Effects of aqueous phase composition upon protein destabilization at water/organic solvent interface

Hongkee Sah, Yogita Bahl

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10 Scopus citations


The objective of this study was to evaluate the effects of an aqueous phase composition upon the destabilization of lactoglobulin toward emulsification. Eight different buffers were used to make aqueous lactoglobulin solutions at pH 5.6-9. Each buffer concentration varied from 20 to 60, 100, and 150 mM. After emulsification of the aqueous solutions in methylene chloride, the aqueous contents of lactoglobulin were determined by a native SEC-HPLC assay. The properties of lactoglobulin precipitates detected at the water/methylene chloride interface were assessed by SDS-PAGE experiments. Dynamic interfacial tension was also monitored to examine the interfacial adsorption of lactoglobulin. When dissolved in plain water, only 41.1 ± 1.4% of lactoglobulin was recovered after emulsification. The remaining lactoglobulin became aggregated at the interface. A series of SDS-PAGE experiments demonstrated that its aggregation was driven by intermolecular covalent linkages and hydrophobic interactions. Interestingly, both the buffer type and its concentration considerably influenced the degree of lactoglobulin recovery. For instance, at 150 mM buffer concentrations, percent recovery of lactoglobulin was as low as 27.3 ± 1.7%. In contrast, the degree of its recovery increased up to 89.4 ± 3.9% at a 20 mM buffer concentration. The dynamic interfacial tension study substantiated that at an optimal concentration buffer species helped lactoglobulin molecules to better withstand interfacial destabilization. Therefore, choosing a suitable buffer and its concentration should deserve special attention, since they could affect the interfacial stability of a protein of interest during emulsification.

Original languageEnglish
Pages (from-to)51-61
Number of pages11
JournalJournal of Controlled Release
Issue number1-2
StatePublished - 18 Aug 2005

Bibliographical note

Funding Information:
This work was supported by a grant R05-2001-000-00667 from the Korea Science and Engineering Foundation.


  • Emulsification
  • Microencapsulation
  • Microspheres
  • Protein stability


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