青枯菌HPLC分析中样品制备方法的优化

Optimization of Ralstonia Solanacearum Preparation Methods by HPLC Analysis

采用高效离子交换色谱(HPLC)和紫外-激光光散射仪在线双检测系统研究了青枯菌细胞的制备方法对细胞生命活力和细胞表面特性的影响。通过青枯菌在不同悬浮缓冲液中菌数变化的统计、不同离心速度和洗涤次数对青枯菌色谱行为影响的分析以及不同色谱峰青枯菌细胞的透射电子显微镜观察,结果表明:采用5000r/min离心10min收集菌体细胞、超纯水(>16MΩ)悬浮和洗涤青枯菌、重复洗涤二次的样品制备方法,既可以避免培养基成分对青枯菌色谱行为造成的干扰,又可以保持青枯菌细胞的生命活力和细胞表面的原有性质。

High performance ion -exchange liquid chromatography （HPLC） equipped with UV and laser scattering light detectors were used to study the influence of cell preparation methods of Ralstonia solanacearum on the cell viability and cell surface properties. By calculating bacterial number in different suspension buffer,analyzing chromatographic graphs at different centrifugation speed and washing times, and observing three chromatographic peaks of Ralstonia solanacearum under electro - microscope. The results showed that the batter method was as follows： bacterial cultures harvested by centrifugation at 5000r/min for 10 min, washed and centrigugated twice, resuspended in millipore water （ 〉 16MΩ ）. Using this method could not only avoid the interference of medium on the chromatographic behavior of Ralstonia solanacearum, but also keep the cell viability and cell surface properties.