高效液相色谱-三重四极杆质谱法测定猪饲料、养殖用水中五氯酚的残留

doi:10.16431/j.cnki.1671-7236.2024.04.012

摘要/Abstract

摘要： 【目的】本试验旨在建立一种测定猪饲料、饲料添加剂、水等养殖环节投入品中五氯酚残留的高效液相色谱-三重四极杆质谱方法。【方法】待测样品经碱性甲醇水溶液提取,使用阴离子固相萃取柱富集净化,经氮吹浓缩后用2%甲酸甲醇复溶,以5 mmol/L乙酸铵溶液-甲醇为流动相,经Poroshell 120 EC-C18(3 mm×100 mm,2.7 μm)色谱柱分离;负离子模式下,采用多反应监测模式(multiple reaction monitoring,MRM)进行检测;使用基质匹配外标法测定五氯酚的含量,并对所建方法的线性关系、检测限(limit of detection,LOD)、定量限(limit of quantitation,LOQ)、回收率、准确度、精密度、基质效应等指标进行考察。【结果】猪饲料和饲料添加剂中五氯酚的检测限和定量限分别为0.1和0.5 μg/kg,养殖用水中五氯酚的检测限和定量限分别为5和12.5 ng/L;饲料和饲料添加剂中五氯酚定量范围为0.5~20 μg/kg,养殖用水中五氯酚定量范围为12.5~500 ng/L,方法线性关系良好,相关系数(R²)均>0.999。该方法在饲料和饲料添加剂以及养殖用水中的平均加标回收率为69.8%~119.5%,相对标准偏差(relative standard deviation,RSD)为1.62%~9.86%,基质效应(matrix effect,ME)为0.62~1.29。【结论】本试验所建方法简便、快捷、灵敏度高、重复性好,适用于猪配合饲料等养殖投入品中五氯酚的测定,可为养殖环节投入品中五氯酚污染筛查提供准确高效的技术手段,从源头保障动物源性产品安全。

关键词: 五氯酚; 饲料; 固相萃取; 高效液相色谱-三重四极杆质谱

Abstract: 【Objective】 A high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS) method was developed for the determination of pentachlorophenol (PCP) in swine feed,feed additives and water.【Method】 The samples were extracted with aqueous alkaline methanolic solution,enriched and cleaned up using an anionic solid phase extraction column,reconstituted with 2% formate methanol after nitrogen drying.With 5 mmol/L ammonium acetate solution-methanol as mobile phase,Poroshell 120 EC-C18 (3 mm×100 mm,2.7 μm) column was used to separate PCP.Under the negative ionization,multiple reaction monitoring mode (MRM) was used to determine the content of PCP.Matrix-matched external quantification method was utilized to quantitate PCP.Linear relationship,limits of detection (LOD),limits of quantification (LOQ),recovery,accuracy,precision,matrix effect and other indicators of the established method were investigated.【Result】 The LOD and LOQ were 0.1 and 0.5 μg/kg in livestock feed,feed ingredients and feed additives,while 5 and 12.5 ng/L in farm water,respectively.The linear range of PCP in livestock feed,feed ingredients and feed additives was from 0.5 to 20 μg/kg,and the linear range of farm water was from 12.5 to 500 ng/L.The linear coefficient (R²) was more than 0.999,showing the good linearity.In addition,the recoveries ranged from 69.8% to 119.5% and the relative standard deviation (RSD) were from 1.62% to 9.86%.The matrix effect (ME) was 0.62-1.29.【Conclusion】 The method established in this study with high sensitivity,high accuracy and good robustness was suitable for the screening of PCP in swine formula feed and other farming inputs to ensure the quality and safety of meat products.

Key words: pentachlorophenol; feed; solid phase extraction; HPLC-MS/MS

中图分类号:

S816

朱季升, 邢维海, 李政, 王力, 张亚伟, 张元庆, 杨丹, 杨悠悠. 高效液相色谱-三重四极杆质谱法测定猪饲料、养殖用水中五氯酚的残留[J]. 中国畜牧兽医, 2024, 51(4): 1438-1447.

