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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 25,
Number 1, January/February,
pp. 69-75
Rapid
Confirmation/Quantitation of Ecgonine Methyl Ester, Benzoylecgonine, and Cocaine
in Urine Using On-Line Extraction Coupled with Fast HPLC and Tandem Mass Spectrometry
Patrick
M. Jeanville[1], Estela S. Estapé[2], Ivette Torres-Negrón[3],
and Arturo Martí[4]
[1]Drug Metabolism Technology Department, Pfizer Inc., Central Research Division,
Eastern Point Road, Bldg. 118N, N211, Box 4133, Groton, Connecticut 06340; [2]College
of Health-Related Professions, Medical Sciences Campus, University of Puerto
Rico, P.O. Box 365067, San Juan, Puerto Rico 00936-5067; [3]Science and Technology
Department, Metropolitan University, P.O. Box 21150, San Juan, Puerto Rico 00928-1150;
and [4]Chemical Forensic Laboratories, School of Pharmacy, University of Puerto
Rico, P.O. Box 365067, San Juan, Puerto Rico 00936-5067
A rapid, rugged,
and highly specific assay for the quantitation of cocaine (COC) and especially
its primary metabolites benzoylecgonine (BZE) and ecgonine methyl ester (EME)
in human urine has been established. Here, we investigated the use of on-line
sample extraction coupled to rapid chromatography systems for tandem mass analysis
of COC, EME, and BZE in human urine. Using this method, sample preparation consisted
of a sole centrifugation step. Combined extraction and chromatographic run times
were < 3.5 min. The lower limits of detection were 0.5 ng/mL, 2.0 ng/mL,
and 0.5 ng/mL for EME, BZE, and COC, respectively. Linear calibration curves
ranging from 7.5 ng/mL to 1000 ng/mL were produced for the test analytes. Within-day
and between-day precision and accuracy of the assay were determined using human
urine quality-control specimens at 5, 10, or 15; 150; and 1000 ng/mL. The analyses
were performed over the course of five days, rendering %CVs < 10% for EME,
BZE, and COC. Percent mean accuracy for the three analytes of 97 to 113% were
obtained. Our data suggest that on-line sample extraction coupled with rapid
high-performance liquid chromatography–tandem mass spectrometry may be
a viable alternative for EME, BZE, and COC analyses in human urine.
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