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Article(id=1217779722902229633, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1217779717386715826, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250307003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1741276800000, receivedDateStr=2025-03-07, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1768270911192, onlineDateStr=2026-01-13, pubDate=1750780800000, pubDateStr=2025-06-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1768270911192, onlineIssueDateStr=2026-01-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1768270911192, creator=13701087609, updateTime=1768270911192, 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=140, endPage=144, ext={EN=ArticleExt(id=1217779723262939792, articleId=1217779722902229633, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Determination of octachlorostyrene residues in milk by high performance liquid chromatography, 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 and optimize conditions for the determination of octachlorostyrene residues in milk by high performance liquid chromatography (HPLC) method. Methods Milk samples were extracted with acetonitrile and cleared up by sodium chloride and anhydrous magnesium sulfate, filtered through a microporous membrane, and analyzed by the high performance liquid chromatography. The separation was performed on a Waters C18 column (250 mm×4.6 mm, 5 μm) with gradient elution by using 0.3% trifluoroacetic acid solution (pH=3.2) and acetonitrile (20:80, V:V) as the mobile phase, and quantified by external standard method. Results The results indicated that the correlations were greater than 0.999 at the range of 0.1 to 2.0 ng/mL. The limit of detection of octachlorostyrene was 0.05 μg/kg, and the limit of quantitation was 0.1 μg/kg. The average recoveries of octachlorostyrene ranged from 70% to 90% at spiled levels of 0.1 to 0.4 μg/kg with relative standard deviation less then 10%. Conclusion The experiment show that the method is simple, rapid, high sensitivity, good reproducibility and suitable for detection of octachlorostyrene residues in milk. The method provide effective technical support to the government’s food and agricultural product quality supervision work.

, correspAuthors=Yong-Heng BO, 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=Shu-Huan LI, Yu-Tong ZHANG, Zhao-Xue ZHANG, Cheng-Jun ZHANG, Feng-Hua ZHAO, Xiao-Fei LIU, Yu-Jiao LI, Yong-Heng BO), CN=ArticleExt(id=1217779724282155714, articleId=1217779722902229633, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=高效液相色谱法定量检测牛奶中八氯苯乙烯残留量, 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 建立高效液相色谱法检测牛奶中八氯苯乙烯残留量检测的分析方法。方法 牛奶中的八氯苯乙烯经乙腈提取, 用氯化钠和无水硫酸镁除脂, 提取液离心、过微孔滤膜后上机分析, 采用Waters C18色谱柱(250 mm×4.6 mm, 5 μm)分离, 以0.3%的三氟乙酸溶液(pH=3.2)-乙腈(20:80, V:V)为流动相进行等度洗脱, 紫外检测器检测, 外标法定量。结果 八氯苯乙烯在0.1~2.0 ng/mL的质量浓度范围内线性关系良好, 相关系数r大于0.999。样品中八氯苯乙烯的检出限为0.05 μg/kg, 定量限为0.10 μg /kg。在0.10~0.40 μg/kg添加浓度范围内, 八氯苯乙烯的回收率均为70%~90%, 相对标准偏差均小于10%。结论 该方法具有简便快捷、灵敏度高和定性准确等特点, 适用于牛奶中八氯苯乙烯残留量的检测, 为政府的食品、农产品质量监管工作提供有效的技术支撑。

, correspAuthors=薄永恒, authorNote=null, correspAuthorsNote=
*薄永恒(1980—), 男, 博士, 高级兽医师, 主要研究方向为畜产品质量安全监测与标准开发。E-mail:
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李淑焕(1983—), 女, 博士, 副教授, 主要研究方向为食品质量检测与分析技术研究。E-mail:

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李淑焕(1983—), 女, 博士, 副教授, 主要研究方向为食品质量检测与分析技术研究。E-mail:

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李淑焕(1983—), 女, 博士, 副教授, 主要研究方向为食品质量检测与分析技术研究。E-mail:

