Study On The Preparation And Characterization Of Rana Bone Flesh Polypeptide Chelated Calcium And Its Effect On Promoting Calcium Absorption

Rana,which is mainly distributed in the three northeastern provinces of China,is a very famous frog family.It has high value,mainly food supplement,health care,beauty and economic medicinal value.However,at present,the processing and utilization of Rana chensinensis is extremely low,which leads to the development and production of industries such as catering,medicine,health care products,which are the raw materials of Rana.The products are still dominated by Rana oil products.Rana oil only accounts for about 15%of female Rana.The remaining 85%of female Rana and all male Rana cannot be effectively utilized.They are only partially used as animal feed,and most of them are discarded as waste.As a result,it caused great waste of resources and serious pollution of the environment.Therefore,this experiment uses the unique Rana bone flesh of Northeast China as the raw material,and optimizes the enzymatic hydrolysis process of Rana sylvestris with the degree of hydrolysis and peptide yield.Then,the chelation rate is used to optimize the chelation of enzymatic hydrolysate and CaCl₂.The process was followed by designing a Caco-2cell model to investigate the absorption characteristics of the Rana bone flesh chelated calcium relative to other calcium supplements from the cytological point of view.Finally,the related products of the Rana bone flesh were comprehensively scored as indicators.Optimize the formulation process of chelated calcium tablets.The main findings are as follows:?1?Study on enzymatic hydrolysis of Rana bone flesh:Based on the degree of hydrolysis and the yield of the peptide,the enzymes of the Rana bone flesh were prepared under the conditions of five enzymes:alkaline protease,neutral protease,trypsin,pepsin and flavor protease.The peptide liquid of Rana bone flesh was prepared,and the hydrolysis of trypsin was the best.The degree of hydrolysis was18.77±0.374%,and the yield of peptide was 34.66±0.348%.The effects of hydrolysis time,hydrolysis temperature,enzyme amount and pH on the hydrolysis of Rana bone flesh polypeptide were studied.L₉?3⁴?orthogonal test was used to optimize the extraction conditions of Rana bone flesh polypeptide.The optimal enzymatic conditions were:The enzyme amount was 5000 U/g,pH 8,temperature 45?,time 4 h,the degree of hydrolysis at this time was 21.34±0.017%,and the peptide yield was40.11±0.043%.Under the optimal process,the Rana bone flesh polypeptide liquid was prepared.The peptide liquid was made into polypeptide powder,and the physicochemical properties of the polypeptide powder were detected.The results showed that there was a UV characteristic absorption peak of the polypeptide at 210nm,and the characteristic functional group of the polypeptide was detected by Fourier infrared spectroscopy at 1538 cm^-1,indicating The Rana bone flesh polypeptide powder was successfully prepared.?2?Preparation and characterization of polypeptide chelating calcium from Rana bone flesh peptide:Using the Rana bone flesh polypeptide liquid and CaCl₂ as raw materials,the peptide chelated calcium was prepared under certain conditions,and the chelation rate was used to investigate the chelation temperature,time,peptide-calcium ratio and ethanol addition ratio.The effect of factors on chelation rate.The single factor response surface test study was used to determine the optimal process.The chelating conditions were:time 30min,temperature 30?,peptide-calcium ratio?The ratio of the volume of the peptide liquid to the mass of calcium chloride added?10:1?ml/g?,ethanol addition ratio 8,and chelation rate reached 79.43±0.065%.The prepared structure of the chelated calcium of the polypeptide was analyzed by Ultraviolet spectroscopy analysis,Fourier infrared analysis,scanning electron microscopy,thereby presuming the structure of the polypeptide chelated calcium.The results show that after analysis by ultraviolet and infrared,it can be clearly seen that the chemical structure of the polypeptide and the chelated calcium are changed.After scanning electron microscopy,more"white crystals"can be clearly seen.The above results indicate that the polypeptide forms a new chelate with calcium.?3?Study on the absorption characteristics of calcium chelating polypeptide in Rana bone:Using the Caco-2 cell model,the cell viability was measured by MTT assay to construct a cell model.Taking CaCl₂,calcium gluconate and CCP-Ca as the control group,the absorbing characteristics of the Rana bone flesh polypeptide chelated calcium relative to other calcium supplements were discussed from the cytological point of view.The Rana bone flesh polypeptide chelate calcium will have a certain effect on the survival of the cells.The excessive addition will increase the growth of the cells.When the amount of the Rana bone flesh polypeptide is more than0.5 mg/ml,the cell survival The rate is significantly reduced.The Rana bone flesh polypeptide chelate calcium is similar to CCP-Ca in cell absorption and is superior to CaCl₂ and calcium gluconate.?4?Study on the preparation technology of Rana bone flesh chelated calcium tablets:Preparation of Rana bone flesh polypeptide chelated calcium tablets by wet granulation.Results The selected excipients of the Rana sylvestris bone polypeptide chelating calcium tablets were:the filler was microcrystalline cellulose,the flavoring agent was sorbitol,and the acidulant was malic acid.Taking the comprehensive score of the tablet as the index,the best formula of the tablet excipient was obtained by the single factor response surface test:the addition amount of the filler?microcrystalline cellulose?was 30%,and the flavoring agent?sorbitol?The amount added was 45%,and the amount of the sour agent?malic acid?was 5%.The other formula added in the preparation of the tablet is:the disintegrant?croscarmellose sodium?is added in an amount of 7%,the lubricant?magnesium stearate?is added in an amount of 3%,and the forest frog bone meat polypeptide is chelated.The amount of calcium added was20%.The obtained tablet has a round appearance,a smooth surface,milky white color and uniform color;no taste on the mouthfeel,suitable milky taste,sweet and sour taste,moderate hardness,no graininess.