口腔细菌终末代谢酸的反相梯度高效液相色谱法分析条件研究

A Conditional Study of Analyzing Oral Bacterial End Products with Gradient Reversed-phase High Performance Liquid Chromatography

目的 :探讨测定口腔细菌终末代谢酸的反相梯度高效液相色谱法最佳分析条件。方法 :采用高效液相色谱技术梯度洗脱程序 ,流速 1 0～ 2 0ml min ,检测波长 2 17nm ,流动相为 6 0 % 0 2mol L磷酸盐甲醇溶液pH 2 8,对常见的细菌终末代谢酸进行不同培养基、pH值、温度、萃取液、沉淀剂等分析条件进行研究。结果 :口腔细菌在牛心脑浸液 (BHI)培养液中经 4 8h厌氧培养 ,上清液在pH 1 7的酸度下用乙醚提取可得到理想的色谱分析结果。结论 :该方法可推广应用于细菌终末代谢酸鉴定。

Objective:The purpose of this study was to optimize the conditions of analyzing oral bacterial end products by using reversed_phase high performance liquid chromatography (RP_HPLC). Methods:Different culture fluid, temperature, solution pH, extract, deposition were used to investigate oral bacterial end products with HPLC. The optimal chromatographic parameters were: flow rate of 1 0 ml/min and 2 0 ml/min, detection UV at 217 nm, 5% methanol in phosphate buffer solution (pH 2 8) as mobile phase, linear gradient wash with 5% to 40% methanol for 10 min, at ambient temperature. Results:Oral bacteria cultured in brain heart infusion broth (BHI); their supernatant was adjusted to pH 1 7, and then extracted by ether. With the above conditions, an optimal result can be obtained by analyzing using HPLC. Conclusion:The results of this paper will facilitate the identification and classification of oral bacteria by analyzing oral bacterial end products.\;