加速溶剂萃取/超高效液相色谱-荧光法检测猪肉中氟苯尼考和氟苯尼考胺残留

doi:10.16431/j.cnki.1671-7236.2019.09.031

中国畜牧兽医 ›› 2019, Vol. 46 ›› Issue (9): 2753-2759.doi: 10.16431/j.cnki.1671-7236.2019.09.031

加速溶剂萃取/超高效液相色谱-荧光法检测猪肉中氟苯尼考和氟苯尼考胺残留

刁志祥^1,2, 张培杨^1,2, 王旭堂^1,2, 谢恺舟^1,2, 王波^2,3, 赵霞^1,2, 郭亚文^1,2, 张跟喜^1,2, 张涛^1,2, 戴国俊^1,2, 刘学忠^2,3

1. 扬州大学动物科学与技术学院, 扬州 225009;
2. 教育部农业与农产品安全国际合作联合实验室, 扬州 225009;
3. 扬州大学兽医学院, 扬州 225009

收稿日期:2019-04-02 出版日期:2019-09-20 发布日期:2019-09-21
通讯作者: 谢恺舟 E-mail:yzxkz168@163.com
作者简介:刁志祥(1996-),男,江苏泰州人,硕士,研究方向:畜产品品质与安全,E-mail:1178257857@qq.com
基金资助:
国家现代农业产业技术体系项目（CARS-41-G23）；扬州大学"高端人才支持计划"项目（201801）；江苏省农业科技自主创新基金项目（CX（18）3022）

Determination of Florfenicol and Florfenicol Amine Residues in Pork by Accelerated Solvent Extraction/Ultra-high Performance Liquid Chromatography-fluorescence

DIAO Zhixiang^1,2, ZHANG Peiyang^1,2, WANG Xutang^1,2, XIE Kaizhou^1,2, WANG Bo^2,3, ZHAO Xia^1,2, GUO Yawen^1,2, ZHANG Genxi^1,2, ZHANG Tao^1,2, DAI Guojun^1,2, LIU Xuezhong^2,3

1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
2. Joint International Research Laboratory of Agriculture & Agri-Product Safety, Yangzhou University, Yangzhou 225009, China;
3. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China

Received:2019-04-02 Online:2019-09-20 Published:2019-09-21

摘要/Abstract

摘要： 为给动物源性食品中同时检测氟苯尼考（FF）及其主要代谢产物氟苯尼考胺（FFA）的残留提供新的安全可靠的方法，本研究建立了猪肉中FF及FFA残留检测的加速溶剂萃取/超高效液相色谱-荧光检测（ASE/UPLC-FLD）法。试验采用乙酸乙酯：氨水（98：2，V/V）为提取剂，样品经加速溶剂萃取后浓缩，用乙酸乙酯饱和正己烷除脂，再经浓缩复溶后进行UPLC-FLD检测。结果表明，FF及其代谢产物FFA在添加浓度为9.8~400和3.2~400 μg/kg的范围内线性关系良好，相关系数（R²）分别为0.9993和0.9999。FF的检测限（LOD）和定量限（LOQ）分别为3.3和9.8 μg/kg；FFA的LOD和LOQ分别为1.2和3.2 μg/kg。样品在LOQ、0.5 MRL、1.0 MRL和2.0 MRL加标水平下的平均回收率为82.92%~96.72%，相对标准偏差（RSD）低于3.11%。综上所述，加速溶剂萃取（ASE）方法省时、环保、提取效率高，超高效液相色谱-荧光检测（UPLC-FLD）检测方法高效、灵敏、回收率高，是符合检测标准的新方法。

关键词: 猪肉; 氟苯尼考(FF); 氟苯尼考胺(FFA); 残留; 加速溶剂萃取(ASE); 超高效液相色谱-荧光检测(UPLC-FLD)

Abstract: In order to provide a new safe and reliable method for simultaneous detection of florfenicol (FF) and its metabolite florfenicol amine (FFA) residues in animal-derived foods,this study established an accelerated solvent extraction/ultra-high performance liquid chromatography-fluorescence detector (ASE/UPLC-FLD) method for the determination of FF and FFA in pork.Samples were concentrated after accelerated solvent extraction,fat was removed by n-hexane saturated with ethylacetate,then concentrated and re-dissolved before analysis in UPLC-FLD system.The results showed that FF and its metabolite FFA had a good linear relationship in the range of 9.8 to 400 and 3.2 to 400 μg/kg,respectively,and the coefficient of determination (R²) were 0.9993 and 0.9999,respectively.The limit of detection (LOD) and the limit of quantification (LOQ) of FF were 3.3 and 9.8 μg/kg,for FFA were 1.2 and 3.2 μg/kg,respectively.The average recoveries of the samples at LOQ,0.5MRL,1.0MRL and 2.0MRL spiked levels ranged from 82.92% to 96.72%,and the relative standard deviations were below 3.11%.In summary,the ASE extraction method had many advantages such as high extraction efficiency,time saving,environmental protection and so on.The UPLC-FLD method had high efficiency,sensitivity and recovery,and it provided a new method for simultaneous determination of FF and FFA residues in animal-derived food and it was a new method fitting on meets testing standards.

