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.