Research On Technology Of Preparing Hemin By Horse Blood

Horse blood is rich in heme which is an important complex with ferriporphyrin. It is a biostateironsupplementary without toxic or side effect. When heme is divorced from globin， it is quickly oxidized intohemin. Therefore, the heme prepared by horse blood is usually the perchloride iron heme. The text took theanti-freezing horse blood of Sinkiang horses as the experimental material. In the aspect of general heminpreparation method, the preparation process of hemin is divided as four independent stages—breaking ofred blood cells, resolution of heme and globin, separation and extraction of hemin and heminpurification—to discuss the optimizing of process conditions at each stage. Research results are shownbelow:(1) Took the yield of hemin as the index, and researched the inlfuences of power, agent, proportionand time of cell breaking on the effect of cell breaking and compared it with the the reagent mixture cellbreaking method. The best process condition for the ultrasonic cell breaking method can be obtained by theorthogonal test: cell breaking power0.8kW, agent (water: alcohol) proportion, cell breaking treatment6min.(2) Took the resolution rate of globin and heme as the test index, researched influences of the PHvalue, additive amount of acetone and treatment time on the resolution rate and determined the processcondition model through single factor test and application of the Box-behnken response surface design. Theresearch shows the best process condition: the additive amount of acetone is5.25times of the red blood cellvolume, the PH value is2.86and the treatment time is3.38h. It is basically in line with the predicted data.(3) Took the purity and yield of hemin as the test index, the best process condition for CMC-Namethod is obtained through the orthogonal test: the pH value is6.0, the additive amount of CMC-Na is0.45g, the extraction is8h and the purity and yield of hemin is29.42%and56.08%respectively; the bestprocess condition for extracting hemin by sodium acetate method is that the additive amount of is2.0%ofthe acetone extraction liquid of hemin, the pH value is4.8, the extraction is15h, and purity and yield ofhemin is36.30%and44.88%respectively. The advantage of the CMC-Na method extraction of hemin isthat the complex of heme and sodium carboxymethylcellulose can be used as the tabletting material forantianemic agents directly, and the advantages of sodium acetate method extraction of hemin is that thepurity of crude hemin is high so that the purification in the later stage is simple, and the overall cost is low.(4) Took the purity of hemin as the test index, separated and purify hemin by the acid-base purification method, researched influences of ammonia concentration, solid-liquid ratio and acidification time on thepurity of hemin and selected the High Performance Liquid Chromatography to identify the purification ofhemin. The best process condition gained through the orthogonal test is that the ammonia concentration is1.5%, the solid-liquid ratio is1:30, the acidification time is10min and the purity of hemin afterpurification can be as high as92.87%.