Article(id=1217529314380271777, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217529305693864468, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250219005, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1739894400000, receivedDateStr=2025-02-19, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768211209148, onlineDateStr=2026-01-12, pubDate=1752508800000, pubDateStr=2025-07-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768211209148, onlineIssueDateStr=2026-01-12, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768211209148, creator=13701087609, updateTime=1768211209148, updator=13701087609, issue=Issue{id=1217529305693864468, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='13', 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=1768211207077, creator=13701087609, updateTime=1768212057891, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1217532874337730593, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217529305693864468, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1217532874337730594, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217529305693864468, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=87, endPage=92, ext={EN=ArticleExt(id=1217529315219132687, articleId=1217529314380271777, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Rapid detection of enrofloxacin residues in complex mixed matrix of Hainan’s special “soil-vegetable” system by ultra performance liquid chromatography-tandem mass spectrometry, 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 an analytical method for the determination of enrofloxacin residues in complex mixed matrix of the “soil-vegetable” system by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The sample was subjected to vortex ultrasonic extraction with acidified acetonitrile extract [containing 5% ethylene diamine tetraacetic acid (EDTA)-Mcllvaine buffer solution], settling with centrifugation, getting solvent conversion and being filtered by 0.22 μm nylon filter membrane. The mobile phase was composed of acetonitrile and 0.1% formic acid aqueous solution (containing 2 mmol/L ammonium acetate), and the mixture was subjected to Waters ACQUITY UPLC ® BEH C18 chromatographic column (2.1 mm×100 mm, 1.7 μm) gradient elution separation, using mass spectrometry as a detector for qualitative and quantitative analysis of enrofloxacin. Results In the mass concentration range of 1-200 ng/mL, the standard curve showed a good linear relationship (correlation coefficient=0.9996), the limit of detection of the method was 0.11 μg/kg, the recovery rates ranged from 87.4% to 103.3%, and the relative standard deviations (RSDs) were 0.4% to 4.0% (n=6). Conclusion This method has fast analysis speed, high accuracy and good repeatability, and is suitable for the detection of enrofloxacin residues in complex mixed matrix of the “soil-vegetable” system. It also has the potential to indirectly reveal the risk of antibiotic resistance genes (ARGs) contamination in the system.

, correspAuthors=Bin WANG, 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=Chao-Feng LIU, Xiao-Yu GUO, Hong-Miao CHEN, Rui KANG, Yu-Ling ZHOU, Bin WANG), CN=ArticleExt(id=1217529316007661935, articleId=1217529314380271777, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=超高效液相色谱-串联质谱法快速检测海南特色“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留, 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 建立基于超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定海南特色“土壤-蔬菜”系统中复杂混合基质恩诺沙星残留的分析方法。方法 样品经酸化乙腈提取液[含5%乙二胺四乙酸(ethylene diamine tetraacetic acid, EDTA)-Mcllvaine缓冲溶液]涡旋超声提取, 离心沉降, 溶剂转换, 0.22 μm尼龙滤膜过滤后, 以乙腈和0.1%甲酸水溶液(含2 mmol/L乙酸铵)为流动相, 经Waters ACQUITY UPLC® BEH C18色谱柱(2.1 mm×100 mm, 1.7 μm)梯度洗脱分离, 使用质谱仪作为检测器对恩诺沙星进行定性定量。结果 在1~200 ng/mL的质量浓度范围中, 标准曲线线性关系良好(相关系数为0.9996), 方法的检出限为0.11 μg/kg, 加标回收率范围为87.4%~103.3%, 相对标准偏差(relative standard deviations, RSDs)为0.4%~4.0% (n=6)。结论 该分析方法快速、准确、重复性良好, 适用于“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留的检测, 拥有间接揭示系统中抗生素耐药性基因(antibiotic resistance genes, ARGs)污染风险的潜力。, correspAuthors=王彬, authorNote=null, correspAuthorsNote=
*王彬(1973—), 女, 高级工程师, 主要研究方向为食品安全及食品检验。E-mail:
, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=iKaaKPhOj8B/jMvWkTwuRw==, magXml=bm6BBHoQRA3KIuBa+ur3og==, pdfUrl=null, pdf=dGNGguUUdjErnodh7lOiXA==, pdfFileSize=651187, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=3y4KS9OZX/mamd5YhGYO4w==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=weQVh1u+c2P7hdEBTf1NDA==, mapNumber=null, authorCompany=null, fund=null, authors=

