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目的：建立一种快速测定胆红素ＩＸα及其异构体的相对含量的方法．方法：采用高效液相色谱法，以ＫｒｏｍａｓｉｌＣ１８（１５０ｍｍ×４．６ｍｍ，５μｍ）为分析柱，以乙腈∶甲醇∶醋酸铵－醋酸ｐＨ＝４．７（１∶７∶２Ｖ／Ｖ／Ｖ）为流动相，检测波长λ：４５０ｎｍ；流速：２．０ｍｌ／ｍｉｎ，可直接分离测定３个胆红素异构体．结果：对４批胆红素样品中３个异构体的相对含量用归一法处理进行了测定，胆红素ＩＸαｔＲ＝１３．１８ｍｉｎ，胆红素ＸⅢαｔＲ＝９．６５ｍｉｎ，胆红素ⅢαｔＲ＝１７．３５ｍｉｎ，胆红素异构体的分离度（Ｒ）分别为Ｒ１＝１．８０，Ｒ２＝２．０４．结论：本法快速、准确、灵敏、重现性好，可适用于胆红素对照品、胆红素的制备纯度鉴定及中药中胆红素的质量控制，对血清及胆汁中胆红素的测定也具有实际应用价值．
Objective: To establish a rapid determination of the relative content of bilirubin IXα and its isomers. Methods: High performance liquid chromatography (HPLC) with Kromasil C18 (150mm × 4.6mm, 5μm) as analytical column and acetonitrile: methanol: ammonium acetate - acetic acid pH = 4.7 (1: 7: 2 V / V / V) Mobile phase, detection wavelength λ: 450nm; flow rate: 2.0ml / min, direct separation and determination of three bilirubin isomers. Results: The relative contents of three isomers in four batches of bilirubin samples were determined by the normal method. The bilirubin IXαtR = 13.18min, the bilirubin XⅢαtR = 9.65min, the bilirubin ⅢαtR = 17.35min, bilirubin isomers of the separation (R) were R1 = 1.80, R2 = 2.04. Conclusion: The method is rapid, accurate, sensitive and reproducible. It can be applied to bilirubin reference substance, the purity of bilirubin preparation and the quality control of bilirubin in Chinese medicine. The bilirubin Determination also has practical value.