Simultaneous Determination of Adenosine Phosphate and Coenzyme Ⅰ in Cells of Saccharomyces cerevisiae by RP-HPLC

Abstract:

In this manuscript,an RP-HPLC method was developed to determine simultaneously adenosine phosphate and coenzyme I in Saccharomyces cerevisiae,in order to monitor metabolic state.The intracellular metabolites extracted by the method of rubbing with liquid nitrogen were separated on an Agilent Zorbax SB-Aq column(5 μm,4.6 mm×150 mm).A 0.025 mmol/L triethylamine-phosphoric acid buffer solution(pH 6.0) and acetonitrile mixture was used as mobile phase,with a flow rate of 1 mL/min.Detection wavelength was set at 254 nm.Under these conditions,three kinds of adenosine phosphate and two kinds of coenzyme I could be well separated.The recoveries were between 98.30% and 103.42%,and the RSD of the method was less than 3%.The results showed that this method is sensitive,accurate,reproducible and can be applied to determinate the content of intracellular adenosine phosphate and coenzyme I of S.cerevisiae.