Abstract: Objective To establish a method for the quantitative determination of adenosine phosphates in mouse brain by high performance liquid chromatography-variable wavelength detector(HPLC-VWD). Methods The protein in mouse brain tissue sample was precipitated with perchloric acid. Separation was performed on a reverse phase Supelco Discovery C18 column(150 mm×4.6 mm, 5 μm), equipped with a pre-column of the same material by Agilent 1200 HPLC-VWD. The wavelength was 254 nm. The mobile phase contained methanol-30 mmol/L potassium dihydrogen phosphate buffer, pH 5.85(5∶95, v/v) . The flow rate was set at 1.0 mL/min and the injection volume was 20 μL. The column temperature was 30 ℃. Results The linear relationship between the range(10~100) mg/L was good. The linear equations were ATP: A=28.924C+14.144, γ=0.999 90; ADP: A=31.504C-12.608, γ=0.999 92; AMP: A=46.539C-5.255, γ=0.999 95. The average recovery rate was 95.3%~103.2%. The RSDs of intra-and interday were 1.14%~1.75% and 2.45%~3.97%, respectively. Conclusion The experimental data proved that the HPLC-VWD method could detect the concentration of ATP, ADP and AMP in the mouse brain on repeated hypoxic exposures accurately, rapidly and simply.

Key words: adenosine phosphates, high performance liquid chromatogram, brain tissue, mouse