华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

短链脂肪酸的梯度高效液相色谱法分析研究

张萍,叶利民,肖晓蓉,朱硃,张玉   

  1. 610041 四川大学华西口腔医学院(张 萍,肖晓蓉,朱 ),四川大学华西药学院(叶利民),成都铁路中心医院 (张 玉)
  • 收稿日期:2001-10-25 修回日期:2001-10-25 出版日期:2001-10-20 发布日期:2001-10-20

A Study of Analyzing Short-chain Fat Acid by Gradient Reversed-phase High-perfermance Liquid Chromatography

Zhang Ping, Ye Liming, Xiao Xiaorong, et al   

  1. Key Laboratory,College of Stomatology,West China University of Medical Scienses
  • Received:2001-10-25 Revised:2001-10-25 Online:2001-10-20 Published:2001-10-20

摘要:

目的:探讨测定甲酸、乙酸、丙酸、丁酸、琥珀酸、戊酸、乳酸的分离条件,为检测细菌终末代谢酸奠定基础。方法:采用反相梯度高效液相色谱技术对同时分离7种短链脂肪酸所需的检测波长、流动相及梯度洗脱程序等条件进行研究。结果:高效液相色谱技术在流速110~210 mlPmin,检测波长217 nm,流动相为60% 012 molPL磷酸盐甲醇溶液pH218的条件下,采用线性梯度洗脱,能同时分离甲酸、乳酸、乙酸、琥珀酸、丙酸、丁酸、戊酸,其保留时间分别为:3154、4131、4161、5182、6150、14172、24175 min。此方法的相对标准差(RSD)为118%,平均回收率为92%,重复性变异系数小于4%。结论:反相梯度高效液相色谱技术可测定同一标本中的乳酸、甲酸、乙酸、丙酸、丁酸、琥珀酸、戊酸7种短链脂肪酸,这一结果为测定细菌终末代谢酸提供了重要的手段。

关键词: 反相梯度高效液相法, 短链脂肪酸, 口腔细菌

Abstract:

Objective:The objective of this study is to obtain the optimized separating conditions of formic acid, acetic acid, propionic acid, butyric acid, valeric acid, lactic acid, succinic acid from bacteria using reversed-phase high performance liquid chromato- graph (RP-HPLC), and to set up a reliable progress in identifying the end-products of oral bacteria.Methods: The conditions of separating seven short-chain fat acidswere optimized using the method of gradient RP-HPLC.Results: The study showed that the optimal chromatographic parameters were: 110 mlPmin and 210 mlPmin of flow rates, and the wavelength of ultraviolet detection was 217 nm. The composition of eluent: the ratio of phosphate buffer(pH218) tomethanol is95B5 (vPv), gradient5%~50%. The liner gradientwash will be performed in 10 minutes at room temperature. The average recovery rate was 92%, and RSDwas 118%.Conclusion:The results of this paperwill be helpful to determine acid composition of end products of oral bacteria.

Key words: reversed-phase high-perfermance liquid chromatography, short-chain fat acid, oral bacteria