水产品中微囊藻毒素的检测方法研究

Detection Technology of Microcystins in Aquatic Product by HPLC

采用富集微囊藻毒素的黑色大头鲢鱼为样品,对微囊藻毒素(microcystin-RR-,-LR)的提取、纯化和HPLC检测方法进行研究.实验建立了固相萃取(SPE)结合高效液相色谱(HPLC)分析水产品中痕量微囊藻毒素的方法,利用二极管阵列检测器在238nm进行检测.其中,微囊藻毒素的提取溶剂分别为5%乙酸、75%甲醇和正丁醇-甲醇-水溶液(1∶4∶15),80%甲醇为洗脱溶剂.HPLC检测中的流动相为含微量三氟乙酸的55%甲醇溶液.研究结果表明,MC-RR的回收率为84.1%,MC-LR的回收率为89.9%.该法具有操作简便、灵敏度高、方法稳定的特点,适用于快速分析环境产品中的两种常见微囊藻毒素MC-LR和MC-RR.

To optimize the solid-phase extraction method of isolation, purification and determination of intracellular Microcystin-RR, -LR （ MC-RR,-LR ） , the sample was a black big head carp which enriched Microcystins. A method for trace level analysis of Microcystins in aquatic product using SPE and HPLC was developed, and the aquatic product was detected by diode array detection （ LC-DAD ） at 238 nm. During extraction, samples which enriched Microcystins were treated with different solvent （ 5 % acetic acid, 75 % methanol and mixed solvents 1-butanol,methanol,and water （ 1 ∶ 4 ∶ 15 ） ,eluted with 80 % methanol. Further, gradient elution was employed to separate microcystins in high performance liquid chromatography （ HPLC ） , the mobile phase consisted of 55 % methanol with small amount of trifluoroacetate acid （ TFA ） .The recoveries of MC-RR and MC-LR were 84.1 % and 89.9 % ,respectively. And the linear range is from 0.25 to 5g/L.The method is simple, sensitive, stable and suitable for the determination of the two most common Microcystins -LR,-RR.