acetylcodeine and Heroin-Dependence

Confirmation or exclusion of recent heroin consumption is still one of the major challenges for forensic and clinical toxicologists. A great variety of biomarkers is available for heroin abuse confirmation, including various opium alkaloids (eg, morphine, codeine), street heroin impurities (eg, 6-acetylcodeine [6-AC], noscapine, papaverine) as well as associated metabolites (eg, 6-monoacetylmorphine [6-MAM], morphine glucuronides). However, the presence of most of these biomarkers cannot solely be attributed to a previous heroin administration but can, among other things, also be due to consumption of poppy seed products ('poppy seed defense'), opium preparations or specific medications, respectively. A reliable allocation is of great importance in different contexts, for instance in the case of DUID (driving under the influence of drugs) investigations, in driving licence re-granting processes, in workplace drug testing (WDT), as well as in post-mortem identification of illicit opiate use. Additionally, differentiation between illicit street heroin abuse and pharmaceutical heroin administration is also important, especially within the frame of heroin-assisted treatments. Therefore, analysis of multiple biomarkers is recommended when illicit opiate consumption is assumed to obtain the most reliable results possible. Beyond that, interpretation of positive opiate test results requires a profound insight into the great variety of biomarkers available and their validity regarding the alleged consumption. This paper aims to provide an overview of the wide variety of heroin abuse biomarkers described in the literature and to review them regarding their utility and reliability in daily routine analysis.

Topics: Biomarkers; Codeine; Glucuronides; Heroin; Heroin Dependence; Humans; Morphine Derivatives; Opium; Reproducibility of Results; Substance Abuse Detection

Only limited data exist concerning the utility of complementary specimens in heroin-related deaths. As such, this report employed a validated LC-MS-MS method to quantify 6-monoacetylmorphine (6-MAM), 6-acetylcodeine (6-AC), and their metabolites morphine and codeine in blood with (BN) and without preservative (B) and the additional unpreserved specimens of vitreous humor, urine, stomach contents, and bile from 20 postmortem cases in which heroin was the primary cause of death. The median concentration of 6-MAM in BN was 0.011 mg/L, B was 0.008 mg/L, urine was 0.186 mg/L, vitreous humor was 0.022 mg/L, stomach contents was 0.147 mg/L, and bile was 0.012 mg/L. Only one case was found to be positive for 6-AC in B (case 6, 0.002 mg/L), and the median concentration of 6-AC was 0.002 mg/L in BN, 0.012 mg/L in urine, 0.003 mg/L in vitreous humor, 0.057 mg/L in stomach contents, and 0.004 mg/L in bile. These findings present new information on the distribution of these analytes in complementary matrices and support their inclusion for accurately determining the role of heroin in opioid-related deaths.

Topics: Adult; Aged; Bile; Biomarkers; Chromatography, Liquid; Codeine; Female; Forensic Toxicology; Gastrointestinal Contents; Heroin Dependence; Humans; Male; Mass Spectrometry; Middle Aged; Morphine; Morphine Derivatives; Substance Abuse Detection; Vitreous Body; Young Adult

A method was developed and validated for analyzing 6-monoacetylmorphine, morphine, 6-acetylcodeine, and codeine in routine postmortem liver and kidney specimens using liquid chromatography-tandem mass spectrometry. Samples were prepared with a Stomacher instrument followed by solid-phase extraction. All calibration curves [0.5-1000 ng/g] were linear with coefficients of determination greater than 0.99 and limits of quantification of 1.0 ng/g. Within-run precision ranged between 2.0% and 8.0%, between-run precision ranged between 1.0% and 9.0%, and accuracy ranged between -5.0% and +3.0%. Matrix effects ranged from -18% to +9%. After matrix effects were excluded, analytical recoveries ranged from 76% to 94%. The distributions of 6-monoacetylmorphine, morphine, 6-acetylcodeine, and codeine were investigated in 31 postmortem cases in which heroin was the primary cause of death. In the current study, the median free morphine ratios were calculated for liver to blood and kidney to blood, which were 2.2 and 4.0, respectively. The current report highlights the importance of testing multiple specimens, including liver and kidney, in heroin-related deaths, especially if no blood samples are available. Furthermore, this work presents new information regarding the distribution of heroin metabolites in liver and kidney.

