6-o-monoacetylmorphine and Opioid-Related-Disorders

The interpretation of toxicological findings is critical for the thorough investigation of the use and abuse of psychoactive substances. A positive analytical result for a sample taken could usually result in criminal proceedings and a punitive outcome for the defendant whose sample was analysed. The detection of markers of illicit opiate misuse is important both in the management of substance misuse and in the postmortem identification of illicit opiate use. The aim of this study was to emphasise the role of opiate biomarkers available at the laboratory and in the clinical environment. Urine remains the biological tool of choice for qualitative detection of illicit drug use in a clinical setting, while quantitative accuracy remains strictly the domain of blood. Accurate interpretation of the screening tests within a clinical setting alongside other relevant information remains the key to the usefulness of any test. Moreover, the finding of a morphine/codeine concentration ratio in blood exceeding unity is a strong evidence that the person had used heroin, as opposed to having taken a prescription analgesic drug containing codeine.

Topics: Biomarkers; Humans; Morphine Dependence; Morphine Derivatives; Opiate Alkaloids; Opioid-Related Disorders; Pain; Substance Abuse Detection

This report presents the different strategies for identifying heroin users. The criteria allowing a clear distinction between an abuse of heroin and a lawful consumption of opiates are deeply discussed. Reliable and sensitive analytical methods are now available for forensic opiate testing. The detection of 6-mono-acetylmorphine (MAM) indicates that heroin was administered within 24 hours or less of specimen collection. In the absence of MAM or after consumption of several opiates, the relative ratios of morphine, codeine and eventually ethylmorphine must be known in order to determine which opiate(s) was (were) taken. A total amount of opiates of less than 0.3 mg/l very often precludes any characterization of the ingested drug(s). Here we have to point out that forensic opiate testing should be done carefully. Interpretation of results requires more than detection of opiates or morphine alone, irrespective of the number of techniques used.

Topics: Forensic Medicine; Heroin Dependence; Humans; Morphine; Morphine Derivatives; Narcotics; Opioid-Related Disorders

To compare the efficacy of slow-release oral morphine (SROM) and methadone as maintenance medication for opioid dependence in patients previously treated with methadone.. Prospective, multiple-dose, open label, randomized, non-inferiority, cross-over study over two 11-week periods. Methadone treatment was switched to SROM with flexible dosing and vice versa according to period and sequence of treatment.. Fourteen out-patient addiction treatment centres in Switzerland and Germany.. Adults with opioid dependence in methadone maintenance programmes (dose ≥50 mg/day) for ≥26 weeks.. The efficacy end-point was the proportion of heroin-positive urine samples per patient and period of treatment. Each week, two urine samples were collected, randomly selected and analysed for 6-monoacetyl-morphine and 6-acetylcodeine. Non-inferiority was concluded if the two-sided 95% confidence interval (CI) in the difference of proportions of positive urine samples was below the predefined boundary of 10%.. One hundred and fifty-seven patients fulfilled criteria to form the per protocol population. The proportion of heroin-positive urine samples under SROM treatment (0.20) was non-inferior to the proportion under methadone treatment (0.15) (least-squares mean difference 0.05; 95% CI = 0.02, 0.08; P > 0.01). The 95% CI fell within the 10% non-inferiority margin, confirming the non-inferiority of SROM to methadone. A dose-dependent effect was shown for SROM (i.e. decreasing proportions of heroin-positive urine samples with increasing SROM doses). Retention in treatment showed no significant differences between treatments (period 1/period 2: SROM: 88.7%/82.1%, methadone: 91.1%/88.0%; period 1: P = 0.50, period 2: P = 0.19). Overall, safety outcomes were similar between the two groups.. Slow-release oral morphine appears to be at least as effective as methadone in treating people with opioid use disorder.