刘超烽(1996—), 男, 硕士, 工程师, 主要研究方向为食品安全及色谱分析。E-mail:

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刘超烽(1996—), 男, 硕士, 工程师, 主要研究方向为食品安全及色谱分析。E-mail:

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刘超烽(1996—), 男, 硕士, 工程师, 主要研究方向为食品安全及色谱分析。E-mail:

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figureFileSmall=gjNnGoOmJ5VVDrP8iB/wwA==, figureFileBig=R6KY1V7GlWUiaEdVBKa2hw==, tableContent=null), ArticleFig(id=1217901255532396963, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217529314380271777, language=EN, label=Table 1, caption=

Program of gradient elution

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时间/min 流动相A/% 流动相B/%
起始 90 10
0.3 90 10
1.0 60 40
2.8 10 90
3.3 10 90
3.4 90 10
4.0 90 10
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梯度洗脱程序

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时间/min 流动相A/% 流动相B/%
起始 90 10
0.3 90 10
1.0 60 40
2.8 10 90
3.3 10 90
3.4 90 10
4.0 90 10
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Parent ion, daughter ion, cone voltage and collision energy of enrofloxacin

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化合物 母离子(m/z) 子离子(m/z) 锥孔电压/V 碰撞能量/eV
恩诺沙星 360 360>245 30 24
360>316* 30 19
恩诺沙星-D5 365 365>321 35 19
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恩诺沙星母离子、子离子、锥孔电压和碰撞能量

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化合物 母离子(m/z) 子离子(m/z) 锥孔电压/V 碰撞能量/eV
恩诺沙星 360 360>245 30 24
360>316* 30 19
恩诺沙星-D5 365 365>321 35 19
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Results of recoveries of enrofloxacin in the “soil-vegetable” system (n=6)

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基质种类 添加浓度
/(μg/kg)
回收率
/%
RSDs
/%
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100 101.9 1.5
500 88.0 1.9
桥头地瓜 10 95.3 3.2
100 100.9 0.4
500 87.4 2.2
土壤 10 100.2 4.0
100 103.3 1.7
500 91.6 4.0
复合基质 10 94.2 1.2
100 100.4 3.6
500 94.3 1.8
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“土壤-蔬菜”系统中恩诺沙星的加标回收结果(n=6)

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基质种类 添加浓度
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回收率
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RSDs
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500 88.0 1.9
桥头地瓜 10 95.3 3.2
100 100.9 0.4
500 87.4 2.2
土壤 10 100.2 4.0
100 103.3 1.7
500 91.6 4.0
复合基质 10 94.2 1.2
100 100.4 3.6
500 94.3 1.8
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超高效液相色谱-串联质谱法快速检测海南特色“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留
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刘超烽 1, 2 , 郭晓宇 1, 2 , 陈虹妙 1, 2 , 康蕊 1, 2 , 周玉玲 2, 3 , 王彬 2, 4, *
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(13): 87-92
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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(13): 87-92
超高效液相色谱-串联质谱法快速检测海南特色“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留
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刘超烽1, 2 , 郭晓宇1, 2, 陈虹妙1, 2, 康蕊1, 2, 周玉玲2, 3, 王彬2, 4, *
作者信息
  • 1 海南省检验检测研究院食品检验检测中心, 海口 570311
  • 2 国家市场监督管理总局重点实验室(热带果蔬质量与安全), 海口 570311
  • 3 海南省检验检测研究院质量技术监督标准与信息所, 海口 570311
  • 4 海南省检验检测研究院食品审评中心, 海口 570311
  • 刘超烽(1996—), 男, 硕士, 工程师, 主要研究方向为食品安全及色谱分析。E-mail:

通讯作者:

*王彬(1973—), 女, 高级工程师, 主要研究方向为食品安全及食品检验。E-mail:
Rapid detection of enrofloxacin residues in complex mixed matrix of Hainan’s special “soil-vegetable” system by ultra performance liquid chromatography-tandem mass spectrometry
Chao-Feng LIU1, 2 , Xiao-Yu GUO1, 2, Hong-Miao CHEN1, 2, Rui KANG1, 2, Yu-Ling ZHOU2, 3, Bin WANG2, 4, *
Affiliations
  • 1 Institute of Food Testing, Hainan Academy of Inspection and Testing, Haikou 570311, China
  • 2 Key Laboratory of Tropical Fruits and Vegetables Quality and Safety, State Administration for Market Regulation, Haikou 570311, China
  • 3 Institute of Quality and Standardization, Hainan Academy of Inspection and Testing, Haikou 570311, China
  • 4 Center for Food Evaluation, Hainan Academy of Inspection and Testing, Haikou 570311, China
出版时间: 2025-07-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250219005
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目的 建立基于超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定海南特色“土壤-蔬菜”系统中复杂混合基质恩诺沙星残留的分析方法。方法 样品经酸化乙腈提取液[含5%乙二胺四乙酸(ethylene diamine tetraacetic acid, EDTA)-Mcllvaine缓冲溶液]涡旋超声提取, 离心沉降, 溶剂转换, 0.22 μm尼龙滤膜过滤后, 以乙腈和0.1%甲酸水溶液(含2 mmol/L乙酸铵)为流动相, 经Waters ACQUITY UPLC® BEH C18色谱柱(2.1 mm×100 mm, 1.7 μm)梯度洗脱分离, 使用质谱仪作为检测器对恩诺沙星进行定性定量。结果 在1~200 ng/mL的质量浓度范围中, 标准曲线线性关系良好(相关系数为0.9996), 方法的检出限为0.11 μg/kg, 加标回收率范围为87.4%~103.3%, 相对标准偏差(relative standard deviations, RSDs)为0.4%~4.0% (n=6)。结论 该分析方法快速、准确、重复性良好, 适用于“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留的检测, 拥有间接揭示系统中抗生素耐药性基因(antibiotic resistance genes, ARGs)污染风险的潜力。
恩诺沙星  /  超高效液相色谱-串联质谱法  /  “土壤-蔬菜”系统  /  复杂混合基质  /  抗生素残留  /  抗生素耐药性基因

Objective To establish an analytical method for the determination of enrofloxacin residues in complex mixed matrix of the “soil-vegetable” system by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The sample was subjected to vortex ultrasonic extraction with acidified acetonitrile extract [containing 5% ethylene diamine tetraacetic acid (EDTA)-Mcllvaine buffer solution], settling with centrifugation, getting solvent conversion and being filtered by 0.22 μm nylon filter membrane. The mobile phase was composed of acetonitrile and 0.1% formic acid aqueous solution (containing 2 mmol/L ammonium acetate), and the mixture was subjected to Waters ACQUITY UPLC ® BEH C18 chromatographic column (2.1 mm×100 mm, 1.7 μm) gradient elution separation, using mass spectrometry as a detector for qualitative and quantitative analysis of enrofloxacin. Results In the mass concentration range of 1-200 ng/mL, the standard curve showed a good linear relationship (correlation coefficient=0.9996), the limit of detection of the method was 0.11 μg/kg, the recovery rates ranged from 87.4% to 103.3%, and the relative standard deviations (RSDs) were 0.4% to 4.0% (n=6). Conclusion This method has fast analysis speed, high accuracy and good repeatability, and is suitable for the detection of enrofloxacin residues in complex mixed matrix of the “soil-vegetable” system. It also has the potential to indirectly reveal the risk of antibiotic resistance genes (ARGs) contamination in the system.

