纤维蛋白原-凝血酶时间法用于水蛭质量控制方法的研究  

Research on the method of fibrinogen-thrombin time by coagulometer for quality control of hirudo

作　　者：苏斌[1] 王志斌[2] 郭玉东[2] 宋程程[1] 胡宇驰[2] 王碧松[2] 金家金 

机构地区：[1]北京中医药大学中药学院,北京100102 [2]北京市药品检验所,北京100035

出　　处：《药物分析杂志》2014年第10期1802-1806,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立一种基于用血液凝固分析仪测定纤维蛋白原-凝血酶时间的水蛭生物活性测定方法(Fibg-TT法),定义效价并建立水蛭标准品。方法:就基源为蚂蟥和日本医蛭的水蛭分别建立了反应体系,水蛭浸提液比空白对照凝固时间延长1 s作为1个效价单位(U),定义并测得水蛭标准品效价。将高、中、低浓度的水蛭标准品和供试品的浸提液与0.5%的纤维蛋白原混合,加入凝血酶,用血凝仪测定凝固时间,结果进行对数转换后,用生物检定统计法中的量反应平行线3.3法测定供试品的效价。结果:本实验测得蚂蟥和日本医蛭标准品的效价分别为5650 U·g-1和4130 U·g-1,当两者浸提液的浓度分别在121.5~1130 U·mL-1和17.3~66.1 U·mL-1范围内时,对数凝固时间与浓度线性较好(r=0.9985;0.9965),准确度(RSD=4.5%,n=6;RSD=3.6%,n=6)、重复性(RSD=3.8%,n=6;RSD=2.4%,n=6)和中间精密度(RSD=1.7%,n=6;RSD=2.4%,n=6)均良好,该方法测定结果可靠。结论:该方法用延长凝固时间来定义水蛭的效价,并建立水蛭标准品,具有可行性。Fibg-TT法,操作简便,人为因素影响小,客观准确,重复性和中间精密度好,能区分和量化不同批次水蛭的效价,可用于水蛭效价的测定,控制其质量。Objective： To establish a new method for hirudo bio - assay based on fibrinogen - thrombin time （ Fibg -TT） and define the titer, and to further establish the standard substance of hirudo. Methods：Firstly, the reaction system for hirudo with base sources of Whitmania pigra Whitman and Hirudo nipponica Whitman was respectively established. One titer unit （U） was defined as one second longer for hirudo extract liquid than the coagulation time of the blank control, and the titer of hirudo standard substance was defined and measured. Secondly, the standard sub- stance of hirudo at high, middle and low concentrations and the extract liquid of the test sample were mixed with 0. 5 % fibrinogen, then added with thrombin and the Fibg - TT was determined by coagulometer. After logistic trans- formation of the obtained results, the titer of the test sample was determined by Method 3.3 parallel line quantitative analysis described in the Biological Test Statistical Methods. Results： The titer of the standard substance of Whitmaniapigra Whitman and Hirudo nipponica Whitman was measured to be 5650 U · g-1 and 4130 U · g-1 Within the range of 121.5 -1130 U · mL-1 and 17.3 -66. 1 U · mL-1, the log coagulation time（ Fibg- q^F）and the Whitmaniapigra Whitman and Hirudo nipponica Whitman concentration showed a good linear relationship respectively（ r = 0. 9985 ;0. 9965 ）. The method of Fibg - TT was accurate and reliable with good accuracy （ RSD = 4.5 % , n = 6 ; RSD = 3.6 %, n = 6 ） and reproducibility （ RSD = 3.8 %, n = 6 ; RSD = 2.4%, n = 6 ） and intermediate precision （ RSD = 1.7%, n = 6 ; RSD = 2.4%, n = 6 ）. Conclusion： The method of Fibg - TT to define the titer of hirudo and establish the standard substance of hirudo based on prolonging coagulation time is practical. The Fibg - TT method is simple, objective, accurate, reproducible, precise and the human influencing factors are few. This meth- od can distinguish and quantify the titer of different batches of hirudo and can be used for

关 键 词：水蛭 标准品 纤维蛋白原-凝血酶时间 效价 生物检定法 方法学考察 

分 类 号：R917[医药卫生—药物分析学]