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 follows 50 mmol/L phosphate buffer (buffer for K2HPO4-KH2PO4, pH 6.60, containing 22% methanol, and 4 mmol/L tetrabutylammonium bisulfate). Shimadzu HPLC system and Agilent C18 column (4.6 mm×250 mm, 5 μm) filled with the same material pre-column (12.5 mm×4.6 mm, 5 μm) 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.