水中微囊藻毒素的超高效液相色谱-四极杆-静电场轨道阱高分辨质谱测定法
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摘要
目的建立水中微囊藻毒素(MC)煮沸浓缩-超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLCquadrupole-Orbitrap MS)的测定方法。方法水样在电热板上加热浓缩后,以0.22μm滤膜过滤,采用BEH C_(18)色谱柱(50mm×2.0 mm,2.1μm),以甲醇-2‰甲酸水溶液作为流动相,梯度洗脱,采用电喷雾电离源正离子模式(HESI+)在t-SIM模式下分析检测,通过测得的精确质量数与理论精确质量数吻合度、目标化合物的元素组成、保留时间进行定性。结果MC-LR、MC-YR、MC-RR、MC-LA、MC-LF、MC-LY在0.2～5μg/L,MC-WR在1～25μg/L之间线性良好,相关系数均>0.998;MC-RR、MC-YR、MC-LR、MC-WR、MC-LA、MC-LY、MC-LF的检出限分别为0.18、0.26、0.40、1.31、0.29、0.29、0.29ng/L;高、中、低浓度加标的RSD为4.11%～10.8%,平均回收率为61.8%～103%。结论该方法简便快捷,选择性好,灵敏度高,重现性好,可满足水中微囊藻毒素检测的要求。
Objective To establish a method for the determination of microcystins in water using ultra high performance liquid chromatography-quadrupole-Orbitrap mass spectrometry. Methods Samples were heated and concentrated on electric heating plate,then filtered through 0.22 μm filter membrane. Sample purification was performed on BEH C_(18) column(50 mm×2.0 mm,2.1 μm) with the gradient elution of methanol and 2‰ formic acid. The samples were analyzed by using the electrospray ionization positive ion source(HESI+) and were scanned in the t-SIM mode,the objective compounds were qualitatively detected through the element compositions of target fragment,retention time and the matching level of accurate mass number and theoretical accurate mass number. Results The calibration curves showed great linearity between 0.2-5 μg/L for MC-LR、MCYR、MC-RR、MC-LA、MC-LF、MC-LY,between 1-25 μg/L for MC-WR,the correlation coefficient of them were all above 0.998. The method detection limits of MC-RR, MC-YR, MC-LR, MC-WR, MC-LA, MC-LY and MC-LF were 0.18, 0.26,0.40, 1.31, 0.29, 0.29 and 0.29 ng/L respectively. The average recoveries at three levels(low,moderate and high concentration)for each species were in the range of 61.8%-103%,the relative standard deviations were in the range of 4.11%-10.8%.Conclusion This method is easy to operate and shows high sensitivity,good selectivity and reproducibility,meets all the detection requirements of microcystins in water.
Objective To establish a method for the determination of microcystins in water using ultra high performance liquid chromatography-quadrupole-Orbitrap mass spectrometry. Methods Samples were heated and concentrated on electric heating plate,then filtered through 0.22 μm filter membrane. Sample purification was performed on BEH C_(18) column(50 mm×2.0 mm,2.1 μm) with the gradient elution of methanol and 2‰ formic acid. The samples were analyzed by using the electrospray ionization positive ion source(HESI+) and were scanned in the t-SIM mode,the objective compounds were qualitatively detected through the element compositions of target fragment,retention time and the matching level of accurate mass number and theoretical accurate mass number. Results The calibration curves showed great linearity between 0.2-5 μg/L for MC-LR、MCYR、MC-RR、MC-LA、MC-LF、MC-LY,between 1-25 μg/L for MC-WR,the correlation coefficient of them were all above 0.998. The method detection limits of MC-RR, MC-YR, MC-LR, MC-WR, MC-LA, MC-LY and MC-LF were 0.18, 0.26,0.40, 1.31, 0.29, 0.29 and 0.29 ng/L respectively. The average recoveries at three levels(low,moderate and high concentration)for each species were in the range of 61.8%-103%,the relative standard deviations were in the range of 4.11%-10.8%.Conclusion This method is easy to operate and shows high sensitivity,good selectivity and reproducibility,meets all the detection requirements of microcystins in water.
引文
[1]黄艺,张郅灏.微囊藻毒素的致毒机理和人体健康风险评价研究进展[J].生态环境学报,2013,22(2):357-364
[2]孟元华,龚燕,朱鹏飞,等.水中微囊藻毒素-RR的量子点荧光猝灭快速测定法[J].环境与健康杂志,2016,33(7):636-639.
[3]Li HY.In vivo study on the efects of microcystin extracts on the expression profiles of proto-oncogenes(c-fos,c-jun and c-myc)in liver,kidney and testis of male Wistar rats injected i.v.with toxins[J].Toxicon,2009,53:169-175.