Key words: pork; FF; FFA; residue; ASE; UPLC-FLD

中图分类号:

S859.84

刁志祥, 张培杨, 王旭堂, 谢恺舟, 王波, 赵霞, 郭亚文, 张跟喜, 张涛, 戴国俊, 刘学忠. 加速溶剂萃取/超高效液相色谱-荧光法检测猪肉中氟苯尼考和氟苯尼考胺残留[J]. 中国畜牧兽医, 2019, 46(9): 2753-2759.

DIAO Zhixiang, ZHANG Peiyang, WANG Xutang, XIE Kaizhou, WANG Bo, ZHAO Xia, GUO Yawen, ZHANG Genxi, ZHANG Tao, DAI Guojun, LIU Xuezhong. Determination of Florfenicol and Florfenicol Amine Residues in Pork by Accelerated Solvent Extraction/Ultra-high Performance Liquid Chromatography-fluorescence[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(9): 2753-2759.

参考文献

[1] 宋志超,张崇威,陈蔷,等.氟苯尼考注射液在鸡体内残留消除研究[J].中国兽药杂志,2018,52(12):35-43. SONG Z C,ZHANG C W,CHEN Q,et al.Residue deption study of florfenicol injection in chicken[J].Chinese Journal of Veterinary Drug,2018,52(12):35-43.(in Chinese)
[2] YANG J,SUN G Z,QIAN M R,et al.Development of a high-performance liquid chromatography method for the determination of florfenicol in animal feedstuffs[J].Journal of Chromatography B,2017,1068-1069:9-14.
[3] SAMS R A.Florfenicol:Chemistry and metabolism of a novel broad-spectrum antbiotic[J].Tieraerztliche Umschau,1995,50(10):703-707.
[4] 曾勇,董文婷,龚波,等.超高效液相色谱-串联质谱法快速测定禽蛋中氯霉素、氟苯尼考及其代谢物残留[J].中国兽药杂志,2017,51(12):42-46. ZENG Y,DONG W T,GONG B,et al.Rapid determination of chloramphenicol,florfenicol and its metabolites in poultry eggs by UPLC-MS/MS[J].Chinese Journal of Veterinary Drug,2017,51(12):42-46.(in Chinese)
[5] AlECHAGA É,MOYANO E,GALCERAN M T.Ultra-high performance liquid chromatography-tandem mass spectrometry for the analysis of phenicol drugs and florfenicol-amine in foods[J].Analyst,2012,137(10):2486-2494.
[6] 仲伶俐,郭灵安,毛建霏,等.UPLC-MS/MS法同时测定牛奶中替米考星、地塞米松、氟苯尼考和氯霉素残留[J].中国畜牧兽医,2018,45(9):307-314. ZHONG L L,GUO L A,MAO J F,et al.Simultaneous determination of tilmicosin,dexamethasone,florfenicol and chloramphenicol residues in milk by UPLC-MS/MS method[J].China Animal Husbandry & Veterinary Medicine,2018,45(9):307-314.(in Chinese)
[7] 周文中,毕可东,邹本革,等.氟苯尼考与盐酸土霉素体外联合抑菌效果研究[J].中国畜牧兽医,2011,38(7):201-203. ZHOU W Z,BI K D,ZOU B G,et al.Studies on the combined antibacterial activity of florfenicol and oxytetracycline hydrochloride in vitro[J].China Animal Husbandry & Veterinary Medicine,2011,38(7):201-203.(in Chinese)
[8] U.S.FOOD AND DRUG ADMINISTRATION.CFR-Code offederal regulations title 21 part 556 tolerances for residue of new animal drugs in food[S].2014.
[9] 谢恺舟,徐东,陈书琴,等.高效液相色谱荧光检测法同时检测鸡肉中甲砜霉素、氟苯尼考及氟苯尼考胺残留[J].分析试验室,2011,30(7):31-35. XIE K Z,XU D,CHEN S Q,et al.Simultaneous determination of residues ofthiamphenicol forfenicol and forfenicol amine in chicken muscle by HPLC with fluorescence detection[J]. Chinese Journal of Analysis Laboratory,2011,30(7):31-35.(in Chinese)
[10] HUANG S,NING G,LIU H,et al.Simultaneous and specific enrichment of several amphenicol antibiotics residues in food based on novel aptamer functionalized magnetic adsorbents using HPLC-DAD[J].Journal of Chromatography B,2017,1060:247-254.
[11] ENDRIS R,CROUCH L S,WRZESINSKI C L.Determination of florfenicol amine in channel catfish muscle by liquid chromatography[J].Journal of Aoac International,2003,86(3):515-520.
[12] 谢恺舟,徐东,陈书琴,等.高效液相色谱荧光检测法检测鸡肉中甲砜霉素残留[J].中国畜牧兽医,2011,38(3):163-166. XIE K Z,XU D,CHEN S Q,et al.Determination of residues of thiamphenicol in chicken muscle by HPLC with fluorescence detection[J].China Animal Husbandry & Veterinary Medicine,2011,38(3):163-166.(in Chinese)
