High-performance liquid chromatographic determination of cocaine and benzoylecgonine by direct injection of human blood plasma sample into an alkyl-diol-silica (ADS) precolumn.
A column-switching high-performance liquid chromatographic method with UV detection for the determination of cocaine (COC) and benzoylecgonine (BZE) in human blood plasma samples is described. The method uses an alkyl-diol-silica ADS-C18 extraction precolumn. A 50- micro L plasma sample was introduced to the ADS precolumn in order to separate the analytes from proteins and endogenous compounds. The fraction containing COC and BZE was back-flushed and transferred to an Alltech mixed-mode C(18)/cation-exchange analytical column for final separation. The validation of the method revealed quantitative recoveries from 95.0 to 99.0% for COC at three different concentrations (0.5, 1.0 and 2.0 micro g mL(-1)), and from 96.0 to 99.0% for BZE at the same concentration levels with coefficients of variation <4.00% (n=5). The detection limit (signal to noise ratio (S/N)>3) was 0.03 micro g mL(-1) for all the compounds with an injection volume of 50 micro L. However, it was possible to enhance the sensitivity further by injecting larger plasma volumes, up to 200 micro L, at the same optimal conditions. The overlap of sample preparation, analysis and reconditioning of the extraction column, increase the overall sample throughput to 5 samples h(-1). The developed method has been applied to human blood plasma samples from subjects suspected of cocaine abuse.
Brunetto
Rosario
R
IVAIQUIM (Venezuelan Andean Institute for Chemical Research), Faculty of Science, University of Los Andes, P.O. Box 3 Ipostel La Hechicera, Mérida 5101-A, Venezuela. brunetto@ula.ve
Gutiérrez
Lubin
L
Delgado
Yelitza
Y
Gallignani
Máximo
M
Burguera
José Luis
JL
Burguera
Marcela
M
D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D003042:Cocaine / Q000031:analogs & derivatives* / Q000097:blood*; D004867:Equipment Design; D006801:Humans; D006863:Hydrogen-Ion Concentration; D012015:Reference Standards; D012822:Silicon Dioxide; D012997:Solvents; D019966:Substance-Related Disorders / Q000097:blood / Q000175:diagnosis
D012997:Solvents; C095072:alkyl-diol silica; C005618:benzoylecgonine; D012822:Silicon Dioxide; D003042:Cocaine