Research And Formulation Design Of CCD - RSM For Increasing Bone Density Health Function Food

The subject mainly studies of biological calcium carbonate, collagen, chondroitin sulfate, glucosamine four main functional component of composite synergistically increased bone mineral density. Cell model selection is MC3T3-E1 osteoblast, by central composite design and response surface optimization method of four factors and five levels of central composite design, through the use of design Expert8.0.6 experimental design software to optimization the optimal experiment scheme, to carry out four main functional component of composite synergistically increased bone density study, through the use of alkaline phosphatase activity detection and mineralized nodule detection, finally obtained the optimal formula of the four major functional components, especially discusses the four main functional component of the best combination of formula, in order to improve the composition of science and clinical, and increased bone mineral density functional foods provide scientific method and experimental evidence.In the selection of the four main functional components of the optimal formula, continue to use CCD-RSM to choose the Maltodextrin, Xylitol, L-malic acid addition to affect calcium collagen chewing the product of comprehensive sensory evaluation factors, a three factor five level central composite design, the design Expert8.0.6 software for experiment design and variance analysis of experimental data results were analyzed, and the effects of collagen calcium chewable tablets of comprehensive sensory quality of primary and secondary factors: Maltodextrin> Xylitol>L-malic acid and between the three factors exist significant interaction. Therefore, the optimal formula of calcium collagen protein was finally determined.The main results of this paper are as follows:1. The 3-(4,5-dimethyl thiazolyl-2)-2.5 diphenyl tetrazolium bromide salt method (3-4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide that MTT method to calculate the cell survival rate and analysis of tested drugs increase bone density.2. Using Expert8.0.6 Design experimental design software, processing experimental results to establish a multivariate regression mathematical model two. According to a multiple quadratic fitting equation draw Fig.2D contour map and 3D effect factors of effect index trend investigation, choosing the best mix proportion according to drawing two-dimensional contour map and 3D effect graph and experimental verification. According to the calculated value of the deviation of the indicators of each experimental data to prove the accuracy of the size of two multiple quadratic regression mathematical model is established to predict the experimental results.3. The prediction value is (62.75%+0.08) and experimentally measured values are (65.69%+0.05) coordination of MC3T3-E1 body into bone cell proliferation and the greatest value obtained by CCD 31 experimental group rate for 65.69%, while the commercially available a collagen chondroitin calcium increase value-added rate of bone mineral density was 60.74%. And through the data obtained from the experiment shows that the city sold a collagen chondroitin calcium in main component role than biological carbonate calcium, collagen, CS, GS four synergistically increased bone mineral density has a significant effect. Among the 31 groups, the first 4 groups were compared with the experimental group which was used in the third chapter.4. The activity of alkaline phosphatase was detected by the modified Gomori’s calcium cobalt method to determine the degree of differentiation and functional status of osteoblasts.5. By using alizarin red staining of mineralized nodules to find out the best formula of the experiment.6. Through 4 and 5 of the experimental study, in the 4 group of the best program in the final determination of the first groups of programs for the best formula, first groups of four main functional components of the dose for the increase in bone density function of the best formula ratio of food.7. According to the obtained optimal formulation verify the effectiveness of multivariate polynomial equations, through repeated experiment and sensory score of measured value was 8.79, prediction of multivariate polynomial equations value 8.73 and second difference between 0.06 close comparison, shows that the model is scientific, reasonable and effective, the equation can better predict and screening theory and a comprehensive sensory score, it is of great practical significance,Can be used for the development of calcium collagen chewing tablets.8. Considering the effect of surface to that examines factors for the respective optimal region:Maltodextrin add amount to 10%, Xylitol was added 3.1%, L-malic acid addition of 0.15%.9. To explore the application of central composite design and response surface optimization method in health food of multiple factors and multiple levels in the experiment.