Development and validation of a liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the determination of ribavirin in human plasma and serum

A liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed for the analysis of antiviral agent ribavirin in human plasma and serum. The samples (0.1 ml) were extracted from the matrix using a simple protein precipitation procedure. The supernatants were evaporated to dryness, reconstituted and injected onto the LC/MS/MS system. The chromatography separation was achieved on a silica column operated with an aqueous–organic mobile phase. The use of a silica column not only provided adequate retention for the extremely polar compound of ribavirin, but also enhanced electrospray ionization sensitivity with the use of high percentage organic solvent in the mobile phase. The method has been validated over the concentration range of 10–10 000 ng/ml ribavirin in human plasma and serum. Bamethan was used as the internal standard. The protein precipitation extraction has been automated based on 96-well format with the use of robotic liquid handlers to improve the overall throughput of the analysis.