Article(id=1153986790378693171, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986777279877909, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241021005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1729440000000, receivedDateStr=2024-10-21, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1753061490863, onlineDateStr=2025-07-21, pubDate=1736870400000, pubDateStr=2025-01-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1753061490863, onlineIssueDateStr=2025-07-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1753061490863, creator=13701087609, updateTime=1753061490863, updator=13701087609, issue=Issue{id=1153986777279877909, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='1', 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=0, createTime=1753061487741, creator=13701087609, updateTime=1757901302572, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1174286432060453412, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986777279877909, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1174286432060453413, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986777279877909, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=22, endPage=27, ext={EN=ArticleExt(id=1153986791276274233, articleId=1153986790378693171, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Simultaneous determination of 317 kinds of pesticide residues in pomelo flowers by ultra performance liquid chromatography-tandem mass spectrometry coupled with QuEChERS, columnId=1153986581653349021, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Application of Modern Analysis Instrument in Food Detection, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a method for simultaneous determination of 317 kinds of pesticide residues in pomelo flowers by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) coupled with QuEChERS extraction. Methods Pomelo flowers samples were extracted with acetonitrile and salted with extraction salt pack, cleaned up by ethylenediamine-N-propylsilane silica gel (PSA) and graphitized carbon blank (GCB), the analytes were separated on an ACQUITY UPLC HSS T3 column to achieve the separation within 30 min. The 317 kinds of pesticides were detected in electron spray ionization with positive ion mode and electron spray ionization with negative ion mode with multiple reaction monitoring (MRM) mode and quantified by matrix matching curve external standard method. Results The 317 kinds of pesticides in pomelo flowers showed good linearity with correlation coefficients (r) greater than 0.995 within the range of 2.0-200.0 µg/L, and the limits of quantification were 0.002-0.010 mg/kg, the average recoveries at 3 spiked levels were 74.3%-120.3%, and the relative standard deviations were 0.36%-10.00%. Conclusion The method is simple, rapid, accurate and sensitive, can meet the rapid detection requirements of 317 kinds of pesticides in pomelo flowers and provide reliable data support for further standardizing for edible safety evaluation of pomelo flowers.

, correspAuthors=Xu-Xia LIANG, 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=Chun-Xia LI, Chun-Li DENG, Cai-Ting YE, Yi-Wen LI, Shi-Yao LI, Hai HONG, Su-Wen CHEN, Zhi LI, Hui LIANG, Xu-Xia LIANG), CN=ArticleExt(id=1153986799274812132, articleId=1153986790378693171, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=QuEChERS-超高效液相色谱-串联质谱法同时测定柚花中317种农药残留量, columnId=1153986778571723567, journalTitle=食品安全质量检测学报, columnName=专题:现代分析仪器在食品检测中的应用, runingTitle=null, highlight=null, articleAbstract=

目的 建立QuEChERS前处理法结合超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)同时测定柚花中317种农药残留量的方法。方法 柚花样品采用乙腈作为提取剂, 经萃取盐盐析处理, 采用乙二胺-N-丙基硅烷化硅胶(ethylenediamine-N-propylsilane silica gel, PSA)和石墨化炭黑(graphitized carbon black, GCB)作为净化剂, 选用ACQUITY UPLC HSS T3色谱柱, 采用梯度洗脱方式在30 min完成317种目标物质的分析。电喷雾源正离子和负离子同时扫描, 采用多重反应监测(multiple reaction monitoring, MRM)模式、基质匹配标准曲线外标法定量。结果 317种农药在该条件下在2.0~200.0 µg/L质量浓度范围内均线性良好, 相关系数r>0.995, 方法定量限为0.002~0.010 mg/kg, 在低、中、高3个添加水平下的平均回收率为74.3%~120.3%, 相对标准偏差在0.36%~10.00%。结论 该方法具有简单、快速、准确、灵敏等特点, 为柚花中多种农药残留的测定和柚花研制的食品安全地方标准提供了可靠的技术支持。

