Li F, Lim C K, Peters T J
J Chromatogr. 1986 Feb 26;353:19-26. doi: 10.1016/s0021-9673(01)87072-2.
A novel high-performance liquid chromatography (HPLC) system is described for the separation of bilirubin and its conjugates in body fluids. Biliary bilirubin mono- and diglucuronide can be analysed directly on a C18 reversed-phase column with acetonitrile-dimethyl sulphoxide-0.1 M ammonium acetate (pH 5.16) (50:50:85, v/v/v) as mobile phase. However, the simultaneous determination of conjugated and unconjugated bilirubins in plasma required conversion of the conjugates into their methyl esters by alkaline methanolysis before HPLC separation of the C18 column eluted with acetonitrile-dimethyl sulphoxide-0.50 M ammonium acetate (pH 4.6) (50:50:40, v/v/v). The method is superior to, and more flexible than, previously described reversed-phase systems by allowing precise control of retention times by adjustment of pH, buffer concentration and the relative proportion of organic modifier in the mobile phase.
本文描述了一种新型高效液相色谱（HPLC）系统，用于分离体液中的胆红素及其共轭物。胆汁中的胆红素单葡萄糖醛酸酯和双葡萄糖醛酸酯可以直接在C18反相柱上进行分析，流动相为乙腈-二甲基亚砜-0.1M醋酸铵（pH 5.16）（50:50:85，v/v/v）。然而，在血浆中共轭胆红素和非共轭胆红素的同时测定需要通过碱性甲醇解将共轭物转化为甲酯，然后在C18柱上用乙腈-二甲基亚砜-0.50M醋酸铵（pH 4.6）（50:50:40，v/v/v）进行HPLC分离。该方法优于先前描述的反相系统，并且更灵活，通过调节pH、缓冲液浓度和流动相中有机改性剂的相对比例，可以精确控制保留时间。