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Journal of Food Safety & Quality, 2024, 15(15): 8-19., articleTitle=Determination of immunoglobulin G content in milk by high performance liquid chromatography, refAbstract=null)], funds=[Fund(id=1217833928807862481, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, awardId=BSQJ201808, language=CN, fundingSource=山东农业工程学院博士科研基金项目(BSQJ201808), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1217833920545084083, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, xref=1, ext=[AuthorCompanyExt(id=1217833920553472691, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, companyId=1217833920545084083, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 College of Food Science and Engineering, Shandong Agriculture and Engineering University, Jinan 250100, China), AuthorCompanyExt(id=1217833920557666997, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, companyId=1217833920545084083, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 山东农业工程学院食品科学与工程学院, 济南 250100)]), AuthorCompany(id=1217833920683496127, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, xref=2, ext=[AuthorCompanyExt(id=1217833920691884736, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, companyId=1217833920683496127, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 Shandong Livestock Product Quality and Safety Center, Jinan 250100, China), AuthorCompanyExt(id=1217833920700273346, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, companyId=1217833920683496127, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 山东省畜产品质量安全中心, 济南 250100)])], figs=[ArticleFig(id=1217833927352438864, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=EN, label=Fig.1, caption=Ultraviolet scanning scan image of octachlorostyrene standard solution, figureFileSmall=46uDVcbzpTHT7mU6MRDX+Q==, figureFileBig=Ad3cqrZDi5i7uMVQaEXoFw==, tableContent=null), ArticleFig(id=1217833927461490778, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=CN, label=图1, caption=八氯苯乙烯标准溶液紫外扫描图, figureFileSmall=46uDVcbzpTHT7mU6MRDX+Q==, figureFileBig=Ad3cqrZDi5i7uMVQaEXoFw==, tableContent=null), ArticleFig(id=1217833927595708518, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=EN, label=Fig.2, caption=Liquid chromatogram of blank milk sample, figureFileSmall=sVsBSPoMWmCH8SFIL9WHdA==, figureFileBig=eG1powBfy/n/jKZvMGx6gw==, tableContent=null), ArticleFig(id=1217833927729926257, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=CN, label=图2, caption=空白牛奶液相色谱图, figureFileSmall=sVsBSPoMWmCH8SFIL9WHdA==, figureFileBig=eG1powBfy/n/jKZvMGx6gw==, tableContent=null), ArticleFig(id=1217833927847366779, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=EN, label=Fig.3, caption=Chromatogram of milk sample with added octachlorostyrene (0.2 μg/kg), figureFileSmall=NdhYweTHod0ya7AFWwSonw==, figureFileBig=zAkCXYnBKSvy0ofQGo+IJQ==, tableContent=null), ArticleFig(id=1217833927973195908, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=CN, label=图3, caption=空白牛奶中添加八氯苯乙烯液相色谱图(0.2 μg/kg), figureFileSmall=NdhYweTHod0ya7AFWwSonw==, figureFileBig=zAkCXYnBKSvy0ofQGo+IJQ==, tableContent=null), ArticleFig(id=1217833928124190864, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=EN, label=Fig.4, caption=Chromatogram of octachlorobenzene standard substance (0.2 ng/mL), figureFileSmall=cNtjOTagb4wAbR7w86VCMA==, figureFileBig=R8KOkQ29rxXgFJDJUFmy8g==, tableContent=null), ArticleFig(id=1217833928229048470, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=CN, label=图4, caption=八氯苯乙烯标准品色谱图(0.2 ng/mL), figureFileSmall=cNtjOTagb4wAbR7w86VCMA==, figureFileBig=R8KOkQ29rxXgFJDJUFmy8g==, tableContent=null), ArticleFig(id=1217833928338100387, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=EN, label=Table 1, caption=

