Nayak Vikram S, Tan Zhijun, Ihnat Peter M, Russell Reb J, Grace Michael J
Analytical Development & Testing, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA.
J Chromatogr Sci. 2012 Jan;50(1):21-5. doi: 10.1093/chromsci/bmr015.
An evaporative light scattering detection (ELSD) based high-performance liquid chromatography (HPLC) method is developed for the determination of polysorbate 80 (tween 80) in therapeutic protein formulations. The method is simple and overcomes the difficulties associated with specificity and sensitivity. The method is suitable for the quantitation of polysorbate 80 in the usual formulation range (0.01-0.1%) as well as in trace amounts ≥13 µg/mL. The analysis is based on the removal of protein first by solid-phase extraction using Oasis HLB cartridges followed by HPLC analysis using Inertsil ODS-3 C 18 column (4.6×150 mm, 5 µm) using reversed-phase conditions. The detector response changes exponentially with an increase in polysorbate concentration. A very good linear fit of log ELSD response against log polysorbate 80 concentration is observed. The specificity, sensitivity, precision, and accuracy of the method are suitable for the quantitation of polysorbate 80 in protein formulations.
建立了一种基于蒸发光散射检测（ELSD）的高效液相色谱（HPLC）方法，用于测定治疗性蛋白质制剂中的聚山梨酯80（吐温80）。该方法简单，克服了特异性和灵敏度方面的难题。该方法适用于常规制剂范围（0.01 - 0.1%）以及痕量≥13 µg/mL的聚山梨酯80的定量分析。分析过程首先使用Oasis HLB柱通过固相萃取去除蛋白质，然后使用Inertsil ODS - 3 C18柱（4.6×150 mm，5 µm）在反相条件下进行HPLC分析。随着聚山梨酯浓度的增加，检测器响应呈指数变化。观察到log ELSD响应与log聚山梨酯80浓度之间具有非常好的线性拟合。该方法的特异性、灵敏度、精密度和准确度适用于蛋白质制剂中聚山梨酯80的定量分析。