Topics: Adolescent; Adult; Aged; Biomarkers; Chromatography, Liquid; Codeine; Female; Forensic Toxicology; Heroin Dependence; Humans; Kidney; Limit of Detection; Liver; Male; Middle Aged; Morphine; Morphine Derivatives; Solid Phase Extraction; Substance Abuse Detection; Tandem Mass Spectrometry; Young Adult

Surveys of current trends indicate heroin abuse is associated with nonmedical use of pain relievers. Consequently, there is an interest in evaluating the presence of heroin-specific markers in chronic pain patients who are prescribed controlled substances. A total of 926,084 urine specimens from chronic pain patients were tested for heroin/diacetylmorphine (DAM), 6-acetylmorphine (6AM), 6-acetylcodeine (6AC), codeine (COD), and morphine (MOR). Heroin and markers were analyzed using liquid chromatography tandem mass spectrometry (LC-MS-MS). Opiates were analyzed following hydrolysis using LC-MS-MS. The prevalence of heroin use was 0.31%, as 2871 were positive for one or more heroin-specific markers including DAM, 6AM, or 6AC (a known contaminant of illicit heroin). Of these, 1884 were additionally tested for the following markers of illicit drug use: 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), methamphetamine (MAMP), 11-nor-9-carboxy-Δ(9)-tetracannabinol (THCCOOH), and benzoylecgonine (BZE); 654 (34.7%) had positive findings for one or more of these analytes. The overall prevalence of heroin markers were as follows: DAM 1203 (41.9%), 6AM 2570 (89.5%), 6AC 1082 (37.7%). MOR was present in 2194 (76.4%) and absent (

Topics: Analgesics, Opioid; Biomarkers; Buprenorphine; Chromatography, Liquid; Chronic Pain; Codeine; Heroin; Heroin Dependence; Humans; Illicit Drugs; Methadone; Morphine Derivatives; Pain Clinics; Tandem Mass Spectrometry

Although self reports of illicit drug use may not be reliable, this information is frequently collected and relied upon by national drug surveys and by counselors in drug treatment programs. The addition of oral fluid testing to these programs would provide objective information on recent drug use.. The goal of this study was to compare oral fluid tests for cocaine, benzoylecgonine, 6-acetylmorphine, morphine, codeine and 6-acetylcodeine to self reports of recent cocaine and heroin use by patients in an outpatient methadone treatment program.. Patients (n=400) provided an oral fluid specimen and completed a short questionnaire on illicit drug use over the last seven days. Oral fluid was collected with the Intercept Oral Fluid Collection device. Oral fluid was analyzed by a validated assay using liquid chromatography coupled with tandem mass spectrometry. The presence of an analyte was confirmed if all identification criteria were met and its concentration (ng/mL) was ≥ LOQ (cocaine, 0.4; benzoylecgonine, 0.4; morphine, 2; codeine, 2; 6-acetylmorphine, 0.4; and 6-acetylcodeine, 1).. Analyses of oral fluid specimens collected from the 400 methadone maintained patients revealed that a majority (95%) of subjects who admitted to recent cocaine use were confirmed positive, whereas slightly more than 50% were confirmed positive who admitted to heroin over the last seven days. For those patients who denied recent cocaine and heroin use, approximately 30% were positive for cocaine and 14% were positive for heroin.. Oral fluid testing provides an objective means of verifying recent drug use and for assessment of patients in treatment for substance use disorders.

Topics: Cocaine; Cocaine-Related Disorders; Codeine; Heroin; Heroin Dependence; Humans; Methadone; Morphine; Morphine Derivatives; Narcotics; Opiate Substitution Treatment; Saliva; Self Report; Substance Abuse Detection