ZHU Jisheng, XING Weihai, LI Zheng, WANG Li, ZHANG Yawei, ZHANG Yuanqing, YANG Dan, YANG Youyou. Determination of Pentachlorophenol Residues in Swine Formula Feed and Farm Water Using High Performance Liquid Chromatography-triple Quadrupole Mass Spectrometry[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1438-1447.

参考文献

[1] 潘胜东,陈晓红,何仟,等.超高效液相色谱-高分辨质谱确证分析泥鳅体内五氯酚代谢物[J].色谱,2017,35(12):1245-1250. PAN S D,CHEN X H,HE Q,et al.Confirmation and analysis of the metabolites of pentachlorophenol in loaches using ultra-performance liquid chromatography-high resolution mass spectrometry[J].Chinese Journal of Chromatography,2017,35(12):1245-1250.(in Chinese)
[2] 黄轶昕.我国灭螺药物研究和现场应用[J].中国血吸虫病防治杂志,2019,31(6):679-684. HUANG Y X.Research and field application of snail killing drugs in China[J].Chinese Journal of Schistosomiasis Control,2019,31(6):679-684.(in Chinese)
[3] XU Q,AI S,GE G,et al.Human health ambient water quality criteria and risk assessment of pentachlorophenol in Poyang lake basin,China[J].Environmental Geochemistry and Health,2022,45(6):3669-3682.
[4] 黄高峰,徐挺.酚类污染物对氧化磷酸化毒性效应的研究进展[J].安徽农业科学,2018,46(8):31-35. HUANG G F,XU T.Research progress on toxic effects of phenolic pollutants on oxidative phosphorylation[J].Journal of Anhui Agricultural Sciences,2018,46(8):31-35.(in Chinese)
[5] 穆应花,邢家溧,沈坚,等.QuEChERS-气相色谱-质谱法检测鱼肉中19种氯酚类化合物[J].色谱,2022,40(5):477-487. MU Y H,XING J L,SHEN J,et al.Detection of 19 chlorophenolic compounds in fish by QuEChERS-gas chromatography-mass spectrometry[J].Chinese Journal of Chromatography,2022,40(5):477-487.(in Chinese)
[6] 刘印平,云鹏,路杨,等.河北省淡水鱼中五氯酚钠的残留调查[J].食品安全质量检测学报,2020,11(20):7642-7646. LIU Y P,YUN P,LU Y,et al.Residue investigation of sodium pentachlorophenolate in freshwater fish in Hebei province[J].Journal of Food Safety and Quality,2020,11(20):7642-7646.(in Chinese)
[7] 孙金影,安丽红,杨敏,等.QuEChERS-高效液相色谱串联质谱法快速筛查分析食品中五氯酚酸钠残留[J].食品安全质量检测学报,2020,11(14):4757-4762. SUN J Y,AN L H,YANG M,et al.Rapid screening and analysis of sodium pentachlorophenolate residues in food by QuEChERS-high performance liquid chromatography tandem mass spectrometry[J].Journal of Food Safety and Quality,2020,11(14):4757-4762.(in Chinese)
[8] ABOU-ELWAFA ABDALLAH M,ZAKY A H,COVACI A.Levels and profiles of organohalogenated contaminants in human blood from Egypt[J].Chemosphere,2017,176:266-272.
[9] 郭剑秋,王铮,张济明,等.江苏省某县学龄期儿童五氯酚暴露水平及影响因素[J].环境与职业医学,2019,36(9):797-802. GUO J Q,WANG Z,ZHANG J M,et al.Pentachlorophenol exposure level and influencing factors in school-age children in a county in Jiangsu province[J].Journal of Environmental and Occupational Medicine,2019,36(9):797-802.(in Chinese)
