Determination of 3H-labelled cytosine arabinoside and eight metabolites in cell extracts by high-performance liquid chromatography and scintillation spectrometry.
We have developed a method for the separation and quantitation of radiolabeled cytosine arabinoside and its eight metabolites in cell extracts by anion-exchange gradient high-performance liquid chromatography. Baseline separation of cytosine arabinoside and uracil arabinoside and their respective 5'-mono-, di- and triphosphates, as well as cytosine arabinoside diphosphocholine was obtained with the shortest interval between peaks being 3 min. This degree of separation was found to be essential for quantitation of 3H-labeled metabolites by scintillation counting of 1-min fractions. Application of this procedure to the quantitation of [3H] cytosine arabinoside and its metabolites from HL60 human leukemia cells is demonstrated.
Wang
L M
LM
Cancer Center Wake Forest University, Winston-Salem, NC 27103.
Woodward
C N
CN
White
J C
JC
Capizzi
R L
RL
D001086:Arabinofuranosyluracil / Q000032:analysis; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003561:Cytarabine / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007938:Leukemia / Q000378:metabolism; D012588:Scintillation Counting; D013056:Spectrophotometry, Ultraviolet; D014407:Tumor Cells, Cultured
D003561:Cytarabine; D001086:Arabinofuranosyluracil