Thacker D L, Abernethy D R, Raines A, Flockhart D A
Department of Pharmacology, Georgetown University Medical Center, Washington, DC 20007, USA.
J Chromatogr B Biomed Sci Appl. 1998 Jun 12;710(1-2):149-55. doi: 10.1016/s0378-4347(98)00096-6.
A method is described for the measurement of 5,5-diphenylbarbituric acid in plasma using high-performance liquid chromatography with UV detection. Briefly, the compounds are separated on a C18 reversed-phase column using a mobile phase of 50 mM sodium acetate (pH 4.5) and methanol. The flow-rate is 1.0 ml/min and 25 microl are injected and detected at 215 nm. The method is specific and sensitive in the range of concentrations tested, with a limit of quantification of 0.25 microg/ml. The calibration curves are linear for concentrations between 0.25 and 10 microg/ml. Intra-day and inter-day coefficients of variation are less than 8.5 and 10.5%, respectively, over the linear range. Intra-day and inter-day bias are less than 7.0 and 8.0%, respectively. A pharmacokinetic study conducted in male Beagle dogs administered 10 mg/kg of 1,3-dimethoxymethyl-5,5-diphenylbarbituric acid or 8 mg/kg of 5,5-diphenylbarbituric acid intravenously demonstrates the utility of this method.
描述了一种使用带紫外检测的高效液相色谱法测量血浆中5,5-二苯基巴比妥酸的方法。简而言之，化合物在C18反相柱上分离，使用50 mM醋酸钠（pH 4.5）和甲醇的流动相。流速为1.0 ml/min，进样25微升，并在215 nm处进行检测。该方法在所测试的浓度范围内具有特异性和灵敏性，定量限为0.25微克/毫升。校准曲线在0.25至10微克/毫升的浓度范围内呈线性。在该线性范围内，日内和日间变异系数分别小于8.5%和10.5%。日内和日间偏差分别小于7.0%和8.0%。在雄性比格犬静脉注射10 mg/kg的1,3-二甲氧基甲基-5,5-二苯基巴比妥酸或8 mg/kg的5,5-二苯基巴比妥酸后进行的药代动力学研究证明了该方法的实用性。