One month before (T-1) and 12 months after (T12) controlled i.v. administration of pharmaceutical heroin-HCl (10-100 mg/day) in the context of a heroin maintenance program (HMP), concentrations of opiates and cocaine as well as its metabolites were determined in head hair (n = 46) using a validated gas chromatographic-mass spectrometric method. In addition, a patient collective of a methadone maintenance program (MMP, daily doses 15-260 mg) was examined (n = 35). The incidence of additional cocaine consumption decreased in both groups during the study period (T-1 to T12): in HMP from 64.6% to 45.8% and in MMP from 71.4% to 60.0%. A significant reduction of cocaine consumption was defined as an at least 30% reduction of analyte concentrations in hair (Deltac > 30%). Accordingly, in HMP, a decrease in 45.8% of initially (T-1) cocaine-positive patients was determined; in MMP, the reduction was 48.6%. In 22.9% of HMP and 37.1% of MMP, an increase of cocaine concentrations was detected. Codeine and acetylcodeine were found in 50.0% and 43.5% (T-1) and 13.0% and 10.9% (T12) of the samples of the HMP, as well as in 45.7% and 25.7% (T-1) and 17.1% and 5.7% (T12) in MMP, respectively. The missing of acetylcodeine, in particular at T-1, questions its applicability as a characteristic marker of a preceding consumption of illicit heroin in hair analysis.

Topics: Cocaine; Codeine; Dopamine Uptake Inhibitors; Gas Chromatography-Mass Spectrometry; Hair; Heroin; Heroin Dependence; Humans; Linear Models; Methadone; Narcotics; Substance Abuse Detection

For a pharmacokinetic-pharmacodynamic study in opioid tolerant patients, who were treated with heroin in combination with methadone, a liquid chromatographic assay with tandem mass spectrometry detection (LC-MS/MS) was developed for the simultaneous determination of heroin, methadone, heroin metabolites 6-monoacetylmorphine, morphine, and morphine-6 and 3-glucuronide and methadone metabolite EMDP. To detect any abuse of substances besides the prescribed opioids the assay was extended with the detection of cocaine, its metabolites benzoylecgonine and norcocaine and illicit heroin adulterants acetylcodeine and codeine. Heroin-d6, morphine-d3, morphine-3-glucuronide-d3 and methadone-d9 were used as internal standards. The sample pre-treatment consisted of solid phase extraction using mixed mode sorbent columns (MCX Oasis). Chromatographic separation was performed at 25 degrees C on a reversed phase Zorbax column with a gradient mobile phase consisting of ammonium formate (pH 4.0) and acetonitrile. The run time was 15 min. MS with relatively mild electrospray ionisation under atmospheric pressure was applied. The triple quadrupole MS was operating in the positive ion mode and multiple reaction monitoring (MRM) was used for drug quantification. The method was validated over a concentration range of 5-500 ng/mL for all analytes. The total recovery of heroin varied between 86 and 96% and of the heroin metabolites between 76 and 101%. Intra-assay and inter-assay accuracy and precision of all analytes were always within the designated limits (< or =20% at lower limit of quantification (LLQ) and < or =15% for other samples). This specific and sensitive assay was successfully applied in pharmacokinetic studies with medically prescribed heroin and toxicological cases.

Topics: Administration, Inhalation; Chromatography, High Pressure Liquid; Cocaine; Codeine; Heroin; Heroin Dependence; Humans; Methadone; Narcotics; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Time Factors

Acetylcodeine (AC), an impurity of illicit heroin synthesis, has been suggested as an interesting biomarker of illicit heroin use.. Procedures were developed for quantification of (a) morphine, 6-monoacetylmorphine (6-AM), and codeine in urine and (b) diacetylmorphine and AC in urine. Solid-phase extraction of the analytes was performed, and the extracted analytes were analyzed by selected-ion monitoring with gas chromatography-mass spectrometry. This procedure required prior derivatization with propionic anhydride.. Different validation parameters were determined, such as linearity, reproducibility, extraction recoveries, and cutoffs. Seventy-one urine specimens of illicit heroin abusers and 44 urine specimens of subjects in a heroin maintenance program were analyzed. AC was detected in 85.9% of the samples of the first group but not in any of the samples from subjects taking medical heroin. In the two groups, there were 94.4% and 84.1% 6-AM positive urine specimens, respectively. Detection times were determined for AC and codeine by parallel administration of heroin containing various percentages of AC to four voluntary patients in a heroin maintenance program. The measured detection times were 8 and 23 h for AC and codeine, respectively.. These results indicate that, together with detection of 6-AM in urine, AC is a suitable marker of illicit heroin use.

