Study On The Preparation And Properties Of Chelated Salt Of Pig Blood Polypeptide-iron

Iron deficiency anemia(IDA),one of the most common anemia in the world,is a kind of cell hypochromic anemia due to insufficient iron which is needed to make hemoglobin.According to WHO,about 10%to 30%of the whole world population are suffered from different degrees of iron deficiency,the incidence of men by about 10%, 20%more than women.At present,prevention and treatment of iron deficiency anemia is mainly achieved through the intake of iron agents such as ferrous sulfate,ferrous chloride,ferrous gluconate,ferrous lactic,ferrous succinate and ferrous fumarate.These agents have good effects in iron supplementation for their high iron content,whereas they can be relatively low utilized in the body,and have side effects and a special taste of metal rust.Polypeptide chelated iron,a biostate iron chelated and generated by polypeptide from albuminolysis and ferrous iron ion,is an ideal agent for it can not be affected by such interferents as carbonate,tannic acid and cellulose,but can always be absorbed directly by intestinal cells without any toxic side effects and irritation.China is one of the biggest pig producing countries,slaughtering 4 - 500,000,000, pigs each year.And each pig has about 2kg blood,so about 8 to 1,000,000,000 kg pig blood will be produced each year,which is equivalent to the protein content of 4,000,000 tons of pork or 4,600,000 tons of eggs.While the pig blood is nutritious,but because of its poor color,heavy blood smell,poor palatability and non-digestible absorption of hemoglobin,it has not been fully utilised.At present,only less than one third of the pig blood in China is processed as food and blood powder,and there are about 70%of it abandoned.That leads to a waste of valuable resources and environmental pollution.So,in this paper the pig blood powder has been used to be raw materials,and been hydrolysed by alkaline proteinase,and then the chelating reaction has been conducted between the polypeptides separated and purified by film from the pig blood and ferrite. And then physicochemical properties and functional properties of the chelated pig blood polypeptide-iron salt have been studied,and its toxicological safety has been evaluated with a view to providing a ideal supplementation iron agent for people who are iron deficiency anemia.Finally,the theoretical basis for its promotion and application and a useful reference have been provided.This paper consists of the following findings:1,Research on preparation of pig blood polypeptides by means of enzymatic hydrolysis1.1 Concerning hydrolysis,2709 alkaline protease had relatively good effects on pig blood powder,and the optimum hydrolysis technology parameters were:pH value,10.5; hydrolysis temperature,45℃;concentration of substrate,2%;the ratio of enzyme and the substrate quality,3.0%;hydrolysis time,6h..1.2 Activated carbon had good decolorization effects on the enzymolysed liquid of pig blood powder,and the best decolorization process parameters were:the dosage of activated carbon,2.5%;pH value,4.0;decolorization temperature,60℃;adsorption time, 1.0h.Under the best decolorization conditions the decolorization rate of the enzymolysed liquid of pig blood powder was 92.20%,while the loss rate of nitrogen was 10.42%.1.3 Ion exchange resins had relatively good demineralization effects on the decolorized enzymolysis liquid of pig blood powder.The demineralization rate was 82.11%when the sample injection speed of decolorized enzymolysis liquid of pig blood powder was 2BV/h,while the loss rate of nitrogen was 6.59%.Polypropylene hollow fiber ultrafiltration membrane had relatively good effects of retention and separation on the demineralized enzymolysis polypeptides liquid of pig blood powder.The optimum operation pressure of ultrafiltration was 0.15MPa,and the optimum temperature of polypeptides solution was 25℃.Under the conditions of optimum operation pressure and optimum temperature the pig blood polypeptides of four relative molecular mass segments were obtained,and they were the segments above 10kDal,5～10kDal,1～5kDal and the segment below 1kDal.The rates of polypeptide yield were 4.38%,13.97 %,74.76%and 6.89%respectively,and the relative molecular mass concentrated on 1～5kDal.1.4 The yielded pig blood polypeptides are in the form of light yellow powder with delicate mouth.feel and taste,slightly bitter but no smell of blood.2,Preparation and component analysis of the chelated salt of pig blood polypeptide-iron2.1 From dissolubility of ferrous sulfate and ferrous chloride in different concentrations of ethanol,we could see that ferrous chloride could be completely dissolved in different concentrations of ethanol solution,and it was more suitable for it to be used as the iron source in the chelation.2.2 The