enrofloxacin  /  ultra performance liquid chromatography-tandem mass spectrometry  /  “soil-vegetable” system  /  complex mixed matrix  /  antibiotic residues  /  antibiotic resistance genes
刘超烽, 郭晓宇, 陈虹妙, 康蕊, 周玉玲, 王彬. 超高效液相色谱-串联质谱法快速检测海南特色“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留. 食品安全质量检测学报, 2025 , 16 (13) : 87 -92 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250219005
Chao-Feng LIU, Xiao-Yu GUO, Hong-Miao CHEN, Rui KANG, Yu-Ling ZHOU, Bin WANG. Rapid detection of enrofloxacin residues in complex mixed matrix of Hainan’s special “soil-vegetable” system by ultra performance liquid chromatography-tandem mass spectrometry[J]. Journal of Food Safety & Quality, 2025 , 16 (13) : 87 -92 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250219005
自1928年青霉素问世以来, 抗生素相关的研究一直在蓬勃发展。但随着各类抗生素的广泛使用乃至滥用, 驱动了抗生素耐药性基因(antibiotic resistance genes, ARGs)在环境中广泛分布, 并成为了世界六大新兴环境问题之一[1]。在“土壤-蔬菜”系统中, 并无直接的抗生素污染引入风险, 但由于养殖业中抗生素的大规模应用, 抗生素污染已通过动物粪肥、污染废水等方式外溢, 间接进入了“土壤-蔬菜”系统中, 进而通过选择压力促进了ARGs广泛分布, 两者间存在强相关性, 现已有多篇相关报导[2-7]
相较于对食用动物的严格抗生素风险监控, 当前对于蔬菜中抗生素风险的监控, 无论是相关的检验标准, 还是有关部门的监管, 均存在一定的空白。与此同时, 相较于肉类的熟食习惯, 蔬菜有更为普遍的生食习惯, 更容易使得抗生素残留以及ARGs通过该方式进入人体, 进而产生相应的健康风险[8-9], 因此, 亟需建立相关方案对此类风险进行监控。
蔬菜属于“土壤-蔬菜”系统的一部分, 该系统不同组分以及各组分的不同部位, 对于抗生素及ARGs的富集程度存在一定差异, 此外抗生素残留及ARGs还可以通过水体、土壤和空气等环境介质传播[2,10-12]。因此, 单独检测系统中某一组分, 如蔬菜的可食用部位, 并不能很好地展示当前系统中抗生素及ARGs的污染程度及潜在风险, 所以有必要建立一种将“土壤-蔬菜”系统作为整体进行抗生素类风险监测的方案。
在当前研究中, 针对“土壤-蔬菜”系统中各个组分(蔬菜、土壤)中的抗生素及ARGs, 均有相应的测定方案。其中, 抗生素的检验方法更为成熟可靠, 包括微生物法、免疫法、理化法等, 其中理化法包括高效液相色谱法、液相色谱-质谱法、传感器法等[13-16]; 而ARGs的检测方法较为局限, 聚合酶链式反应(polymerase chain reaction, PCR)技术、基因芯片等方法可以进行检测[17-21], 但是此类方法样品前处理、实验步骤复杂、仪器普及率差, 难以实现大范围、高效率的检验, 实现风险监测目的。而且由于基因突变的随机性, ARGs的检测法均具有一定的滞后性[22], 不适合作为抗生素类风险的前瞻性监控指标。因此, “土壤-蔬菜”系统中的抗生素残留是较好的抗生素类风险监控指标。