Topics: Biomarkers; Codeine; Gas Chromatography-Mass Spectrometry; Heroin Dependence; Humans; Immunoassay; Reproducibility of Results; Substance Abuse Detection

Acetylcodeine (AC), which is an impurity of illicit heroin synthesis, was suggested as a marker of heroin abuse. A procedure for simultaneous quantitation of 6-monoacetylmorphine (6-MAM), which is the major metabolite of heroin, morphine, codeine, and AC in hair was developed. Fifty-milligram hair samples were incubated in 0.01 M HCl overnight at 60 degrees C. The resulting hydrolyzed solutions were extracted by an automated solid-phase extraction procedure and drugs were analyzed by gas chromatography-mass spectrometry in selected ion monitoring mode (SIM). This required prior derivatization with propionic anhydride. Different validation parameters, such as linearity, intra-assay accuracy, extraction recoveries, and limit of quantitation, were described. Seventy-three hair samples from heroin abusers and 43 hair samples from subjects who had completed a heroin-maintenance program were analyzed. AC was detected in 92% of the first sample group and in only 12% of the second sample group. In the two groups, about 98% of AC-positive samples were found. These results prove that AC can be considered as a suitable marker of illicit heroin use, along with 6-MAM detection.

Topics: Adolescent; Adult; Biomarkers; Codeine; Female; Gas Chromatography-Mass Spectrometry; Hair; Heroin Dependence; Humans; Male; Middle Aged; Morphine Derivatives; Pilot Projects; Substance Abuse Detection

The planned introduction of a prescription heroin program in Germany created a need for differentiation between non-prescription and prescribed diamorphine use. The following substances were chosen as markers of non-prescription heroin: acetylcodeine (AC); its metabolites codeine (C) and codeine 6-glucuronide (C6G); papaverine (P); and noscapine (N). Typical heroin markers diamorphine (DAM) and its metabolites monoacetylmorphine (MAM) and morphine (M) were also determined. The drugs were extracted from urine samples with solid-phase extraction (C18) using standard 200-mg columns and 96-well microplates (100 mg). The extracts were examined with liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (positive ionization) in two isocratic systems. Selected ion monitoring procedures were applied for protonated molecular masses and characteristic fragments of drugs involved. The limits of detection were in the range of 0.5-1 ng/mL urine. The occurrence of selected heroin markers was investigated in 25 urine samples collected from heroin abusers (road traffic offenders and overdosed patients). C6G was found in all samples, C in 24 samples, N in 22 samples, MAM in 16 samples, P in 14 samples, DAM in 12 samples, and AC in 4 samples. The appearance of these compounds in urine reflects their pharmacokinetic properties and the composition of non-prescription heroin.

Topics: Adult; Biomarkers; Chromatography, Liquid; Codeine; Drug Prescriptions; Heroin; Heroin Dependence; Humans; Mass Spectrometry; Narcotics; Papaverine; Sensitivity and Specificity; Vasodilator Agents

Acetylcodeine (AC), an impurity of illicit heroin synthesis, was investigated as a urinary biomarker for detection of illicit heroin use. One hundred criminal justice urine specimens that had been confirmed positive by GC/MS for morphine at concentrations > 5000 ng/ml were analyzed for AC, 6-acetylmorphine (6AM), codeine, norcodeine and morphine. The GC/MS analysis was performed by solid phase extraction and derivatization with propionic anhydride. Total codeine and morphine concentrations were determined by acid hydrolysis and liquid/liquid extraction. AC was detected in 37 samples at concentrations ranging from 2 to 290 ng/ml (median, 11 ng/ml). 6AM was also present in these samples at concentrations ranging from 49 to 12 600 ng/ml (median, 740 ng/ml). Of the 63 specimens negative for AC, 36 were positive for 6AM at concentrations ranging from 12 to 4600 ng/ml (median, 124 ng/ml). When detected, the AC concentrations were an average of 2.2% (0.25 to 10.2%) of the 6AM concentrations. There was a positive relationship between AC concentrations and 6AM concentrations (r = 0.878). Due to its very low concentration in urine, AC was found to be a much less reliable biomarker for illicit heroin use than 6AM in workplace or criminal justice urine screening programs. However, AC detection could play an important role in determining if addicts in heroin maintenance programs are supplementing their supervised diacetylmorphine doses with illicit heroin.

