免疫亲和柱净化-液相色谱-串联质谱法测定水环境中微囊藻毒素

Determination of Microcystins in Water Environment by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry and Cleaned Up by Immunoaffinity Column

建立了水环境中微囊藻毒素-LR(microcystin-LR, MC-LR)和微囊藻毒素-RR(microcystin-RR, MC-RR)的免疫亲和柱净化-超高效液相色谱-串联质谱(IAC-UPLC-MS/MS)的分析方法。过滤好的水质加入抗坏血酸防止氧化,混合均匀后将pH调节至中性,经过免疫亲和柱富集和净化后,采用LC-MS/MS测定,采用外标方法定量。以乙腈-0.1%甲酸水溶液(含5 mmol·L~(-1)乙酸铵)为流动相,梯度洗脱分离,电喷雾正离子多反应监测模式监测。结果显示,MC-LR、MC-RR在0.50~20.00 ng·mL~(-1)浓度范围内呈良好线性,线性相关系数均>0.999 5,定量限为0.30 ng·mL~(-1)和0.50 ng·mL~(-1),在2.00、5.00和10.00 ng·mL~(-1) 3种添加水平下的加标回收率分别为88.52%~94.83%、84.71%~91.59%,相对标准偏差分别为0.54%~2.27%,0.39%~5.94%。结果表明本方法重现性好,灵敏度高,适合水环境中MC的实际测定。

Established immunoaffinity column purification-ultra-high performance liquid chromatography-tandem for microcystin-LR(microcystin-LR,MC-LR)and microcystin-RR(MC-RR)in water environment Mass spectrometry(IAC-UPLC-MS/MS)analysis method.Filtered water was added with ascorbic acid to prevent oxidation,following mixing adjusted the pH to neutral.After cleaned up and concentrated by immunoaffinity column,the samples were analyzed by LC-MS/MS and quantitatively determined by external standard method.Used acetonitrile-0.1%formic acid aqueous solution(containing 5 mmol·L-1 ammonium acetate)as mobile phase,gradient elution separation,electrospray positive ion multiple reaction monitoring mode monitoring.The results showed that MC-LR and MC-RR showed good linearity in the concentration range of 0.50-20.00 ng·mL-1,the linear correlation coefficients were both>0.9995,and the quantification limits were 0.30 ng·mL-1 and 0.50 ng·mL-1.The recoveries of standard addition at the three addition levels of 2.00,5.00 and 10.00 ng·mL-1 were 88.52%-94.83%,84.71%-91.59%,and the relative standard deviations respectively were 0.54%-2.27%,0.39%-5.94%.The results show that this method has good reproducibility and high sensitivity,and is suitable for the actual determination of MC in water environment.