针对此问题, 本研究选择了海南特色的桥头地瓜作为“土壤-蔬菜”系统作为代表。桥头地瓜作为海南特色经济作物, 其中地上茎叶部分可作为蔬菜食用, 地下根茎部分同样属于可食用部分, 相较于常见蔬菜仅有地上部分或地下部分为可食部分, 其基质更为丰富, 更具代表性。在抗生素残留类别上, 喹诺酮类药物在人类医学史上极具重要性, 被世界卫生组织列为“最重要的抗菌药物”[23], 恩诺沙星是喹诺酮类抗生素的典型代表, 结构见图1, 其在养殖业得到广泛的应用, 是国家指定的动物专用药之一, 在近年的《食品安全监督抽检实施细则》中多次被列为检验项目, 在畜禽肉、蔬菜、土壤中常被检出[24], 是具有代表性的大量使用抗生素之一, 以其建立方法, 更具有实际意义, 因此, 本研究选择恩诺沙星作为研究对象。
基于超高效液相色谱-串联质谱法的高准确性、高选择性及高效性[12-14,16,25-28], 本研究以其为基础, 建立了一种能够快速检测海南特色的桥头地瓜“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留的分析方法。本研究突破了常规研究以单一基质作为研究对象的局限性, 为动态监测“土壤-蔬菜”系统抗生素风险提供了新的监控策略, 由于抗生素污染与ARGs的强关联性, 本研究为快速、前瞻性地揭示系统中ARGs污染程度提供了潜在的风险评估新范式, 为相关监管领域提供了助力, 为保障海南自贸港生态安全, 发展高效农业提供了技术支撑。
桥头地瓜、地瓜叶、土壤均为海南本地田间直接获得。
Waters ACQUITY UPLC I-Class Plus/Xevo TQ-XS超高效液相-三重四极杆质谱联用仪、Waters ACQUITY UPLC® BEH C18色谱柱(2.1 mm×100 mm, 1.7 μm)(美国Waters公司); MS3002TS电子天平(精度0.01 mg)、FE20K酸度计(瑞士梅特勒托利多公司); SK7200超声波清洗机(上海科导超声仪器有限公司); IKA涡旋混合器(德国IKA集团); Milli-Q超纯水机(美国Millipore公司); 5804R冷冻离心机(德国艾本德公司); 0.22 μm 滤膜(上海安谱实验科技股份有限公司); AutoEVA-60 氮吹仪(厦门睿科集团股份有限公司)。
乙腈、甲酸(质谱纯)、乙酸铵、乙腈(色谱纯)(德国默克公司); 恩诺沙星(质量浓度1000 μg/mL)、恩诺沙星-D3(质量浓度1000 μg/mL)、恩诺沙星(纯度≥98%)(德国Dr.Ehrenstorfer GmbH公司); 乙二胺四乙酸二钠、磷酸氢二钠、柠檬酸、盐酸、氢氧化钠、乙腈、甲酸(分析纯, 广州化学试剂厂); 实验用水为Milli-Q超纯水机制备的超纯水。
(1)样品制备、称量、提取
桥头地瓜、地瓜叶分别进行匀浆, 获得相应待检测样; 土壤样品直接研磨后过筛获得待检测样; 将部分制备好的地瓜、地瓜叶、土壤待检测样按照5:5:90的比例混合均匀, 获得“土壤-蔬菜”系统复合基质的待检测样。
准确称取1 g均匀样品于50 mL塑料离心管中, 加入恩诺沙星内标溶液, 加入10 mL提取液[乙二胺四乙酸(ethylene diamine tetraacetic acid, EDTA)-Mcllvaine缓冲溶液:酸化乙腈=0.5:9.5, V:V] (EDTA-Mcllvaine缓冲溶液: 称取71.63 g磷酸氢二钠配制成1000 mL水溶液; 称取21.01 g柠檬酸配制成1000 mL水溶液; 将1000 mL柠檬酸溶液与625 mL磷酸氢二钠溶液混合, 调pH至4.00±0.05; 称取60.5 g乙二胺四乙酸二钠放入上述混合溶液, 配制成EDTA-Mcllvaine缓冲溶液), 涡旋5 min混匀后超声提取15 min, 将样品混合物以10000 r/min离心5 min, 取上层清液1 mL备用。
(2)样品溶剂转换、净化、上机
将上层清液经氮气吹干(40 ℃), 以1 mL甲醇复溶, 若存在溶剂效应, 可使用初始流动相进行复溶, 可根据实际浓度调整复溶体积, 经0.22 μm尼龙微孔滤膜过滤净化, 获得待测定样液, 使用超高效液相色谱-串联质谱仪器上机进行测定。
在样品需要快速获得结果的情况下, 可将上层清液经0.22 μm尼龙微孔滤膜净化后直接上机进行测定, 实验完成后需及时清洗系统。
(1)液相色谱条件
