[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.


CONCLUSIONS
This method is rapid, precise, accurate and suitable for the determination of the high energy nucleotides in hydrocephalus tissues.
Sun
Ai-min
AM
Analytical Test Center, Sichuan University, Chengdu 610041, China.
Wang
En-ren
ER
Mao
Bo-yong
BY
Chao
Ruo-bing
RB
Huang
Xun
X
Xu
Xiu-cheng
XC
D000244:Adenosine Diphosphate / Q000032:analysis; D000249:Adenosine Monophosphate / Q000032:analysis; D000255:Adenosine Triphosphate / Q000032:analysis*; D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003401:Creatine / Q000032:analysis; D004285:Dogs; D006849:Hydrocephalus / Q000378:metabolism*; D008297:Male; D010725:Phosphocreatine / Q000032:analysis*; D011897:Random Allocation; D012680:Sensitivity and Specificity
D010725:Phosphocreatine; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D003401:Creatine