高效液相色谱-串联质谱法测定废水中5种喹诺酮类抗生素
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摘要
建立高浓度有机废水中5种喹诺酮类抗生素的高效液相色谱-串联质谱测定方法。水样经HLB固相萃取小柱富集净化,12 ml甲醇洗脱、浓缩并加入内标溶液后,定容至1 mL待测。以C18柱为分离柱,含0.01%甲酸的甲醇-含0.01%甲酸的水溶液为流动相,目标物质在10 min内分离。在0.25~1 250 ng/mL范围内,目标物质线性关系良好(R2>0.99)。基质加标试验结果表明,纯水中的回收率为61.40%~91.92%,废水中的回收率为54.92%~101.87%,检出限为0.25~2.5 ng/L,方法定量限为0.36~3.99 ng/L。应用该方法对21家猪场的64份废水样品进行分析,5种喹诺酮类抗生素的检出频率为47%~95%,平均检出浓度为980~5 734 ng/L。该方法快速、准确,适用于高浓度有机废水中喹诺酮类抗生素的同时测定。
A method was established for determination of 5 fluoroquinolone antibiotics in high concentration of organic wastewater using solid phase extraction(SPE) followed by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) in this paper.Aqueous samples were developed with Oasis HLB solid-phase extraction(SPE) cartridges and the targeted antibiotics were subsequently eluted by 12 mL methanol.After concentration,the elutes were added internal standards and determined by HPLC-MS/MS equipped with a reverse-phase C18 column.The results of method development showed that the 5 kinds of fluoroquinolone could be efficiently separated within 10 minutes by HPLC-MSMS;the calibration curve ranged 0.25-1 250 ng/mL and showed high relationship(R2>0.99).The recoveries for all analytes in pure water matrices were above 60%,and the recoveries of target compounds in real wastewater samples ranged 54.92%-101.87%;detection limits varied between 0.25-2.5 ng/L and quantization limits were 0.36-3.99 ng/L.The presented method was successfully applied to quantify the five fluoroquinolones in 64 wastewater samples from 21 pig farms.The detection frequency of five quinolones in real samples was 47%-95% and the average detection concentration was found to be 980 to 5 734 ng/L.All the results indicate that this method is suitable for rapid determination of fluoroquinolone antibiotics in high concentration of organic wastewater.
A method was established for determination of 5 fluoroquinolone antibiotics in high concentration of organic wastewater using solid phase extraction(SPE) followed by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) in this paper.Aqueous samples were developed with Oasis HLB solid-phase extraction(SPE) cartridges and the targeted antibiotics were subsequently eluted by 12 mL methanol.After concentration,the elutes were added internal standards and determined by HPLC-MS/MS equipped with a reverse-phase C18 column.The results of method development showed that the 5 kinds of fluoroquinolone could be efficiently separated within 10 minutes by HPLC-MSMS;the calibration curve ranged 0.25-1 250 ng/mL and showed high relationship(R2>0.99).The recoveries for all analytes in pure water matrices were above 60%,and the recoveries of target compounds in real wastewater samples ranged 54.92%-101.87%;detection limits varied between 0.25-2.5 ng/L and quantization limits were 0.36-3.99 ng/L.The presented method was successfully applied to quantify the five fluoroquinolones in 64 wastewater samples from 21 pig farms.The detection frequency of five quinolones in real samples was 47%-95% and the average detection concentration was found to be 980 to 5 734 ng/L.All the results indicate that this method is suitable for rapid determination of fluoroquinolone antibiotics in high concentration of organic wastewater.
引文
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[2]王云鹏,马越.养殖业抗生素的使用及潜在危害[J].中国抗生素杂志,2008,33(9):519-521.WANG Yunpeng,MA Yue.Potential public hazard of using antibiotics in livestock industry[J].Chinese Journal of Antibiotics,2008,33(9):519-521.
[3]王慧慧,王淑平.畜禽排泄物中抗生素残留与控制技术研究进展[J].土壤通报,2011,42(1):250-256.WANG Huihui,WANG Shuping.Residue antibiotics in livestock and poultry excretion and related control technologies:a review[J].Chinese Journal of Soil Science,2011.42(1):250-256.
