Ganes D A, Wagner J G
College of Pharmacy, University of Michigan, Ann Arbor 48109.
J Chromatogr. 1988 Nov 18;432:233-42. doi: 10.1016/s0378-4347(00)80648-9.
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).
开发了一种使用高效液相色谱（HPLC）的灵敏且特异的方法，用于定量测定血浆中的5-溴-2'-脱氧尿苷（BUdR）和5-溴尿嘧啶（BU）。首先用乙酸乙酯和2-丙醇的混合物从用固体硫酸铵预饱和的血浆中提取BUdR和BU。有机提取物蒸发后，将剩余残渣用饱和硫酸铵溶液复溶，用正戊烷 - 二氯甲烷混合物洗涤，再用原溶剂混合物重新提取。有机提取物蒸发后，用流动相复溶，并在常规孔径的ODS HPLC柱上进行色谱分析，采用紫外吸收检测。BUdR和BU的定量限均为0.1 microM，批内变异系数均值分别为5.0%和5.6%，批间变异系数均值分别为5.4%和10.7%。该方法用于测定一名接受连续静脉输注BUdR（每天20 mg/kg）的患者的稳态股动脉和肝静脉血浆中BUdR和BU的浓度。
