An ion-pair reversed-phase HPLC method for analyzing the ATP, ADP and AMP contents in hippocampus of mice
10.13220/j.cnki.jipr.2016.06.023
- Author:
Hong-Chao ZHANG
1
Author Information
1. Guangzhou University of Traditional Chinese Medicine
- Publication Type:Journal Article
- Keywords:
ADP;
AMP;
ATP;
HPLC;
Senescence accelerated mouse
- From:
Journal of International Pharmaceutical Research
2016;43(6):1151-1155
- CountryChina
- Language:Chinese
-
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.
