的快速、高灵敏、高通量且低成本的农药残留分析方法已经刻不容缓。
本研究采用昆虫酯酶结合荧光显色, 与农药的结合能力更强[27], 样品制备简单, 检测速度快, 使农药残留检测技术能够延伸到田间、地头和交易市场, 对于提升水果和蔬菜农药残留的快速检测能力, 提高农产品质量, 具有重要社会效益和经济效益[28-30]
根据农业农村部第1490号公告作物分类, 选取蔬菜水果各类别代表作物16种: 砂糖橘、苹果、桃子、葡萄、香蕉、哈密瓜、洋葱、花椰菜、大白菜、茄子、黄瓜、菜豆、莴苣、萝卜、莲藕、绿豆芽, 均采购于北京市大型连锁超市, 经仪器检测确证为无农药残留的阴性样品。农贸市场随机采集砂糖橘、苹果、葡萄、花椰菜、圆白菜、韭菜、黄瓜、豇豆样本用于与参比方法的一致性实验部分。
昆虫酯酶(酶活0.85 U)、荧光素酶受体和荧光底物(北京中检葆泰生物技术有限公司); 地虫硫磷、毒虫畏、对硫磷、庚烯磷、甲拌磷、氯唑磷、杀虫畏、速灭磷、特丁硫磷、治螟磷、丁硫克百威、苯线磷、灭线磷、氧乐果、乙酰甲胺磷、内吸磷、克百威、甲基硫环磷、久效磷、磷胺、杀扑磷、蝇毒磷、灭多威、蚜灭磷、喹硫磷、硝虫硫磷、保棉磷、混灭威、甲萘威、2,4-滴丁酸、六六六、滴滴涕、甲氰菊酯、除草定、杀虫脒标准品溶液(质量浓度1 mg/L, 德国Dr. Ehrenstorfer公司); 无水硫酸钠(纯度99.0%, 中国医药集团有限公司); 甲醇、乙腈、乙酸乙酯、丙酮(纯度99.9%, 美国Thermo Fisher Scientific公司)。
NovaLUM II-X荧光检测仪、ER-INC-2×20型孵育器(北京中检葆泰生物技术有限公司); Xevo TQ-S液相色谱串联高分辨质谱仪(liquid chromatography-tandem mass spectrometry, LC-MS/MS)(美国Waters公司); 7890A气相色谱串联质谱仪(gas chromatography-tandem mass spectrometry, GC-MS/MS)、Eclipse Plus C18安捷伦色谱柱(2.1 mm×100 mm, 1.8 µm)、ZORBAX SB-C18安捷伦色谱柱(2.1 mm×100 mm, 3.5 µm)(美国安捷伦科技有限公司); SH-Rtx-1701岛津气相色谱柱(30 m×0.25 mm, 0.25 µm)(日本岛津公司); JC-SNS空气吹干仪(青岛精诚仪器仪表有限公司); Lab Dancer微型振荡器(德国IKA公司); Milli-Q Advantage A10纯水仪(德国默克公司); ML104分析天平(精度0.1 mg, 瑞士Mettler Toledo公司)。
根据市场调查和文献调研情况, 选取水果和蔬菜中的23种常用农药(地虫硫磷、毒虫畏、对硫磷、庚烯磷、甲拌磷、氯唑磷、杀虫畏、速灭磷、特丁硫磷、治螟磷、丁硫克百威、苯线磷、灭线磷、氧乐果、乙酰甲胺磷、内吸磷、克百威、甲基硫环磷、久效磷、磷胺、杀扑磷、蝇毒磷和灭多威)作为目标物。依据GB 2763—2021和GB 2763.1—2022规定的16种典型代表作物中前述农药的最大残留限量(表1), 采取空白基质加标的方式分别制备16种代表作物23种农药加标浓度为0.5倍检出限和1倍检出限的加标样品。
取5 g样品与10 mL 20%硫酸钠溶液混匀制备样品液。吸取2 mL样品液, 加入2 mL硫酸钠溶液和2 mL提取溶剂, 混匀, 4500 r/min离心2 min。取100 μL上清液至离心管中, 在45 ℃下吹至近干。加入1 mL去离子水复溶, 即为待测溶液。
准确移取100 μL待测溶液至检测微管中, 加入50 μL昆虫酯酶溶液, 混匀并孵育。向微管底部加入50 μL酶受体溶液, 混匀并孵育。取出微管, 加入1 mL荧光底物溶液, 用NovaLUM II-X荧光检测仪读取结果。
