Simultaneous Separation and Determination of Thirteen Antibiotics in Soil and Water by High Performance Capillary Electrophoresis with Pretreatment by Solid Phase Extraction
摘 要
采用固相萃取(SPE)纯化、富集,高效毛细管电泳(HPCE)分离和紫外光谱检测同时分离并测定水体和土壤样品中13种抗生素的含量。所采得的土壤样品需先经规定方法制成固态分析样品,并取此样品4.000 g,用二乙胺四乙酸二钠0.8 g、Mcllvacine缓冲溶液和乙腈(1+1)混合液提取样品3次(每次加入混合液20.0 mL)。合并3次所得提取液(上清液),用0.22 μm滤膜过滤后,按与水的体积比为1:2.5加水稀释。此溶液待SPE纯化及富集。所采集的水样经0.22 μm滤膜过滤后,用0.1 mol·L-1 HCl溶液调节pH至5.0。此溶液继续进行SPE纯化及富集。分取上述土壤(或水样)样品溶液150 mL,流经HLB SPE柱,用甲醇-水(10+90)混合液淋洗SPE柱除去杂质,随即用甲醇-乙腈(1+1)混合液2 mL洗脱柱上吸附的抗生素,收集洗脱液,吹氮至近干,加入水300 μL溶解残渣,所得溶液进入HPCE柱进行电泳分离,选择由65.0 mmol·L-1硼砂和50.0 mmol·L-1硼酸组成的pH 9.0的缓冲溶液和甲醇及异丙醇(88+10+2)的混合液作为电泳介质,在分离电压为19 kV,柱温为23℃,压力为3.45 kPa条件下进样7 s,13种抗生素可在25 min内完全分离。选择在波长210 nm处进行检测。13种抗生素的线性范围均在150 μg·L-1以内,检出限(3S/N)在0.40~1.0 μg·L-1之间。以空白基质进行加标回收试验,测得回收率在78.5%~107%之间。
Abstract
13 antibiotics in soil or water samples were enriched and purified on SPE column and then separated and determined simultaneously by HPCE with UV-spectrometric detector. Soil sample collected should be pretreated according to a definite process to give analytical sample in solid state, from which 4.000 g were taken and extracted thrice with Na2EDTA 0.8 g and a mixture of mcllvacine buffer and acetonitrile (1+1), using 20.0 mL in each extraction. The extracts (i.e., the supernatants) were combined and filtered through 0.22 μm filtering membrane. The filtrate was diluted with water according to the ratio of volume of the filtrate to water of 1 to 2.5. This solution was ready to be used for SPE treatment. Water sample collected was first filtered through 0.22 μm filtering membrane, and the filtrate was acidified to pH 5.0 with 0.1 mol·L-1 HCl solution. This water sample solution was ready for SPE treatment. A portion of 150 mL of the soil sample solution or the water sample solution (as obtained above) was passed through HLB SPE column, and the column was rinsed with CH3OH-H2O (1+9) to eliminate impurities, and then eluted with 2 mL of a mixture of CH3OH and CH3CN (1+1) to elute the antibiotics from the column. The eluate was collected and evaporated to near dryness by N2-blowing. 300 μL of water were added to dissolve the residue, and the solution was introduced into the HPCE system for electrophoresis. A mixture of (A) pH 9.0 buffer solution composed of 65.0 mmol·L-1 of borax and 50.0 mmol·L-1 of boric acid, (B) methyl alcohol and (C) iso-propyl alcohol mixed in the ratio of A:B:C=88:10:2, was used as electrophoretic medium. The 13 antibiotics could be well separated within 25 min under the following conditions, (a) separation voltage:19 kV; (b) column temperature:23℃; (c) pressure of sample introduction:3.45 kPa; (d) time of sample injection:7 s. UV-detection was performed at 210 nm. Linearity ranges for the 13 analytes were obtained all within 150 μg·L-1. Detection limits (3S/N) found were in the range of 0.40-1.0 μg·L-1. Tests for recovery were carried out by addition of standard solution to matrixes of blank solutions, values of recovery found were ranged from 78.5% to 107%.
中图分类号 O657.8 DOI 10.11973/lhjy-hx201907005
所属栏目 工作简报
基金项目 国家自然科学基金资助项目(81773481)
收稿日期 2018/10/8
修改稿日期
网络出版日期
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备注李兴华,硕士研究生,主要从事环境抗生素理化检测分析
引用该论文: LI Xinghua,MIAO Junjie,KANG Kai,WANG Wei,SHI Hongmei. Simultaneous Separation and Determination of Thirteen Antibiotics in Soil and Water by High Performance Capillary Electrophoresis with Pretreatment by Solid Phase Extraction[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2019, 55(7): 769~777
李兴华,苗俊杰,康凯,王玮,石红梅. 固相萃取-高效毛细管电泳法同时分离测定水体和土壤中13种抗生素[J]. 理化检验-化学分册, 2019, 55(7): 769~777
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参考文献
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【21】檀笑昕,李享,李楠,等.在线固相萃取-HPLC法测定公共绿地土壤中新烟碱类农药残留[J].环境监测管理与技术, 2016,28(5):48-51.
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【24】秦丽婷,童蕾,刘慧,等.环境中磺胺类抗生素的生物降解及其抗性基因污染现状[J].环境化学, 2016,35(5):875-883.