Topics: Biomarkers; Codeine; Gas Chromatography-Mass Spectrometry; Heroin Dependence; Humans; Morphine; Morphine Derivatives; Narcotics; Reproducibility of Results; Substance Abuse Detection

In addition to acetylmorphine (6-AM), acetylcodeine (AC) has been suggested as a marker for the use of illicit heroin. Because no procedure was available for AC testing in hair, a new method was developed for the simultaneous identification and quantitation of morphine (MOR), codeine (COD), 6-AM, and AC. After decontamination, each hair specimen was cut into 1-mm pieces. A 50-mg aliquot was incubated overnight at 50 degrees C in 1 mL Soerensen buffer (pH 7.6) in presence of 200 ng of MOR-d3, COD-d3, 6-AM-d3, and AC-d3. After pH adjustment to 8.4, the analytes were extracted in 5 mL of chloroform/isopropanol/n-heptane (25:10:65, v/v/v). The organic phase was removed and evaporated to dryness, and the residue was derivatized by silylation (BSTFA + 1% TMCS). Drugs were analyzed by gas chromatography-mass spectrometry in electron impact mode. Limits of quantitation were set to 0.1 ng/mg. Fifty hair specimens obtained from subjects who died from fatal opiate overdose were analyzed. AC was detected in 22 samples in concentrations ranging from 0.17 to 5.60 ng/mg with a mean value of 1.04 ng/mg. 6-AM was also present in these samples at concentrations ranging from 1.35 to 41.10 ng/mg with a mean value of 7.79 ng/mg. Of the 28 specimens negative for AC, 21 were positive for 6-AM at concentrations ranging from 0.18 to 7.13 ng/mg. When detected, the AC concentrations were an average of 15.5% (2.8 to 32.6%) of the 6-AM concentrations. There was a positive relationship between AC concentrations and 6-AM concentrations (r = 0.915, p = 0.001). Neither AC nor COD was identified in hair specimens collected from 20 subjects taking part in a heroin-maintenance program in Switzerland and receiving pure pharmaceutical heroin hydrochloride daily. Although it is indicative of illicit heroin use, AC would not make a suitable biomarker in place of 6-AM because of its low concentration in hair compared with that of 6-AM and its absence in about 50% of the specimens that tested positive for 6-AM.

Topics: Biomarkers; Codeine; Gas Chromatography-Mass Spectrometry; Hair; Heroin; Heroin Dependence; Humans; Morphine; Morphine Derivatives; Narcotics; Substance Abuse Detection

In addition to 6-monoacetylmorphine (6-MAM), acetylcodeine (AC) has been suggested as a marker for the use of illicit heroin. We report a sensitive opiate gas chromatographic-mass spectrometric assay that detects AC, diacetylmorphine, and the propionylated derivatives of codeine, morphine, 6-MAM, and norcodeine. The analytes were extracted by solid phase with recoveries from 62 to 98%. The limits of detection (LOD) and quantitation (LOQ) for AC was 0.5 and 1.0 microgram/L. The LOD of the other analytes was 2.0 micrograms/L and the LOQs ranged from 2 to 10 micrograms/L. The assay was linear for each analyte from the LOQ to 200 micrograms/L or 400 micrograms/L (morphine and codeine) with r > or = 0.996, except for diacetylmorphine which was linear to 100 micrograms/L with r = 0.994. The within-run and between-run precision were below 10% CV for all analytes. There was no significant hydrolysis of AC to codeine in urine (pH 4.7 and 8.0) after 23 weeks of refrigeration or freezing. After storage at room temperature in urine of pH 8.0, AC was completely hydrolyzed after 5 weeks, but at pH 4.7, 58% of the AC remained after 15 weeks of storage at room temperature. The sensitivity of this assay was adequate to detect AC in the urine of heroin abusers. In preliminary studies, AC was detected in 6 of 69 opiate positive urines. Concentrations ranging from 1 to 48 micrograms/L were observed. These concentrations were found to be low when compared with the concentrations of 6-MAM, codeine, and morphine also detected in the urines.

Topics: Biomarkers; Calibration; Codeine; Drug Stability; Gas Chromatography-Mass Spectrometry; Heroin Dependence; Humans; Morphine Derivatives; Narcotics; Reproducibility of Results