僵蚕抗凝活性部位中化学成分的初步研究

目的探索僵蚕水煎液、醇沉液和凝胶色谱馏分各部位中的抗凝活性成分。方法用水煎煮、60％醇沉处理和凝胶色谱方法制备抗凝活性部位，用化学反应鉴别、氮测定法、电导、紫外吸收光谱和高效液相色谱方法检测各部位中的蛋白质或多肽和氨基酸类及草酸铵等成分。结果僵蚕的水煎煮提取、醇沉和凝胶色谱分离所得抗凝活性部位中，多肽、氨基酸类和草酸铵都是其中的主要成分，其含量在80％以上。醇沉处理使草酸铵含量下降约52％，而多肽、氨基酸类含量仅降低约8％。凝胶色谱净化后，多肽、氨基酸类含量与水煎部位比约提高28％，与醇沉处理部位比约提高39％；而草酸铵含量与醇沉液部位相当，但比水煎煮部位降低约1倍。各部位的固形物中，15种氨基酸含量在40％～50％，占定氮法测定的蛋白质含量的60％～76％；多肽类成分的分子量为1．0～4．4kDa。结论本研究为僵蚕抗凝活性成分的进一步分离纯化提供了实验基础。

Primary Study on the Chemical Constituents in Anticoagulating Active Fractions of Bombyx Batryticatus

Objective To explore the anticoagulating active components in water extraction, alcohol sedimentation and fraction separated by gelfiltration chromatography of Bombyx Batryticatus. Methods Anticoagulating active fractions were prepared by the methods of water extraction, 70% alcohol sedimentation and gelfiltration chromatography. Components （such as porteins or polypeptide, amino acids, oxalic acid ammonium） in each fraction were determined by chemical reaction indentification, Kjeldahl method, conductance, UV and HPLC. Results Polypeptide, amino acids and oxalic acid ammonium were the main components in anticoagulating active fractions, content of which reached more than 80%. The content of oxalic acid ammonium decreased about 52% processed by alcohol sedimentation, while the content of polypeptide and amino acids only decreased about 8%. The content of peptides and amino acids increased about 28% purified by gelfiltration chromatography compared with the one in water extraction and about 39% compared with the one in alcohol sedimentation. The content of oxalic acid ammonium was the same as the one in alcohol sedimentation, but decreased about 1 time compared with the one in water extraction. The content of 15 kinds of amino acids were 40%-50% in the solids of each fractions, which maked up to 60%-76% of the content of proteins determined by Kjeldahl method. Molecular weight range of peptides is 1.0-4.4 kDa. Conclusion This study provides a experimental basis for further separation and purification of anticoagulating active components from Bombyx Batryticatus.