Polysorbates are widely used as non-ionic surfactant in biopharmaceutical formulations. Recently, the degradation of polysorbate moved into the focus of attention, because in several published studies it was described, that stability issues in polysorbate containing formulations were observed leading to the formation and appearance of sub-visible and visible particles. For this reason, monitoring of polysorbate and its degradation products is of importance throughout the development of parenterals. The aim of the study was to develop a method for the selective marker-based quantification of adequate polysorbate 20 components of interest without the need to apply derivatization or complex detection techniques. A single quadrupole mass (QDa) detector was used coupled to an ultra-high performance liquid chromatography (UHPLC) system.

Method development was based on a developed reversed phase-high performance liquid chromatography assay coupled to a charge aerosol detector (RP-HPLC CAD). Instead of a charge aerosol detector (CAD) a QDa detector was used in order to significantly improve the selectivity. The focus of this study is the development of the QDa based method for the analysis of polysorbate 20. Modifications of the mobile phase and the type of chromatography column allowed the separation of several components of polysorbate 20 from polar non-esterified to apolar higher order species. In addition, a multitude of components could be quantified by their individual m/z values. The peak assignment identified 676 compounds which originated from polysorbate 20. Some of these were selected and defined as marker components. It was shown that the developed method is capable to determine polysorbate 20 in different biopharmaceutical formulations. The proposed assay is based on an artful sample preparation as well as a unique calibration procedure that make the determination of several selected components achievable. Furthermore, it was successfully demonstrated that the analytical procedure is valid to reliably quantify several polysorbate 20 components at its 100% level (corresponds to 0.4 mg/mL intact polysorbate 20) and even at lower concentrations that occur e.g. in case of polysorbate 20 degradation. In conclusion, the method is beneficial to determine selected polysorbate 20 species during formulation development of biopharmaceuticals as well as during stability testing and trouble shooting.

聚山梨酯被广泛用作生物药物制剂中的非离子表面活性剂。近来，聚山梨酯的降解成为关注的焦点，因为在一些公开的研究中描述了在含聚山梨酯的制剂中观察到稳定性问题，导致了亚可见和可见颗粒的形成和外观。因此，在肠胃外药物开发过程中，对聚山梨酯及其降解产物的监测至关重要。该研究的目的是开发一种无需选择性应用衍生化或复杂检测技术即可对感兴趣的适当聚山梨酯20成分进行基于标记的定量的方法。使用单个四极杆质量（QDa）检测器与超高效液相色谱（UHPLC）系统耦合。

方法的开发基于与电荷气溶胶检测器（RP-HPLC CAD）耦合的反相高效液相色谱分析方法。代替电荷气溶胶检测器（CAD），使用了QDa检测器，以显着提高选择性。这项研究的重点是开发基于QDa的聚山梨酯20的分析方法。对流动相的修改和色谱柱的类型允许将聚山梨酯20的几种组分从极性非酯化至非极性高阶物质中分离出来。 。此外，可以通过其各自的m / z来量化多个分量价值观。峰归属确定了676种源自聚山梨酯20的化合物。选择其中一些并将其定义为标记物成分。结果表明，开发的方法能够测定不同生物药物制剂中的聚山梨酯20。提出的测定方法基于巧妙的样品制备以及独特的校准程序，可以确定几种选定的组分。此外，已经成功地证明了该分析程序有效地可靠地定量了几种聚山梨酯20成分，其含量为100％（相当于0.4 mg / mL完整聚山梨醇酯20），甚至在较低浓度下（例如在聚山梨醇酯20降解的情况下）。结论，