色谱柱型号及规格: Waters ACQUITY UPLC® BEH C18色谱柱(2.1 mm×100 mm, 1.7 μm); 流动相组成: A为0.1%甲酸溶液(含2 mmol/L乙酸铵溶液), B为乙腈; 流速: 0.300 mL/min; 进样体积为1 μL; 梯度洗脱程序见表1
(2)质谱条件
电离方式: 电喷雾离子源(electron spray ionization, ESI); 扫描方式: 正离子扫描; 检测方式: 多反应离子监测(multiple reaction monitoring, MRM); 毛细管电压: 3.5 kV; 离子源温度: 500 ℃; 脱溶剂气温度: 550 ℃; 脱溶剂气(N2)流量: 800 L/Hr; 碰撞气: 氩气。
监测离子参数情况见表2
本研究采用了MassLynx V4.2.lnk软件进行超高效液相色谱-串联质谱仪原始数据处理, 进而得到上机浓度、标准工作曲线、曲线相对标准偏差等数据, 之后的数据分析使用了WPS 2025软件, 获得回收率、相对标准偏差等结果。
本研究以桥头地瓜“蔬菜-土壤”系统整体作为研究对象, 因而样品基质需要覆盖系统各个组分, 包括: 地上部分植株(地瓜叶)、地下部分根茎(地瓜)、土壤(含土壤本体及附属杂质)。与传统研究不同(土壤基质通常需经过采样、除杂、烘干、研磨、过筛等处理步骤, 以去除植物根系、水分等成分), 本研究为了保障基质的整体代表性, 直接采用未干燥处理的原始土壤样品。本研究首先对3种基质单独进行初步加标回收测试, 结果显示回收率均为良好(相对回收率均大于87%)。为了设计具备整体代表性的基质选择配比, 本研究首先考虑到基质之间可能存在相互作用[29-30], 先设计了均等基质组分(地瓜:地瓜叶:土壤=1:1:1, m:m:m, 下同); 其次, 地瓜叶与地瓜均为有机质为主的植物成分, 而土壤主要以无机质为主, 考虑到有机质与无机质之间存在巨大物理化学性质差异, 本研究再设计了有机质-无机质均等组分(地瓜:地瓜叶:土壤=1:1:2); 最后, 在实际的田间地头, 桥头地瓜“蔬菜-土壤”系统整体中各组分质量比悬殊, 以地瓜根茎深入土壤30~40 cm估算, 系统中土壤质量占比超99%, 但考虑到地瓜叶、地瓜具有丰富的表面积, 是系统中物质交换过程中关键要素, 直接将待检测样按照质量比混合可能会使得样品不具备整体代表性, 因此, 本研究设计了贴近实际情况的高土壤比例组分(地瓜:地瓜叶:土壤=5:5:90)。对3种质量配比的复合样品基质初步加标回收测试结果显示, 各配比回收率均为良好(相对回收率均大于90%), 初步证明该系统各基质相互作用对本研究的检测方法无明显影响, 因此, 为了更贴近实际桥头地瓜“蔬菜-土壤”系统的组份比例, 本研究最终选择了高土壤含量复合基质样品进行进一步分析。
在常规提取溶剂中, 恩诺沙星微溶于甲醇, 易溶于乙腈, 在酸性条件下溶解度较高。因而, 本研究采用酸化乙腈(含1%甲酸)作为基础提取溶剂。在考察了酸化乙腈对“土壤-蔬菜”系统各组分中恩诺沙星的提取效果后, 因土壤中含有的金属元素(如Ca²⁺、Mg²⁺、Fe³⁺等)与恩诺沙星络合, 对于含土壤组分的样品, 酸化乙腈并不能完成提取, 因此加入了EDTA-Mcllvaine缓冲溶液, 用于结合土壤中的金属成分, 各比例提取效果见图2。因存在内标校正, 加入EDTA-Mcllvaine缓冲溶液后, 回收率均达到了较高水平, 而响应强度随着缓冲液的加入逐渐升高, 由于缓冲溶液为水相, 高水相提取溶液在氮吹时耗时过长, 综合考虑实验效率、响应强度以及质谱对不挥发性盐容纳阈值较低等因素, 本研究采用了含5% EDTA-Mcllvaine缓冲溶液的酸化乙腈作为提取溶剂, 因其在较少量的引入不挥发性盐的同时, 获得了较高的相应强度和优秀的回收率并保证了较高的实验效率。
尼龙材质、聚醚砜材质(polyethersulfone, PES)和聚四氟乙烯(polytetrafluoroethylene, PTFE)的滤膜是当前实验室常用的3种滤膜。本研究使用10 ng/mL恩诺沙星标准工作液进行过膜测试(n=6), 结果显示聚醚砜对恩诺沙星具有强吸附作用(相对回收率为39.75%), 尼龙滤膜与PTFE滤膜的恩诺沙星通过率均接近100%, 但PTFE滤膜的效果(相对回收率为99.42%)略差于尼龙滤膜, 因此, 本研究采取尼龙滤膜。