Topics: Administration, Oral; Adult; Codeine; Cross-Over Studies; Delayed-Action Preparations; Female; Humans; Maintenance Chemotherapy; Male; Medication Adherence; Methadone; Middle Aged; Morphine; Morphine Derivatives; Narcotics; Opiate Substitution Treatment; Opioid-Related Disorders; Treatment Outcome

Several studies have shown an association between asthma and opiate abuse. This retrospective study aims to analyse the demographic, toxicological, and seasonal differences in asthmatic and non-asthmatic subjects who died of opiates. In addition, the relationship between toxicological levels of opiates and histologic grade of lung inflammation is examined. Deaths from 2013 to 2018 involving opiates as the primary cause of death in Cook County, Illinois (USA) were reviewed. Twenty-six cases of opiate deaths of individuals with a history of asthma and lung histology slides available were identified. In comparison, 40 cases of deaths due to opiates only were analysed. A check-list system for the evaluation of the grade of microscopic inflammation in asthma was developed. We found statistically significant differences between the asthmatics and the non-asthmatics regarding demography (age and race) and toxicology (6-MAM presence). In particular, the "opiate and asthma group" was mainly composed of African-American subjects, in contrast with the "opiate group", consisting mostly of Caucasian. The mean age was significantly higher in the "opiate and asthma group" compared with the "opiate group". A greater presence of 6-MAM was detected in the "opiate group" compared with the "opiate and asthma group". While we expected to find that low opiate levels would lead to deaths in asthmatics and, in particular, that lower opiate concentrations would cause deaths in subjects with higher grades of histologic inflammation, our study suggests that the quantity of drug and the level of inflammation are not statistically significant in the determination of death. We, therefore, recommend histologic examination of the lungs to evaluate for asthma, particularly in suspected low-level opiate-related deaths, to help further clarify any relationship between asthma and opiate use.

Topics: Adult; Age Distribution; Asthma; Black or African American; Coroners and Medical Examiners; Female; Heroin Dependence; Humans; Inflammation; Lung; Male; Middle Aged; Morphine; Morphine Derivatives; Opiate Alkaloids; Opioid-Related Disorders; Organ Size; Pulmonary Edema; Retrospective Studies; United States; White People; Young Adult

We have previously demonstrated that heroin's first metabolite, 6-acetylmorphine (6-AM), is an important mediator of heroin's acute effects. However, the significance of 6-AM to the rewarding properties of heroin still remains unknown. The present study therefore aimed to examine the contribution of 6-AM to heroin-induced reward and locomotor sensitization. Mice were tested for conditioned place preference (CPP) induced by equimolar doses of heroin or 6-AM (1.25-5 μmol/kg). Psychomotor activity was recorded during the CPP conditioning sessions for assessment of drug-induced locomotor sensitization. The contribution of 6-AM to heroin reward and locomotor sensitization was further examined by pretreating mice with a 6-AM specific antibody (anti-6-AM mAb) 24 hours prior to the CPP procedure. Both heroin and 6-AM induced CPP in mice, but heroin generated twice as high CPP scores compared with 6-AM. Locomotor sensitization was expressed after repeated exposure to 2.5 and 5 μmol/kg heroin or 6-AM, but not after 1.25 μmol/kg, and we found no correlation between the expression of CPP and the magnitude of locomotor sensitization for either opioid. Pretreatment with anti-6-AM mAb suppressed both heroin-induced and 6-AM-induced CPP and locomotor sensitization. These findings provide evidence that 6-AM is essential for the rewarding and sensitizing properties of heroin; however, heroin caused stronger reward compared with 6-AM. This may be explained by the higher lipophilicity of heroin, providing more efficient drug transfer to the brain, ensuring rapid increase in the brain 6-AM concentration.