[10] SCHMIED-TOBIES M I H,MURAWSKI A,SCHMIDT L,et al.Pentachlorophenol and nine other chlorophenols in urine of children and adolescents in Germany—Human biomonitoring results of the German environmental survey 2014-2017(GerES V)[J]. Environmental Research,2021,196:110958.
[11] 尚颐斌,杨建磊,王伟锋.食品链中五氯酚酸钠风险梳理及对策[J].食品安全导刊,2021,23:55-56. SHANG Y B,YANG J L,WANG W F.Risk sorting and countermeasures of sodium pentachlorophenolate in food chain[J].China Food Safety Magazine,2021,23:55-56.(in Chinese)
[12] 胡浩光,刘少彬,郭楚仪,等.气相色谱法测定水产品中五氯苯酚及其钠盐残留量[J].广州化工,2021,49(4):72-74. HU H G,LIU S B,GUO C Y,et al.Determination of pentachlorophenol and its sodium salt residue in aquatic products by gas chromatography[J].Guangzhou Chemical Industry,2021,49(4):72-74.(in Chinese)
[13] 刘坤,王淑婷,李伟红,等.我国畜肉产品主要药物风险因素分析及控制[J].农产品质量与安全,2022,4:85-90. LIU K,WANG S T,LI W H,et al.Analysis and control of major drug risk factors of livestock and meat products in China[J].Quality and Safety of Agro-products,2022,4:85-90.(in Chinese)
[14] 王玉东,宋翠平,戴廷灿,等.我国近年来禽肉产品主要药物风险因素分析及控制[J].中国动物检疫,2022,39(1):42-47. WANG Y D,SONG C P,DAI T C,et al.Analysis and control of major drug risk factors of poultry meat products in China in recent years[J].China Animal Health Inspection,2022,39(1):42-47.(in Chinese)
[15] 张秀芹,王玉晶,李胤楠,等.动物源产品中五氯酚来源探讨及危害分析[J].食品安全质量检测学报,2021,12(19):7840-7845. ZHANG X Q,WANG Y J,LI Y N,et al.Sources of pentachlorophenol in animal-derived products and hazard analysis[J].Journal of Food Safety and Quality,2021,12(19):7840-7845.(in Chinese)
[16] 田志梅,马现永,崔艺燕,等.饲料中五氯酚钠溯源监测分析[J].广东畜牧兽医科技,2018,43(5):30-33. TIAN Z M,MA X Y,CUI Y Y,et al.Traceability monitoring and analysis of sodium pentachlorophenolate in feed[J]. Guangdong Journal of Animal and Veterinary Science,2018,43(5):30-33.(in Chinese)
[17] 万译文,黄向荣,伍远安,等.高效液相色谱串联质谱法测定渔业水体及渔业饲料中的五氯苯酚[J].中国饲料,2019,5:78-81. WAN Y W,HUANG X R,WU Y A,et al.Determination of pentachlorophenol in fishery water and fishery feed by high performance liquid chromatography tandem mass spectrometry[J].China Feed,2019,5:78-81.(in Chinese)
[18] 何正和,魏云计,朱臻怡,等.气相色谱-串联质谱法测定饲料中五氯酚及其钠盐的研究[J].饲料研究,2021,44(14):120-122. HE Z H,WEI Y J,ZHU Z Y,et al.Determination of pentachlorophenol and sodium salt in feed by gas chromatography-tandem mass spectrometry[J].Feed Research,2021,44(14):120-122.(in Chinese)
[19] 余明霞,曹秀,冯慧慧,等.液液萃取-QuEChERS净化-高效液相色谱-串联质谱法测定恩诺沙星可溶性粉中五氯酚酸钠含量[J].现代农业科技,2022(24):156-159. YU M X,CAO X,FENG H H,et al.Determination of sodium pentachlorophenolate in enrofloxacin soluble powder by liquid-liquid extraction-QuEChERS purification-high performance liquid chromatography-tandem mass spectrometry[J].Modern Agricultural Sciences and Technology,2022(24):156-159.(in Chinese)
