Determination of adenosine phosphates in L.bulgaricus by RP-HPLC method and its application

To evaluate the quenching effect of the quencheing solutions in the metabolomics research, a RP- HPLC method was developed for the determination of adenosine phosphates in L.bulgaricus. The chromategrap-hic separation was achieved on an SinoChrom ODS- BP C 18 column and a mobile phase of methanol30mmol /L phosphate ( V/V = 5 ∶ 95 ) buffer solution ( pH7 ) with a flow rate of 0.7mL /min. The column temperature and UV detection wavelength were set as 25℃ and 254nm, respectively. The method had the good linearity and higher accuracy.The recovery rates of adenosine phosphates were in the range of 94.12% ～ 109.6% .The intracellular metabolite leakages of L.bulgaricus was evaluated by the RP- HPLC method, when 60% ( V/V) methanol /water at -40℃ that was the most popular method to quench the metabolism.The results showed that the leak rate of ATP was as high as 70.43% ± 2.3% , EC value was 0.42 ± 0.015. It was inferred that the quenching fluid could not only cause severe leakage of intracellular metabolites but also not completetly quench the metabolism .The quenching procotol was not suitable for quenches L.bulgaricus.