Perfusion chromatography: an emergent technique for the analysis of food proteins

Perfusion chromatography is a technique arised to overcome the problem asso ciated with mass transfer in the separation of large molecules such as prot eins by high-performance liquid chromatography (HPLC). Perfusion media are constituted by two set of pores: throughpores (6000-8000 Angstrom) and diff usive pores (800-1500 Angstrom) which enable better access of macromolecule s to the inner of the particle by the combination of convective and diffusi ve flow. As a consequence, times required for a chromatographic separation are reduced. Perfusion media are available in different chromatographic mod es: reversed-phase, ion-exchange, hydrophobic interaction, and affinity. Fr om the theoretical models developed to explain the dynamic of retention of solutes in perfusive supports, it was derived that efficiency of a separati on was independent of the flow-rate and only depended slightly on the parti cle diameter. Furthermore, loading capacity was also independent of the sup erficial velocity. All these advantages have promoted the use of this chrom atographic technique for the separation of biomolecules both in analytical and preparative chromatography. Characteristics of perfusion chromatography make this technique very interesting for the analysis of food proteins. Pe rfusion chromatography enables the assessment of protein composition of a f oodstuff at sufficient speed and low cost to be suitable in routine analysi s. (C) 2000 Elsevier Science B.V. All rights reserved.