Results of reovery and precision of the method (%)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品组织 添加浓度/(µg/kg) 批内平均回收率 批内相对标准偏差 批间相对标准偏差
1 2 3 1 2 3
牛奶 0.1 85.02 74.02 75.15 2.8 3.9 2.9 7.2
0.2 83.09 76.65 84.71 2.7 2.4 4.0 5.3
0.4 76.50 82.10 82.23 2.7 3.0 3.1 4.4
), ArticleFig(id=1217833928501678256, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1217779722902229633, language=CN, label=表1, caption=

回收率和精密度实验结果(%)

, figureFileSmall=null, figureFileBig=null, tableContent=
样品组织 添加浓度/(µg/kg) 批内平均回收率 批内相对标准偏差 批间相对标准偏差
1 2 3 1 2 3
牛奶 0.1 85.02 74.02 75.15 2.8 3.9 2.9 7.2
0.2 83.09 76.65 84.71 2.7 2.4 4.0 5.3
0.4 76.50 82.10 82.23 2.7 3.0 3.1 4.4
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高效液相色谱法定量检测牛奶中八氯苯乙烯残留量
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李淑焕 1 , 张雨桐 1 , 张兆雪 1 , 张呈军 2 , 赵丰华 2 , 刘霄飞 2 , 李玉姣 2 , 薄永恒 2, *
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): 140-144
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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): 140-144
高效液相色谱法定量检测牛奶中八氯苯乙烯残留量
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李淑焕1 , 张雨桐1, 张兆雪1, 张呈军2, 赵丰华2, 刘霄飞2, 李玉姣2, 薄永恒2, *
作者信息
  • 1 山东农业工程学院食品科学与工程学院, 济南 250100
  • 2 山东省畜产品质量安全中心, 济南 250100

    • 李淑焕(1983—), 女, 博士, 副教授, 主要研究方向为食品质量检测与分析技术研究。E-mail:

通讯作者:

*薄永恒(1980—), 男, 博士, 高级兽医师, 主要研究方向为畜产品质量安全监测与标准开发。E-mail:
Determination of octachlorostyrene residues in milk by high performance liquid chromatography
Shu-Huan LI1 , Yu-Tong ZHANG1, Zhao-Xue ZHANG1, Cheng-Jun ZHANG2, Feng-Hua ZHAO2, Xiao-Fei LIU2, Yu-Jiao LI2, Yong-Heng BO2, *
Affiliations
  • 1 College of Food Science and Engineering, Shandong Agriculture and Engineering University, Jinan 250100, China
  • 2 Shandong Livestock Product Quality and Safety Center, Jinan 250100, China
出版时间: 2025-06-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250307003
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摘要
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目的 建立高效液相色谱法检测牛奶中八氯苯乙烯残留量检测的分析方法。方法 牛奶中的八氯苯乙烯经乙腈提取, 用氯化钠和无水硫酸镁除脂, 提取液离心、过微孔滤膜后上机分析, 采用Waters C18色谱柱(250 mm×4.6 mm, 5 μm)分离, 以0.3%的三氟乙酸溶液(pH=3.2)-乙腈(20:80, V:V)为流动相进行等度洗脱, 紫外检测器检测, 外标法定量。结果 八氯苯乙烯在0.1~2.0 ng/mL的质量浓度范围内线性关系良好, 相关系数r大于0.999。样品中八氯苯乙烯的检出限为0.05 μg/kg, 定量限为0.10 μg /kg。在0.10~0.40 μg/kg添加浓度范围内, 八氯苯乙烯的回收率均为70%~90%, 相对标准偏差均小于10%。结论 该方法具有简便快捷、灵敏度高和定性准确等特点, 适用于牛奶中八氯苯乙烯残留量的检测, 为政府的食品、农产品质量监管工作提供有效的技术支撑。

