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 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 :3 5 4、4 31、4 6 1、5 82、6 5 0、14 72、2 4 75min。此方法的相对标准差 (RSD)为 1 8% ,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 。
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 chromatograph (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 acids were optimized using the method of gradient RP_HPLC. Results: The study showed that the optimal chromatographic parameters were: 1 0 ml/min and 2 0 ml/min of flow rates, and the wavelength of ultraviolet detection was 217 nm. The composition of eluent: the ratio of phosphate buffer (pH 2 8) to methanol is 95∶5 (v/v), gradient 5%～50%. The liner gradient wash will be performed in 10 minutes at room temperature. The average recovery rate was 92%, and RSD was 1 8%.Conclusion:The results of this paper will be helpful to determine acid composition of end products of oral bacteria.\;
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2001年第5期294-295,299,共3页
West China Journal of Stomatology
关键词
反相梯度高效液相色谱法
短链脂肪酸
口腔细菌学
reversed_phase high_perfermance liquid chromatography
short_chain fat acid
oral bacteria
