Metcalf J S, Codd G A
Department of Biological Sciences, University of Dundee, Dundee, UK.
FEMS Microbiol Lett. 2000 Mar 15;184(2):241-6. doi: 10.1111/j.1574-6968.2000.tb09021.x.
Low-cost, straightforward methods for the extraction of microcystins and nodularins from cyanobacterial cells were developed using a microwave oven and boiling waterbath. The use of organic solvents, such as methanol, which can interfere with sensitive analytical procedures, e.g. immunoassays, can thus be avoided. Analysis by protein phosphatase inhibition assay and high performance liquid chromatography indicated that purified microcystin-LR was unaffected by the microwave oven and boiling waterbath treatments. Four microcystins of differing hydrophobicities were successfully extracted from Microcystis PCC 7813 by both treatments at yields equivalent to those obtained by longer protocols using methanol. Assessment of the microwave oven and boiling waterbath extraction methods with laboratory strains and environmental samples of cyanobacteria showed good correlation with results from lyophilisation and methanol extraction, when extracts were analysed by high performance liquid chromatography with diode array detection (R(2)>/=0.92). The microwave and boiling waterbath extraction methods also sterilised the environmental bloom samples, as evidenced by the abolition of heterotrophic bacterial growth.
利用微波炉和沸水浴开发了从蓝藻细胞中提取微囊藻毒素和节球藻毒素的低成本、简便方法。这样就可以避免使用可能干扰灵敏分析程序（如免疫测定）的有机溶剂，例如甲醇。通过蛋白磷酸酶抑制试验和高效液相色谱分析表明，纯化的微囊藻毒素-LR不受微波炉和沸水浴处理的影响。通过这两种处理方法，成功地从铜绿微囊藻PCC 7813中提取了四种不同疏水性的微囊藻毒素，其产量与使用甲醇的较长方案所获得的产量相当。当用二极管阵列检测的高效液相色谱分析提取物时，对实验室菌株和蓝藻环境样品的微波炉和沸水浴提取方法的评估显示，与冻干和甲醇提取的结果具有良好的相关性（R²≥0.92）。微波炉和沸水浴提取方法还对环境水华中的样品进行了消毒，异养细菌生长的消除证明了这一点。
