摘要
目的:采用多种方法对地龙酶解液进行分离,得到抗血栓肽。方法:以尿激酶作为对照药物,建立体外活性测定方法;同时采用folin法进行多肽含量测定,采用电渗析法、超滤法、DA201-C树脂法、sephadexG15凝胶分离法、HPLC分离法对地龙酶解液进行分离。结果:采用尿激酶为对照药物进行体外纤溶活性测定,得出以溶圈面积为纵坐标,以尿激酶活力(U)的对数为横坐标的回归方程为 Y=1.560 2 X-1.833 1(R^2=0.996 4),尿激酶活性在100~50 000 U范围内线性良好;同时建立多肽测定方法,回归方程为Y=2.028 3 X+0.008 6(R^2=0.999 1),肽在0~0.2 mg/ml范围内线性良好。并通过多种方法地龙酶解液进行分离,最后HPLC分离得到5个组分,活性大小为F 3>F 2>F 1>F 5>F 4。结论:采用多种分离方法对地龙酶解液进行分离,方法可靠。
Objective: To obtain antithrombotic peptides, enzymatic hydrolysate of earthworm was separated by various methods. Methods: By using urokinase as a control drug, in vitro activity assay was established. Meanwhile, folin method was used to establish the determination method of polypeptide content. Electrodialysis method, ultrafiltration method, DA201-C resin method, sephadex G15 gel separation method and HPLC separation method were used to separate the earthworm enzymatic hydrolysate. Results: The in vitro fibrinolytic activity of urokinase was determined by using urokinase as a control drug, and the area of the dissolving ring was plotted on the ordinate. The regression equation with the logarithm of urokinase activity(U) as the abscissa was Y=1.560 2 X-1.833 1(R^2 = 0.996 4), and the linearity was good. At the same time, the polypeptide determination method was established, and the regression equation was Y=2.028 3 X+0.008 6(R^2 = 0.999 1), indicating a good linearity. The enzymatic hydrolysate of earthworm was separated by various methods, and five components were separated by high performance liquid chromatography. The order of activity was F3 >F2>F1> F5>F4. Conclusion: It is reliable to separate the enzymatic hydrolysate of earthworm by various methods.
作者
王佳茜
王少平
刘万卉
Wang Jiaxi;Wang Shaoping;Liu Wanhui(School of Pharmacy in Yantai University,Shandong 264000;Yantai Center for Food and Drug Control,Shandong 264000;BinZhou Medical University,Shandong 264000)
出处
《天津药学》
2019年第3期1-4,共4页
Tianjin Pharmacy
关键词
分离
半制备
地龙
活性测定
separation
semi-preparation
Lumbricus
activity determination
