Determination of esterase-catalyzed cocaine metabolite formation by high-performance liquid chromatography with ultraviolet detection.
A reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of cocaine metabolites produced in vitro by serum and liver esterases is described. Hydrolysis of cocaine at the methyl ester bond produces benzoylecgonine and methanol, whereas hydrolysis at the benzoyl ester bond produces ecgonine methyl ester and benzoic acid. This method quantitates benzoic acid (as a measure of ecgonine methyl ester formation) and benzoylecgonine, which can be determined simultaneously and with great sensitivity by HPLC and ultraviolet detection. Cocaine is found to be hydrolyzed to both ecgonine methyl ester and benzoylecgonine; hepatic microsomes exhibit the highest specific activity.
Kim
K Y
KY
Department of Cellular and Molecular Pharmacology, University of California, San Francisco 94143-0450, USA.
Bornheim
L M
LM
D000818:Animals; D001565:Benzoates / Q000032:analysis / Q000097:blood / Q000652:urine; D019817:Benzoic Acid; D001711:Biotransformation; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D003042:Cocaine / Q000031:analogs & derivatives / Q000032:analysis / Q000097:blood / Q000493:pharmacokinetics* / Q000652:urine; D003600:Cytosol / Q000201:enzymology / Q000378:metabolism; D004950:Esterases / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism; D009294:Narcotics / Q000493:pharmacokinetics*; D013056:Spectrophotometry, Ultraviolet
D001565:Benzoates; D009294:Narcotics; C005618:benzoylecgonine; D019817:Benzoic Acid; D004950:Esterases; D003042:Cocaine; C034524:ecgonine methyl ester