CONCENTRIC ENCAPSULATION OF LYCOPENE AND OMEGA FATTY ACIDS

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Date
2019
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DIVISION OF FOOD SCIENCE AND POSTHARVEST TECHNOLOGY ICAR-INDIAN AGRICULTURAL RESEARCH INSTITUTE NEW DELHI
Abstract
A study was undertaken to achieve concentric encapsulation of lycopene and ω-fatty acid (rice bran oil) using suitable techniques. Entrapment of lycopene from grapefruit and papaya into rice bran oil (RBO) was achieved using two methods, (i) Supercritical-CO2 (SC-CO2) extraction of lycopene from grapefruit and papaya matrices using rice bran oil as a co-solvent and (ii) Supercritical-CO2 extraction of lycopene from grapefruit and papaya matrices using ethanol as a co-solvent and subsequently encapsulating it into rice bran oil using high pressure homogenization. Rice bran oil encapsulating the lycopene obtained from both the aforesaid methods were subsequently encapsulated using a combination of polysaccharide (from jackfruit seed & tapioca) and protein concentrate (from whey & soybean) using spray drying technique. SC-CO2 extraction of lycopene from grapefruit (cv. Red Blush) and papaya (cv. Red Lady) matrices with RBO as a co-solvent was separately optimized with temperature, pressure, and time of extraction as independent variables. The flow rates of SC-CO2 and RBO were fixed at 35g/min. and 1g/min. respectively. Extraction of lycopene from lyophilized powder of grapefruit and papaya were optimized. Each of the independent variables were used with five levels (pressure 250, 300, 375, 450 & 500 bars; temperature 55, 60, 70, 80 & 85°C; and extraction time 60, 90, 135, 180 & 210 min.) for experimentation using a central composite rotatable design (CCRD) of response surface methodology. Twenty experimental runs were carried out for each of the two lycopene matrices i.e. grapefruit and papaya. Responses were analyzed using Design Expert software for maximization of lycopene extraction efficiency and retention of γ-oryzanol content in oil. For the extraction of lycopene from grapefruit the optimum combination of independent variables were 325 bars, 64°C, and 143 min. and for that of papaya the same were 354 bar, 67°C, and 130 min. At the optimum conditions lycopene recoveries of 70.52 ± 3.65% and 79.27 ± 1.69% and γ-oryzanol content of 11154 ± 148 ppm and 11079 ± 146 ppm were predicted. For validation of the predicted values two set of experiments at the optimum condition with three replications were carried out. Average of the experimental lycopene recoveries obtained were 70.66 ± 3.02% and 78.81 ± 1.78%; and those for γ-oryzanol content were 11099 ± 186 ppm and 10984 ± 190 ppm. Ethanolic extract of lycopene from grapefruit and papaya matrices using SCCO2 were separately encapsulated using high pressure homogenization (HPH) with whey protein concentrates (WPC) or soy protein concentrates (SPC) respectively as emulsifiers. Oil in water emulsion formulation for both the cases were designed using central composite design (CCD) with five levels each of respective emulsifier concentration (WPC/SPC) 0.25, 0.5, 0.75, 1.0 and 1.25% w/v; pressure of homogenization 10000, 15000, 20000, 25000 and 30000 psi; and number of passes through the homogenizer 1, 2, 3,4 and 5). A total of 20 experimental runs were conducted for each of the two sets and conditions were optimized based on maximization of emulsification efficiency and γ-oryzanol content; minimization of average particle size, polydispersity index. Oil content in pre-emulsions was kept constant at 6%, w/v in all the experimental runs. The optimum combinations of independent variables for encapsulation of lycopene extracted from grapefruit into RBO were 0.9% emulsifier concentration, 21000 psi homogenization pressure and 2 passes through the homogenizer. The same for encapsulation of lycopene extracted from papaya into RBO were 1.0%, 19000 psi and 3 respectively. At the optimum conditions emulsification efficiencies were 92.18 ± 1.89% and 87.37 ± 3.12%; γoryzanol content were 11141 ± 162 ppm and 10985 ± 144 ppm; average particle size 167 ± 25 nm and 390 ± 18 nm; and polydispersity index 0.20 ± 0.015 and 0.24 ± 0.032; and ζ-potential were -36.7 ± 1.25 mV and -39.2 ± 1.29 mV respectively. In fact, ζ-potential for all forty experimental runs was found to be below -30 mV indicating excellent emulsion stability. The optima obtained for emulsification of lycopene extracted from grapefruit using WPC as an emulsifier was used for the emulsification of lycopene extracted from papaya. The emulsified lycopene extracted from papaya using WPC as an emulsifier had an emulsification efficiency of 92.09 ± 2.06%, γ-oryzanol of 11128 ± 183 ppm, average particle size of 161 ± 27 nm, polydispersity index of 0.20 ± 0.016 and ζ-potential of -36.41 ± 1.41 mV. Similarly, emulsification