Preparation And Evaluation Of Lutein Ternary Composite Nanoparticles

Objective:Lutein（LUT）is a natural antioxidant with prevents macular degeneration,visual fatigue-reducing and immune-boosting properties.However,its use is limited due to its low water solubility and poor stability.Therefore,in this study,Lutein-soy protein isolate nanoparticles（LUT-SPI NPs）and Lutein-soy protein isolate-fucoidan nanoparticles（LUT-SPI-FUD NPs）was constructed to improve solubility and stability,providing research basis for the development of subsequent preparations of LUT.Methods:A method for the in vitro analysis of LUT was established by high performance liquid chromatography.and the solubility of LUT in different p H buffers and water was determined.The influence of LUT stability under the condition of light,temperature and oxygen was studied.Fristly,LUT-SPI NPs were prepared using encapsulation rate,drug loading rate and leakage rate as study parameters.The preparation process of LUT-SPI NPs was optimized using univariate studies combined with Box-Behnken design experiments.The formation mechanism of LUT-SPI NPs was investigated using UV spectral scanning,Fourier transform infrared spectroscopy（FTIR）,differential scanning calorimetry（DSC）and X-ray diffraction（XRD）.LUT-SPI-FUD NPs were prepared by electrostatic self-assembly method,their stability,solubility and in vitro simulated digestion under different conditions were studied,and conduct corresponding characterization analysis on it.The in vitro antioxidative activity of LUT-SPI NPs and LUT-SPI-FUD NPs was investigated by using DPPH radical scavenging,ABTS radical scavenging and Fe³⁺total reduction force measurement,while the in vivo antioxidative activity was investigated using an ethanol oxidative damage model.Results:The established method for in vitro and in vivo analysis of LUT was stable and feasible.Best prescription and process of LUT-SPI NPs:drug load ratio 1:7.587,SPI mass concentration 0.83 mg·m L^-1,stirring time 1.493 h,The encapsulation rate（EE%）,drug loading（DL%）and leakage rate（LR%）of LUT-SPI NPs were（91.52±1.05）%,（8.80±0.09）%and（1.00±0.14）%,respectively;The optimal SPI to FUD mass ratio of LUT-SPI-FUD NPs was determined to 4:1,and the average particle size,polydispersion coefficient（PDI）and Zeta potential were（139.8±1.2）nm,0.15±0.02 and（-31.9±1.7）m V,respectively.The formation of LUT-SPI NPs and LUT-SPI-FUD NPs was further verified by UV spectroscopy,FTIR,DSC and XRD.The solubility experiments showed that LUT-SPI NPs and LUT-SPI-FUD prepared by Lutein were 350.74 and 432.42 times higher in purified water,respectively.The stability study results showed that LUT-SPI NPs and LUT-SPI-FUD NPs both improved the stability of lutein under UV light,high temperature and oxygen conditions,and the stability of LUT-SPI-FUD was better than that of LUT-SPI NPs.In vitro simulated digestion studies,Comparing with free LUT,the cumulative release rates of LUT-SPI NPs and LUT-SPI-FUD NPs were improved;The antioxidant activity of Lut-SPI NPs and LUT-SPI-FUD was higher than that of LUT.In vitro antioxidant capacity experiments demonstrated that LUT-SPI NPs and LUT-SPI-FUD NPs were better than LUT in terms of DPPH radical scavenging,ABTS radical scavenging and Fe³⁺total reduction force measurement;In vivo antioxidant experiment,SOD and CAT activities in serum and liver of LUT-SPI NPs and LUT-SPI-FUD NPs significantly increased,while MDA content decreased（p<0.05 or p<0.01）compared with model group and LUT group,indicating that the they can enhance antioxidant activity of LUT,and at the same dose,The antioxidant activity of LUT-SPI-FUD NPs group was better than that of LUT-SPI NPs group（p<0.05 or p<0.01）.Conclusion:The Lutein-soy protein isolate nanoparticles and Lutein-soy protein isolate-fucoidan nanoparticles drug delivery system were constructed in this study,effectively improving the solubility of LUT,enhanced its stability and antioxidant activity in vitro and in vivo,providing a new choice for LUT drug use.