牛奶  /  八氯苯乙烯  /  残留量  /  高效液相色谱法

Objective To establish and optimize conditions for the determination of octachlorostyrene residues in milk by high performance liquid chromatography (HPLC) method. Methods Milk samples were extracted with acetonitrile and cleared up by sodium chloride and anhydrous magnesium sulfate, filtered through a microporous membrane, and analyzed by the high performance liquid chromatography. The separation was performed on a Waters C18 column (250 mm×4.6 mm, 5 μm) with gradient elution by using 0.3% trifluoroacetic acid solution (pH=3.2) and acetonitrile (20:80, V:V) as the mobile phase, and quantified by external standard method. Results The results indicated that the correlations were greater than 0.999 at the range of 0.1 to 2.0 ng/mL. The limit of detection of octachlorostyrene was 0.05 μg/kg, and the limit of quantitation was 0.1 μg/kg. The average recoveries of octachlorostyrene ranged from 70% to 90% at spiled levels of 0.1 to 0.4 μg/kg with relative standard deviation less then 10%. Conclusion The experiment show that the method is simple, rapid, high sensitivity, good reproducibility and suitable for detection of octachlorostyrene residues in milk. The method provide effective technical support to the government’s food and agricultural product quality supervision work.

milk  /  octachlorostyrene  /  residue  /  high performance liquid chromatography
李淑焕, 张雨桐, 张兆雪, 张呈军, 赵丰华, 刘霄飞, 李玉姣, 薄永恒. 高效液相色谱法定量检测牛奶中八氯苯乙烯残留量. 食品安全质量检测学报, 2025 , 16 (12) : 140 -144 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250307003
Shu-Huan LI, Yu-Tong ZHANG, Zhao-Xue ZHANG, Cheng-Jun ZHANG, Feng-Hua ZHAO, Xiao-Fei LIU, Yu-Jiao LI, Yong-Heng BO. Determination of octachlorostyrene residues in milk by high performance liquid chromatography[J]. Journal of Food Safety & Quality, 2025 , 16 (12) : 140 -144 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250307003
