Improved high-performance liquid chromatographic assay for the quantification of 5-bromo-2'-deoxyuridine and 5-bromouracil in plasma.
A sensitive and specific procedure using high-performance liquid chromatography (HPLC) was developed for the quantification of 5-bromo-2'-deoxyuridine (BUdR) and 5-bromouracil (BU) in plasma. BUdR and BU were first extracted with a mixture of ethyl acetate and 2-propanol from plasma presaturated with solid ammonium sulfate. Following evaporation of the organic extract, the remaining residue was reconstituted in saturated ammonium sulfate solution, washed with a mixture of n-pentane-methylene chloride and re-extracted with the original solvent mixture. The organic extract was evaporated, reconstituted in mobile phase and chromatographed on a regular-bore ODS HPLC column using ultraviolet absorbance detection. The BUdR and BU quantification limits were both 0.1 microM, the mean intra-assay coefficients of variation were 5.0 and 5.6%, respectively, and the mean inter-assay coefficients of variation were 5.4 and 10.7%, respectively. This method was used to determine steady-state femoral arterial and hepatic venous plasma concentrations of BUdR and BU in a patient receiving a continuous intravenous infusion of BUdR (20 mg/kg per day).
Ganes
D A
DA
College of Pharmacy, University of Michigan, Ann Arbor 48109.
Wagner
J G
JG
D000368:Aged; D001973:Bromodeoxyuridine / Q000097:blood*; D001976:Bromouracil / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D003110:Colonic Neoplasms / Q000097:blood / Q000188:drug therapy / Q000473:pathology; D006801:Humans; D007202:Indicators and Reagents; D007262:Infusions, Intravenous; D008113:Liver Neoplasms / Q000097:blood / Q000556:secondary; D008297:Male; D014498:Uracil / Q000031:analogs & derivatives / Q000097:blood
D007202:Indicators and Reagents; D001976:Bromouracil; D014498:Uracil; D001973:Bromodeoxyuridine; C048747:5-iodouracil