铜绿微囊藻中磷酸腺苷的提取及分析

Extraction and Analysis of Adenosine Phosphate in Cells of Microcystis Aeruginosa

比较了酸提取、有机溶剂提取、MgSO4水溶液加热提取以及MgSO4水溶液加热超声波提取4种方式对磷酸腺苷（ATP、ADP和AMP）的提取效果，确定以MgS04水溶液加热超声波提取效果最佳。采用MgSO4加热超声波提取时，2mL提取液对ATP、ADP和AMP的提取效果较好。将ATP、ADP和AMP的混合标准溶液放于沸水浴中保温时，随着保温时间的延长，对ATP和AMP的影响较大，而对ADP的影响相对较小。实验结果证明，以MgSO4水溶液为提取液，用100℃加热10min后，在超声波细胞粉碎机中超声破碎10min的提取效率最高，既简单又无毒。用反相高效液相色谱等强度洗脱分离与紫外检测分析藻细胞中的ATP、ADP和AMP的含量，在较短的时间内（10min）实现了较好的分离，分析准确而快速，是一较好的定性和定量分析方法。ATP、ADP和AMP的回收率分别为88％～97％、103％～107％和109％～115％，均在80％～120％之间，并且标准偏差和相对标准偏差均小于10％，证实了可以用加热超声波破碎提取，HPLC分析ATP、ADP和AMP的方法来提取和分析藻细胞中的ATP、ADP和AMP。

Four methods, perchloric acid extraction, extraction and boiling bitter salt solution with ultrasonic organic solvent extraction, boiling bitter salt solution extraction, were used to extract adenosine triphosphate （ATP）, denosine diphosphate （ADP） and adensine monophosphate （AMP） from the cells of Microcystis aeruginosa. Bitter salt solution with ultrasonic extraction was testified to be the optimum method. The optimum volume of extracting solution was 2 mL by the method of boiling bitter salt solution with ultrasonic. When ATP, ADP and AMP were put in boiling water bar, the concentration of ATP and AMP changed largely but the content of ADP changed smaller. In this study, the cells were put into boiling water bar for 10 min and then were staved with ultrasonic for 10 min with bitter salt solution as extractive solution. This method was simple and nonpoisonous. A reversed phase high-performance liquid chromatography with UV detection was used for the determination of ATP, ADP and AMP. They were distinctly separated within 10 min. Moreover, the method has the advantage of high sensitivity, rapidity and accurcy. The recovery of ATP, ADP and AMP was 88% -97%, 103% -107%, 109% -115% respectively. The relative standard deviations were less than 10%