小鼠肠道菌群HPLC分析中色谱分离条件的优化

Optimization of Separating Conditions of Mouse Gut Flora by High Performance Ion Exchange Chromatography Analysis

研究了在小鼠肠道菌群的高效离子交换色谱(HPLC)分析中,不同色谱分离条件对分离效果的影响,确立了最佳色谱条件:样品上样于Toyopearl TSK gel SuperQ-650c强阴离子交换树脂柱,以0.02mol/L哌嗪-HCl缓冲液(pH8.0)平衡,洗脱盐浓度为1.0mol/L NaCl,洗脱梯度为0.1～0.5mol/L NaCl线性梯度洗脱80min,再经0.5～0.75mol/L NaCl线性梯度洗脱25min,流速定为1ml/min,进样量1ml。小鼠肠道菌群HPLC分析方法的建立,为深入研究小鼠肠道菌群的组成和动态变化奠定了基础。

The influence of separating effect of different chromatographic conditions of mouse gut flora by high performance ion exchange chromatography analysis was studied.The optimum chromatographic conditions for separating gut bacteria were determined.The sample was applied to the chromatography column packed with Toyopearl SuperQ-650c anion resin,equilibrated with 0.02mol/L piperazin-hydrochloric acid buffer （ pH 8.0）,and elution salt 1 mol/L NaCl,eluted with the gradient of 0-50% NaCl/80 min,then 50% - 75% NaCl/25 min at the flow rate 1 ml/min,and injecting volume was 1 ml. Under these conditions,intestinal flora were separated into several fractions. The establishment of HPLC analysis method will lay a foundation of further research on the components of mouse gut flora and their dynamic changes.