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

Perfusion chromatography is a technique arised to overcome the problem associated with mass transfer in the separation of large molecules such as proteins by high-performance liquid chromatography (HPLC). Perfusion media are constituted by two set of pores: throughpores (6000–8000 Å) and diffusive pores (800–1500 Å) which enable better access of macromolecules to the inner of the particle by the combination of convective and diffusive flow. As a consequence, times required for a chromatographic separation are reduced. Perfusion media are available in different chromatographic modes: reversed-phase, ion-exchange, hydrophobic interaction, and affinity. From the theoretical models developed to explain the dynamic of retention of solutes in perfusive supports, it was derived that efficiency of a separation was independent of the flow-rate and only depended slightly on the particle diameter. Furthermore, loading capacity was also independent of the superficial velocity. All these advantages have promoted the use of this chromatographic 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. Perfusion chromatography enables the assessment of protein composition of a foodstuff at sufficient speed and low cost to be suitable in routine analysis.