, correspAuthors=梁旭霞, authorNote=null, correspAuthorsNote=
*梁旭霞(1972—), 女, 博士, 主任技师, 主要研究方向为食品药品质量标准。E-mail:
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李春霞(1985—), 女, 硕士, 主管技师, 主要研究方向为水及涉水产品、食品理化检验。E-mail:

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李春霞(1985—), 女, 硕士, 主管技师, 主要研究方向为水及涉水产品、食品理化检验。E-mail:

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Journal of Instrumental Analysis, 2023, 42(12): 1537-1546., articleTitle=Screening of 166 pesticide residues in tea based on ultra high performance liquid chromatography- quadrupole/ electrostatic field orbitrap high resolution mass spectrometry, refAbstract=null)], funds=[Fund(id=1174369398241505936, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, awardId=C2022016, language=CN, fundingSource=广东省医学科学技术研究基金项目(C2022016), fundOrder=null, country=null), Fund(id=1174369398317003410, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, awardId=2023C0301001, language=CN, fundingSource=梅州市科技计划项目(2023C0301001), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1174369394160448015, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, xref=null, ext=[AuthorCompanyExt(id=1174369394168836624, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, companyId=1174369394160448015, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Guangdong Provincial Institute of Biological Products and Materia Medica, Guangzhou 510440, China), AuthorCompanyExt(id=1174369394177225233, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, companyId=1174369394160448015, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=广东省生物制品与药物研究所, 广州 510440)])], figs=[ArticleFig(id=1174369397750772356, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, language=EN, label=Fig.1, caption=Recovery rate distribution of different purification systems for pesticides added at a concentration of 20 μg/L, figureFileSmall=mVN1hbaBwQ8Q6hbBUrdI+g==, figureFileBig=3AnvlCNOsxfvho9T8Cr1lg==, tableContent=null), ArticleFig(id=1174369397813686917, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, language=CN, label=图1, caption=不同净化剂体系对添加质量浓度为20 μg/L农药物质的加标回收率分布

注: ①~⑤分别为150 mg PSA、150 mg PSA+150 mg C18、150 mg PSA+ 150 mg C18+15 mg GCB、150 mg PSA+15 mg GCB、300 mg PSA+300 mg C18+150 mg GCB。

, figureFileSmall=mVN1hbaBwQ8Q6hbBUrdI+g==, figureFileBig=3AnvlCNOsxfvho9T8Cr1lg==, tableContent=null), ArticleFig(id=1174369397868212871, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, language=EN, label=Table 1, caption=

Mobile phases and recommend separation conditions of UPLC

, figureFileSmall=null, figureFileBig=null, tableContent=
时间
/min
流速/(mL/min) 水(含2 mmol/L甲酸铵和0.01%甲酸)/% 甲醇(含2 mmol/L甲酸铵和0.01%甲酸)/%
0.0 0.3 97 3
1.0 0.3 97 3
1.5 0.3 85 15
2.5 0.3 50 50
18.0 0.3 30 70
23.0 0.3 2 98
27.0 0.3 2 98
27.1 0.3 97 3
30.0 0.3 97 3
), ArticleFig(id=1174369397931127433, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, language=CN, label=表1, caption=

UPLC流动相及参考分离条件

, figureFileSmall=null, figureFileBig=null, tableContent=
时间
/min
流速/(mL/min) 水(含2 mmol/L甲酸铵和0.01%甲酸)/% 甲醇(含2 mmol/L甲酸铵和0.01%甲酸)/%
0.0 0.3 97 3
1.0 0.3 97 3
1.5 0.3 85 15
2.5 0.3 50 50
18.0 0.3 30 70
23.0 0.3 2 98
27.0 0.3 2 98
27.1 0.3 97 3
30.0 0.3 97 3
), ArticleFig(id=1174369397994041995, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, language=EN, label=Table 2, caption=

