HIGH-THROUGHPUT LIQUID CHROMATOGRAPHIC DETERMINATION OF ROFECOXIB IN HUMAN PLASMA USING A FULLY AUTOMATED ON-LINE SOLID-PHASE EXTRACTION SYSTEM^‡

Abstract

A fully automated on-line solid-phase extraction method combined with high-performance liquid chromatography-fluorescence detection for the determination of rofecoxib in human plasma is described. A Spark Holland PROSPEKT system was used to perform all the automated on-line sample extraction and quantitation tasks, including sample loading, cartridge conditioning and washing, analyte elution, and start of a data acquisition system. Under postcolumn ultraviolet–irradiation conditions, the analytes were photochemically converted to highly fluorescent products and then monitored by a fluorescence detector. The assay was validated in the concentration range of 0.5–100 ng/mL using 0.25 mL of human plasma. The utility of this method was demonstrated by analysis of plasma samples from human subjects receiving oral doses of rofecoxib.

‡ This work was presented at the Fifteenth Annual International Symposium on Laboratory Automation and Robotics, October 1997, Boston, MA, USA.

ACKNOWLEDGMENTS

The authors thank Dr. C. Ferguson for directing and monitoring the clinical program from which the human plasma samples were obtained for the analyses. The contribution of Ms. C. Mathews during the determination of I using the manual HPLC assay is gratefully acknowledged.

Notes

‡ This work was presented at the Fifteenth Annual International Symposium on Laboratory Automation and Robotics, October 1997, Boston, MA, USA.