|
|
Optimization of HPLC conditions for determination of biological active compounds.
Kotíková, Eva ; Kučera, Radim (advisor) ; Mokrý, Milan (referee)
Optimalization of HPLC conditions for determination of biological active compounds Diploma thesis Eva Kotíková Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Chemistry and Drug Control Adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosin monophosphate (AMP), creatine (C) and phosphocreatine (CP) are polar substances of great importance not only for the human body. In this work HPLC / UV method which allows the separation of these substances in isocratic elution was developed. As a suitable stationary phase zirconia column was chosen. The main advantage in comparison with the traditional silica columns is stability in the whole range of pH as well as at temperatures up to 200 žC. The best separation was achieved on a ZirChrom® - PHASE column by using a binary mobile phase with flow rate 1 ml/min - Part A: 10 mM hydrogenphosphate buffer, pH 7 and Part B: 100% acetonitrile. The detection was performed by using a UV detector at wavelengths of 254 nm for ATP, ADP, AMP, cAMP and 215 nm for C and CP. The final retention time valves under these conditions were: cAMP = 2.1 min, C = 3.7 min, AMP = 4.7 min, CP = 6.4 min, 10.7 min = ADP, ATP = 27.9 min. In the analysis of cardiac tissue the sample was mechanically homogenised with 57 µl of 100mM...
|
guest ::
RSS feed.