[4]张艳,李彦文,莫测辉,等.高效液相色谱-荧光测定蔬菜中喹诺酮类抗生素[J].广东农业科学,2009(6):176-180.ZHANG Yan,LI Yanwen,MO Cehui,et al.Determination of quinolones antibiotics in vegetable using high performance liquid chromatography fluorescence[J].Journal of Guang Dong Agricultural Sciences,2009(6):176-180.
[5]张树清,张夫道,刘秀梅,等.规模化养殖畜禽粪主要有害成分测定分析研究[J].植物营养与肥料学报,2005,11(6):822-829.ZHANG Shuqing,ZHANG Fudao,LIU Xiumei,et al.Determi-nation and analysis on main harmful composition in excrement of scale livestock and poultry feedlo-ts[J].Plant Nutrition and Fertilizer Science,2005,11(6):822-829.
[6]张树清.规模化养殖畜禽粪有害成分测定及其无害化处理效果[D].北京:中国农业科学院,2004.
[7]邓玉,倪福全.水环境中抗生素残留及其危害[J].南水北调与水利科技,2011,9(3):96-100.DENG Yu,NI Fuquan.Research on antibiotics residues and hazardous in aquatic environment:a review[J].South-to-North Water Diversion and Water Science&Tech-nology,2011,9(3):96-100.
[8]孙刚,袁守军,计峰,等.畜禽粪便中抗生素残留危害及其研究进展[J].环境与健康杂志,2009,26(3):277-279.SUN Gang,YUAN Shoujun,JI Feng,et al.Environmental impact of antibiotics contamination from livestock and poultry dejecta:a review of recent[J].Journal of Environment and Health,2009,26(3):277-279.
[9]季秋洁.3种氟喹诺酮类抗生素在猪粪中的残留与降解[D].合肥:安徽农业大学,2012.
[10]邰义萍,罗晓栋,莫测辉,等.广东省畜牧粪便中喹诺酮类和磺胺类抗生素的含量与分布特征研究[J].环境科学,2011,32(4):1 188-1 193.TAI Yiping,LUO Xiaodong,MO Cehui.et al.Occurrence of quin-olone and sulfonamide antibiotics in swine and cattle manures from large-scale feeding operations of Guangdong province[J].Environmental Science,2011,32(4):1 188-1 193.
[11]刘锋,廖德润,李可,等.畜禽养殖基地磺胺类喹诺酮类和大环内酯类抗生素污染特征[J].农业环境科学学报,2013,32(4):847-853.LIU Feng,LIAO Derun,LI Ke,et al.Pollution characteristics of the sulfonamides,quinolones and macrolides in the samples collected from livestock and poultry feedlots[J].Journal of Agro-Environment Science,2013,32(4):847-853.
[12]李佩佩,张小军,梅光明,等.超高效液相色谱-串联质谱检测养殖水体中喹诺酮类抗生素[J].浙江大学学报(理学版),2015,42(3):334-338.LI Peipei,ZHANG Xiaojun,MEI Guangming,et al.Determination of quinolones in aquaculture waters by ultra performance liquid chromatography-tandem mass spectrometry[J].Journal of Zhejiang University(Science Edition),2015,42(3):334-338.
[13]中华人民共和国农业部.第2292号公告[BE/OL].(2015-09-08)[2017-01-05].http://www.moa.gov.cn/sydw/stzz/ybfk/dt/201509/t20150908_4820832.htm.
[14]孙艳侠.高效液相色谱在乳品抗生素残留检测中的应用[J].山西农业科学,2010.38(3):10-13.SUN Yanxia.Review on determination of antibiotics residues in dairy products by high performance liquid chromatography[J].Journal of Shanxi Agricultural Sciences,2010.38(3):10-13.
[15]钟东莲,丁明,汤富彬,等.高效液相色谱-电喷雾串联质谱法测定畜禽粪便中四环素类抗生素[J].分析科学学报,2014,30(3):433-436.ZHONG Donglian,DING Ming,TANG Fubin,et al.Determination of tetracycline’s antibiotics residues in anim-al manur by liquid chromatography with tandem mass spectrometry[J].Journal of Analytical Science,2014,30(3):433-436.