Mass spectrogram parameters, and retention time datas of typical pesticides

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 中文
名称
电离
模式
母离子 子离子 锥孔
电压/V
碰撞能
/eV
保留
时间/min
有机磷类 乙酰甲
胺磷
ESI+ 184.0 143.0*/
125.0
18 8/20 3.11
杀虫剂 啶虫脒 ESI+ 223.1 126.0*/
56.1
34 20/20 4.07
除草剂 莠灭净 ESI+ 228.1 186.1*/
68.1
38 18/36 9.45
杀菌剂 嘧菌酯 ESI+ 404.1 372.0*/
329.0
28 15/30 10.35
杀螨剂 苯螨特 ESI+ 364.1 199.1*/
105.0
20 18/20 18.28
生物
农药
除虫菊
素I
ESI+ 329.1 161.1*/
133.1
25 10/15 22.39
植物生长调节剂 多效唑 ESI+ 294.1 70.2*/
125.1
20 15/40 11.32/
11.60
杀虫剂 氟虫腈 ESI- 434.9 330.0*/
250.0
24 15/30 15.51
), ArticleFig(id=1174369398115676813, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986790378693171, language=CN, label=表2, caption=

部分代表性农药的质谱参数、保留时间

, figureFileSmall=null, figureFileBig=null, tableContent=
类别 中文
名称
电离
模式
母离子 子离子 锥孔
电压/V
碰撞能
/eV
保留
时间/min
有机磷类 乙酰甲
胺磷
ESI+ 184.0 143.0*/
125.0
18 8/20 3.11
杀虫剂 啶虫脒 ESI+ 223.1 126.0*/
56.1
34 20/20 4.07
除草剂 莠灭净 ESI+ 228.1 186.1*/
68.1
38 18/36 9.45
杀菌剂 嘧菌酯 ESI+ 404.1 372.0*/
329.0
28 15/30 10.35
杀螨剂 苯螨特 ESI+ 364.1 199.1*/
105.0
20 18/20 18.28
生物
农药
除虫菊
素I
ESI+ 329.1 161.1*/
133.1
25 10/15 22.39
植物生长调节剂 多效唑 ESI+ 294.1 70.2*/
125.1
20 15/40 11.32/
11.60
杀虫剂 氟虫腈 ESI- 434.9 330.0*/
250.0
24 15/30 15.51
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QuEChERS-超高效液相色谱-串联质谱法同时测定柚花中317种农药残留量
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李春霞 , 邓春丽 , 叶彩婷 , 李亦文 , 李诗瑶 , 洪海 , 陈素文 , 李志 , 梁辉 , 梁旭霞 *
食品安全质量检测学报 | 专题:现代分析仪器在食品检测中的应用 2025,16(1): 22-27
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食品安全质量检测学报 | 专题:现代分析仪器在食品检测中的应用 2025, 16(1): 22-27
QuEChERS-超高效液相色谱-串联质谱法同时测定柚花中317种农药残留量
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李春霞 , 邓春丽, 叶彩婷, 李亦文, 李诗瑶, 洪海, 陈素文, 李志, 梁辉, 梁旭霞*
作者信息
  • 广东省生物制品与药物研究所, 广州 510440
  • 李春霞(1985—), 女, 硕士, 主管技师, 主要研究方向为水及涉水产品、食品理化检验。E-mail:

通讯作者:

