尿液结合性胆红素的纯化  

Purification of Conjugated Bilirubin from Human Urine Guo Yuanbiao

作　　者：郭元彪[1] 陈曼玲[1] 涂屹[2] 罗红权[3] 李胜利[2] 严大刚[2] 牛敏[2] 

机构地区：[1]华西医科大学基础医学院生物化学教研室,610041 [2]攀枝花市中心医院 [3]四川省妇幼保健院

出　　处：《华西医科大学学报》2001年第4期579-581,共3页Journal of West China University of Medical Sciences

摘　　要：目的　探索一种来源方便 ,具有延续性 ,利于胆红素测定校准物或质控物制备 ,且方法简便、实用、产率和纯度可靠的结合性胆红素 (Bc)纯化方法。方法　应用 CHCl3 ∶ CH3 CH2 OH(1∶ 1,V/V)萃取与 CNBractivated Sepharose4B- HSA亲和层析相结合的方法对胆道梗阻性高胆红素尿液进行分离纯化 ,并以 Doum as J- G法和 HPL C分析产品。结果　由 CHCl3 ∶ CH3 CH2 OH(1∶ 1,V/V)萃取所得的粗纯品经 J- G法检测产率为 90 .2±5 .4% (n=3) ,胆红素含量占总干重的 6 7.9± 6 .4% (n=3)。亲和层析的洗脱峰间的分离度为 1.5、1.737。层析后再萃取所得精纯品经 HPL C分析 ,产率为 46 .9% ,纯度 94.7% ,其中 BDG76 .2 % ,BMG18.5 %。结论　采用有机物萃取和 CNBr activated Sepharose4B- HSA亲和层析相结合的方法纯化结合性胆红素 (BMG& BDG)Objectives To develop a simple, practical method with fine yield and purity for separating conjugated bilirubin(Bc) from a sustainable and facile material in order that it could serve as a calibrator for total and direct bilirubin determination. Methods Isolating Bc from urine of patients with obstructive jaundice by CHCl 3∶CH 3CH 2OH(1∶1,V/V) followed by CNBr activated Sepharose 4B HAS affinity chromatography, we analyzed the products by the use of Doumas J G diazo method and HPLC, respectively. Results Determined by J G diazo method, the crude product extracted by CHCl 3∶CH 3CH 2OH(1∶1,V/V) yields 90.2±5.4%( n =3) of urine Bc, the ratio of Bc to total dry weight being 67.9±6.4%( n =3). The final product purified by affinity chromatography with the ratio of separation of 1.5, 1.737 and analyzed by HPLC has the yield of 46.9% and purity of 94.7%(BDG of 76.2%, BMG of 18.5%). Conclusion The above data demonstrate that urine conjugated bilirubin can be successfully purified by CHCl 3∶CH 3CH 2OH extraction followed by Sephrose HSA affinity chromatography with sufficient purity and output. [

关 键 词：结合性胆红素 亲和层析 尿液 纯化方法 

分 类 号：R313[医药卫生—基础医学]