Chem. Pharm. Bull. 55(1) 128—132 (2007)

material, was first recorded in ‘Shennong Bencao Jing’ and has been used clinically in China for 2000 years. It is the dry gallstone of Bos taurus domesticus GMELIN and, according to the theory of traditional Chinese medicine (TCM), has the effects of sedation, anti-hyperspasmia, relieving fever, diminishing inflammation and normalizing function of the gallbladder. Because of its scarcity of natural resource and high price, many studies have been carried out in order to find substitutes for Calculus Bovis, and some fruitful progresses have been achieved. For instance, artificial synthesized and in-vitro cultured Calculus Bovis have already been developed and recently used in clinic and medicine preparations. However, due to the different developmental conditions, chemical constituents of substitutes might be different from those of natural Calculus Bovis, which thus may lead to the variation of therapeutic effects. Therefore, to ensure the quality of Calculus Bovis and its substitutes, efficient quality control approach is urgently needed. Previous studies have also proven that the most important bioactive constituents in Calculus Bovis are bile acids and bilirubin (see the chemical structures in Fig. 1), and hence the quality control of Calculus Bovis and its substitutes should depend on the concentration of bile acids and bilirubin. So, it is necessary to comparatively study these major bioactive constituents in natural, artificial and in-vitro cultured Calculus Bovis. Several techniques are available to analyze bile acids and bilirubin in Calculus Bovis, such as thin layer chromatography (TLC), capillary electrophoresis (CE), high-performance liquid chromatography (HPLC). These methods gener128 Vol. 55, No. 1 Chem. Pharm. Bull. 55(1) 128—132 (2007)