Muhammad Huzaifa Salem, Akram Pezeshki, Babak Ghanbarzade, Hamed Hamishehkar, Maryam Mohammadi
In this study, the effect of three different stabilizers; cholesterol (CHO), beta-sitosterol (BSITO) and gamma-oryzanol (GO) on the production of nanoliposomes (NLs)-loaded with hemp protein hydrolysate (HPH) and their antioxidant activity was investigated. HPHs produced using Pepsin from hemp protein concentrate showed improved antioxidant potential (two- to three-fold increase in several evaluated parameters). The findings showed that the particle size of the NLs was monomodal and in the nanoscale (<150 nm). GO NLs with an average particle size of 64.5 nm, dispersion index of 0.183 and zeta potential of -14.2 mV; had the smallest size and the highest encapsulation efficiency (EE%) (95 ± 2.1%) with better extended release profile than the similar samples. Scanning electron microscopy (SEM) images showed particles with uniform size and spherical shape, Fourier transform infrared spectroscopy (FTIR) confirmed the proper entrapment of HPH, while X-ray diffraction (XRD) patterns revealed an amorphous structural arrangement and differential scanning calorimetry (DSC) analysis showed that GO-stabilized formulations had better thermal stability. Antioxidant evaluations using DPPH assays showed that GO NLs retained significantly higher antioxidant capacity (44.3 ± 1.8%) after 28 day of storage. Overall, the results indicate that incorporation of HPH into nanoliposomal matrices significantly improves their stability, bioavailability and overall delivery efficiency. Among the set of stabilizers, GO was selected as the most efficient stabilizer in enhancing the physical stability of HPH NLs as well as its bioactivity; thus, the promising application of GO in the fields of developing functional foods and nutritious products was highlighted.