*梁旭霞(1972—), 女, 博士, 主任技师, 主要研究方向为食品药品质量标准。E-mail:
Simultaneous determination of 317 kinds of pesticide residues in pomelo flowers by ultra performance liquid chromatography-tandem mass spectrometry coupled with QuEChERS
Chun-Xia LI , Chun-Li DENG, Cai-Ting YE, Yi-Wen LI, Shi-Yao LI, Hai HONG, Su-Wen CHEN, Zhi LI, Hui LIANG, Xu-Xia LIANG*
Affiliations
  • Guangdong Provincial Institute of Biological Products and Materia Medica, Guangzhou 510440, China
出版时间: 2025-01-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241021005
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目的 建立QuEChERS前处理法结合超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)同时测定柚花中317种农药残留量的方法。方法 柚花样品采用乙腈作为提取剂, 经萃取盐盐析处理, 采用乙二胺-N-丙基硅烷化硅胶(ethylenediamine-N-propylsilane silica gel, PSA)和石墨化炭黑(graphitized carbon black, GCB)作为净化剂, 选用ACQUITY UPLC HSS T3色谱柱, 采用梯度洗脱方式在30 min完成317种目标物质的分析。电喷雾源正离子和负离子同时扫描, 采用多重反应监测(multiple reaction monitoring, MRM)模式、基质匹配标准曲线外标法定量。结果 317种农药在该条件下在2.0~200.0 µg/L质量浓度范围内均线性良好, 相关系数r>0.995, 方法定量限为0.002~0.010 mg/kg, 在低、中、高3个添加水平下的平均回收率为74.3%~120.3%, 相对标准偏差在0.36%~10.00%。结论 该方法具有简单、快速、准确、灵敏等特点, 为柚花中多种农药残留的测定和柚花研制的食品安全地方标准提供了可靠的技术支持。

QuEChERS  /  超高效液相色谱-串联质谱法  /  柚花  /  农药残留

Objective To establish a method for simultaneous determination of 317 kinds of pesticide residues in pomelo flowers by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) coupled with QuEChERS extraction. Methods Pomelo flowers samples were extracted with acetonitrile and salted with extraction salt pack, cleaned up by ethylenediamine-N-propylsilane silica gel (PSA) and graphitized carbon blank (GCB), the analytes were separated on an ACQUITY UPLC HSS T3 column to achieve the separation within 30 min. The 317 kinds of pesticides were detected in electron spray ionization with positive ion mode and electron spray ionization with negative ion mode with multiple reaction monitoring (MRM) mode and quantified by matrix matching curve external standard method. Results The 317 kinds of pesticides in pomelo flowers showed good linearity with correlation coefficients (r) greater than 0.995 within the range of 2.0-200.0 µg/L, and the limits of quantification were 0.002-0.010 mg/kg, the average recoveries at 3 spiked levels were 74.3%-120.3%, and the relative standard deviations were 0.36%-10.00%. Conclusion The method is simple, rapid, accurate and sensitive, can meet the rapid detection requirements of 317 kinds of pesticides in pomelo flowers and provide reliable data support for further standardizing for edible safety evaluation of pomelo flowers.

