[Determination of phosphocreatine in muscular tissues by high performance liquid chromatography]

The quantitation of energy stores(phosphocreatine, PCr) is of paramount importance in the study of living-tissues metabolism. This is more so in the heart, which depends to a very great extent on an uninterrupted aerobic metabolism to maintain its normal function. Availability of data on myocardial energy stores is, therefore, indispensable for assessing the responses of heart to drugs or stresses such as ischemia and hypoxemia. A simple and rapid method for the determination of phosphocreatine in muscular tissues by RP-HPLC has been developed. The chromatographic conditions were as follows: Zorbax XDB-C18 column(150 mm x 4.6 mm i.d., 5 microns), V(20 mmol/L KH2PO4 and 2 mmol/L tetrabutyl ammonium phosphate): V(acetonitrile) = 96:4(pH 5.8) mobile phase and UV detector at 215 nm. PCr in muscular tissues was extracted with 0.4 mol/L HClO4. The calibration curve showed a good linearity in 5 mg/L-100 mg/L(r = 0.9992). The average recovery was 99.34%. The limit of detection was 2 mg/L. The verified results demonstrated that this method is precise, accurate and can be used for determination of PCr in muscular tissue.