Can Nafiz O, Arli Goksel
Anadolu University, Faculty of Pharmacy, Department of Analytical Chemistry, Yunusemre Campus, Eskisehir 26470, Turkey.
J AOAC Int. 2010 Jul-Aug;93(4):1077-85.
Development and validation of an RP-HPLC method for determination of levetiracetam in pharmaceutical tablets is described. The separation and quantification of levetiracetam and caffeine (internal standard) were performed using a single analytical procedure with two different types of stationary phases, conventional Phenomenex Gemini C18 (100 x 4.6 mm, 5 microm) and Merck Chromolith Performance RP18e (100 x 4.6 mm, macropore size 2 mm, micropore size 13 nm) monolithic silica. Five-microliter aliquots of samples were injected into the system and eluted using water-acetonitrile (90 + 10, v/v) mobile phase pumped at the rate of 1 mL/min. The analyte peaks were detected at 200 nm using a diode array detector with adequate resolution. Validation studies were performed using the method recommended by the International Conference on Harmonization, the U.S. Pharmacopeia, and AOAC INTERNATIONAL, which includes accuracy, precision, range, limits, robustness, and system suitability parameters. Levetiracetam and caffeine were detected in about 7 min using the conventional column, whereas less than 5 min was required when the monolithic column was used. Calibration plots had r values close to unity in the range of 0.8-8.0 microg/mL. Assay of levetiracetam in a tablet formulation was demonstrated as an application to real samples.
本文描述了一种用于测定药物片剂中左乙拉西坦的反相高效液相色谱（RP-HPLC）方法的开发与验证。左乙拉西坦和咖啡因（内标）的分离与定量通过单一分析程序进行，使用两种不同类型的固定相：传统的菲罗门双子座C18柱（100×4.6 mm，5μm）和默克整体硅胶Chromolith Performance RP18e柱（100×4.6 mm，大孔尺寸2 mm，微孔尺寸13 nm）。将5微升的样品等分试样注入系统，使用以1 mL/min流速泵送的水-乙腈（90 + 10，v/v）流动相进行洗脱。使用二极管阵列检测器在200 nm处检测分析物峰，具有足够的分辨率。按照国际协调会议、美国药典和美国官方分析化学师协会推荐的方法进行验证研究，并包含准确性、精密度、范围、限度、稳健性和系统适用性参数。使用传统色谱柱时，左乙拉西坦和咖啡因在约7分钟内被检测到，而使用整体柱时所需时间不到5分钟。校准曲线在0.8 - 8.0μg/mL范围内的r值接近1。作为对实际样品的应用，展示了片剂制剂中左乙拉西坦的含量测定。