optimum technology parameters of the preparation of the chelated salt of pig blood polypeptide-iron were:pH value,5;the mass ratio of polypeptides and ferrous chloride is 4:1;the polypeptides solution concentration 3.0%.2.3 From the qualitation measurement of the chelating reaction products by sodium sulfide method we could see that the pig blood polypeptides iron existed in the form of polypeptides chelate compound,and it was a new material,and it was different from the pig blood polypeptides and ferrous chloride,and its major components were polypeptide and iron,the mass percentages of the two were 70.02%and 12.23% respectively.3,Study on the physicochemical properties of the chelated salt of pig blood polypeptide-iron3.1 The chelated salt of pig blood polypeptide-iron could be soluble in water,and under the neutral conditions,it had the minimum solubility,and with the increase of acidity or alkalinity,the solubility rised;and the chelated salt of pig blood polypeptide-iron could not be dissolved in organic solvents.3.2 The chelated salt of pig blood polypeptide-iron were more stable in appearance and the state of solid and liquid than ferrous sulfate.3.3 Through a preliminary scan of the ultraviolet and infrared spectroscopy analysis,we could see that the chelated salt of pig blood polypeptide-iron was different from pig blood polypeptide and ferrous chloride,and it was a new type of chelate.4,The study on the antioxidant and anti-anemia function of the chelated salt of pig blood polypeptide-iron4.1 The chelated salt of pig blood polypeptide-iron could remove superoxide radical anion(O₂.) and hydrogen peroxide(H₂O₂).The remove ability of the chelated salt of pig blood polypeptide-iron was weaker than that of vitamin C,but stronger than that of pig blood polypeptides,so the chelated salt of pig blood polypeptide-iron had relatively strong ability of removing superoxide radical anion(O₂) and hydrogen peroxide (H₂O₂).4.2 To establish the model of rat iron deficiency anemia successfully,it involved bloodletting caudal vein each week and feeding it with low iron feed for four weeks. The rat's hemoglobin,the red blood cell count and serum iron levels increased significantly(P＜0.05),fed with the chelated salt of pig blood polypeptide-iron after 4 weeks.It was obvious that the chelated salt of pig blood polypeptide-iron could improve the rat's iron deficiency anemia,and its anti-anemia function was better than that of ferrous gluconate and ferrous chloride.5,Safety evaluation of the chelated salt of pig blood polypeptide-iron5.1 Acute oral toxicity(LD₅₀) test showed that the mice had no toxic reaction after being fed with the chelated salt of pig blood polypeptide-iron by mouth.Counted by the improved formula of Karber method the LD₅₀ of the chelated salt of pig blood polypeptide-iron was 30 998 mg / kg,and it was more than 15 000mg/kg,so it could be concluded that the chelated salt of pig blood polypeptide-iron was innocuous substance. 5.2 Ames test showed that under the conditions of activating the five groups of different dose by adding S-9 or not,the number of anaplasia bacterial colony of the 4 strains, TA97,TA98,TA100 and TA102,was close to that of the negative control group,and less than 2 times.And compared with positive control group the number was much lower.So it could be concluded that the chelated salt of pig blood polypeptide-iron could not lead to back mutation between the Salmonella typhimurium mutation strain and base replacement mutant,and it had no mutagenicity.Micronucleus and abnormality test showed that the difference in the rate of cell micronuclear and abnormal sperm between the positive control group(fed with cyclophosphamide)and the negative control group was significant(p＜0.01),and that between the negative control group and the groups fed with different dose of the chelated salt of pig blood polypeptide-iron was not significant(p＞0.05),and there's no dose - effect relationship between them.So the chelated salt of pig blood polypeptide-iron had no abruption effect on mice bone marrow cells chromosome,and had no effect on mice abnormal sperm.5.3 The test of feeding the mice with the chelated salt of pig blood polypeptide-iron for 30 days showed that the growth development and behaviour of mice of the three groups of different dose were normal,and compared with that of the negative control group the differences in body weight and food utilization were not significant(p＞0.05).Compared with that of the negative control group,the differences in blood biochemical indicator were not significantly different(P＞0.05),and there's no dose-effect relationship between them.Compared with that of the negative control group,the differences in the ratio of viscus and corpora were not significant(p＞0.05),and there is no abnormality in the test of organ histopathology.