猪胆汁中胆红素(游离型)与重氮化试剂发生偶联反应必须借助于乙醇、甲醇等反应促进剂的作用。但是在胆汁中加入乙醇、甲醇最大的问题是产生沉淀,并造成测定误差。本实验方法改用先加入氯仿,充分振摇,再加入甲醇,可提高测定精度。实验还考察了使用不同介质的样品液的紫外吸收波长。本改进方法经用三批胆汁为底物测得回收率在95～103%之间。 The coupling reaction of bilirubin (free type) and diazotization reagent in pig bile must take the help of reaction accelerators such as ethanol and methanol. However, the biggest problem with the addition of ethanol and methanol to bile is the precipitation, which causes measurement errors. In this experiment method, chloroform was added first, shaken sufficiently, and then methanol was added to improve the measurement accuracy. The experiment also examined the ultraviolet absorption wavelengths of sample solutions using different media. The improved method has a recovery rate of 95-103% measured using three batches of bile as a substrate.