• Measurement and interpretation of heroin, 6-acetylmorphine and morphine concentrations in biological tissues obtained from heroin users and heroin-related deaths

      Goldberger, Bruce A.; Caplan, Yale H. (1993)
      Understanding the role of heroin in heroin-related deaths was facilitated by the development of an assay for heroin and its metabolites in biological specimens. Recognizing heroin's susceptibility to rapid chemical and metabolic hydrolysis, procedures were developed for the efficient recovery of heroin, 6-acetylmorphine and morphine from tissues utilizing liquid-liquid and solid phase extraction. Aprotic solvents, mild elution solvents and an enzyme inhibitor were employed to ensure maximum analyte stability. Extracts were analyzed by gas chromatography/mass spectrometry. The assay was highly sensitive and specific for heroin and its metabolites. The limit of detection for each analyte was approximately 1 ng/mL. Specimens were collected and analyzed from twenty-one cases of death due to acute narcotic intoxication. The mode of death in these cases was categorized as rapid, delayed or undetermined. Compared to delayed deaths, rapid deaths were characterized by the following trends: higher mean concentrations of 6-acetylmorphine, free morphine and total morphine in blood; higher ratio of free to total morphine concentration in blood; lower mean concentrations of 6-acetylmorphine and morphine in urine; greater likelihood to detect 6-acetylmorphine in blood; and lesser likelihood to detect heroin in urine. In a detailed study of two heroin overdose cases, heroin was present only in urine specimens. Concentrations of 6-acetylmorphine in cerebrospinal fluid, spleen and brain were substantially higher than in blood, liver, lung and kidney. All specimens were positive for morphine. Hair analysis was used to corroborate prior heroin exposure. Hair specimens, collected from twenty heroin users and two heroin-related deaths, contained 6-acetylmorphine and morphine. Heroin was found in 7 of 20 heroin user specimens. Generally, the 6-acetylmorphine concentration in hair was higher than that of heroin or morphine. The identification of heroin and 6-acetylmorphine in biological tissues effectively established heroin use in cases of acute narcotic intoxication. These studies demonstrated that measurement of heroin and its metabolites provides useful information in the differential diagnosis of heroin-related deaths.
    • The role of ecgonine methyl ester in the interpretation of cocaine concentrations in postmortem blood.

      Isenschmid, Daniel Stephan; Caplan, Yale H. (1991)
      The metabolism and hydrolysis of cocaine (COC) was studied in vitro, in vivo and postmortem to determine if measuring concentrations of the COC metabolites, benzoyl-ecgonine (BE) and ecgonine methyl ester (EME) would facilitate the differential diagnosis of COC related Medical Examiner cases. In vitro, in unpreserved whole blood, COC hydrolyzed to EME. In blood preserved by a pseudocholinesterase inhibitor, a solution of acetylcholinesterase, and phosphate buffer controls, COC hydrolyzed to BE. COC incubated in a solution of pseudocholinesterase and carboxylesterase hydrolyzed to EME. The rate of hydrolysis of COC increased as the pH and temperature of the specimen increased. COC was stable in blood adjusted to pH 5 and preserved with sodium fluoride or organophosphates for at least 150 days at 4C or lower. An in vivo study in human subjects given intravenous and/or smoked COC confirmed that BE is the principal metabolite of COC in blood. All COC was accounted for by BE. EME, when present, did not exceed 5% of the BE concentration.;EME arises in postmortem blood mainly as a result of non-metabolic hydrolysis of COC. Thus, in unpreserved blood specimens, the EME concentration can be added to the COC concentration to estimate the blood COC concentration prior to in vitro hydrolysis, while BE would be attributable to prior COC metabolism. Application of this theory to a study of Medical Examiner cases supported this conclusion. COC in blood was determined between 1 and 8 days following death and again 10 to 70 days after further storage (N = 10). The COC lost was accounted for by the EME formed. Good correlation (r = 0.9677) was observed when the blood COC concentrations in Medical Examiner cases were compared to blood COC concentrations predicted by the addition of blood COC and EME concentrations; hence, analysis for EME and estimation of perimortem COC concentrations can assist in defining deaths associated with COC use.