QuEChERS  /  ultra performance liquid chromatography-tandem mass spectrometry  /  pomelo flowers  /  pesticide residues
李春霞, 邓春丽, 叶彩婷, 李亦文, 李诗瑶, 洪海, 陈素文, 李志, 梁辉, 梁旭霞. QuEChERS-超高效液相色谱-串联质谱法同时测定柚花中317种农药残留量. 食品安全质量检测学报, 2025 , 16 (1) : 22 -27 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241021005
Chun-Xia LI, Chun-Li DENG, Cai-Ting YE, Yi-Wen LI, Shi-Yao LI, Hai HONG, Su-Wen CHEN, Zhi LI, Hui LIANG, Xu-Xia LIANG. Simultaneous determination of 317 kinds of pesticide residues in pomelo flowers by ultra performance liquid chromatography-tandem mass spectrometry coupled with QuEChERS[J]. Journal of Food Safety & Quality, 2025 , 16 (1) : 22 -27 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241021005
柚子是我国南方地区农业农村经济发展的重要支柱产业之一。近代医学研究证明, 柚子对防治冠心病等心血管病有一定的疗效, 柚花的食药用价值也不容忽视, 其不仅有柚子的保健药理功效, 还有生发、润燥、美容的作用。《本草纲目》[1]《民间常用草药汇编》[2]《广西中药志》[3]等文献记载: 柚花具有行气、除痰、镇痛作用, 治胃脘胸膈间痛。人们还将柚花作为菜肴食用, 例如《肠胃病中医辩证有效食疗》[4]记载治虚寒性胃痛, 采用猪肚和柚花一起煮汤, 饮汤食猪肚。柚花的芳香气味及功效成分的研究, 使其营养价值和保健功效近年来备受关注, 食用柚花成为花类的一个重要应用方向, 在食品领域有较高的开发潜能。因此, 南方省份在积极制定柚花相关的食品安全地方标准[5-6]
在柚花期, 病虫害的发生可导致柚子的成花率低、座果率下降, 目前主要防控措施是喷洒化学农药, 因此, 有效监控柚花中农药残留量对柚花的食用推广、保障人民健康有着极其重要的意义。目前国内对农药残留的测定, 多采用气相色谱法[7-8]、气相色谱-质谱法[9-11]、气相色谱-串联质谱法[12-16]、高效液相色谱法[17-18]、高效液相色谱-串联质谱法[19]。超高效液相色谱-串联质谱(ultra performance liquid chromatography-tandem mass spectrometry, UPLC- MS/MS)[20-25]特别适用于热不稳定、难挥发的农药组分如克百威、吡虫啉等农药的定性定量分析, 尤其适合多农药残留分析, 本研究依据GB 2763—2021《食品安全国家标准 食品中农药最大残留限量》、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》和相关文献报道[26-28], 针对冻干柚花中含有大量挥发性芳香成分、较高蛋白质含量、丰富膳食纤维和低脂肪的特点, 本研究优化QuEChERS前处理方法, 建立了UPLC-MS/MS同时测定柚花中317种农药残留量的方法。为柚花中多农药残留的测定和柚花研制食品安全地方标准提供可靠的技术支持, 对促进柚花产业的高质量发展有着非常重要的意义。
Waters TQ-S Micro超高效液相色谱-串联质谱仪、Waters ACQUITY HSS T3色谱柱(2.1 mm×100 mm, 1.8 μm)(美国Waters公司); IKA V3 S025涡旋振荡器[艾卡(广州)仪器设备有限公司]; Thermo Sorvall ST8R高速离心机(美国Thermo Fisher Scientific公司); METTLER AE 240电子天平(感量0.1 mg, 梅特勒-托利多仪器上海有限公司)。
