Study On Synthesis Of Phosphocreatine Disodium Salt

Creatine phosphate disodium salt also called phosphocreatine disodium salt, is an important energy storage substance of body, which can serve as nutriment and clinicl medicine directly. It has been widely used preventing and treating different heart disease and also as biochemistry reagent on clinical diagnosis.The synthesis of creatine phosphate disodium salt by the phosphorylation of creatine with POCl3 in the alkaline solution was studied. The appropriate reaction conditions were determined by changing the temperature and reaction time, the amount of alkaline and raw material.Cation-exchange resin was introduced instead of traditional Na2SO4 precipitation method, thus phosphocreatine barium changed to phosphocreatine disodium salt, the appropriate resin was selected and the good performance condition was determined. The reaction liquid was isolated with anion-exchange resin instead of traditional adding organic solvent method, the different types of strong-base ion-exchange resins were compared and then the best was chosen out .The suitable concentration and flow rate of sample solution was determined, 0.2mol/L CaCl2 was selected as eluent and suitable elution flowrate is 2BV/h .The yield of three methods including precipitation, precipitation combining to resin and resin are almost consistent, but resin method can increase the purity of product from 70.9% to 90.1%, and the excessive SO42-remained in product could be avoided.The crystallization condition of phosphocreatine disodium was investigated. The best conditions were:ethanol as crystallization solvent, crystallization temperature is -10℃,concentration of phosphocreatine disodium salt is 5%,adding four volume ethanol, using ultrasonic nucleation make crystallization time shorten much. The purity of the refining product can exceed 99% and yield reachs to 93.4%.A method to determin the contents of phosphocreatine disodium salt by high performance liquid chromatography was established. The influential factor such as detective wavelength, the concentration of inorganic salt, the proportion of organic solvent, pH value, temperature, theconcentration of ion-pairing agent was studied. The optimal chromatographic parameters were deceided. On these conditions, the linear range of phosphocreatine disodium salt is 12.3mg/L-98.2mg/L, the detection limit is 0.8mg/L.Another method of determinating its content with phosphomolybdate blue was studied. The selected reducing agent is SnCl2, the maximum absorptive wavelengh of phosphomolybdate blue is 706 nm and the suitable colour-developed time is 30 minutes .The linear regression line is well. Compared to high-performance liquid chromatography method, the method is simple and rapid, the results show relative good accuracy and the standard deviation is under 0.6%.The quality of product reaches to foreign standard by analysis of NMR, IR and HPLC.