将零质控读数的相对光单位(relative light unit, RLU)乘以系数获得每次检测的质控点。当样品数值的RLU高于质控点时, 结果为阴性; 当样品的RLU数值低于或等于质控点时, 结果为阳性。
取加标浓度为1倍检出限的各基质阳性样品, 每种样品设3个重复, 分别使用甲醇、乙腈、乙酸乙酯、乙酸乙酯-丙酮、丙酮作为提取溶剂, 进行样品前处理和测定, 根据灵敏度(检出阳性结果的阳性样品数占总阳性样品数的百分比)选择最佳提取溶剂。
取加标浓度为1倍检出限的各基质阳性样品, 每种样品设3个重复, 按表2中的条件进行正交实验, 时间1为提取液与酶溶液的孵育反应时间, 时间2为荧光素酶受体孵育反应时间。根据灵敏度确定最佳孵育温度和孵育时间。
16种代表作物的空白样品50个, 23种农药加标浓度为0.5倍检出限和1倍检出限的加标样品各50个, 采取优化后的方法进行样品前处理和检测。根据《市场监管总局关于规范食品快速检测使用的意见》(国市监食检规〔2023〕1号)中“食品快速检测产品符合性评价技术要求”计算灵敏度、假阳性率和假阴性率。
以不含农药的空白苹果样品为基质, 加入结构类似农药(蚜灭磷、喹硫磷、硝虫硫磷、保棉磷、混灭威、甲萘威)和其他常见类农药(2,4-滴丁酸、六六六、滴滴涕、甲氰菊酯、除草定、杀虫脒), 制备加标浓度为2~200 mg/kg的样品, 采取优化后的方法进行样品前处理和检测, 并计算交叉反应率/%=目标物质检出限/干扰物质检出阳性时的最小浓度。
市售砂糖橘、苹果、葡萄、花椰菜、圆白菜、韭菜、黄瓜、豇豆样本, 分别用酶受体生物荧光法与参比方法GB 23200.121—2021《食品安全国家标准 植物源性食品中331种农药及其代谢物残留量的测定 液相色谱-质谱联用法》、GB 23200.113—2018《食品安全国家标准 植物源性食品中208种农药及其代谢物残留量的测定 气相色谱-质谱联用法》和GB 23200.13—2016《食品安全国家标准 茶叶中448种农药及相关化学品残留量的测定 液相色谱-质谱法》检测23种农药残留, 并对结果进行对比。
实验结果以百分比表示, 数据使用Excel 2016统计分析软件对检测方法的灵敏度、假阳性率与假阴性率进行分析, 图表使用Excel 2016软件制成。
使用5种提取液分别对添加浓度为1倍检出限的16种基质样品提取后检测, 计算灵敏度。结果如图1所示, 使用甲醇作为提取液, 灵敏度为71.74%~78.26%; 乙腈作为提取液, 灵敏度为71.74%~76.09%; 乙酸乙酯作为提取液, 灵敏度为76.09%~89.13%; 乙酸乙酯-丙酮作为提取液, 灵敏度为100.00%; 丙酮作为提取液, 灵敏度84.78%~86.96%。因此, 选择乙酸乙酯-丙酮作为最佳提取溶剂。
不同测定条件的灵敏度结果见表3。在各孵育温度条件下, 随着孵育时间的延长, 灵敏度逐渐升高; 在相同的孵育时间条件下, 灵敏度开始随着温度升高逐渐升高, 后续随着孵育温度的继续升高有所下降, 说明温度过高会引起酶的失活, 与文献报道一致[27]。当孵育温度为35 ℃, 孵育时间为10 min+5 min时, 灵敏度达到100.0%, 因此选择孵育温度35 ℃, 提取液与酶溶液孵育反应时间10 min, 荧光素酶受体孵育反应时间5 min为最佳测定条件。
采用1.5.1中获得的数据, 计算灵敏度、假阳性率及假阴性率。对于所有基质, 空白样品检测结果均为阴性, 加标浓度为1倍检出限的阳性样品检测结果均为阳性, 加标浓度为0.5倍检出限的样品中, 香蕉、洋葱、茄子、莴苣、绿豆芽各有一个样品检出阳性。经过计算, 本研究建立的酶受体生物荧光法检测23种农药残留的灵敏度为100%, 假阴性率为0%, 假阳性率<5%。