[4]Zhao Y Y,Xie P.In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit[J].Comp Biochem Physiol Part C,2008,148:204-210.
[5]袁春阳,谢平,刘颖,等.微囊藻毒素-LR急性呼吸道暴露对小鼠生殖细胞凋亡的影响及其作用机制[J].环境与健康杂志,2017,34(11):961-965
[6]李砚,舒为群.微囊藻毒素对人群健康影响的流行病学研究进展[J].环境与健康杂志,2010,27(8):730-733.
[7]IARC Monographs on the evaluation of carcinogenic risks to humans.Ingested nitrate and nitrite and cyanobacterial peptide toxins[R].WHO,2010.
[8]中华人民共和国卫生部,中国国家标准化管理委员会.GB5750-2006生活饮用水标准检验方法[S].北京:中国标准出版社,2007.
[9]钮伟民,肖付刚,戴维杰,等.液相色谱-串联质谱法同时检测水中5种微囊藻毒素,安徽农业科学,2008,36(29):12554.
[10]王超,吕怡兵,许人骥,等.液相色谱-二极管阵列检测器/离子阱质谱法检测水中的5种微囊藻毒素,色谱,2011,29(3):212.
[11]张明,唐访良,陈峰,等.固相萃取-超高效液相色谱-电喷雾串联质谱法同时测定地表水中9种微囊藻毒素[J].色谱,2012,30(1):51-55
[12]唐琳,孔芳,罗苹,等.生活饮用水中痕量微囊藻毒素-LR和-RR的免疫亲和净化-高效液相色谱-串联四极杆质谱测定法[J].环境与健康杂志,2017,34(9):832-835
[13]Fischer WJ,Garthwaite I,Miles CO,et al.Congener-independent immunoassay for microcystins and nodularins[J].Environ Sci Technol,2001,35:4849-4856.
[14]张艺,黄飚,金坚,等.微囊藻毒素-LR的生物素-亲和素-时间分辨荧光免疫分析法[J].环境与健康杂志,2015,32(1):58-60
[15]郭浩免疫方法在藻类和贝毒素检测中的应用[J].卫生研究,1999,28(2):22-144.
[2]孟元华,龚燕,朱鹏飞,等.水中微囊藻毒素-RR的量子点荧光猝灭快速测定法[J].环境与健康杂志,2016,33(7):636-639.
[3]Li HY.In vivo study on the efects of microcystin extracts on the expression profiles of proto-oncogenes(c-fos,c-jun and c-myc)in liver,kidney and testis of male Wistar rats injected i.v.with toxins[J].Toxicon,2009,53:169-175.
[4]Zhao Y Y,Xie P.In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit[J].Comp Biochem Physiol Part C,2008,148:204-210.
[5]袁春阳,谢平,刘颖,等.微囊藻毒素-LR急性呼吸道暴露对小鼠生殖细胞凋亡的影响及其作用机制[J].环境与健康杂志,2017,34(11):961-965
[6]李砚,舒为群.微囊藻毒素对人群健康影响的流行病学研究进展[J].环境与健康杂志,2010,27(8):730-733.
[7]IARC Monographs on the evaluation of carcinogenic risks to humans.Ingested nitrate and nitrite and cyanobacterial peptide toxins[R].WHO,2010.
[8]中华人民共和国卫生部,中国国家标准化管理委员会.GB5750-2006生活饮用水标准检验方法[S].北京:中国标准出版社,2007.
[9]钮伟民,肖付刚,戴维杰,等.液相色谱-串联质谱法同时检测水中5种微囊藻毒素,安徽农业科学,2008,36(29):12554.
[10]王超,吕怡兵,许人骥,等.液相色谱-二极管阵列检测器/离子阱质谱法检测水中的5种微囊藻毒素,色谱,2011,29(3):212.
[11]张明,唐访良,陈峰,等.固相萃取-超高效液相色谱-电喷雾串联质谱法同时测定地表水中9种微囊藻毒素[J].色谱,2012,30(1):51-55
[12]唐琳,孔芳,罗苹,等.生活饮用水中痕量微囊藻毒素-LR和-RR的免疫亲和净化-高效液相色谱-串联四极杆质谱测定法[J].环境与健康杂志,2017,34(9):832-835
[13]Fischer WJ,Garthwaite I,Miles CO,et al.Congener-independent immunoassay for microcystins and nodularins[J].Environ Sci Technol,2001,35:4849-4856.
[14]张艺,黄飚,金坚,等.微囊藻毒素-LR的生物素-亲和素-时间分辨荧光免疫分析法[J].环境与健康杂志,2015,32(1):58-60
[15]郭浩免疫方法在藻类和贝毒素检测中的应用[J].卫生研究,1999,28(2):22-144.