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Mutual Influence of Mannitol and Trehalose on Crystallization Behavior in Frozen Solutions

Abstract

Purpose

Phase separation of trehalose during freeze-drying could render it ineffective as a lyoprotectant. The bulking agent, mannitol, on the other hand, should crystallize readily upon freezing. It is therefore imperative to understand the mutual interaction of these sugars during freezing to ensure preservation of the API during freeze-drying.

Methods

We investigated the effect of mannitol to trehalose ratio (R) on the crystallization behavior of both solutes using Differential Scanning Calorimetry, X-Ray Crystallography and FTIR Spectrosopy during controlled freezing and annealing.

Results

When R = 1, crystallization of both mannitol (as hemihydrate) and trehalose (as α-anhydrate) were observed. When R ≥ 1, extent of mannitol crystallization was directly proportional to the value of R. When R < 1, trehalose completely suppressed mannitol crystallization. At R > 1, the freeze concentrate was heterogeneous and characterized by two glass transitions – the lower temperature transition (Tg”) attributed to systems containing “extra” unfrozen water. When heated above Tg”, crystallization of mannitol and the associated unfrozen water resulted in Tg’ (glass transition temperature of the freeze-concentrate).

Conclusions

R and not the total solute concentration, dictates the composition of the freeze concentrate as well as the physical stability of the excipients

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Abbreviations

API:

Active pharmaceutical ingredient

DSC:

Differential scanning calorimetry

FCL:

Freeze-concentrated liquid

FTIR:

Fourier transform infrared spectroscopy

IR:

Infrared

XRD:

Powder X-ray Diffractometry

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ACKNOWLEDGMENTS AND DISCLOSURES

This research was funded by an NSF grant (CBET-1335936) to A.A. The authors are grateful to Dr. Seema Thakral for guidance while performing the powder X-ray diffraction. Parts of this work were carried out in the Characterization Facility, University of Minnesota, which receives partial support from NSF through the MRSEC program.

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Correspondence to Alptekin Aksan.

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Jena, S., Suryanarayanan, R. & Aksan, A. Mutual Influence of Mannitol and Trehalose on Crystallization Behavior in Frozen Solutions. Pharm Res 33, 1413–1425 (2016). https://doi.org/10.1007/s11095-016-1883-7

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Key Words

  • crystallization
  • frozen
  • IR spectroscopy
  • mannitol
  • microscopy
  • trehalose
  • X-ray diffractometry