高效液相色谱法测定肌肉组织中的磷酸肌酸

Determination of Phosphocreatine in Muscular Tissues by High Performance Liquid Chromatography

采用反相离子对液相色谱法测定肌肉组织中的磷酸肌酸 (PCr)的含量。样品经高氯酸溶液沉淀蛋白 ,匀浆后离心 ,上清液用KOH溶液中和后用反相液相色谱法分离测定。方法的最低检测限为 2mg/L(S/N≥ 3) ,PCr在 5mg/L～ 10 0mg/L范围内与其峰面积呈良好的线性关系 (r =0 .9992 ) ;PCr的平均回收率为 99.34 % ;测定结果的日内RSD <4.42 % ,日间RSD <8.6 7%。方法准确、灵敏、快速 ,适用于动物和人体肌肉组织中PCr含量的测定。

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 C 18 column(150 mm×4.6 mm i.d.,5 μm), V (20 mmol/L KH 2PO 4 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 HClO 4. The calibration curve showed a good linearity in 5 mg/L 100 mg/L( r =0.999?2). 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.