甲醇(色谱纯)、甲酸(质谱纯)、乙酸(质谱纯)、甲酸铵(质谱纯)(美国Fisher Chemical公司); 无水硫酸镁(MgSO4)、氯化钠(NaCl)、柠檬酸钠二水合物、柠檬酸氢二钠(分析纯, 广州化学试剂厂); 乙腈(色谱纯)、乙二胺-N-丙基硅烷化硅胶(ethylenediamine-N-propylsilane silica gel, PSA)、石墨化炭黑(graphitized carbon black, GCB)(粒径40~60 μm, 批号DX22440)(上海安谱实验科技股份有限公司); 0.22 μm微孔滤膜(天津市津腾实验设备公司); Genin PURIST超纯水(上海乐枫生物科技有限公司); 317种农药标准品(质量浓度100 μg/mL, 纯度≥99%)(天津阿尔塔标准物质研究院有限公司)。
柚花样品来源: 我国南方若干沙田柚种植点, 采摘柚树盛花期不同位置柚花, 每份样品取不少于2 kg新鲜柚花, 冷冻干燥后打成粉, 密封冷冻保存。
混合标准中间工作溶液: 分别移取适量体积各组农药标准溶液, 用乙腈溶解定容配制成1.0 μg/mL的标准中间工作溶液, -18 ℃冰箱中保存, 可使用1个月。移取适量混合标准中间工作溶液用空白柚花基质溶液配成1.00 mL不同质量浓度(2.0~200.0 μg/L)的基质混合标准工作溶液, 用于做基质标准工作曲线。
称取1.0 g(精确到0.1 mg)试样于50.0 mL塑料离心管中, 加9.00 mL水涡旋混匀, 静置30 min。加入10.00 mL乙腈及1颗陶瓷均质子, 剧烈振荡1 min, 加入萃取盐(4 g MgSO4、1 g NaCl、1 g柠檬酸钠二水合物、0.5 g柠檬酸氢二钠), 剧烈振荡1 min后8000 r/min离心5 min。定量吸取6.00 mL上清液至内含除水剂和净化材料的固相萃取净化管中(900 mg MgSO4、150 mg PSA、15 mg GCB), 涡旋混匀1 min, 8000 r/min离心5 min, 吸取上清液过0.22 μm有机相微孔滤膜, 供UPLC-MS/MS测定。
(1)色谱条件:
色谱柱为ACQUITY HSS T3 (2.1 mm×100 mm, 1.8 μm), 柱温40 ℃, 流速0.3 mL/min, 进样量2 μL, 流动相梯度见表1
(2)质谱条件:
电离方式: 电喷雾源正离子模式(electron spray ionization with positive ion mode, ESI+)和负离子模式(electron spray ionization with negative ion mode, ESI-)同时扫描; 毛细管电压: ESI+ 0.5 kV, ESI- 2.5 kV; 脱溶剂气温度: 550 ℃; 脱溶剂气流量: 1000 L/hr; 雾化气、干燥气: 氮气, 碰撞气: 氩气; 检测方式: 多反应监测(multiple reaction monitoring, MRM)。部分代表性农药的监测离子对、相关电压参数、扫描离子模式等参数见表2
采用Waters MassLynx软件进行原始数据处理, Microsoft 2010系列软件制表、制图, 样品做6次平行实验, 并且利用OriginPro 8进行统计学数据处理分析。
由于柚花富含芳香类物质, 冻干柚花蛋白质含量达到16%以上, 膳食纤维含量超过38%, 矿物质含量为6.5%左右, 脂肪含量在1%~2%, 因此, 需要针对这些特质, 考虑合适的提取溶剂与净化条件。
本研究中柚花为冻干粉状干燥试样, 试样与提取试剂的料液比会影响到目标农药残留化合物的回收率。选取空白柚花样品, 分别称取1 g试样4份, 加9 mL水涡旋混匀, 静置30 min, 分别加入10 μg/kg的农药混标溶液, 然后再分别加入乙腈3.0、5.0、7.5和10.0 mL, 考察料液比(g:mL)为1:3、1:5、1:7.5和1:10时对农药残留物质回收率的影响。当料液比为1:3和1:5时, 有机磷类等极性较强的化合物回收率小于50%; 当料液比为1:7.5时, 基质样品中约84%农药物质的回收率为70%~120%; 当料液比为1:10时, 基质样品中农药物质回收率均在74.3%~120.3%范围内; 因此, 确定本研究中柚花试样与提取试剂料液比为1:10 (g:mL)。