正文
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0 引言
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八氯苯乙烯是一种具有独特结构的多氯代芳香化合物, 分子式是C8Cl8, 相对分子质量为379.71, 含氯量74.7%, CAS号为29082-74-4。八氯苯乙烯主要来源于盐溶液中使用石墨阳极的电解过程, 特别是氯碱工业, 其他过程还包括电解熔融氯化镁制镁、含氯溶剂的生产、等离子刻蚀铝(用于生产微电子元件)、铝铸造业中用六氯乙烷的脱气过程、钛金属和铌/钽矿石的氯化以及废弃物的燃烧等[1], 其工业产废水在我国水污染控制中被列为重点解决的有害废水之一[2]。其作为一种非故意生产的工业副产物, 被美国环境保护局指定为12种I级优先控制的持久的生物富集性有毒化学品之一[3], 同时也被列为《世界野生动物基金会环境内分泌干扰物分类表》的一类环境内分泌干扰物[4]。八氯苯乙烯具有四性, 即环境持久存在性、生物富集性、环境长距离迁移性以及影响人体代谢等毒性[1]。一方面八氯苯乙烯具有强亲脂性可在动物脂肪组织内累积, 并沿食物链逐步放大, 使其通过食物链延伸富集至食链终端, 使人类健康受到威胁; 另一方面, 八氯苯乙烯凭借半挥发特征在大气中广泛迁移, 形成的“蚱蜢跳效应”和“全球蒸馏效应”可扩散到地球的极地地区, 可导致其在全球范围内污染传播[5-7]。
长期食用八氯苯乙烯的食物将导致器官病变[1]、细胞凋亡[8], 八氯苯乙烯可与人体内甲状腺激素转运蛋白的结合能力是甲状腺素的1.1倍, 可能会干扰甲状腺激素作用及视黄醇的转运[9]。有研究表明, 对长期暴露在八氯苯乙烯环境下工作的工人进行采血, 检测结果表明其血液中的八氯苯乙烯含量明显超标[10]。除此之外, 在鱼类、鸟类, 甚至在北极熊和南极海鸟的组织中都发现了八氯苯乙烯[11]。近年来各国各行业越来越重视这种持久性污染物对于全球环境和人类健康的危害性, 其造成的全球性环境问题已逐渐受到世界各国关注[12]。
我国东部工业发达, 但随着工业污染的不断加剧, 东部工业产业大量转移至西部、西北部地区[13], 而这些地区是我国奶牛饲养和生长的重要天然牧场[14], 工业中的废气颗粒、废水将会导致草原牧草、饲料和水源的污染, 进而被奶牛采食, 而泌乳是奶牛体内药物代谢的重要途径, 从而造成牛奶中八氯苯乙烯残留, 八氯苯乙烯通过牛奶迁移到人体, 并不断富集, 给人类健康造成潜在的危险。牛奶作为居民一种重要的高蛋白饮品, 特别是作为婴幼儿的重要饮食来源, 其质量安全意义尤为重要[15]。而目前, 国内外鲜见牛奶中八氯苯乙烯残留量检测方法的文献报道。
目前, 八氯苯乙烯检测方法主要有仪器检测法和生物检测法, 仪器检测法主要是涉及气相色谱法、高效液相色谱法、超临界流体色谱法、毛细管电泳法、气相色谱-质谱法、液相相色谱-质谱法等使用大型仪器设备的方法[16-23], 生物检测法是近年来兴起的免疫分析法[24-25]。仪器检测法具有良好的准确度和较高的灵敏度[26-27], 因此本研究以高效液相色谱仪这一常用仪器作为检测基础, 从样品的前处理、色谱方法等方面进行研究并优化条件, 建立了高效液相色谱检测方法用于牛奶中八氯苯乙烯残留量的测定, 为政府的食品、农产品质量监管工作提供有效的技术支撑。
1 材料与方法
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1.1 材料与试剂
本研究所用牛奶均购自于电商平台, 于4 ℃冷藏保存, 待检测。
八氯苯乙烯标准溶液(质量浓度100 μg/mL, 天津阿尔塔科技有限公司); 乙腈(色谱纯, 美国Thermo Fisher Scientific公司); 氨水、三氟乙酸、氯化钠、无水硫酸镁(分析纯, 国药集团化学试剂有限公司)。
1.2 仪器与设备
E2695高效液相色谱仪(配紫外检测器、二级阵列管检测器)、Waters C18色谱柱(250 mm×4.6 mm, 5 μm)、PRiME HLB固相萃取柱、XHLB固相萃取柱(200 mg/6cc)(美国Waters公司); XPE205天平(感量0.01 g)(瑞士梅特勒-托利多仪器有限公司); Syncore台式高速冷冻离心机(美国Thermo Fisher Scientific公司); VX-III多管涡旋振荡器(北京踏锦科技有限公司); PEP-2固相萃取柱(200 mg/6cc)(美国Cleanert公司)。
