两步液液萃取-固相萃取净化结合高效液相色谱-串联质谱法测定猪肉中11种磺胺类兽药残留

doi:10.3724/SP.J.1123.2019.04005

摘要/Abstract

摘要： 建立了猪肉中11种常见的磺胺类兽药残留的两步液液萃取-固相萃取净化-高效液相色谱-串联质谱（LC-MS/MS）检测方法。猪肉样品经乙酸乙酯（含2%（v/v）甲酸）及丙酮分两步液液萃取，正己烷脱脂，Oasis MCX混合阳离子固相萃取小柱净化，氮吹浓缩，定容，过滤膜后进行高效液相色谱-串联质谱分析。采用多反应监测正离子模式进行检测，以基质校准曲线外标法定量。结果表明，在20~400 μg/L范围内11种磺胺类药物均呈现良好的线性关系（相关系数（r²）≥ 0.99），检出限（LODs）（S/N=3）和定量限（LOQs）（S/N=10）分别为0.1~1.0 μg/kg和0.2~3.0 μg/kg。对阴性猪肉样品，在50、100、200 μg/kg 3个水平下分别进行加标回收试验，测得各待测物的平均回收率为79.3%~105.5%之间，相对标准偏差为1.3%~11.6%（n=6）。该方法比采用一步液液萃取法具有更高的提取效率，同时结合固相萃取净化方法进一步富集目标化合物，降低了基质干扰，提高了检测灵敏度。

关键词: 高效液相色谱-串联质谱法, 两步液液萃取, 固相萃取, 磺胺, 兽药残留, 猪肉

Abstract: A method based on two-step liquid-liquid extraction-solid phase extraction purification coupled with high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the determination of 11 common sulfonamides in pork. The samples were extracted first with ethyl acetate containing 2%(v/v) formic acid and then with acetone for the second step. The supernatant was degreased with n-hexane and cleaned using an Oasis MCX SPE column. Finally, nitrogen was blown over the obtained mixture, and the mixture was filtered for conducting HPLC-MS/MS analysis. The sulfonamides were detected in the multiple reaction monitoring positive ion mode, and quantification was performed by employing the external standard method with matrix calibration curves. All the 11 sulfonamides demonstrated good linearity in the range of 20-400 μg/L, with the correlation coefficient (r²) higher than 0.99. The limits of detection (LODs; S/N=3) and limits of quantification (LOQs; S/N=10) were found to be in the ranges of 0.1-1.0 μg/kg and 0.2-3.0 μg/kg, respectively. Furthermore, the results indicated that the average recoveries of the 11 sulfonamides ranged from 79.3% to 105.5% at three spiked levels of 50, 100, and 200 μg/kg, with the relative standard deviations (RSDs) in the range of 1.3%-11.6% (n=6). This method showed higher extraction efficiency than the one-step liquid-liquid extraction, and the detection sensitivity was highly improved.

Key words: high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), two-step liquid-liquid extraction, solid phase extraction (SPE), sulfonamides, veterinary drug residues, pork

中图分类号:

O658

刘培勇, 张惠, 米之金, 张良成, 张光仁. 两步液液萃取-固相萃取净化结合高效液相色谱-串联质谱法测定猪肉中11种磺胺类兽药残留[J]. 色谱, 2019, 37(10): 1098-1104.

LIU Peiyong, ZHANG Hui, MI Zhijin, ZHANG Liangcheng, ZHANG Guangren. Determination of 11 sulfonamides in pork by two-step liquid-liquid extraction-solid phase extraction purification coupled with high performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, 2019, 37(10): 1098-1104.

参考文献

[1] Sun Y Z, Xu Y J, Liu H H, et al. Food Science, 2010, 31(2):120 孙玉增, 徐英江, 刘慧慧, 等. 食品科学, 2010, 31(2):120
[2] Zhuang J, Chi Y H, Li K A, et al. Acta Chimica Sinica, 1998, 56(8):827 庄稼, 迟燕华, 李克安, 等. 化学学报, 1998, 56(8):827
[3] Zhang Y, Li W Q, Zhou W E, et al. Food Science, 2015, 36(23):340 张元, 李伟青, 周伟娥, 等. 食品科学, 2015, 36(23):340
[4] Spielmeyer A, Ahlborn J, Hamscher G, et al. Anal Bioanal Chem, 2014, 406(11):2513
[5] Papadopoulou M V, Bloomer W D, Rosenzweig H S, et al. Antimicrob Agents CH, 2014, 58(11):6828
[6] Baran W, Adamek E, Ziemianska J, et al. J Hazard Mater, 2011, 196:1
[7] Li X J, Yu H, Yang R, et al. Journal of Instrumental Analysis, 2016, 35(10):1255 李晓晶, 于鸿, 杨蓉, 等. 分析测试学报, 2016, 35(10):1255
[8] Cao H, Chen X Z, Zhu Y, et al. New Carbon Materials, 2015, 30(6):572 曹慧, 陈小珍, 朱岩, 等. 新型炭材料, 2015, 30(6):572
[9] Song W, Zhao M Y, Han F, et al. Chinese Journal of Chromatography, 2018, 36(12):1261 宋伟, 赵暮雨, 韩芳, 等. 色谱, 2018, 36(12):1261
[10] Wang Z B, Wang Y, Meng Q J, et al. Journal of Instrumental Analysis, 2013, 32(9):1044 王志兵, 王月, 孟庆娟, 等. 分析测试学报, 2013, 32(9):1044
[11] Zhong W Y, Wang X R, Xu D K, et al. Food Science, 2016, 37(2):193 钟文英, 王兴如, 许丹科, 等. 食品科学, 2016, 37(2):193
[12] Qu B. Food Science, 2013, 34(5):327 曲斌. 食品科学, 2013, 34(5):327
[13] Economou A, Petraki O, Tsipi D, et al. Talanta, 2012, 97:32
[14] Liu T Z, Wang R, Liu W R, et al. Food Science, 2005, 26(11):183 刘铁铮, 王冉, 柳伟荣, 等. 食品科学, 2005, 26(11):183
[15] Wang H, Xu Y, Song W, et al. J Sep Sci, 2011, 34(18):2489
[16] Maryam I, Ali E, Mohammad B, et al. J Sep Sci, 2018, 41(4):910
[17] Gavilán R, Nebot N, Patyra E, et al. Food Addit Contam A, 2018, 35(6):1093
[18] Fang C R, Gao J, Wang Y X, et al. Chinese Journal of Chromatography, 2018, 36(11):1119 方从容, 高洁, 王雨昕, 等. 色谱, 2018, 36(11):1119
[19] Chen J, Yang L Z, Zhang J J, et al. Analytical Instrumentation, 2018, 35(3):51 陈建, 杨灵芝, 张金金, 等. 分析仪器, 2018, 35(3):51
[20] Wang L, Guan D, Zhou X, et al. Journal of Agricultural University of Hebei, 2017, 28(3):100 王玲, 管笛, 周欣, 等. 河北农业大学学报, 2017, 28(3):100
[21] Yang W Q, Zhang X Y, Yao Q, et al. Chinese Journal of Chromatography, 2019, 37(1):46 杨雯筌, 张晓燕, 姚倩, 等. 色谱, 2019, 37(1):46
[22] Huang J,Gui Q W,Gao L,et al. Chinese Journal of Chromatography, 2019, 37(1):2 黄娟, 桂茜雯, 高玲, 等. 色谱, 2019, 37(1):2