湖南水华微囊藻中藻毒素提取方法对比

The Comparison of Microcystins Extraction Methods from Microcystis in Hunan Province during Cyanobacterial Blooms

摘要: 于湖南省某富营养化水体采集一定量的微囊藻，在5%冰乙酸萃取、煮沸处理、研磨处理、反复冻融四种方法中选择出一种效率最高的藻毒素提取方法。利用某天然水华水体的微囊藻为原料，设计实验对比了5%冰乙酸萃取、煮沸处理、研磨处理和反复冻融四种提取方法对微囊藻毒素的提取效果，用高效液相色谱法(HPLC)测量藻毒素浓度。5%冰乙酸萃取获得的三种藻毒素(MC-RR、MC-YR、MC-LR)浓度分别为：0.73 ± 0.03、0.43 ± 0.04、0.85 ± 0.06 μg/mL；煮沸处理获得的三种藻毒素浓度分别为：0.97 ± 0.03、0.76 ± 0.12、0.77 ± 0.03 μg/mL，研磨处理获得的三种藻毒素浓度分别为：1.07 ± 0.02、0.46 ± 0.09、1.17 ± 0.22 μg/mL，反复冻融获得的三种藻毒素浓度分别为：1.20 ± 0.15、0.93 ± 0.01、1.83 ± 0.17 μg/mL，四种方法所获藻毒素总浓度分别为2.01 ± 0.01 μg/mL、2.49 ± 0.06 μg/mL、2.7 ± 0.07 μg/mL、3.96 ± 0.31 μg/mL。因此，4种藻毒素提取方法的效率由高到低依次为反复冻融、研磨处理、煮沸处理和5%冰乙酸萃取法。

Abstract: Using Microcystis in Hunan Province during cyanobacterial blooms as raw materials, four micro-cystins (MCs) extraction methods (5% acetic acid method, boiling method, grinding method and freeze-thaw method) were compared in order to find the most efficient one. The concentrations of MCs were measured by high performance liquid chromatography (HPLC). The concentrations of three MCs (MC-RR, MC-YR and MC-LR, the same below) obtained from 5% acetic acid extraction were 0.73 ± 0.03, 0.43 ± 0.04 and 0.85 ± 0.06 μg/mL. The concentrations of three MCs obtained by boiling method were 0.97 ± 0.03, 0.76 ± 0.12 and 0.77 ± 0.03 μg/mL. The concentrations of three MCs obtained by grinding method were 1.07 ± 0.02, 0.46 ± 0.09 and 1.17 ± 0.22 μg/mL. The con-centrations of three MCs obtained by freeze-thaw method were 1.20 ± 0.15, 0.93 ± 0.01 and 1.83 ± 0.17 μg/mL. The total concentrations of MCs obtained by the four methods were 2.01 ± 0.01 μg/mL, 2.49 ± 0.06 μg/mL, 2.7 ± 0.07 μg/mL and 3.96 ± 0.31 μg/mL. Therefore, the highest MCs extraction efficiency method is freeze-thaw method, which can obtain more MCs than grinding method, boiling method and 5% acetic acid extraction.