1.3 实验方法
1.3.1 标准溶液配制
标准储备液: 精密量取八氯苯乙烯标准溶液100 μL, 用乙腈定容至10 mL, 制成1 µg/mL的溶液, 作为标准储备液。
1.3.2 色谱条件
色谱柱: C18柱(250 mm×4.6 mm, 5 μm); 流动相为0.3%的三氟乙酸溶液(临用前氨水调pH=3.2)-乙腈(20:80, V:V); 流速1.0 mL/min; 进样量20 μL; 柱温30 ℃。
1.3.3 样品前处理
精密称取5.00 g牛奶样品, 置于50 mL具塞离心管中, 加乙腈5.0 mL, 涡旋30 s, 放入振荡器中, 振荡提取5 min, 加入0.5 g氯化钠和0.5 g无水硫酸镁, 振荡提取5 min; 8000 r/min离心5 min, 用一次性吸管吸取上层乙腈提取液于另一10 mL具塞离心管中, 用一次性注射器吸取1.0 mL提取液, 过0.22 μm微孔滤膜到进样瓶中, 上机测定。
1.3.4 标准曲线的绘制
分别精密量取标准储备液适量, 用流动相稀释成质量浓度分别为0.1、0.2、0.5、1.0、2.0 ng/mL标准溶液, 经0.22 μm微孔滤膜过滤后, 高效液相色谱仪检测。以八氯苯乙烯峰面积为纵坐标(Y), 相应浓度为横坐标(X, ng/mL), 绘制标准曲线。
1.3.5 灵敏度
将八氯苯乙烯标准溶液加入到空白牛奶中, 分别制成0.05、0.10、0.20、0.40 μg/kg等4个浓度空白添加的待测样品, 按1.3.3方法提取并依1.3.2条件检测, 计算目标峰的信噪比并记录对应浓度, 确定其检出限和定量限。
1.3.6 准确度与精密度
回收率实验采用标准添加法, 在空白的牛奶中添加3个不同浓度的八氯苯乙烯, 每个浓度, 开展5个样品平行实验, 重复3个批次, 求批内、批间的相对标准偏差。
1.4 数据处理
实验结果采用Waters Empower 3数据处理软件对样品中八氯苯乙烯残留量定性定量分析, 各平行实验结果采用Excel 2012进行统计分析。
2 结果与分析
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2.1 样品处理方法的优化
样品提取步骤本着快速便捷的目的进行优化。实验摈弃1 g和2 g牛奶的称样量, 采取5 g牛奶的称样量, 提升了目标物富集量。乙腈提取即可以有效提取样品中八氯苯乙烯, 又可以沉淀牛奶样品中的蛋白以及油脂, 而甲醇沉淀蛋白效果比乙腈差[28-29]。为充分沉淀蛋白与油脂, 实验选择安捷伦QuECHERs盐包(4 g无水硫酸钠, 1 g氯化钠)和现称0.5 g氯化钠、0.5 g无水硫酸镁。实验表明, 两种方法在蛋白沉淀效果、柱子通过性、实验重复性、峰形等差异较小, 因此选择经济性强0.5 g氯化钠、0.5 g无水硫酸镁作为蛋白、油脂的沉淀剂。
经一次性提取, 空白样品添加回收率均在70%~90%之间, 符合残留检验实验的要求, 因此实验未再进行二次提取, 以提升回收率。
为有效去除样品杂质, 选取PRiME HLB、PEP-2、XHLB固相萃取柱和不进行固相萃取柱纯化, 发现乙腈的提取液经固相萃取柱萃取和不经萃取, 对上机样品出峰影响不大, 分析原因是乙腈提取液经0.22 μm微孔滤膜, 可达到净化效果, 为减少提取步骤, 直接取1.0 mL乙腈提取液, 过0.22 μm微孔滤膜后上机即可。
2.2 色谱条件的优化
在确定用乙腈提取八氯苯乙烯后, 选定乙腈与水为流动相主要溶剂。乙腈与水不同比例作为流动相测定发现八氯苯乙烯峰形较差, 因此将流动相水相改为酸性以改善峰形。因磷酸二氢钠与水相混合宜析出, 易堵色谱柱, 摒弃磷酸二氢钠的酸性水相作为流动相[30], 首选0.3%三氟乙酸水相降低pH[31], 然后用氨水调节pH至3~4之间, 经测定, pH约为3.2时, 峰形尖、面积大, 为最优。调节乙腈与0.3%三氟乙酸水溶液比例, 结果表明在乙腈与0.3%三氟乙酸水溶液(pH 3.2)体积比为80:20时, 出峰时间快, 峰形好, 且经样品提取后测定, 该比例流动相对样品杂质峰与目标峰分离较好。因此确定流动相为0.3%的三氟乙酸溶液(临用前氨水调pH=3.2)-乙腈(20:80, V:V)。
