| Rana sylvatica is a valuable nutrient commonly found in the northeastern region of China.It is used in the preparation of Rana sylvatica oil and other products.Rana sylvatica collagen polypeptide is a highly active biological product obtained by enzymatic hydrolysis of Rana sylvatica,which is a by-product of the production of Rana sylvestris oil.This study starts from the collagen peptide and chelates it with ferrous ions.A functional peptide iron supplement was synthesized to improve the current status of iron deficiency anemia that is widespread in today's society.Compared with the previous generations of iron supplements,this product does not produce irritating effects on the human gastrointestinal tract,has good palatability,and is easy to store and transport.The main findings of this paper are as follows:1.Select Rana sylvatica collagen peptide and ferrous chloride to chelate to prepare Rana sylvatica collagen peptide iron chelate;calculate the standard curve of iron by phenanthroline colorimetric method to calculate chelation.The content of iron in the material.The effects of Rana henryi collagen peptide concentration,p H value,ascorbic acid concentration,reaction time and reaction temperature on the chelation rate of Rana sylvatica collagen peptide ferrous chelate were examined in turn through a single-factor test.The four factors that have a greater impact on the four-factor three-level orthogonal optimization test.The optimum technological parameters for preparing the ferrous protein chelate of Rana sylvatica collagen peptide were as follows: frog collagen peptide concentration 9%,p H value 5,ascorbic acid concentration 2.5%,reaction time 80 min,reaction temperature 50?.2.In order to improve the antioxidation of the product and enhance the fluidity of cell membrane,glycosylation modification was performed on collagen protein polypeptide of raw Rana sylvatica,and then the modified Rana henryi collagen peptide was chelated with ferrous ion to prepare peptide iron chelates.The chelation rates of glycosylation-modified Rana chensinensis collagen polypeptide iron chelates were examined by single factor tests,followed by the following five factors: sugar type,ratio of peptides and sugars,p H value,reaction temperature,and reaction time.The four factors and three levels of orthogonal optimization experiments were selected for the influence of the four factors.The optimum technological parameters for preparation of glycosylated Rana sylvatica collagen peptide iron chelate were selected through orthogonal optimization experiments.The optimal parameters were as follows: glucose was selected for glycosylation modification and the ratio of peptide to sugar was 1:1,p H 7,reaction temperature 70?,reaction time 18 h.3.Sulfide method was used to qualitatively study the composition of iron frog collagen protein peptide ferrous chelate.It was confirmed that there was no free polypeptide in chelate,and it was confirmed that the peptide and ferrous ion were fully chelated;Infrared spectroscopy analysis of the chelated materials before and after the structure were identified,according to the different peaks and their changing characteristics to determine the formation of chelating products;through a series of experiments on the product of the determination of anti-oxidation.The scavenging capacities of DPPH,hydroxyl radicals,superoxide anion and their reducing ability were measured and compared before and after glycosylation.The experimental results showed that the ferrous chelates of Rana henryi collagen peptide modified by glycosylation were significantly higher than the ferrous chelates of Rana henryi collagen peptides that were not modified by glycosylation in the above aspects.Glycosylation is beneficial to improve the antioxidant properties of a substance.At the same time,it was concluded that when the glycosylation-modified chelate concentration is 4 mg/m L,its DPPH,hydroxyl free radical and superoxide anion scavenging ability is the strongest;its reducing ability is proportional to the concentration.4.Rapain collagen peptide polyferric chelate microcapsules were prepared by an orifice-coagulation bath method.Firstly,the effects of four factors,namely concentration of sodium alginate,ratio of core to wall,reaction temperature,and concentration of calcium chloride,on the embedding rate of collagen conjugated iron chelate microcapsules of Rana chensinensis were investigated in a single-factor test.Based on the single factor experiment,the response surface method was used to optimize the preparation process of Rana chensinensis collagen peptide ferrous chelate microcapsules.The concentration of sodium alginate,core wall ratio,reaction temperature,and calcium chloride concentration were used as response factors.In order to establish a quadratic regression equation,the embedding rate of Rana sylvatica collagen polypeptide iron chelate microcapsules was the response value.Get the following equation:Y=85.36-0.29A+0.88B-0.76C+1.09D+0.45AB+1.15AC+0.72AD-1.19BC-0.21BD+0.23CD-11.42A2-6.28B2-5.05C2-7.38D2.It can be seen that the model selected in this experiment is significant(P<0.05),the fitting degree is good,and the correlation between the predicted value and the experimental value is very good.The optimum process conditions obtained by the response surface software were as follows:sodium alginate concentration of 2.96%,core wall ratio of 0.41,reaction temperature of 45.8°C,and calcium chloride concentration of 4.28%.Polypeptides were prepared by the regression equation.The highest encapsulation efficiency of iron chelate microcapsules is 85.46%,and the confidence is 99.3%,which has good simulation. |
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