TY - JOUR
T1 - High-performance liquid chromatography method for the determination and quantitation of arabinosylguanine triphosphate and fludarabine triphosphate in human cells
AU - Rodriguez, Carlos O.
AU - Plunkett, William
AU - Paff, Melanie T.
AU - Du, Min
AU - Nowak, Billie
AU - Ramakrishna, Prameen
AU - Keating, Michael J.
AU - Gandhi, Varsha
N1 - Funding Information:
This work was supported in part by grants CA57629 and CA32839 from the National Cancer Institute, Department of Health and Human Services and by a sponsored agreement with Glaxo Wellcome, Inc. The authors are thankful to Dr. Maureen Goode for critically editing the manuscript.
PY - 2000/8/18
Y1 - 2000/8/18
N2 - A gradient anion-exchange high-performance liquid chromatographic assay was developed for the simultaneous determination and quantitation of the cytotoxic triphosphates of arabinosylguanine (ara-GTP) and fludarabine (F- ara-ATP). The method was validated with respect to selectivity, recovery, linearity, precision, and accuracy using authentic standards. To test this assay in a more complex biological matrix, perchloric acid extracts of circulating human leukemia cells spiked with known concentrations of ara-GTP and F-ara-ATP were examined. Finally, to assess the clinical utility of our method, perchloric acid extracts of circulating human leukemia cells isolated from patients treated with fludarabine and nelarabine were analyzed. The range of quantitation was 0.0125-10 nmol for the ara- and native NTPs in cellular extracts. This assay should be helpful in establishing the mechanistic rationales for drug scheduling and combinations of nelarabine and fludarabine, and for correlating the therapeutic efficacy and levels of the cytotoxic triphosphates in target cells. (C) 2000 Elsevier Science B.V.
AB - A gradient anion-exchange high-performance liquid chromatographic assay was developed for the simultaneous determination and quantitation of the cytotoxic triphosphates of arabinosylguanine (ara-GTP) and fludarabine (F- ara-ATP). The method was validated with respect to selectivity, recovery, linearity, precision, and accuracy using authentic standards. To test this assay in a more complex biological matrix, perchloric acid extracts of circulating human leukemia cells spiked with known concentrations of ara-GTP and F-ara-ATP were examined. Finally, to assess the clinical utility of our method, perchloric acid extracts of circulating human leukemia cells isolated from patients treated with fludarabine and nelarabine were analyzed. The range of quantitation was 0.0125-10 nmol for the ara- and native NTPs in cellular extracts. This assay should be helpful in establishing the mechanistic rationales for drug scheduling and combinations of nelarabine and fludarabine, and for correlating the therapeutic efficacy and levels of the cytotoxic triphosphates in target cells. (C) 2000 Elsevier Science B.V.
KW - 506U78
KW - Arabinosylguanine
KW - Fludarabine
KW - Nelarabine
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U2 - 10.1016/S0378-4347(00)00303-0
DO - 10.1016/S0378-4347(00)00303-0
M3 - Article
C2 - 11043760
AN - SCOPUS:0343826903
SN - 1387-2273
VL - 745
SP - 421
EP - 430
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
IS - 2
ER -