细菌活细胞分离的HPLC研究

HPLC Study on Separation of Viable Bacterium Cells

文章研究了利用离子交换高效液相色谱法对细菌活细胞进行分析过程中,不同色谱条件对分离效果的影响,确定了利用HPLC分离细菌活细胞的最佳分离条件:采用YOPEARL SuperQ-650C强阴离子交换树脂,0.02 mol/L哌嗪盐酸缓冲液(pH7.0)平衡;0.05~0.7 mol/L NaCl线性洗脱40 min,再用1.0 mol/L NaCl洗脱10 min;流速:1 mL/L;并对大肠杆菌、变形杆菌、植物乳杆菌三种菌株组成的肠道混合菌株进行色谱分离表征,建立了利用高效液相色谱分离细菌活细胞的最佳色谱条件。

The influence of isolation effect of different chromatographic conditions of viable bacterium cells by high performance ion-exchange liquid chromatograph was studied.The optimal separation conditions for separating bacterial living cells by HPLC were determined:YOPEARL SuperQ-650C anion resin,equilibrated 0.02mol/L piperazin-hydrochloric acid buffer(pH7.0),eluted with gradient of 0.05~0.75%NaCl/40min,then eluted with 100%NaCl/10min at the flow rate:1ml/L.Under these conditions,mixed intestinal mixed strains composed of Escherichia coli,Protens vulgaris,Lactobacillus plantarum were properly isolated.The optimal chromatographic conditions for the separation of living bacterial cells by high performance liquid chromatography have been established,which provides a simple and efficient method for further research on the number changes and interactions of mixed strains in the micro-ecological system.