A Rapid, Sensitive Assay for Cocaine and Its Metabolites in Biological Fluids using Solid-Phase Extraction and High-Performance Liquid Chromatography

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Published: Journal of Analytical Toxicology, ISSN 0146-4760, Volume 24, Number 6, September 2000, pp. 438-441

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A Rapid, Sensitive Assay for Cocaine and Its Metabolites in Biological Fluids using Solid-Phase Extraction and High-Performance Liquid Chromatography
Subhash C. Jamdar, Carol B. Pantuck, Jaime Diaz, and Berend Mets
Department of Anesthesiology, College of Physicians and Surgeons, Columbia University, 630 W. 168th Street, New York, New York 10032

An improved method for the simultaneous determination of cocaine and its metabolites, benzoylecgonine (BE), norcocaine, and ecgoninemethylester (EME), in rat plasma and urine is described. Following derivatization of EME to p-fluorococaine, chromatography was performed on two high-performance liquid chromatography (HPLC) columns in series (5-µm spheric C⁸ and 5-µm cyanopropyl) using a mobile phase containing acetonitrile/HPLC water/trifluoroacetic acid (28:72:0.1) with bupivacaine as an internal standard. Quantitation limits were 25 ng/mL for cocaine, BE, and norcocaine and 50 ng/mL for EME using 300–500 µL rat plasma and 500 µL of rat urine. The assay was linear from the limit of quantitation to 2000 ng/mL for cocaine and its metabolites in both plasma and urine samples. Because this method uses a small amount of sample (300 µL plasma or 500 µL of urine), it is applicable to study of the pharmacokinetics and disposition of cocaine and its major metabolites.

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