[20] ZHAO D.Determination of pentachlorophenol residue in meat and fish by gas chromatography-electron capture detection and gas chromatography-mass spectrometry with accelerated solvent extraction[J].Journal of Chromatographic Science,2014,52(5):429-435.
[21] 潘明迪.液相色谱-三重四极杆串联质谱结合QuEChERS测定牛百叶中五氯酚酸钠的含量[J].食品安全导刊,2023,12:45-47. PAN M D.Determination of sodium pentachlorophenolate in bovine louvers by liquid chromatography-triple quadrupole tandem mass spectrometry combined with QuEChERS[J].China Food Safety Magazine,2023,12:45-47.(in Chinese)
[22] 陆阳,朱鹏飞,姚誉阳,等.固相萃取-高效液相色谱法测定纸质食品包装材料中五氯酚含量[J].食品安全质量检测学报,2022,13(1):65-71. LU Y,ZHU P F,YAO Y Y,et al.Determination of pentachlorophenol content in paper food packaging materials by solid phase extraction-high performance liquid chromatography[J].Journal of Food Safety and Quality,2022,13(1):65-71.(in Chinese)
[23] 杨霄,李小玲,刘伶俐,等.大体积固相萃取/高效液相色谱-串联质谱法测定地表水中的五氯酚[J].湖南师范大学自然科学学报,2021,44(4):119-125. YANG X,LI X L,LIU L L,et al.Determination of pentachlorophenol in surface water by large-volume solid phase extraction,high performance liquid chromatography-tandem mass spectrometry[J].Journal of Natural Sciences of Hunan Normal University,2021,44(4):119-125.(in Chinese)
[24] 张文韬,罗晓飞.顶空固相微萃取-气相色谱法同时测定饮用水中2,4-二氯酚、2,4,6三氯酚、五氯酚[J].预防医学情报杂志,2021,37(8):1159-1163. ZHANG W T,LUO X F.Simultaneous determination of 2,4-dichlorophenol,2,4,6-trichlorophenol and pentachlorophenol in drinking water by headspace solid phase microextraction-gas chromatography[J].Journal of Preventive Medicine Information,2021,37(8):1159-1163.(in Chinese)
[25] 胡博,刘韫祺.顶空固相微萃取气相色谱法测定水产品中五氯酚残留量[J].食品工业,2015,36(1):271-274. HU B,LIU Y Q.Determination of pentachlorophenol residues in aquatic products by headspace solid phase microextraction by gas chromatography[J].The Food Industry,2015,36(1):271-274.(in Chinese)
[26] ANNA O,DAVID P,JORDI G,et al.Determination of pentachlorophenol in certified waste waters,soil samples and industrial effluents using ELISA and liquid solid extraction followed by liquid chromatography[J].Analytica Chimica Acta,1997,346(1):49-59.
[27] 龚蕾,周陶鸿,彭青枝,等.免疫层析法快速检测动物源性食品中五氯酚酸钠含量[J].食品安全质量检测学报,2022,13(3):888-893. GONG L,ZHOU T H,PENG Q Z,et al.Rapid detection of sodium pentachlorophenolate in foods of animal origin by immunochromatography[J].Journal of Food Safety and Quality,2022,13(3):888-893.(in Chinese)
[28] 马成孝.衍生气相色谱法测定水中2,4,6-三氯酚、五氯酚[J].中国资源综合利用,2022,40(12):14-16. MA C X.Determination of 2,4,6-trichlorophenol and pentachlorophenol in water by derivatization gas chromatography[J].China Resources Comprehensive Utilization,2022,40(12):14-16.(in Chinese)