因柚花粉中蛋白质含量较高, 选择提取溶剂需考虑其对大多数农药有良好溶解性外, 还需考虑其沉淀蛋白的性能, 以减少上机干扰。乙腈作为强极性溶剂, 对样品基质的渗透力较强, 且乙腈在沉淀水溶性蛋白和高等级蛋白成分方面较甲醇等其他有机溶剂更完全, 考虑到乙腈不能溶解膳食纤维, 柚花粉中高含量的膳食纤维可通过高速离心除去。考察了用乙腈、甲醇、丙酮、乙酸乙酯、0.1%乙酸-乙腈做为提取溶剂对同一柚花样品进行前处理, 发现样品提取液的颜色甲醇最深, 呈棕黄色; 其次是丙酮和乙酸乙酯, 呈黄绿色; 乙腈和0.1%乙酸-乙腈颜色最浅, 呈浅绿色, 说明乙腈提取的上清液中色素类共提物较少, 基质干扰相对较少, 且采用乙腈提取时回收率在70%~110%的化合物数量最多。综上, 选取乙腈作为提取剂进行后续实验。
实验选取QuEChERS方法的常用净化剂对基质样品进行净化除杂, 从而降低干扰物对目标分析物的干扰, 提高检测的灵敏度和准确性[29-30]。柚花中糖类、有机酸和黄酮类物质含量丰富, 净化剂首先考虑使用PSA去除极性干扰物。另外, 干柚花中色素含量虽然不高, 但黄酮类物质柚皮苷含量超过8.5%[28], 本研究的样本含量更是高达14%。柚皮苷和柚皮素都是具有平面芳香环结构以及具有六元环结构的分子, GCB对其具有很好的选择性, 所以在净化剂中加入GCB可以较好地吸附它们。由于柚花粉中的脂肪含量很低, 仅有1%左右, 为防止某些脂溶性的农药被吸附损失, 故不考虑使用C18或C8吸附剂。
本研究考察了不同净化剂的种类和用量对农药残留目标分析物加标回收率的影响, 分别采150 mg PSA、150 mg PSA+150 mg C18、150 mg PSA+150 mg C18+15 mg GCB、150 mg PSA+15 mg GCB、300 mg PSA+300 mg C18+150 mg GCB 5种方案对乙腈提取液进行净化除杂处理, 考察同一添加质量浓度(20 μg/L)下317种农药的回收率差异。各净化体系对于柚花样品提取液的脱色效果明显, 增加净化剂用量可提高目标物的净化效果, 但从图1中317种农药物质的加标回收率分布情况来看, 净化剂用量增加后, 甲胺磷、乙酰甲胺磷等有机磷农药、除虫菊酯类等农药化合物的回收率降低了。实验表明, 针对柚花特性进行的净化剂选择分析是合理的, 选择150 mg PSA+15 mg GCB净化除杂可以获得更多目标分析物更佳的回收率。
比较317种农药混合标准溶液在ZORBAX RRHD SB-C18 (2.1 mm×150 mm, 1.8 μm)和ACQUITY UPLC HSS T3 (2.1 mm×100 mm, 1.8 μm)色谱柱上的分离效果, 通过实验表明, 2种型号的色谱柱均可以实现317种农药物质的有效分离, 但ACQUITY UPLC HSS T3柱具有较尖锐的色谱峰和更高的响应灵敏度, 更适合农药多残留分析。其原因可能是ACQUITY UPLC HSS T3色谱柱先进的T3键合技术提高了柱效、改善了峰形和稳定性, 独特的封尾技术确保了对极性化合物有很好地保留, 与纯水相流动相兼容, 平衡极性与非极性化合物的保留和分离具有更佳的表现。
流动相通常有甲醇和乙腈两种体系, 但它们对目标化合物响应值与峰形的影响是不同的, 本研究中选用甲醇比选用乙腈更能提高加氢峰母离子化合物或加铵峰母离子化合物的响应值。在正离子扫描模式下, 加入少量甲酸铵和甲酸可提升目标化合物的离子化程度, 从而增强其质谱峰响应值[22]。本研究通过实验, 选择水(含2 mmol/L甲酸铵和0.01%甲酸)和甲醇(含2 mmol/L甲酸铵和0.01%甲酸)作为流动相, 采用梯度洗脱方式(见表1), 30 min可完成317种目标物质的分析。
配制质量浓度为50 µg/L的317种农药混合标准溶液,在MS1 Scan模式下分别对农药进行一级质谱分析,通过优化锥孔电压获得每种农药具有最高响应信号的母离子。在Daughter Scan模式下对母离子进行二级质谱分析,优化碰撞能量,选择响应信号最强的两个子离子分别作为定量离子和定性离子。部分代表性农药的保留时间和质谱参数见表2
