[Separation of natural ribonucleoside triphosphates and their determination in an acid-soluble cell fraction by anion exchange high performance liquid chromatography].
An anion-exchange HPLC procedure for CTP, UTP, ATP and GTP determination in the acid-soluble fraction of cells is described. Ribonucleoside triphosphates are separated on LiChrosorb AN isocratically with NH4H2PO4-acetonitrile. The dependence of the separation efficiency on the salt and acetonitrile concentrations and pH was analyzed and the optimal conditions were chosen. The range, wherein the linearity between the ribonucleoside triphosphate amount and the area of the corresponding peak is observed, was defined and the regression equations were derived. The CTP, UTP, ATP and GTP content in the ovarian cancer cells CaOv in culture was found to be 418 +/- 32, 1122 +/- 21, 9262 +/- 442 and 1036 +/- 49 pmole/10(6) cells, respectively. After 2 hr incubation with 6-mercaptopurine (10(-4) M) the level of ATP and GTP is reduced by 55%, and after 24 hr incubation--by 73% for ATP and 85% for GTP. At the same time the UTP and CTP content is decreased by 12-31%.
D000255:Adenosine Triphosphate / Q000032:analysis; D002458:Cell Fractionation; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003570:Cytidine Triphosphate / Q000032:analysis; D005260:Female; D006160:Guanosine Triphosphate / Q000032:analysis; D006801:Humans; D010051:Ovarian Neoplasms / Q000032:analysis*; D012265:Ribonucleotides / Q000032:analysis*; D014544:Uridine Triphosphate / Q000032:analysis
D012265:Ribonucleotides; D003570:Cytidine Triphosphate; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D014544:Uridine Triphosphate
Razdelenie prirodnykh ribonukleozidtrifosfatov i ikh opredelenie v kislotorastvorimoĭ fraktsii kletok metodom anionoobmennoĭ zhidkostnoĭ khromatografii vysokogo razresheniia.