Abdel-Fattah L, Abdel-Aziz L, El-Kosasy A, Gaied M
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, Ain Shams University, Cairo, Egypt.
Drug Discov Ther. 2010 Dec;4(6):418-22.
A simple, rapid, and sensitive method of reversed-phase high-performance liquid chromatography with fluorescence detection has been developed and validated for use in determining levels of nebivolol•HCl in human plasma. Sample preparation involves a simple single-step protein precipitation procedure and extraction of nebivolol in acetonitrile. The separation was performed on a Kromasil® RP-C18 column (Ф 4.6 mm × 250 mm, 5 μm) with a mobile phase consisting of 0.05 M potassium dihydrogen phosphate buffer/acetonitrile (40:60, v/v) adjusted to pH 3 using orthophosphoric acid. Analysis was carried out under isocratic conditions at a flow rate of 1.5 mL/min and at room temperature using a fluorescence detector with excitation at 288 nm and emission at 310 nm. The chromatographic run was 4 min. The calibration curve was linear over the concentration range 0.2-20 ng/mL. The method was validated in terms of its accuracy, precision, and specificity. The assay enabled the measurement of nebivolol with a minimum quantification limit of 0.16 ng/mL. The average recovery of nebivolol from spiked human plasma was 98.4 ± 3.3%. This method was successfully used in a pharmacokinetic study of oral administration of 5-mg tablets to healthy human volunteers.
已开发并验证了一种简单、快速且灵敏的反相高效液相色谱荧光检测法，用于测定人血浆中盐酸奈必洛尔的含量。样品制备包括简单的一步蛋白沉淀程序和用乙腈萃取奈必洛尔。分离在Kromasil® RP-C18柱（Ф 4.6 mm × 250 mm，5 μm）上进行，流动相由0.05 M磷酸二氢钾缓冲液/乙腈（40:60，v/v）组成，用正磷酸调节至pH 3。分析在等度条件下进行，流速为1.5 mL/min，在室温下使用荧光检测器，激发波长为288 nm，发射波长为310 nm。色谱运行时间为4分钟。校准曲线在0.2 - 20 ng/mL的浓度范围内呈线性。该方法在准确性、精密度和特异性方面得到了验证。该测定法能够测定奈必洛尔，最低定量限为0.16 ng/mL。从加标的人血浆中回收奈必洛尔的平均回收率为98.4 ± 3.3%。该方法已成功用于对健康人类志愿者口服5毫克片剂的药代动力学研究。
