Protein Analytical Chemistry Department, Genentech, 1 DNA Way, South San Francisco, California 94080, USA.
Anal Chem. 2011 May 15;83(10):3934-42. doi: 10.1021/ac2005789. Epub 2011 Apr 27.
Polysorbates (PS) are widely used as oil-in-water emulsifiers, stabilizers, wetting agents, solubilizers, and dispersants in the agricultural, food, personal care, and pharmaceutical industries due to their cost effectiveness, biocompatibility, formulation flexibility, low toxicity, and good stabilizing and protecting properties. The polysorbates are often pictured as polyoxyethylated sorbitan monoesters of saturated and/or unsaturated fatty acids. In reality, polysorbates are complex mixtures of multiple components, as follows from the reactions involved in their production. In this work, we report a novel application of liquid chromatography-mass spectrometry (LC-MS) for the characterization of polysorbates. This method takes advantage of accurate mass measurements and information on the identity of a fatty acid from "in-source" generated characteristic dioxolanylium ions. The method allowed us to perform quick profiling of fatty acids in PS 20 and 80 which, combined with a computer-aided peak assignment algorithm, facilitated detailed characterization of their constituents. As a major finding, we determined that different samples of PS 20 varied from 0% to 15% in relative amounts of unsaturated oleic acid. Although the consequences of this difference were not fully evaluated in this work, one might expect that PS 20 with larger amounts of oleic acid will be more prone to autoxidation, thus potentially having greater impact on the oxidative degradation of the biotherapeutics it formulates.
聚山梨酯（PS）由于其成本效益、生物相容性、配方灵活性、低毒性以及良好的稳定和保护性能，被广泛用作农业、食品、个人护理和制药行业的水包油型乳化剂、稳定剂、润湿剂、增溶剂和分散剂。聚山梨酯通常被描绘为聚氧乙烯化山梨糖醇单酯的饱和和/或不饱和脂肪酸。实际上，聚山梨酯是多种成分的复杂混合物，这是由其生产过程中涉及的反应所决定的。在这项工作中，我们报告了液相色谱-质谱（LC-MS）在聚山梨酯表征中的一种新应用。该方法利用准确的质量测量和来自“源内”生成的特征二氧戊环阳离子的脂肪酸身份信息。该方法允许我们对 PS 20 和 80 中的脂肪酸进行快速分析，结合计算机辅助的峰分配算法，有助于对其成分进行详细表征。作为一个主要发现，我们确定 PS 20 的不同样品中，不饱和油酸的相对含量在 0%到 15%之间变化。尽管在这项工作中没有充分评估这种差异的后果，但人们可能会认为含有更多油酸的 PS 20 更容易自动氧化，从而可能对其配制的生物治疗药物的氧化降解产生更大的影响。
