Comparison of NMR and Dynamic Light Scattering for Measuring Diffusion Coefficients of Formulated Insulin: Implications for Particle Size Distribution Measurements in Drug Products

Abstract

Particle size distribution, a measurable physicochemical quantity, is a critical quality attribute of drug products that needs to be controlled in drug manufacturing. The non-invasive methods of dynamic light scattering (DLS) and Diffusion Ordered SpectroscopY (DOSY) NMR can be used to measure diffusion coefficient and derive the corresponding hydrodynamic radius. However, little is known about their use and sensitivity as analytical tools for particle size measurement of formulated protein therapeutics. Here, DLS and DOSY-NMR methods are shown to be orthogonal and yield identical diffusion coefficient results for a homogenous monomeric protein standard, ribonuclease A. However, different diffusion coefficients were observed for five insulin drug products measured using the two methods. DOSY-NMR yielded an averaged diffusion coefficient among fast exchanging insulin oligomers, ranging between dimer and hexamer in size. By contrast, DLS showed several distinct species, including dimer, hexamer, dodecamer and other aggregates. The heterogeneity or polydisperse nature of insulin oligomers in formulation caused DOSY-NMR and DLS results to differ from each other. DLS measurements provided more quality attributes and higher sensitivity to larger aggregates than DOSY-NMR. Nevertheless, each method was sensitive to a different range of particle sizes and complemented each other. The application of both methods increases the assurance of complex drug quality in this similarity comparison.

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Abbreviations

DLS:

Dynamic light scattering

DOSY:

Diffusion Ordered SpectroscopY

NMR:

Nuclear magnetic resonance

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Acknowledgements

We thank Darón Freedberg, Marcos Battistel, and Hugo Azurmendi for the assistance in setting up the DOSY-NMR experiments and for their helpful discussions. Support for this work comes from the US FDA CDER Critical Path funds and is gratefully acknowledged.

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Correspondence to Kang Chen.

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This article reflects the views of the author and should not be construed to represent US FDA’s views or policies.

Electronic Supplementary Material

ESM 1

Bruker 2D DOSY-NMR pulse sequence employed for data acquisition. Individual diffusion coefficients obtained from NMR and DLS. (DOCX 66 kb)

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Patil, S.M., Keire, D.A. & Chen, K. Comparison of NMR and Dynamic Light Scattering for Measuring Diffusion Coefficients of Formulated Insulin: Implications for Particle Size Distribution Measurements in Drug Products. AAPS J 19, 1760–1766 (2017). https://doi.org/10.1208/s12248-017-0127-z

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KEY WORDS

  • diffusion
  • particle size distribution
  • physicochemical equivalence
  • protein drug product
  • similarity