采用1.5.2中获得的数据,以目标物质检出限0.2 mg/kg为基准计算交叉反应率。结果显示, 结构类似农药(蚜灭磷、喹硫磷、硝虫硫磷、保棉磷、混灭威、甲萘威)的检出限为200 mg/kg, 交叉反应率为0.10%, 其他常用农药(2,4-滴丁酸、六六六、滴滴涕、甲氰菊酯、除草定、杀虫脒)的检出限为2000 mg/kg, 交叉反应率为0.01%。
为进一步评价该方法, 分别用酶受体生物荧光法与国家标准方法检测8种常见市售蔬菜水果中23种农药残留的含量。结果显示, 酶受体生物荧光法检测出豇豆样品为阳性, 其他均为阴性, 用国家标准方法对相同样品进行检测, 结果显示豇豆样品中氧乐果含量为0.03 mg/kg, 其他样品均为阴性(表4)。两种方法的检测结果完全一致。
本研究通过对样品前处理方法和检测条件进行优化, 确定乙酸乙酯-丙酮为提取液, 35 ℃作为最佳孵育温度, 10 min+5 min作为最佳孵育时间, 建立了可用于蔬菜水果中多农药残留快速检测的酶受体生物荧光法。性能指标验证结果显示, 当以23种常见农药国标限量标准为检出限时, 该方法的灵敏度均为100%, 假阴性率均为0%, 假阳性率均<5%; 与结构类似物农药和其他常用农药的交叉反应率分别为0.10%和0.01%。8种蔬菜水果真实样品检测显示本方法和国家标准方法检测结果一致。由上述数据可知, 本研究建立的基于酶受体生物荧光法可同时检测水果和蔬菜中的多种农药残留, 检出限满足GB 2763—2021中规定的限量要求; 灵敏度高、特异性强、操作简单、检测速度快。因此本研究建立的酶受体生物荧光法能满足蔬菜和水果中农药多残留快速测定的要求, 易于推广应用。对于指导、规范农户合理施药, 提升基层监管部门的农药残留监测能力, 提高水果和蔬菜的质量安全水平, 有着重要的现实意义。快速检测向着适用于市场和产地环境的便携式高通量快速筛查的方向发展, 将是未来农药残留现场检测的趋势。
  • 中国质量检验检测科学研究院基本科研业务费项目(2024JK029)
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2025年第16卷第12期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250403001
  • 接收时间:2025-04-03
  • 首发时间:2026-01-13
  • 出版时间:2025-06-25
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  • 收稿日期:2025-04-03
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中国质量检验检测科学研究院基本科研业务费项目(2024JK029)
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    1 中国质量检验检测科学研究院食品安全研究所, 北京 100176
    2 北京中检葆泰生物技术有限公司, 北京 100176

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*凌云(1979—), 女, 博士, 研究员, 主要研究方向为营养与食品卫生学。E-mail:
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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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