Robotic Sample Preparation and HPLC Determination of the Major Component of Ivermectin in Human Plasma

Abstract

A robotic sample preparation method combined with high performance liquid chromatography (HPLC)‐fluorescence detection for the determination of the major component of ivermectin in human plasma is presented. A Cyberlab C‐300 workstation equipped with customized software and hardware was utilized to perform all the semi‐automated liquid–solid phase extraction (LSPE) steps including acetonitrile (ACN) and water addition, sample loading, and hexane elution on Chem Elut^TM cartridges. Under the automated pre‐column derivatization conditions, both ivermectin and internal standard of the extracted samples were chemically converted to highly fluorescent derivatives and then monitored by a fluorescence detector. The assay was validated in the concentration range of 0.5–40 ng/mL, using 1 mL of human plasma. The application of this assay was demonstrated by analysis of plasma samples from human subjects receiving oral doses of ivermectin.

Keywords:

• Robotic sample preparation
• HPLC
• Ivermectin
• Automated pre‐column derivatization

Acknowledgments

This work was presented, in part, at the LabAutomation 2002, Palm Spring, CA, January 26–30, 2002, Poster #1 of Applications section. The authors wish to thank Drs. Kenneth Lasseter and Cynthia Guzzo for directing and monitoring the clinical program from which the blood samples were obtained for the analyses.