华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

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

张萍,朱硃,肖晓蓉,张玉,周松华   

  1. 610041 四川大学华西口腔医学院重点实验室(张 萍,朱 ,肖晓蓉),成都铁路中心医院口腔科(张 玉),上海铁路中心医院口腔科(周松华)
  • 收稿日期:2002-04-25 修回日期:2002-04-25 出版日期:2002-04-20 发布日期:2002-04-20

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

Zhang Ping, Zhu Zhu, Xiao Xiaorong et al   

  • Received:2002-04-25 Revised:2002-04-25 Online:2002-04-20 Published:2002-04-20

摘要:

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110~210 mlPmin,检测波长217 nm,流动相为60% 012 molPL磷酸盐甲醇溶液pH218,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48 h厌氧培养,上清液在pH 117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。

关键词: 细菌终末代谢酸, 高效液相色谱法, 口腔细菌, 分析条件

Abstract:

Objective:The purpose of this studywas 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 productswithHPLC. The optimal chromatographic parameterswere: flowrate of 110 mlPmin and 210 mlPmin, detection UV at 217 nm, 5% methanol in phosphate buffer solution (pH218) as mobile phase, linear gradientwashwith 5% to 40% methanol for10 min, at ambient temperature.Results:Oral bacteria cultured in brain heart infusion broth (BHI); their supernatantwas adjusted to pH117, and then extracted by ether. With the above conditions, an opti- mal 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.

Key words: bacterial end acid products, reversed-phase high performance liquid chromatography, oral bacteria, analytic conditions