Glycine Crystallization During Freezing: The Effects of Salt Form, pH, and Ionic Strength

Abstract

Purpose. The purpose of the study is to characterize glycine crystallization during freezing of aqueous solutions as a function of the glycine salt form (i.e., neutral glycine, glycine hydrochloride, and sodium glycinate), pH, and ionic strength.

Methods. Crystallization was studied by thermal analysis, microscopy, x-ray diffraction, and pulsed Fourier transform nmr spectroscopy.

Results. A solution of neutral glycine with no additives undergoes rapid secondary crystallization during freezing, forming the β polymorph, with a eutectic melting temperature of −3.4°C. Glycine hydrochloride solutions undergo secondary crystallization relatively slowly, and the eutectic melting temperature is −28°C. Sodium glycinate crystallizes from frozen solution at an intermediate rate, forming a eutectic mixture with a melting temperature of −17.8°C. Where secondary crystallization does not occur rapidly, a complex glass transition is observed in the −70° to − 85°C temperature range in the DSC thermograms of all systems studied. Rates of secondary crystallization and the type of crystal formed are influenced by solution pH relative the the pKs of glycine, and also by the change in ionic strength caused by adjustment of pH. Increased ionic strength significantly slows the crystallization of neutral glycine and promotes formation of the γ polymorph. Thermal treatment or extended holding times during the freezing process may be necessary in order to promote secondary crystallization and prevent collapse during freeze drying.

Conclusions. The results underscore the importance of recognizing that seemingly minor changes in formulation conditions can have profound effects on the physical chemistry of freezing and freeze drying.