Metcalf J S, Beattie K A, Saker M L, Codd G A
Division of Environmental and Applied Biology, School of Life Sciences, University of Dundee, DD1 4HN, Dundee, UK.
FEMS Microbiol Lett. 2002 Nov 5;216(2):159-64. doi: 10.1111/j.1574-6968.2002.tb11430.x.
The effect of organic solvents on the high performance liquid chromatography (HPLC) analysis of cylindrospermopsin using photodiode array detection was examined since organic solvents are commonly used to extract this toxin from cyanobacteria and in the mobile phase compositions used in HPLC. Increasing concentrations of methanol resulted in an increase in the UV absorbance of purified cylindrospermopsin according to spectrometry, but to a marked decrease during HPLC analysis when the concentration of this solvent was greater than 50% methanol, or when acetonitrile concentrations exceeded 30% (v/v). Precipitation of cylindrospermopsin at these high concentrations of organic solvents was not observed. Solid phase extraction methods were developed to recover the toxin from spent extracellular growth medium after laboratory culture of Cylindrospermopsis raciborskii strain CR3 as an aid to toxin purification and from spiked environmental water samples. Using C18 and polygraphite carbon cartridges in series, 100% recoveries of cylindrospermopsin were achieved for lake waters spiked at 1 micro g l(-1).
由于有机溶剂常用于从蓝藻中提取这种毒素以及用于高效液相色谱（HPLC）分析的流动相组成中，因此研究了有机溶剂对使用光电二极管阵列检测的柱孢藻毒素高效液相色谱分析的影响。根据光谱分析，甲醇浓度的增加会导致纯化的柱孢藻毒素的紫外吸光度增加，但在HPLC分析中，当该溶剂浓度大于50%甲醇或乙腈浓度超过30%（v/v）时，紫外吸光度会显著降低。未观察到柱孢藻毒素在这些高浓度有机溶剂中沉淀。开发了固相萃取方法，用于从实验室培养的拉氏柱孢藻菌株CR3的细胞外生长培养基中回收毒素，以辅助毒素纯化，以及从加标的环境水样中回收毒素。串联使用C18和多石墨碳柱，对于加标浓度为1微克/升的湖水，柱孢藻毒素的回收率达到100%。
