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Multivariate Analysis of Phenol in Freeze-Dried and Spray-Dried Insulin Formulations by NIR and FTIR

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Abstract

Dehydration is a commonly used method to stabilise protein formulations. Upon dehydration, there is a significant risk the composition of the formulation will change especially if the protein formulation contains volatile compounds. Phenol is often used as excipient in insulin formulations, stabilising the insulin hexamer by changing the secondary structure. We have previously shown that it is possible to maintain this structural change after drying. The aim of this study was to evaluate the residual phenol content in spray-dried and freeze-dried insulin formulations by Fourier transform infrared (FTIR) spectroscopy and near infrared (NIR) spectroscopy using multivariate data analysis. A principal component analysis (PCA) and partial least squares (PLS) projections were used to analyse spectral data. After drying, there was a difference between the two drying methods in the phenol/insulin ratio and the water content of the dried samples. The spray-dried samples contained more water and less phenol compared with the freeze-dried samples. For the FTIR spectra, the best model used one PLS component to describe the phenol/insulin ratio in the powders, and was based on the second derivative pre-treated spectra in the 850–650 cm−1 region. The best PLS model based on the NIR spectra utilised three PLS components to describe the phenol/insulin ratio and was based on the standard normal variate transformed spectra in the 6,200–5,800 cm−1 region. The root mean square error of cross validation was 0.69% and 0.60% (w/w) for the models based on the FTIR and NIR spectra, respectively. In general, both methods were suitable for phenol quantification in dried phenol/insulin samples.

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ACKNOWLEDGEMENTS

The authors would like to acknowledge funding by Apotekerfonden af 1991 for the Bomem FTIR spectrometer

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Authors and Affiliations

  1. Department of Pharmaceutics and Analytical Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark

    Morten Jonas Maltesen, Marco van de Weert & Holger Grohganz

  2. Department of Oral Protein Delivery, Novo Nordisk A/S, Novo Nordisk Park, 2760, Måløv, Denmark

    Simon Bjerregaard

  3. Department of Oral Formulation Development, Novo Nordisk A/S, Novo Nordisk Park, 2760, Måløv, Denmark

    Lars Hovgaard

  4. Department of Diabetes Protein Engineering, Novo Nordisk A/S, Novo Nordisk Park, 2760, Måløv, Denmark

    Svend Havelund

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  1. Morten Jonas Maltesen
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  2. Simon Bjerregaard
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Correspondence to Morten Jonas Maltesen.

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Maltesen, M.J., Bjerregaard, S., Hovgaard, L. et al. Multivariate Analysis of Phenol in Freeze-Dried and Spray-Dried Insulin Formulations by NIR and FTIR. AAPS PharmSciTech 12, 627–636 (2011). https://doi.org/10.1208/s12249-011-9618-x

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