[16]吴小莲,包艳萍,向垒,等.固相萃取-高效液相色谱-串联质谱法同时测定蔬菜中8种磺胺类抗生素[J].环境化学,2013,32(6):1 038-1 044.WU Xiaolian,BAO Yanping,XIANG Lei,et al.Simultaneous dete-rmination of eight sulfonamide antibiotics in vegetab-les using solid phase extraction and high performance liquid chromatography coupled with mass spectrome-try[J].Environmental Chemistry,2013,32(6):1 038-1 044.
[17]李刚,颜智勇,谭秀益,等.畜禽养殖废水中抗生素检测技术研究进展[J].绿色科技,2011(11):97-99.LI Gang,YAN Zhiyong,TAN Xiuyi,et al.Advances of detect-ion technology for antibiotics from livestock and poultry breeding wastewater[J].Journal of Green Science and Technology,2011(11):97-99.
[18]YE Z Q,HOWARD S,WEINBERG.Trace analysis of trimethoprim and sulfonamide,macrolide,quinolone,and tetracycline antibiotics in chlorinated drinking water using liquid chromatography electrospray tandem mass pectrometry[J].Anal Chem,2007,79(3):1 135-1 144.
[19]CAHILL J,FURLONG E T,BURKHARDT M R,et al.Determination of pharmaceutical compounds in surface-water and ground-water samples by solid-phase extraction and high-performance liquid chromatography electrospray ionization mass spectrometry[J].Journal of Chromatography A,2004,1 041(1/2):171-180.
[20]秦宏兵,顾海东.超高效液相色谱-串联质谱法测定饮用水源水中磺胺类抗生素[J].中国环境监测,2013,29(1):98-102.QIN Hongbing,GU Haidong.Determination of sulfonamide antibiotics in drinking source water using ultra performance liquid chromatography tandem mass spectrometry[J].Environmental Monitoring in China,2013,29(1):98-102.
[21]万位宁,陈熹,居学海,等.固相萃取-超高效液相色谱串联质谱法同时检测畜禽粪便中多种抗生素残留[J].分析化学研究报告,2013,41(7):993-999.WAN Weining,CHEN Xi,JU Xuehai,et al.Simultaneous determination of residual antibiotics in livestock manure by solid phase extraction-ultra-high performance liquid chromatography tandem mass spectrometry[J].Chinese Journal of Analytical Chemistry,2013,41(7):993-999.
[22]刘佳佳,佘永新,刘洪斌,等.高效液相色谱-串联质谱法同时测定动物源食品中24种磺胺类药物残留[J].分析实验室,2011,30(2):9-13.LIU Jiajia,SHE Yongxin,LIU Hongbin,et al.Simultaneous determination of 24 sulfonamides residues in animal pro-ducts by high performance liquid chromatography tandem mass spectrometry[J].Chinese Journal of Analysis Laboratory.2011,30(2):9-13.
[23]BENITO-PEA E,PARTAL-RODERA A I,LEN-GONZLEZ M E,et al.Evaluation of mixed mode solid phase extraction cartridges for the preconcentration of beta lactam antibiotics in wastewater using liquid chromatography with UV-DAD detection[J].Analytical Chemical Acta,2006,556(2):415-422.
[24]薄海波,雒丽丽,曹彦忠,等.超高效液相色谱-串联质谱法测定牛奶和奶粉中6种聚醚类抗生素残留量[J].分析化学研究报告,2009,37(8):1 161-1 166.BO Haibo,LUO Lili,CAO Yanzhong,et al.Determination of six polyether antibiotics residues in milk and milk powder by ultra performance liquid chromatography-tandem mass spectrometry[J].Chinese Journal of Analytical Chemistry.2009,37(8):1 161-1 166.
[25]张俊,罗阳,潘曼曼,等.水体中磺胺、四环素、喹诺酮类抗生素检测方法[J].中国环境监测,2013,29(4):111-115.ZHANG Jun,LUO Yang,PAN Manman,et al.Analysis of sulfonamide,tetracycline and quinolones antibiotics in surface water[J].Environmental Monitoring in China,2013,29(4):111-115.
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