[29] 唐吉旺,何浩,王淑霞,等.固相萃取-高效液相色谱-串联质谱法同时测定动物源性食品中五氯酚和游离棉酚[J].食品科技,2020,45(7):364-371. TANG J W,HE H,WANG S X,et al.Simultaneous determination of pentachlorophenol and free gossypol in animal-derived foods by solid phase extraction-high performance liquid chromatography-tandem mass spectrometry[J].Food Science and Technology,2020,45(7):364-371.(in Chinese)
[30] IDA K,NATASA N,MAJA L Š,et al.Non-destructive detection of pentachlorophenol residues in historical wooden objects[J].Polymers,2021,13(7):1052.
[31] 徐玲珍,胡明友,姚建花.QuEChERS-混合型吸附剂分散固相萃取-气相色谱-三重四级杆串联质谱法测定鸡肉和猪肉中的五氯酚[J].实用预防医学,2022,29(11):1351-1353. XU L Z,HU M Y,YAO J H.Determination of pentachlorophenol in chicken and pork by QuEChERS-mixed adsorbent dispersed solid phase extraction-gas chromatography-triple quadrupole tandem mass spectrometry[J].Practical Preventive Medicine,2022,29(11):1351-1353.(in Chinese)
[32] 夏宝林,严秋钫,杨娜,等.增强型除脂固相萃取技术结合超高效液相色谱-串联质谱法测定动物源性食品中五氯酚残留量[J].食品安全质量检测学报,2019,10(14):4698-4705. XIA B L,YAN Q F,YANG N,et al.Determination of pentachlorophenol residues in animal-derived foods by enhanced degreasing solid-phase extraction technology combined with ultra-performance liquid chromatography-tandem mass spectrometry[J].Journal of Food Safety and Quality,2019,10(14):4698-4705.(in Chinese)
[33] 何平,郑铖,张友萍.全自动样品制备系统-超高效液相色谱-串联质谱法测定动物源食品中五氯酚钠[J].农产品质量与安全,2023,2:69-73. HE P,ZHENG C,ZHANG Y P.Determination of sodium pentachlorophenolate in animal-derived foods by ultra-performance liquid chromatography-tandem mass spectrometry by automatic sample preparation system[J].Quality and Safety of Agro-products,2023,2:69-73.(in Chinese)
[34] 张美,朱莉莎,徐飘飘,等.高效液相色谱-串联质谱法检测鸡蛋中五氯酚钠药物残留研究[J].现代畜牧兽医,2023,3:19-22. ZHANG M,ZHU L S,XU P P,et al.Detection of drug residues of sodium pentachlorophenolate in eggs by high performance liquid chromatography-tandem mass spectrometry[J].Modern Journal of Animal Husbandry and Veterinary Medicine,2023,3:19-22.(in Chinese)
[35] 沈登辉,章清荷,王冰霜,等.饮用水中五氯酚的固相萃取-气相色谱串联质谱测定法[J].环境与健康杂志,2014,31(1):51-53. SHEN D H,ZHANG Q H,WANG B S,et al.Solid phase extraction-gas chromatography tandem mass spectrometry determination of pentachlorophenol in drinking water[J].Journal of Environment and Health,2014,31(1):51-53.(in Chinese)
[36] 李丽,王建宇,王蕴平,等.在线固相萃取-超高效液相色谱-三重四极杆串联质谱法检测水中痕量有机磷农药和五氯酚[J].环境化学,2019,38(9):2166-2169. LI L,WANG J Y,WANG Y P,et al.Detection of trace organophosphorus pesticides and pentachlorophenol in water by online solid phase extraction-ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry[J].Environmental Chemistry,2019,38(9):2166-2169.(in Chinese)
[37] 陈晓红,潘胜东,金米聪.固相萃取-超快速液相色谱串联质谱测定食品中五氯酚[J].卫生研究,2018,47(3):440-445. CHEN X H,PAN S D,JIN M C.Determination of pentachlorophenol in food by solid phase extraction-ultrafast liquid chromatography tandem mass spectrometry[J].Journal of Health Research,2018,47(3):440-445.(in Chinese)