| The methods of preparing silkworm pupa amino acid powder for transfusion by hydrolyzing degreased silkworm pupa with HC1,sublimating the hydrolyzed solution through ion exchange and adsorption with cation colophony,removing pyrogen from amino acid solution with active carbon and diatomite,condensing and freezing desiccation are introduced in this article . The Limulus Amoebocyte Lysate(LAL) test and rabbit test which are used to examine the pyrogen in amino acid solution are also narrated in this thesis. The results not only showed that the content of pyrogen in silkworm pupa amino acid powder was satisfied with the demands but also proved that the feasibility to produce amino acid transfusion from silkworm pupae protein. Moreover,the content and composition of silkworm pupa amino acid powder are mensurated with several kinds of instruments,including Hitachi 835-50 automatic amino acid analyzer,Foss SOXTEC Auto Extraction Unit(2050),Tector Automatic Nitrogen analyzer(2300)and Absorption Flame Emisson Spectrophotometer.The whole experiment has four parts. The first part:Preparation of silkworm pupa amino powder.The degreased silkworm pupae was hydrolyzed by 6 mol/L HC1 for 24 hours between 105Cand 110C. The hydrolysate was cleansed acid and purified with exchanger Resion. There were other procedures which were used to obtain the amino acids powder from the previous purified solution,such as condense,freeze drying and so on. The experimental results showed that the silkworm pupae protein could be hydrolyzed completely with hydrochloric acid under the conditions reported above and the obtained amino acids could keep their biological activity as well. The experiment also indicated that this type of amino acid powder contained 7 kinds of essential amino acids for human beings with high contents (tryptophan has been destroyed during the course of hydrolysate with hydrochloric acid) and its obtainment rate is high to 40%. The second part:Removing of pyrogen in silkworm pupae amino acids powder.The mainly way to elimination pyrogen in silkworm pupae mixed amino acid powder in this study is adsorption with active carbon. The approaches are as followings:adding 25% worked active carbon (according to the quality of silkworm pupae) to the purified solution;then mixing it up as heating;last,filtrating it and obtained the adsorbed solution of silkworm pupae amino acid . The test to the pyrogen in the above solution indicated that the content of pyrogen in it was satisfying with our goals. The third part:Test of pyrogen in the amino acids powder of silkworm pupa .It was two ways to test the pyrogen in the amino acid powder of silkworm pupae in this experiment. One is LAL test,the other is Rabbit test.LAL test is a method to estimate whether the content of pyrogen is eligibility or not by forming gelatin. During the course of LAL test,the LAL reagent sensitivity was confirmed and the interference test was done before using the reagent for examining the concentration of bacterial endotoxins. The confirmation of LAL reagent sensitivity and interference are all carried out under the regulation of pharmaceopoeia of China. It was found that the 1:4,1:8,1:16 diluted solution,adsorbed by active carbon,with the content of 10% amino acid have interferences;while the 1:32 diluted solution with the same content of amino acid did not occur the interference. However,all of the four kinds of diluted solutions which were adsorbed by 25% diatomite had interferences. After the LAL reagent sensitivity test and the interference test,the following test was the examination of pyrogen. The process of it is the same with the two texts reported previously. The results of this examination showed that the content of the pyrogen in the solution (1:32) adsorbed by active carbon was unqualified.During the course of pyrogen examination,if the LAL test proves the content of pyrogen to be eligible,we can judge the qualification of pyrogen . If the LAL test reaches a contrary result,we have to use Rabbit test to identify this result and...
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