A highly automated 96-well solid phase extraction and liquid chromatography/tandem mass spectrometry method for the determination of fentanyl in human plasma

A high-throughput bioanalytical method based on automated sample transfer, automated solid phase extraction, and fast liquid chromatography/tandem mas s spectrometry (LC/MS/MS) analysis, has been developed for the determinatio n of the analgesic fentanyl in human plasma. Samples were transferred into 96-well plates using an automated sample handling system. Automated solid p hase extraction (SPE) was carried out using a 96-channel programmable liqui d-handling workstation using a mixed-mode sorbent. The extracted samples we re then dried down, reconstituted and injected onto a silica column using a n aqueous/organic mobile phase with tandem mass spectrometric detection. Th e method has been validated over the concentration range 0.05-100 ng/mL fen tanyl in human plasma, based on a 0.25-mL sample size. The assay is sensiti ve, specific and robust. More than 2000 samples have been analyzed using th is method. The automation of the sample preparation steps not only increase d the analysis throughput, but also facilitated the transfer of the method between different bioanalytical laboratories of the same organization. Copy right (C) 2001 John Wiley & Sons, Ltd.