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. 2022 Jul;43(13-14):1408-1414.
doi: 10.1002/elps.202100189. Epub 2022 Apr 26.

Polysorbate 80 and histidine quantitative analysis by NMR in the presence of virus-like particles

Richard R Rustandi  1
Affiliations

Polysorbate 80 and histidine quantitative analysis by NMR in the presence of virus-like particles

Richard R Rustandi. Electrophoresis. 2022 Jul.

Abstract

Polysorbate-80 (PS80) and histidine are common excipients in vaccine and therapeutic protein formulation. A simple quantitative NMR method to measure both PS80 and histidine in human papillomavirus (HPV) virus-like particle (VLP) vaccine for aqueous and alum-containing samples is described. The new NMR method is compared to current colorimetric methods for PS80 and RP HPLC for histidine. The new NMR method is comparable to current assays with an advantage of a simpler sample treatment for PS80. The efficiency is also increased because one method can now provide two assay results instead of two separate methods. Furthermore, the NMR method can detect PS80 stability due to hydrolysis and oxidation when PS80 is stored in a stainless steel container by observing a change of its NMR line shape profile.

Keywords: HPV; NMR; histidine; polysorbate-80; stainless steel; vaccine.

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Conflict of interest statement

The author has declared no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Polysorbate‐80 structure
FIGURE 2
FIGURE 2
(A) NMR spectrum of PS80 and histidine in the presence of HPV VLP samples illustrating that no HPV VLP is observed; (B) the enlarged of PS80 and histidine peaks at 3.70 and 7.21 ppm, respectively used for quantitation. (C) Standard curve for histidine. (D) Standard curve for PS80
FIGURE 3
FIGURE 3
Comparison of NMR peak at 3.70 ppm of PS80 (0.015%) in histidine buffer stored in glass (blue trace) and stainless steel (red trace) containers. The PS80 containing HPV VLP sample stored in stainless steel (green trace) and PS80 reference standard control at 0.02% (black trace)
FIGURE 4
FIGURE 4
NMR peak at 3.70 ppm of PS80‐treated with NaOD (red trace) and compared with PS80‐containing HPV VLP stored in stainless steel (green trace) and PS80 reference standard as control (black trace)
See this image and copyright information in PMC

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