A Highly Sensitive Method for the Quantitation of Polysorbate 20 and 80 to Study the Compatibility between Polysorbates and m-Cresol in the Peptide Formulation

Abstract

A highly sensitive method has been developed for the quantitation of polysorbate 20 (PS20) and 80 (PS80) in therapeutic peptide formulations. A mixed-mode HPLC column was used to separate polysorbates from the peptide and other excipients, and a charged aerosol detector (CAD) was used for the detection. The method was capable of reporting polysorbates as low as 5 ppm, and the sensitivity could be further improved on a needed basis. The method has been used to study the compatibility between polysorbates and m-cresol in the peptide formulation. It was found that both PS20 and PS80 are compatible with m-cresol (at 2.8 mg/ml) when their levels were not greater than 20 ppm. Significant losses of polysorbates were observed when PS20 and PS80 concentrations were above 50 ppm. Furthermore, the agitation study demonstrated that even trace levels of PS20 and PS80 (e.g., 20 ppm) could stabilize the peptide against fibrillation and aggregation.

Antimicrobial Preservatives for Protein and Peptide Formulations: An Overview

“…Furthermore, PS20 and PS80 were spiked to the peptide formulation at 20 ppm PS20 (0.002%) and 50 ppm PS80 (0.005%). In conclusion, Shi et al (2015) confirmed that higher levels of polysorbate 20 and 80 (50 ppm, 100 ppm, 800 ppm) are incompatible with m-cresol leading to increased turbidity of the solution and loss of polysorbate content due to PS degradation. In comparison, PS80 was less compatible with m-cresol than PS20.…”

“…Even though these incompatibility issues are known, many protein formulations including preservatives and polysorbates do exist on the market. Recently, studies were conducted focusing on interactions between preservatives and polysorbates [112,113]. These studies differed from earlier study as they did not analyze the impact of polysorbates on the antimicrobial activity of preservatives but focused on the impact of preservatives on the stability of polysorbates [114].…”

“…These studies differed from earlier study as they did not analyze the impact of polysorbates on the antimicrobial activity of preservatives but focused on the impact of preservatives on the stability of polysorbates [114]. For example, Shi et al (2015) studied whether mcresol is compatible with trace levels of PS20 and PS80 focusing on whether the known incompatibility issues are dependent on the polysorbate concentration [113]. To study the polysorbate-m-cresol interactions, they used HPLC-CAD (charged aerosol detector).…”

Corona charged aerosol detector non-uniform response factors of purified alcohol ethoxylated homologues using liquid chromatography

“…Comparison with a mass spectrometer could be considered. Today, only a few studies have hypothesized that homologues of the same type have a different response, but none have offered proof or explanations [46,50]. In order to deepen the explanation of this phenomenon, molecular modeling could be considered to understand how molecules are organized together during the formation of particles.…”

“…In their work, Shi et al highlighted that the correlation between peak area and polysorbate concentration were not the same between two similar molecules polysorbate 20 and polysorbate 80 4 [46]. They explained this difference by the length of fatty acid chains present in PS80, which are longer than in PS20, and induced higher responses on HPLC-CAD.…”