【2】YAO L L, WANG Y X, TONG L, et al. Seasonal variation of antibiotics concentration in the aquatic environment:A case study at Jianghan Plain, central China[J]. Science of the Total Environment, 2015,527/528:56-64.
【3】ZHANG Q Q, YING G G, PAN C G, et al. Comprehensive evaluation of antibiotics emission and fate in the river basins of China:Source analysis, multimedia modeling, and linkage to bacterial resistance[J]. Environmental Science & Technology, 2015,49(11):6772-6782.
【4】ZHANG X X, ZHANG T. Occurrence, abundance, and diversity of tetracycline resistance genes in 15 sewage treatment plants across China and other global locations[J]. Environmental Science & Technology, 2011,45(7):2598-2604.
【5】林涛,陈燕秋,陈卫.水体中磺胺嘧啶对斑马鱼的生态毒性效应[J].安全与环境学报, 2014,14(3):324-327.
【6】刘曦薇,吴玲玲.氯霉素对斑马鱼早期发育的毒性效应[J].环境科学学报, 2010,30(8):1649-1657.
【7】方淑霞,王大力,朱丽华,等.抗生素对微生物的联合与低剂量毒性研究进展[J].生态毒理学报, 2015,10(2):69-75.
【8】LI P, LI J, WU C Z, et al. Synergistic antibacterial effects of β-lactam antibiotic combined with silver nanoparticles[J]. Nanotechnology, 2005,16(9):1912-1917.
【9】DU L F, LIU W K. Occurrence, fate, and ecotoxicity of antibiotics in agro-ecosystems. A review[J]. Agronomy for Sustainable Development, 2012,32(2):309-327.
【10】KUMAR K, THOMPSON A, SINGH A K, et al. Enzyme-linked immunosorbent assay for ultratrace determination of antibiotics in aqueous samples[J]. Journal of Environment Quality, 2004,33(1):250-256.
【11】FENG Y, WEI C J, ZHANG W J, et al. A simple and economic method for simultaneous determination of 11 antibiotics in manure by solid-phase extraction and high-performance liquid chromatography[J]. Journal of Soils and Sediments, 2016,16(9):2242-2251.
【12】CHÁFER-PERICÁS C, MAQUIEIRA Á, PUCHADES R, et al. Multiresidue determination of antibiotics in feed and fish samples for food safety evaluation. Comparison of immunoassay vs LC-MS-MS[J]. Food Control, 2011,22(6):993-999.
【13】王浩,赵丽,杨红梅,等.液相色谱-串联质谱法测定牛奶中35种四环素类、磺胺类、青霉素类、大环内酯类、氯霉素类抗生素残留[J].色谱, 2015,33(9):995-1001.
【14】李鹏,邱月明,蔡慧霞,等.气相色谱-质谱联用法测定动物组织中氯霉素、氟甲砜霉素和甲砜霉素的残留量[J].色谱, 2006,24(1):14-18.
【15】TAKATSUKI K, USHIZAWA I, SHOJI T. Gas chromatographic-mass spectrometric determination of macrolide antibiotics in beef and pork using single ion monitoring[J]. Journal of Chromatography A, 1987,391:207-217.
【16】李爱梅,黄茁,卢文平,等.毛细管电泳法测定水体中四环素类抗生素的基质效应及场放大进样技术的应用[J].色谱, 2014,32(8):897-903.
【17】左艳丽,孙汉文,魏立静.快速溶剂萃取一毛细管电泳法测定土壤和底泥中磺胺类药物残留[J].分析试验室, 2012,32(2):62-66.
【18】HORKÁ M, TESAROVÁ M, KARÁSEK P, et al. Determination of methicillin-resistant and methicillin-susceptible staphylococcus aureus bacteria in blood by capillary zone electrophoresis[J]. Analytica Chimica Acta, 2015,868:67-72.
【19】ZHOU X, XING D, ZHU D, et al. Development and application of a capillary electrophoresis-electrochemiluminescent method for the analysis of enrofloxacin and its metabolite ciprofloxacin in milk[J]. Talanta, 2008,75(5):1300-1306.
【20】QUESADA-MOLINA C, GARCÍA-CAMPAÑA A M, OLMO-IRUELA L, et al. Large volume sample stacking in capillary zone electrophoresis for the monitoring of the degradation products of metribuzin in environmental samples[J]. Journal of Chromatography A, 2007,1164(1/2):320-328.
【21】檀笑昕,李享,李楠,等.在线固相萃取-HPLC法测定公共绿地土壤中新烟碱类农药残留[J].环境监测管理与技术, 2016,28(5):48-51.
【22】MITCHELL S M, ULLMAN J L, TEEL A L, et al. PH and temperature effects on the hydrolysis of three β-lactam antibiotics:Ampicillin, cefalotin and cefoxitin[J]. Science of the Total Environment, 2014,466/467:547-555.
【23】章强,辛琦,朱静敏,等.中国主要水域抗生素污染现状及其生态环境效应研究进展[J].环境化学, 2014,33(7):1075-1083.
【24】秦丽婷,童蕾,刘慧,等.环境中磺胺类抗生素的生物降解及其抗性基因污染现状[J].环境化学, 2016,35(5):875-883.
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