Lippold Steffen, Koshari Stijn H S, Kopf Robert, Schuller Rudolf, Buckel Thomas, Zarraga Isidro E, Koehn Henning
F. Hoffmann-La Roche, Grenzacherstrasse 124, 4070 Basel, Switzerland.
Department of Chemical and Biomolecular Engineering, University of Delaware, 150 Academy Street, Newark, DE 19716, USA; Late Stage Pharmaceutical Development, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
J Pharm Biomed Anal. 2017 Jan 5;132:24-34. doi: 10.1016/j.jpba.2016.09.033. Epub 2016 Sep 26.
Determination of excipient content in drug formulation is an important aspect of pharmaceutical formulation development and for analytical testing of the formulation. In this study, the influence of polysorbate subspecies, in particular mono- and poly-esters, for determining polysorbate (PS) content were investigated by comparing three of the most widely used PS quantitation approaches, the Fluorescence Micelle Assay (FMA) and Mixed-Mode High Performance Liquid Chromatography coupled with Charged Aerosol Detection (MM-CAD) or Evaporative Light Scattering Detection (MM-ELSD). FMA and MM-CAD were employed to investigate the quantitation behavior of PS20 and PS80 subspecies and corresponding degradation products in placebo formulations using forced degradation conditions at 40°C for up to 12 weeks. While both methods allowed accurate and comparable quantification of neat PS at the beginning of stress studies, pronounced differences in content determination between the methods were observed at later time points, which were attributable to substantial differences in the contribution of individual mono- and poly-esters to the overall quantitation results. It was particularly surprising to find that the main component of PS20, polyoxyethylene sorbitan monolaurate, did not show a signal at the studied concentration using FMA. Moreover, the degradation of polysorbate poly-esters, was reflected much stronger in FMA than MM-CAD results. Additional experiments employing chemical oxidation and base hydrolysis to degrade PS20, quantified by FMA and MM-ELSD, also show preferential reduction in certain subspecies depending on the degradation pathway involved. For PS20 degraded by chemical oxidation, quantitation results were lower for FMA than MM-ELSD, while the opposite trend was observed with base hydrolysis.
确定药物制剂中的辅料含量是药物制剂开发以及制剂分析检测的一个重要方面。在本研究中，通过比较三种最广泛使用的聚山梨酯定量方法，即荧光胶束分析法（FMA）以及与带电气溶胶检测（MM-CAD）或蒸发光散射检测（MM-ELSD）联用的混合模式高效液相色谱法，研究了聚山梨酯亚种，特别是单酯和多酯，对聚山梨酯（PS）含量测定的影响。采用FMA和MM-CAD研究了在40°C强制降解条件下长达12周时，PS20和PS80亚种以及相应降解产物在安慰剂制剂中的定量行为。虽然在应激研究开始时，这两种方法都能对纯PS进行准确且可比的定量，但在后期时间点观察到两种方法在含量测定上存在明显差异，这归因于各个单酯和多酯对总体定量结果的贡献存在显著差异。特别令人惊讶的是，发现使用FMA时，PS20的主要成分聚氧乙烯山梨醇酐单月桂酸酯在所研究的浓度下未显示信号。此外，聚山梨酯多酯的降解在FMA中的反映比MM-CAD结果要强得多。采用化学氧化和碱水解降解PS20并通过FMA和MM-ELSD进行定量的额外实验也表明，根据所涉及的降解途径，某些亚种会优先减少。对于通过化学氧化降解的PS20，FMA的定量结果低于MM-ELSD，而在碱水解时则观察到相反的趋势。
