Preliminary Study on Contents and Ratios of Different Types of Bilirubin in Natural and in Vitro Cultured Calculus Bovis / 中国药学杂志

ABSTRACT

OBJECTIVE: To develop a method for determination of different types of bilirubin in bezoar and explore the differences in their contents or ratios between natural and in vitro cultured Calculus Bovis to provide reference for identification of Chinese patent drugs containing bezoar. METHODS: Free bilirubin was determined directly by HPLC, while unconjugated and total bilirubin in bezoar were converted into free bilirubin respectively using different chelating agents. Converted bilirubin was eluted on a Diamonsil C18 column(4.6 mm×150 mm, 5 μm)using isocratic mobile phase composed of acetonitrile-1% acetic acid (95∶5) and detected at 450 nm. RESULTS: A small amount of free bilirubin was detected in Calculus Bovis but not found in in-vitro cultured Calculus Bovis. The content of total bilirubin in Calculus Bovis from different sources varied greatly while being similar in in vitrocultured samples. Bilirubin in covalent form occupied a great part in Calculus Bovis, and bilirubin in conjugated form hold a great proportionin in vitro cultured Calculus Bovis. CONCLUSION: The contents or ratios of different types of bilirubin can be used as the quality control parameters of naturaland in vitro cultured Calculus Bovis. The established method is easy to operate with high effectiveness in quality control, suggesting a new way of identifying bezoar.