对恩诺沙星标准工作溶液在1~200 ng/mL范围内进行测定, 得到标准曲线方程为: Y=5.23444X+1.1242 (r2=0.9996), 线性关系良好。检出限一般定义为信噪比(S/N)等于3:1时的浓度, 本研究对恩诺沙星质量浓度为1 ng/mL的空白基质加标点重复进样6次, 测得信噪比为273, 由此可得, 检出限为0.11 μg/kg, 能够满足实际检验需求。
分别于地瓜叶、桥头地瓜、土壤、复合基质的空白基质中加入恩诺沙星标准溶液, 制备得到质量浓度为10、100、500 μg/kg 3个浓度梯度的样品, 样品分别同时检测6次, 各基质综合回收率为87.4%~103.3%, 相对标准偏差(relative standard deviations, RSDs)为0.4%~4.0%, 本方法回收率、精密度良好, 能够满足检测要求, 具体数据见表3
在海南澄迈本地多个桥头地瓜“土壤-蔬菜”体系中进行样品收集并检测, 结果均未检出恩诺沙星。恩诺沙星属于喹诺酮类抗生素, 其在环境中检出通常是养殖业抗生素滥用外溢的结果。据《2024年度桥头镇人民政府工作报告》得知, 桥头镇当前农业和养殖业主要推进方向为桥头地瓜种植及深海养殖产业, 两者在空间上隔绝, 因而降低了养殖业抗生素污染外溢的风险。但根据T/QTDGXH 02—2022《桥头地瓜技术规程》, 桥头地瓜种植时需要施加大量的有机肥(如牛粪、羊粪等), 养殖业抗生素残留存在通过有机肥传导至桥头地瓜“土壤-蔬菜”体系中的潜在风险, 除此, 抗生素残留可通过水系传导, 进入到桥头地瓜灌溉用水中, 进而进入系统中, 因此, 该系统仍存在抗生素暴露风险, 需保持动态风险监测。
本研究建立了对“土壤-蔬菜”体系中整体复杂混合基质和各单独基质中恩诺沙星残留的超高效液相色谱-串联质谱法快速检验方法。该方法仅需25 min即可完成除溶剂转换、过膜外的前处理步骤, 在紧急需求时, 可在30 min内完成前处理步骤, 一次上机检测只需4 min, 方法回收率为87.4%~103.3%, RSDs为0.4%~4.0%, 检出限为0.11 μg/kg。本研究方法检验速度快、检验结果准确、检出限灵敏, 适用于海南特色的桥头地瓜“土壤-蔬菜”系统复杂混合基质中恩诺沙星残留的批量快速测定, 可以直接反应系统整体抗生素污染程度, 并具有揭示区域ARGs污染的潜力, 能够为前瞻性监管提供技术支撑, 具有较高的实际应用价值, 后续重点研究方向为扩展抗生素种类、“土壤-蔬菜”体系类别、完善区域抗生素整体污染程度与耐药性基因问题间的关联分析, 进一步助力监管。
  • 国家市场监督管理总局重点实验室(热带果蔬质量与安全)自主研究课题项目(ZZ-2024007)
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2025年第16卷第13期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250219005
  • 接收时间:2025-02-19
  • 首发时间:2026-01-12
  • 出版时间:2025-07-15
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  • 收稿日期:2025-02-19
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国家市场监督管理总局重点实验室(热带果蔬质量与安全)自主研究课题项目(ZZ-2024007)
作者信息
    1 海南省检验检测研究院食品检验检测中心, 海口 570311
    2 国家市场监督管理总局重点实验室(热带果蔬质量与安全), 海口 570311
    3 海南省检验检测研究院质量技术监督标准与信息所, 海口 570311
    4 海南省检验检测研究院食品审评中心, 海口 570311

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*王彬(1973—), 女, 高级工程师, 主要研究方向为食品安全及食品检验。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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