An ion-pair reversed-phase HPLC method for analyzing the ATP, ADP and AMP contents in hippocampus of mice / 国际药学研究杂志

ABSTRACT

Objective To establish an ion-pair reverse-phased high-performance liquid chromatography(RP-HPLC)method for simultaneous determination of ATP,ADP and AMP in the hippocampus of mice. Methods The protein of mouse hippocampus was precipitated with perchloric acid,and neutralized with potassium carbonate-methanol mixture. Mobile phase was as follows50 mmol/L phosphate buffer(buffer for K2HPO4-KH2PO4,pH 6.60,containing 22%methanol,and 4 mmol/L tetrabutylammonium bisul?fate). Shimadzu HPLC system and Agilent C18 column(4.6 mm×250 mm,5μm)filled with the same material pre-column(12.5 mm× 4.6 mm,5μm)were used. The contents of ATP,ADP and AMP in mouse hippocampus were analyzed at a wavelength of 254 nm,the flow rate of 0.6 ml/min and room column temperature. Results Stability tests showed that intra-day and inter-day precision of the method were 1.27%-3.42%and 0.88%-3.52%,respectively,and recovery rates were 95.67%-104.05%. Conclusion The HPLC method established in this study is simple,accurate and efficient in detecting the levels of ATP,ADP,and AMP in mice hippocampus.