基质效应通常在样品离子化过程中出现, 会影响分析结果的准确性和可靠性, 主要表现为基质抑制或基质增强。不同的农药种类、不同的样品基质的基质效应不相同, 基质效应的相对强度=(基质标准溶液的峰面积/溶剂标准溶液的峰面积)×100%。
柚花样品基质较为复杂, 本研究通过分别配制质量浓度为2.0、5.0、10.0、20.0、50.0、100.0和200.0 μg/L的317种农药混合标准的试剂空白和空白柚花基质系列标准溶液, 上机测定, 考察基质效应的影响。结果显示, 317种农药物质的相关系数r均大于0.995, 表明在2.0~200.0 µg/L范围内溶剂曲线和柚花基质曲线均具有良好的线性关系, 其中柚花基质中有240种物质存在弱基质效应(小于20%)、37种存在中等基质效应(20%~50%)、40种存在强基质效应影响(大于50%)。为避免基质干扰, 采用基质标准曲线法来消除基质效应带来的影响。
各物质在空白柚花基质加标0.002 mg/kg的浓度下进行测定, 以信噪比(S/N)为3和10时分别计算检出限和定量限; 除了吲唑磺菌胺、禾草灵、苯醚甲环唑、吡氟酰草胺、氟苯虫酰胺、甲胺磷、丙炔噁草酮的定量监测离子对在此浓度下的信噪比小于10但大于3, 其余各物质在该浓度定量监测离子对的信噪比均远远大于10, 得出各物质的定量限为0.002~ 0.010 mg/kg, 均符合国内外法规关于农药残留限量的要求。
为考察方法适用性, 分别将低(0.01 mg/kg)、中(0.05 mg/kg)、高(0.08 mg/kg) 3个不同浓度水平的农药混合标准溶液添加到空白柚花样品中进行加标回收实验研究, 各浓度水平做6次平行, 结果表明, 柚花基质中317种农药物质在上述加标水平下的回收率在74.3%~120.3%范围内, 相对标准偏差(relative standard deviation, RSD)在0.36%~10.00%, 表明该方法的准确性和精密度满足农药多残留的分析要求。
对南方沙田柚种植区域选择了36个种植园, 采集了36个柚花样品进行检测分析, 部分样品检出农药, 检出频次较多的农药主要有噻虫胺、乙螨唑、多菌灵、多效唑、苯醚甲环唑等, 都是低毒和微毒农药,与走访和调查了解到的柚树花期及生长期经常使用的杀虫药、防病杀菌药、植物生长调节剂用药基本一致[26-28]。六六六和滴滴涕被列为持久性有机污染物, 是制定食品安全地方标准时必检的项目, 它们在本研究36份样品中均未检出。
本研究利用QuEChERS前处理方法结合UPLC-MS/MS, 建立了柚花中317种农药残留的快速测定方法。317种农药化合物在2.0~200.0 μg/L质量浓度范围内线性良好, 相关系数r>0.995, 定量限为0.002~0.010 mg/kg, 在低、中、高3个浓度水平的加标回收率范围为74.3%~120.3%, RSD在0.36%~10.00%, 说明本方法前处理简单, 灵敏度高、抗基质干扰能力强, 结果准确, 能满足柚花样品多农药残留检测需求, 可以为制定柚花食品安全地方标准提供可靠的技术支持。
  • 广东省医学科学技术研究基金项目(C2022016)
  • 梅州市科技计划项目(2023C0301001)
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2025年第16卷第1期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241021005
  • 接收时间:2024-10-21
  • 首发时间:2025-07-21
  • 出版时间:2025-01-15
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  • 收稿日期:2024-10-21
基金
广东省医学科学技术研究基金项目(C2022016)
梅州市科技计划项目(2023C0301001)
作者信息
    广东省生物制品与药物研究所, 广州 510440

通讯作者:

*梁旭霞(1972—), 女, 博士, 主任技师, 主要研究方向为食品药品质量标准。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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