Concentration profiles of cocaine, pyrolytic methyl ecgonidine and thirteen metabolites in human blood and urine: determination by gas chromatography-mass spectrometry

When cocaine is smoked, a pyrolytic product, methyl ecgonidine (anhydroecgonine methyl ester), is also consumed with the cocaine. The amount of methyl ecgonidine formed depends on the pyrolytic conditions and composition of the illicit cocaine. This procedure describes detection of cocaine and 10 metabolites—cocaethylene, nor‐cocaine, nor‐cocaethylene, methyl ecgonine, ethyl ecgonine, benzoylecgonine, nor‐benzoylecgonine, m‐hydroxybenzoylecgonine, p‐hydroxybenzoylecgonine and ecgonine—in blood and urine. In addition, the detection of pyrolytic methyl ecgonidine and three metabolites—ecgonidine (anhydroecgonine), ethyl ecgonidine (anhydroecgonine ethyl ester) and nor‐ecgonidine (nor‐anhydroecgonine)—are included. The newly described metabolites, ethyl ecgonidine and nor‐ecgonidine, were synthesized and characterized by gas chromatography–mass spectrometry (GC‐MS). All 15 compounds were extracted from 3 mL of blood or urine by solid‐phase extraction and identified by a GC‐MS method. The overall recoveries were 49% for methyl ecgonine, 35% for ethyl ecgonine, 29% for ecgonine and more than 83% for all other drugs. The limits of detection were between 0.5 and 4.0 ng[sol ]mL except for ecgonine, which was 16 ng[sol ]mL. Linearity for each analyte was established and in all cases correlation coefficients were 0.9985–1.0000. The procedure was applied to examine the concentration profiles of analytes of interest in post‐mortem (PM) blood and urine, and in urine collected from living individuals (LV). These specimens previously were shown to be positive for the cocaine metabolite, benzoylecgonine. Ecgonidine, the major metabolite of methyl ecgonidine, was present in 77% of PM and 88% of the LV specimens, indicating smoking as the major route of cocaine administration. The new pyrolytic metabolites, ethyl ecgonidine and nor‐ecgonidine, were present in smaller amounts. The urine concentrations of nor‐ecgonidine were 0–163 ng[sol ]mL in LV and 0–75 ng[sol ]mL in PM specimens. Ethyl ecgonidine was found only in PM urine at concentrations 0–39 ng[sol ]mL. Ethanol‐related cocaine metabolites, ethyl ecgonine or cocaethylene, were present in 69% of PM and 53% of cocaine‐positive LV specimens, implying alcohol consumption with cocaine use. The four major metabolites of cocaine—benzoylecgonine, ecgonine, nor‐benzoylecgonine and methyl ecgonine—constituted approximately 88 and 97% of all metabolites in PM and LV specimens, respectively. The concentrations of nor‐cocaine and n