05). The shelf-life Ruxolitinib in vivo of the edible films (in terms of L. rhamnosus GG survival) ranged from 63 to 100 days and 17 to 30 days for the systems stored at chilled (4 °C)

or room temperature (25 °C) conditions ( Table 2). Extrinsic factors such as water activity, temperature and presence of oxygen are known to adversely influence the viability of encapsulated probiotic living cells ( Fu & Chen, 2011). Moreover, the molecular mobility of solutes driven by the structural and physical state of the immobilising matrix can also influence the stability of probiotics. Thus, the acquirement of low residual water–glassy matrices with low permeability to gases containing free radical scavenging agents (to control lipid oxidation of cellular membranes) has been reported as an efficient strategy for improving probiotics viability in food systems ( Dong et al., 2013 and Soukoulis et al., 2013). In the case of intermediate moisture systems (including edible films) the presence of high amounts of solutes together with Cilengitide nmr the rubbery

physical state (solutes’ increased molecular mobility) facilitates the occurrence of enzymatic and chemical reactions that damage essential cellular structures e.g. phospholipid membrane bilayers ( Fu & Chen, 2011). The stability of the prebiotic films at room temperature is generally comparable to that of anhydrobiotics (e.g. spray dried powders) stored at the same relative humidity conditions ( Ying, Sun, Sanguansri, Weerakkody, & Augustin, 2012). Although a full mechanistic understanding of probiotics stability in biopolymer matrices during storage is not available, it appears that factors such as steric hindrance of solutes and the matrix translational diffusion of oxygen (both

associated with the T − Tg difference), the presence of nutrients and free radical scavenging agents as well as the interaction via hydrogen bonding with the polar head groups of membranes phospholipids can be possible explanations for the stability of probiotics in prebiotic films ( Ananta et al., 2005, Kanmani and Lim, 2013, Semyonov et al., 2011 and Soukoulis et al., 2013). Prebiotics such as inulin, fructo-oligosaccharides find more and polydextrose are able to enhance the stability of probiotics primarily through their impact on the glass transition phenomena albeit no clear evidence on their specific protective action has been provided for each type of probiotics ( Ananta et al., 2005, Corcoran et al., 2004 and Fritzen-Freire et al., 2012). In a first attempt to provide some evidence on the impact of physical state of the matrix on the inactivation rates of L. rhamnosus GG, we have calculated the glass transition temperatures of the systems with the highest and lowest inactivation rate at room temperature viz. inulin and polydextrose using the Couchman–Karasz equation (Eq. (5)) assuming a quaternary system comprising fibre, gelatine, water and glycerol equation(5) Tg=Tg.fibre∗ΔCP,fibre∗Xfibre+Tg.