of lycopene extracted from grapefruit was tried using the optima obtained for emulsification of lycopene extracted from papaya with SPC as an emulsifier. The emulsion so obtained was found to have an emulsification efficiency of 87.93 ± 3.50%, γ-oryzanol of 11010 ± 162 ppm, average particle size of 390 ± 20 nm, polydispersity index of 0.24 ± 0.036 and ζ-potential of -38.69 ± 1.47 mV. Non-significant differences between all the responses for lycopene extracted from both grapefruit and papaya with use of similar emulsifier (WPC/SPC) clearly indicated that the source of lycopene did not have any effect on the emulsification process. Lycopene extracted from grapefruit and papaya matrices using SC-CO2 as solvent and RBO as co-solvent were each separately encapsulated with four different combinations of protein and polysaccharide i.e. Whey Protein Concentrate (WPC) & Jackfruit Seed Polysaccharide (JSP); Soy Protein Concentrate (SPC) & JSP; WPC & Tapioca Polysaccharide (TP); and SPC & TP by spray drying. Additionally, emulsion obtained using lycopene from grapefruit with WPC as an emulsifier was encapsulated using two different wall materials (WPC+JSP and WPC+TP); emulsion obtained using lycopene from grapefruit with SPC as an emulsifier was encapsulated using two different wall materials (SPC+JSP and SPC+TP); emulsion obtained using lycopene from papaya with WPC as an emulsifier was encapsulated using two different wall materials (WPC+JSP and WPC+TP); and emulsion obtained using lycopene from papaya with SPC as an emulsifier was encapsulated using two different wall materials (SPC+JSP and SPC+TP). A total of 16 spray dried products were obtained each with three replications (a total of 48 experimental runs). Protein to polysaccharide ratio was kept at 1:2.6, 1:2.2, 1:3 & 1:2.6 for combination of WPC+JSP, WPC+TP, SPC+JSP & SPC+TP respectively. The total solids (lycopene + oil + wall material) in the spray feed was kept constant at 30% w/v, oil concentration at 20% of the total solids and inlet air temperature at 140°C for all the spray drying experimental runs. The final product was evaluated with respect to their lycopene retention, γ-oryzanol retention, average particle size, peroxide value and antioxidant activity. Lycopene retention in microcapsules was varied from 89.53 to 97.02%, γ-oryzanol retention from 91.75 to 99.47%, average particle size from 31 to 72 µm, peroxide value from 0.99 to 1.37 meq/kg and antioxidants activity from 461 to 498 µmol TEAC/g. In vitro release behaviour of γ-oryzanol and lycopene from microcapsules prepared using spray drying was studied under simulated gastro-intestinal conditions for a total period of 8 h {1 h each at pH 4.0 (upper stomach condition) and pH 2.0 (lower stomach condition); and 6 h at pH 7 (small intestine condition)}. γ-oryzanol and lycopene release from microcapsules at the end of eight hours varied between 48 to 72%, and 44 to 69% respectively. The final concentrically encapsulated lycopene and γ-oryzanol were mainly obtained through two processes, (i) A 2 step Process : SC-CO2 extraction of lycopene using RBO as a co-solvent where the end product is lycopene encapsulated within RBO and subsequently encapsulated within the protein-polysaccharide wall matrix using spray drying; and (ii) A 3 step Process : SC-CO2 extraction of lycopene using ethanol as a co-solvent, the extracted lycopene is encapsulated within RBO using high pressure homogenization and spray drying to encapsulate the lycopene encapsulated within RBO into protein-polysaccharide matrix. If we compare the two processes for its effectiveness towards encapsulation and delivery of lycopene as well as γ-oryzanol considering the initial contents extracted and those which were delivered into the gut, the two step process was found to be more effective compared to the three step process. Encapsulation efficiencies of lycopene and γ-oryzanol varied between 87.6 ± 0.52% to 93.6 ± 0.47% and 94.4 ± 0.82% to 99.5 ± 0.56% respectively for the two step process whereas the same for the three step process varied between 80.5 ± 0.74% to 89.5 ± 0.45% and 91.8 ± 0.78% to 97.7 ± 0.60%. The delivery efficiencies for lycopene and γ-oryzanol for the two step process varied between 44.9 ± 1.26% to 60.5 ± 1.08% and 49.2 ± 0.94% to 65.2 ± 1.05% which for the three step process varied between 39.3 ± 1.13% to 54.1 ± 1.17% and 45.4 ± 1.20% to 57.9 ± 1.24%. If we compare the four different combinations of protein-polysaccharide wall matrices encapsulation efficiencies of those containing JSP were significantly higher than those containing TP. However, the delivery efficiency into the simulated gut conditions of wall materials containing TP was found to significantly higher than th
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T-10270
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