Li X, Yao T W, Zeng S
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, China.
J Chromatogr B Biomed Sci Appl. 2000 Jun 9;742(2):433-9. doi: 10.1016/s0378-4347(00)00178-x.
A reversed-phase high-performance liquid chromatographic method for the determination of the enantiomers of atenolol in rat hepatic microsome has been developed. Racemic atenolol was extracted from alkalinized rat hepatic microsome by ethyl acetate. The organic layer was dried with anhydrous sodium sulfate and evaporated using a gentle stream of air. Atenolol racemic compound was derivatized with 2,3,4,6-tetra-O-acetyl-beta-D-glycopyranosyl isothiocyanate at 35 degrees C for 30 min to form diastereomers. After removal of excess solvent, the diastereomers were dissolved in phosphate buffer (pH 4.6)-acetonitrile (50:30). The diastereomers were separated on a Shimadzu CLC-C18 column (10 microm particle size, 10 cm x 0.46 cm I.D.) with a mobile phase of phosphate buffer-methanol-acetonitrile (50:20:30, v/v) at a flow-rate of 0.5 ml/min. A UV-VIS detector was operated at 254 nm. For each enantiomer, the limit of detection was 0.055 microg/ml (signal-to-noise ratio 3) and the limit of quantification (signal-to-noise ratio 10) was 0.145 microg/ml (RSD <10%). In the range 0.145-20 microg/ml, intra-day coefficients of variation were 1.0-7.0% and inter-day coefficients of variation were 0.4-16.5% for each enantiomer. The assay was applied to determine the concentrations of atenolol enantiomers in rat hepatic microsome as a function of time after incubation of racemic atenolol.
已开发出一种反相高效液相色谱法，用于测定大鼠肝微粒体中阿替洛尔对映体。外消旋阿替洛尔通过乙酸乙酯从碱化的大鼠肝微粒体中萃取出来。有机层用无水硫酸钠干燥，并用温和的气流蒸发。外消旋阿替洛尔化合物在35℃下与2,3,4,6-四-O-乙酰基-β-D-吡喃葡萄糖基异硫氰酸酯反应30分钟，形成非对映异构体。除去过量溶剂后，将非对映异构体溶解于磷酸盐缓冲液（pH 4.6）-乙腈（50:30）中。非对映异构体在岛津CLC-C18柱（粒径10μm，10cm×0.46cm内径）上分离，流动相为磷酸盐缓冲液-甲醇-乙腈（50:20:30，v/v），流速为0.5ml/min。紫外-可见检测器在254nm波长下运行。对于每种对映体，检测限为0.055μg/ml（信噪比3），定量限（信噪比10）为0.145μg/ml（相对标准偏差<10%）。在0.145-20μg/ml范围内，每种对映体的日内变异系数为1.0-7.0%，日间变异系数为0.4-16.5%。该测定法用于测定外消旋阿替洛尔孵育后大鼠肝微粒体中阿替洛尔对映体的浓度随时间的变化。