[13] 杨秋红,艾晓辉,李荣,等.固相萃取-气相色谱法同时检测水产品中的氯霉素、甲砜霉素、氟苯尼考和氟苯尼考胺[J].分析试验室,2015,34(5):533-537. YANG Q H,AI X H,LI R,et al.Simultaneous determination of chloramphenicol,thiamphnicol,florfenicol and forfenicol-amine in aquatic products by gas chromatographic method with solid phase extraction[J].Chinese Journal of Analysis Laboratory,2015,34(5):533-537.(in Chinese)
[14] 胡红美,郭远明,雷科,等.分散固相萃取净化-气相色谱法测定水产品中氯霉素和氟苯尼考[J].食品科学,2014,35(8):231-235. HU H M,GUO Y M,LEI K,et al.Determination of chloramphenicol and florfenicol in fishery products by using dispersive solid phase extraction and gas chromatography[J].Food Science,2014,35(8):231-235.(in Chinese)
[15] 梁晓聪,王玮,田丽,等.气相色谱-串联质谱法测定淡水鱼中氯霉素残留量[J].中国卫生检验杂志,2015,25(7):940-942. LIANG X C,WANG W,TIAN L,et al.Determination of chloramphenicol residues in freshwater fish by GC-MS/MS[J].Chinese Journal of Health Laboratory Technology,2015,25(7):940-942.(in Chinese)
[16] AZZOUZ A,BALLESTEROS E.Multiresidue method for the determination of pharmacologically active substances in egg and honey using a continuous solid-phase extraction system and gas chromatography-mass spectrometry[J].Food Chemistry,2015,178(3):63-69.
[17] CORNEJO J,POKRANT E,RIQUELME R,et al.Single-laboratory validation of an LC-MS/MS method for determining florfenicol (FF) and florfenicol amine (FFA) residues in chicken feathers and application to a residue-depletion study[J].Food Additives & Contaminants:Part A,2017,34(4):469-476.
[18] LIU H Y,LIN S L,FUH M R.Determination of chloramphenicol,thiamphenicol and florfenicol in milk and honey using modified QuEChERS extraction coupled with polymeric monolith-based capillary liquid chromatography tandem mass spectrometry[J].Talanta,2016,150:233-239.
[19] PAN X D,WU P G,JIANG W,et al.Determination of chloramphenicol,thiamphenicol,and florfenicol in fish muscle by matrix solid-phase dispersion extraction (MSPD) and ultra-high pressure liquid chromatography tandem mass spectrometry[J].Food Control,2015,52:34-38.
[20] BARRETO F,RIBEIRO C,BARCELLOS H R,et al.Determination of chloramphenicol,thiamphenicol,florfenicol and florfenicol amine in poultry,swine,bovine and fish by liquid chromatography-tandem mass spectrometry[J].Journal of Chromatography A,2016,1449(4):48-53.
[21] 张静,高玉时,周倩,等.高效液相色谱-电喷雾电离串联质谱法测定鸡肉、鸡蛋中氯霉素、甲砜霉素和氟苯尼考药物残留量[J].食品安全质量检测学报,2017,8(6):2211-2219. ZHANG J,GAO Y S,ZHOU Q.et al.Determination of chlorampheincol,thiamphenicol and florfenicol residues in chicken and egg by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry[J].Journal of Food Safety and Quality,2017,8(6):2211-2219.(in Chinese)
[22] 罗昭军,陈幸,单乃荣,等.高效液相色谱法检测鸡蛋中氯霉素类药物残留[J].中国畜牧兽医文摘,2018,34(3):72-74. LUO Z J,CHEN X,SHAN N R,et al.Determination of chloramphenicol residues in eggs by high performance liquidchromatography[J].Chinese Abstract of Animal Husbandry and Veterinary Medicine,2018,34(3):72-74.(in Chinese)
[23] WANG L,LI Y Q.Simultaneous determination of ten antibiotic residues in milk by UPLC[J].Chromatographia,2009,70(1-2):253-258.
[24] XIE K,JIA L,YAO Y,et al.Simultaneous determination of thiamphenicol,florfenicol and florfenicol amine in eggs by reversed-phase high-performance liquid chromatography with fluorescence detection[J].Journal of Chromatography B,2011,879(23):2351-2354.

加速溶剂萃取/超高效液相色谱-荧光法检测猪肉中氟苯尼考和氟苯尼考胺残留

Determination of Florfenicol and Florfenicol Amine Residues in Pork by Accelerated Solvent Extraction/Ultra-high Performance Liquid Chromatography-fluorescence

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