Rapid method for the determination and quantification of bromosulphophthalein and metabolites in cultured hepatocytes, culture media and bile by high-performance liquid chromatography.

An ion-pair high-performance liquid chromatographic method for the rapid, selective and sensitive analysis of samples containing bromosulphophthalein (BSP) and its conjugates is presented. The method is useful for analysis in bile, culture media and cultured hepatocytes. Two sample preparation methods are described. Even though BSP recovery from albumin binding is complete, only a small percentage of free BSP can be detected in cells, possibly owing to a conjugation-related pool of BSP in cells. As BSP-glutathione recovery is complete, the method offers a useful tool to investigate impairment of glutathione conjugation.