[Determination of creatine, phosphocreatine and adenosinephosphates in experimental hydrocephalus tissue by reversed-phase high performance liquid chromatography].

Objective: To investigate the state of impaired cerebral energy metabolism of the hydrocephalus tissue.

Methods: Adult male dogs were used for establishing the model of kaolin-induced hydrocephalus. A simple and rapid method for the simultaneous determination of creatine (Cr), phosphocreatine (Crp), and adenosinephosphates (AMP, ADP, ATP) in different stages of experimental hydrocephalus tissue by reversed-phase high performance liquid chromatography (RP-HPLC) has been established. The chromatographic conditions were as follows: Inertsil ODS-3 C18 column (4.6 mm x 250 mm i.d. 5 microns), the mobile phase being composed of KH2PO4 buffer (330 mmol/L)-acetonitrile-TBA (45 mmol/L) (94:5.5:0.5) (pH = 6.27) and detector at 210 nm.

Results: The calibration curve showed a good linearity in the mass concentration range of 5.69-3640.50 mumol/L (r = 0.9993) for Cr, 3.47-555.50 mumol/L (r = 0.9999) for Crp, 2.69-1723.00 mumol/L (r = 0.9993) for AMP, 2.66-1704.00 mumol/L (r = 0.9999) for ADP and 2.94-1883.50 mumol/L (r = 0.9999) for ATP. The recoveries ranged from 90.10% to 107.00% with relative standard deviations from 1.58% to 3.88%. The detection limits of this method were 3.55-5.84 mumol/L. By means of this method, the Cr, Crp, AMP, ADP and ATP in different stages of 16 dogs experimental hydrocephalus tissue were determined with satisfactory results.

Conclusion: This method is rapid, precise, accurate and suitable for the determination of the high energy nucleotides in hydrocephalus tissues.