Kopf Robert, Paschen Christoph, Müller Lavinia, Kocar Berk, Wolfring Martin, Vincent Mathilde, Klemm Denis, Bell Christian, Pinto Cosimo
F. Hoffmann-La Roche AG, Grenzacherstrasse 24, CH-4070 Basel, Switzerland.
F. Hoffmann-La Roche AG, Grenzacherstrasse 24, CH-4070 Basel, Switzerland.
J Pharm Biomed Anal. 2023 Nov 30;236:115651. doi: 10.1016/j.jpba.2023.115651. Epub 2023 Aug 15.
Non-ionic surfactants are commonly used in parenteral protein formulations and include polysorbate 20, polysorbate 80 and poloxamer188. Recently, quantification and characterization of surfactants has generated considerable interest due to their connection to visible particle formation, a critical quality attribute for parenteral formulations. Typically, surfactant quantification is performed by mixed mode chromatography with evaporative light scattering detection (ELSD) or charged aerosol detection (CAD). However, these methods often suffer from loss of specificity in highly concentrated protein formulations. Here we present a mixed mode chromatography method using single quad mass detection, overcoming current limitations for highly concentrated proteins. In addition to content determination of intact surfactants, this method allows to quantify and characterize the predominant degradation patterns of polysorbates within a single measurement. Formulations with up to 200 mg/mL active pharmaceutical product (API) containing surfactant levels between 0.16 and 0.64 mg/mL were tested during method qualification. The obtained results for linearity (r > 0.99), precision (max. 3.8 % RSD) and accuracy (96-116 % recovery) meet current requirements for pharmaceutical products as defined in ICH Q2.
非离子表面活性剂常用于肠胃外蛋白质制剂中，包括聚山梨酯20、聚山梨酯80和泊洛沙姆188。最近，由于表面活性剂与可见颗粒形成相关，而可见颗粒形成是肠胃外制剂的一个关键质量属性，因此对表面活性剂的定量和表征引起了相当大的关注。通常，表面活性剂的定量是通过带有蒸发光散射检测（ELSD）或带电气溶胶检测（CAD）的混合模式色谱法进行的。然而，这些方法在高浓度蛋白质制剂中常常存在特异性丧失的问题。在此，我们提出一种使用单四极杆质量检测的混合模式色谱法，克服了高浓度蛋白质的现有局限性。除了完整表面活性剂的含量测定外，该方法还允许在单次测量中对聚山梨酯的主要降解模式进行定量和表征。在方法验证期间，对含有0.16至0.64mg/mL表面活性剂水平、活性药物成分（API）浓度高达200mg/mL的制剂进行了测试。获得的线性（r>0.99）、精密度（最大相对标准偏差3.8%）和准确度（回收率96-116%）结果符合ICH Q2中规定的当前药品要求。
