在线固相萃取/液相色谱-线性离子阱质谱法同时定性定量检测全血和尿液中7种抗凝血杀鼠药
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摘要
建立了同时测定全血和尿液样品中7种抗凝血杀鼠药的在线固相萃取/液相色谱-线性离子阱质谱(on-line SPE/LC-LIT/MS)分析方法。用乙腈沉淀样品中的蛋白质,于稀释、离心、过滤后直接进样。经在线固相萃取柱富集纯化;以C18柱为分析柱,甲醇-乙酸铵水溶液(0.02 mol/L)为流动相进行梯度洗脱;在电喷雾电离(ESI)负离子模式下,记录目标母离子在锁定保留时间窗口内的二级全扫描信号,通过自建数据库进行定性确证,挑选高灵敏度与专属性的二级子离子进行定量,从而实现7种杀鼠药的同时定性和定量分析。7种杀鼠药在各自质量浓度范围内线性关系良好,相关系数为0.994 6~0.999 7,检出限和定量限分别为0.02~1.00 ng/mL和0.10~4.00 ng/mL,3个添加水平的回收率为81.0%~113.9%,相对标准偏差为0.1%~6.2%(n=6)。该方法简单方便,灵敏度高,能够满足全血和尿液样品中7种抗凝血杀鼠药的快速筛查和准确定量。
A new and sensitive analytical method has been developed for the simultaneous determination of seven anticoagulant rodenticides in whole blood and urine samples by liquid chromatography-linear ion trap mass spectrometry(LC-LIT / MS) with on-line solid phase extraction(on-line SPE). The samples were treated with acetonitrile,followed by dilution,centrifugation,and filtration. The resulting solution was injected into the LC system directly and processed by on-line SPE column for enrichment and purification. Separation was performed on a C18 column with mixed mobile phases of methanol and 0. 02 mol / L ammonium acetate aqueous solution for gradient elution. The analytes were detected by the mass spectrometer with electrospray ionization(ESI)in negative mode. MS2 full scan signals of the target parent ions within the locked retention time window were recorded. Self-built database searchingwas performed for qualitative confirmation,and MS2 fragment ions with high sensitivity and specificity were selected for quantification. Simultaneous qualitative and quantitative analyses of the seven rodenticides were achieved in this way. Good linearities were obtained within the investigated mass concentration ranges of the seven rodenticides,with r2≥0. 995 8 in blood and r2≥0. 994 6 in urine. The LODs varied from 0. 02 ng/ mL to 1. 00 ng/ mL,and the LOQs varied from 0. 10 ng / mL to 4. 00 ng / mL. The recoveries at three spiked levels in blood and urine samples ranged from 81. 0% to 113. 9%,with RSDs of 0. 1%-6. 2%(n = 6). The developed method is simple,sensitive,and can be used for the rapid detection and accurate quantification of the seven anticoagulant rodenticides in whole blood and urine samples.
A new and sensitive analytical method has been developed for the simultaneous determination of seven anticoagulant rodenticides in whole blood and urine samples by liquid chromatography-linear ion trap mass spectrometry(LC-LIT / MS) with on-line solid phase extraction(on-line SPE). The samples were treated with acetonitrile,followed by dilution,centrifugation,and filtration. The resulting solution was injected into the LC system directly and processed by on-line SPE column for enrichment and purification. Separation was performed on a C18 column with mixed mobile phases of methanol and 0. 02 mol / L ammonium acetate aqueous solution for gradient elution. The analytes were detected by the mass spectrometer with electrospray ionization(ESI)in negative mode. MS2 full scan signals of the target parent ions within the locked retention time window were recorded. Self-built database searchingwas performed for qualitative confirmation,and MS2 fragment ions with high sensitivity and specificity were selected for quantification. Simultaneous qualitative and quantitative analyses of the seven rodenticides were achieved in this way. Good linearities were obtained within the investigated mass concentration ranges of the seven rodenticides,with r2≥0. 995 8 in blood and r2≥0. 994 6 in urine. The LODs varied from 0. 02 ng/ mL to 1. 00 ng/ mL,and the LOQs varied from 0. 10 ng / mL to 4. 00 ng / mL. The recoveries at three spiked levels in blood and urine samples ranged from 81. 0% to 113. 9%,with RSDs of 0. 1%-6. 2%(n = 6). The developed method is simple,sensitive,and can be used for the rapid detection and accurate quantification of the seven anticoagulant rodenticides in whole blood and urine samples.
引文
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[3]Cai X X,Zhang X Y.Chinese Journal of Health Laboratory Technology(蔡欣欣,张秀尧.中国卫生检验杂志),2008,18(12):2470
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[5]Rong W G,Liu H L,Chen B,et al.Chinese Journal of Analysis Laboratory(荣维广,刘华良,陈蓓,等.分析试验室),2013,32(1):73
[6]Song W,Yang B C,Zhao Y B,et al.Journal of Food Science and Technology(宋薇,杨柏崇,赵永彪,等.食品科学技术学报),2014,32(3):65
[7]Zhu F,Liu H L,Chen B,et al.Chinese Journal of Chromatography(朱峰,刘华良,陈蓓,等.色谱),2013,31(5):473
[8]Wang X H,Zhao W S,Gu Y B,et al.Chinese Journal of Forensic Medicine(王学虎,赵武生,顾月保,等.中国法医学杂志),2010,25(1):17
[9]Vandenbroucke V,Desmet N,De Backer P,et al.J Chromatogr B,2008,869(1/2):101
[10]Jin M C,Ren Y P,Xu X M,et al.Forensic Sci Int,2007,171(1):52
[11]Hernándeza A M,Bernal J,Bernal J L,et al.J Sep Sci,2013,36(16):2593
[12]Hernándeza A M,Bernal J,Bernal J L,et al.J Chromatogr B,2013,925:76
[13]Fourel I,Hugnet C,Goy-Thollot I,et al.J Anal Toxicol,2010,34(2):95
[14]Jin M C,Xu G Z,Ren Y P,et al.J Appl Toxicol,2008,28(5):621
[15]Marek L J,Koskinen W C.J Agric Food Chem,2007,55(3):571
[16]Schaff J E,Montgomery M A.J Anal Toxicol,2013(6),37:321
[17]Chen J,Liu Z J,Dai Z Y,et al.Chinese Journal of Chromatography(陈静,刘召金,戴振宇,等.色谱),2013,31(9):894
[18]Xiao W B,Jian Y,Li H.Chinese Journal of Analytical Chemistry(萧伟斌,蹇阳,李桦.分析化学),2014,42(12):1851
[19]Gu X,Wu J P,Zhang X,et al.Chinese Journal of Analytical Chemistry(顾欣,吴剑平,张鑫,等.分析化学),2014,42(12):1759
[20]Li L F,Su M,Shi X L,et al.Chinese Journal of Chromatography(李龙飞,苏敏,石晓蕾,等.色谱),2014,32(2):194