Rothuizen J, Heirwegh K P, van Kouwen A M
Small Animal Clinic, University of Utrecht, The Netherlands.
J Chromatogr. 1988 May 13;427(1):19-28. doi: 10.1016/0378-4347(88)80101-4.
A method for the separation and quantitation of ethyl anthranilate or p-iodoaniline azo derivatives of bile pigments was developed using reversed-phase high-performance liquid chromatography. A convenient separation was achieved in 15 min, permitting the quantitation of the unconjugated azo-dipyrrole (alpha o) and its glucuronide (delta), xyloside (alpha 2) and glucoside (alpha 3) conjugates. The pathological beta- and gamma-azo pigments, derived from bilirubin glucuronide isomers that occur in cholestatic bile or plasma, are also detected in this system. The results of this method as applied to bile from 25 healthy dogs were in excellent agreement with the values obtained by reversed-phase chromatography of bilirubin and its mono- and dimethyl esters produced from the corresponding conjugates by alkaline methanolysis. This system permits the sensitive and convenient determination of bilirubin and its conjugation pattern in biological fluids.
采用反相高效液相色谱法开发了一种分离和定量胆汁色素邻氨基苯甲酸乙酯或对碘苯胺偶氮衍生物的方法。在15分钟内实现了便捷的分离，可对未结合的偶氮二吡咯（α o）及其葡糖醛酸苷（δ）、木糖苷（α 2）和葡糖苷（α 3）共轭物进行定量。在胆汁淤积性胆汁或血浆中出现的源自胆红素葡糖醛酸苷异构体的病理性β-和γ-偶氮色素，在该系统中也能被检测到。将该方法应用于25只健康犬的胆汁，其结果与通过碱性甲醇ysis从相应共轭物生成的胆红素及其单甲酯和二甲酯的反相色谱法获得的值高度一致。该系统可对生物体液中的胆红素及其共轭模式进行灵敏且便捷的测定。
