NMR Method for Accurate Quantification of Polysorbate 80 Copolymer Composition

Abstract

(13)C NMR spectroscopic integration employing short relaxation delays and a 30° pulse width was evaluated as a quantitative tool for analyzing the components of polysorbate 80. (13)C NMR analysis revealed that commercial polysorbate 80 formulations are a complex oligomeric mixture of sorbitan polyethoxylate esters and other intermediates, such as isosorbide polyethoxylate esters and poly(ethylene glycol) (PEG) esters. This novel approach facilitates the quantification of the component ratios. In this study, the ratios of the three major oligomers in polysorbate 80 were measured and the PEG series was found to be the major component of commercial polysorbate 80. The degree of polymerization of -CH2CH2O- groups and the ratio of free to bonded -CH2CH2O- end groups, which correlate with the hydrophilic/hydrophobic nature of the polymer, were analyzed, and were suggested to be key factors for assessing the likelihood of adverse biological reactions to polysorbate 80. The (13)C NMR data suggest that the feed ratio of raw materials and reaction conditions in the production of polysorbate 80 are not well controlled. Our results demonstrate that (13)C NMR is a universal, powerful tool for polysorbate analysis. Such analysis is crucial for the synthesis of a high-quality product, and is difficult to obtain by other methods.

Synthesis of nano-sized polyindole via emulsion polymerization and doping

“…1 H NMR spectra of PIn with surfactants (SDS, CTAB and TW80) fixed at 1:0.05 of N indole /N surfactant mole ratio were analyzed. The main 1 H NMR spectra of these surfactants were also present namely the À ÀOCH 2 CH 2 OÀ À of TW80 at 3.45 ppm [37], the (CH 2 ) n of SDS located at 1.22 ppm [38], and the CH 3 À ÀN of CTAB present at 3.00 ppm [39]. These results indicate that the PIn structure is comprised of surfactant molecules.…”

Simple NMR methods for evaluating higher order structures of monoclonal antibody therapeutics with quinary structure

“…Nearly all the methods [9,12,14,30] were demonstrated on 1 or 2 proprietary clinical formulations, with sucrose [9,30], trehalose [14] or histidine [12] as excipients, but no polysorbate 80 was used. The presence of polysorbate 80, frequently used in commercial mAb formulations, interferes with NMR protein evaluation due to the high concentration relative to the protein drug, binding to protein and a 5 ppm chemical shift range of NMR signals (1–6 ppm) [42]. Here, the intact and the in-situ IdeS digestion NMR was demonstrated to be applicable on 4 mAb drug products, Remicade ® , Rituxan ® , Ristova ® and Reditux ® after deformulation using SEC-FPLC.…”

Small‐angle X‐ray scattering as a useful supplementary technique to determine molecular masses of polyelectrolytes in solution

“…Obtaining the number ( M n ) and mass‐average ( M w ) molecular masses is frequently a challenging task and this is especially true for charged polymers, as the strong intermolecular interactions may influence the results. Some of the most employed methods to obtain absolute masses are mass spectroscopy, multiangle laser light scattering, membrane osmometry, and NMR, as well as size exclusion techniques such as size exclusion chromatograph and asymmetric flow field‐flow fractionation (AF4) which, combined with multidetector and refractive index analysis, can give information on masses and polydispersity indexes (PDIs) . However, most of those methods are time consuming (especially that based on osmometry), sample consuming or may not be applied to all types of polymers.…”