用二级阵列管检测器对2.0 ng/mL八氯苯乙烯标准溶液扫描, 扫描范围为190~400 nm, 图1结果显示, 在本研究中的流动相下, 波长205.3 nm时吸收值最大、色谱图峰面积最大, 因此将205 nm作为检测波长。
色谱柱采用了不同品牌的C18柱, 其中参数为250 mm× 4.6 mm, 5μm, 柱子较长, 对多杂质的分离效果较好, 品牌间差异小, 说明本方法适用性强; 设置柱温25~35 ℃时, 对分离影响较小, 利于节能, 条件定为室温; 进样量20 µL, 有利于降低检出限; 流速设置为1.0 mL/min时出峰时间较短, 峰形良好。
2.3 方法验证
2.3.1 方法的线性关系
八氯苯乙烯质量浓度在0.1~2.0 ng/mL范围内线性良好, 线性方程为Y=774625X+6502.6, 相关系数r大于0.999。
2.3.2 方法灵敏度
按1.2.5方法, 空白牛奶中在0.05 μg/kg的添加浓度时, 八氯苯乙烯色谱峰的信噪比(S/N)均>3, 确定为本方法的检出限, 0.10 μg/kg的添加浓度时, 八氯苯乙烯色谱峰的信噪比(S/N)均>10, 确定为本方法的定量限, 满足牛奶中八氯苯乙烯残留检测需求。空白牛奶中添加0.20 μg/kg八氯苯乙烯及相应的空白样品的典型高效液相色谱图, 见图2~3。八氯苯乙烯标准品典型色谱见图4。
2.3.3 方法准确度与精密度
空白牛奶中八氯苯乙烯在0.10 μg/kg(定量限)、0.20 μg/kg (2倍定量限)、0.40 μg/kg (4倍定量限) 3个添加水平进行加标实验, 平均回收率为70%~90%, 批内相对标准偏差≤10%, 批间相对标准偏差≤10%, 能够满足样品残留检测的要求。结果见表1。
2.4 实际样品检测
随机在电商平台购买的10种牛奶作为样品, 利用上述实验方法进行检测, 验证方法的可靠性和适用性。结果显示, 八氯苯乙烯的测定值均低于方法检出限。也说明作为这批牛奶奶源的养殖场未受到八氯苯乙烯污染, 牛奶产品质量安全。
3 结论
收起
本研究根据目前养殖环境污染现状, 聚焦环境污染严重的牧场养殖奶牛的乳产品中环境激素残留问题, 选定八氯苯乙烯为目标残留物, 制定了牛奶中八氯苯乙烯残留量液相色谱仪测定方法。实验创建并优化了提取条件及液相色谱条件, 本方法结果表明0.05 μg/kg为八氯苯乙烯在牛奶中的检出限, 0.10 μg/kg设定为定量限, 在0.1~0.4 μg/kg空白样品添加八氯苯乙烯浓度范围内, 八氯苯乙烯回收率为70%~90%, 符合残留实验室质量控制规范。本方法简单快速, 灵敏度高, 重现性好, 适合牛奶中八氯苯乙烯残留的测定, 为政府部门下一步对牛奶中质量安全监测提供了技术支撑。
基金
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  • 山东农业工程学院博士科研基金项目(BSQJ201808)
文献
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250307003
  • 接收时间：2025-03-07
  • 首发时间：2026-01-13
  • 出版时间：2025-06-25
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  • 收稿日期：2025-03-07
基金
山东农业工程学院博士科研基金项目(BSQJ201808)
作者信息
    1 山东农业工程学院食品科学与工程学院, 济南 250100
    2 山东省畜产品质量安全中心, 济南 250100

通讯作者:

*薄永恒(1980—), 男, 博士, 高级兽医师, 主要研究方向为畜产品质量安全监测与标准开发。E-mail:
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2种不同金属材料的力学参数
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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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