In this work, a waxy barley starch-PEG aqueous two-phase system (ATPS) phase diagram was constructed, and starch microsphere preparation was explored at different phase diagram positions. The aim was to investigate starch-PEG ATPS phase behaviour and relate this to starch crystallisation and microsphere formation. The hypothesis was that phase diagram position would influence the starch microsphere preparation and the properties of the microspheres. The microsphere formation process was investigated with regard to microsphere development and starch crystallisation kinetics. Microsphere physicochemical properties and their development during different stages of the preparation were studied by examining freshly produced, freeze-dried, and redispersed microspheres. Enzymatic hydrolysis of redispersed microspheres was also investigated. It was possible to produce microspheres from different positions in the phase diagram using 24 h incubation at 25 °C. However, the operational area for the used production conditions was relatively small compared to the biphasic region of the phase diagram. The main findings were that the starch-PEG ATPS phase behaviour can affect the rate of microsphere formation and particle size, but the additional properties of the dried and redispersed microspheres did not differ to a considerable extent. Thus, we have identified a robust production space where production parameters such as time to obtain microspheres can be considerably influenced by the ATPS system phase diagram position.
We acknowledge the Swedish Foundation for Strategic Research for founding of Zandra Gidlöf (Grant number FID18-0026). This research was also financed through Competence Centre NextBioForm, funded by Vinnova Swedish Governmental Agency for Innovation and The Swedish Research Council under grant number 2018-04730.