Research Paper

Pharmaceutical Research

, Volume 23, Issue 10, pp 2368-2374

First online:

Solute Crystallization in Frozen Systems–Use of Synchrotron Radiation to Improve Sensitivity

  • Dushyant B. VarshneyAffiliated withDepartment of Pharmaceutics, College of Pharmacy, University of MinnesotaEli Lilly and Company, Lilly Corporate Center
  • , Satyendra KumarAffiliated withDepartment of Physics, Kent State University
  • , Evgenyi Y. ShalaevAffiliated withPfizer Groton Laboratories
  • , Shin-Woong KangAffiliated withDepartment of Physics, Kent State University
  • , Larry A. GatlinAffiliated withPfizer Groton Laboratories
  • , Raj SuryanarayananAffiliated withDepartment of Pharmaceutics, College of Pharmacy, University of Minnesota Email author 

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To demonstrate the sensitivity of low temperature synchrotron X-ray diffractometry (SXRD) for detecting solute crystallization in frozen sodium phosphate buffer solutions. To determine the effect of annealing on solute crystallization in frozen solutions.

Materials and Methods

Sodium phosphate buffer solutions, at initial buffer concentrations ranging from 1 to 100 mM (pH 7.4) were cooled to −50°C. The crystallization of disodium hydrogen phosphate dodecahydrate (Na2HPO4 •12H2O) was monitored using a laboratory as well as a synchrotron source. At selected concentrations, the effect of annealing (at −20°C) was investigated.


With the laboratory source, solute crystallization, based on the appearance of one diagnostic peak with a d-spacing of 5.4 Å, was evident only when the initial buffer concentration was at least 50 mM. In contrast, using SXRD, crystallization was detected at initial buffer concentrations down to 1 mM. In addition, the use of a high-resolution 2D detector enabled the visualization of numerous diffraction rings of the crystalline solute. At both 10 and 100 mM buffer concentration, there was no increase in solute crystallization due to annealing.


By using synchrotron radiation, solute crystallization was detected with substantially increased sensitivity, making the technique useful for freeze-drying cycles of practical and commercial importance. Since numerous peaks of the crystalline solute appeared, the technique has potential utility in complex, multi-component systems.

Key words

annealing buffer salt disodium hydrogen phosphate dodecahydrate solute crystallization synchrotron XRD