Topics: Analgesics, Opioid; Animals; Brain; Conditioning, Psychological; Disease Models, Animal; Heroin; Locomotion; Male; Mice; Mice, Inbred C57BL; Morphine Derivatives; Opioid-Related Disorders; Reward

In some forensic autopsies blood is not available, and other matrices are sampled for toxicological analysis. The aims of the present study were to examine whether heroin metabolites can be detected in different post-mortem matrices, and investigate whether analyses in other matrices can give useful information about concentrations in peripheral blood. Effects of ethanol on the metabolism and distribution of heroin metabolites were also investigated. We included 45 forensic autopsies where morphine was detected in peripheral blood, concomitantly with 6-acetylmorphine (6-AM) detected in any matrix. Samples were collected from peripheral blood, cardiac blood, pericardial fluid, psoas muscle, lateral vastus muscle, vitreous humor and urine. Opioid analysis included 6-AM, morphine, codeine, and morphine glucuronides. The 6-AM was most often detected in urine (n = 39) and vitreous humor (n = 38). The median morphine concentration ratio relative to peripheral blood was 1.3 (range 0-3.6) for cardiac blood, 1.4 (range 0.07-5.3) for pericardial fluid, 1.2 (range 0-19.2) for psoas muscle, 1.1 (range 0-1.7) for lateral vastus muscle and 0.4 (range 0.2-3.2) for vitreous humor. The number of 6-AM positive cases was significantly higher (P = 0.03) in the ethanol positive group (n = 6; 86%) compared to the ethanol negative group (n = 14; 37%) in peripheral blood. The distribution of heroin metabolites to the different matrices was not significantly different between the ethanol positive and the ethanol negative group. This study shows that toxicological analyses of several matrices could be useful in heroin-related deaths. Urine and vitreous humor are superior for detection of 6-AM, while concentrations of morphine could be assessed from peripheral or cardiac blood, pericardial fluid, psoas muscle and lateral vastus muscle.

Topics: Alcohol Drinking; Cadaver; Codeine; Forensic Toxicology; Glucuronides; Heroin; Humans; Morphine; Morphine Derivatives; Narcotics; Norway; Opioid-Related Disorders; Pericardial Fluid; Psoas Muscles; Quadriceps Muscle; Substance Abuse Detection; Tissue Distribution; Toxicokinetics; Vitreous Body

Recent advances in analytical capabilities allowing for the identification and quantification of drugs and metabolites in small volumes at low concentrations have made oral fluid a viable matrix for drug testing. Oral fluid is an attractive matrix option due to its relative ease of collection, reduced privacy concerns for observed collections and difficulty to adulterate. The work presented here details the development and validation of a liquid chromatography tandem mass spectrometry (LC-MS-MS) method for the quantification of codeine, morphine, 6-acetylmorphine, hydrocodone, hydromorphone, oxycodone and oxymorphone in neat oral fluid. The calibration range is 0.4-150 ng/mL for 6-acetylmorphine and 1.5-350 ng/mL for all other analytes. Within-run and between-run precision were <5% for all analytes except for hydrocodone, which had 6.2 %CV between runs. Matrix effects, while evident, could be controlled using matrix-matched controls and calibrators with deuterated internal standards. The assay was developed in accordance with the proposed mandatory guidelines for opioid confirmation in federally regulated workplace drug testing. The use of neat oral fluid, as opposed to a collection device, enables collection of a single sample that can be split into separate specimens.

Topics: Analgesics, Opioid; Calibration; Chromatography, Liquid; Codeine; Humans; Hydrocodone; Hydromorphone; Morphine; Morphine Derivatives; Opioid-Related Disorders; Oxycodone; Oxymorphone; Predictive Value of Tests; Reference Standards; Reproducibility of Results; Saliva; Spectrometry, Mass, Electrospray Ionization; Substance Abuse Detection; Tandem Mass Spectrometry

6-Monoacetylmorphine (6-MAM), being a unique metabolite of heroin, is routinely tested in urine samples to monitor heroin use. However, detection of 6-MAM-related opiates such as morphine is known to be affected by in vitro urine adulteration using oxidizing adulterants such as potassium nitrite. This study aimed to investigate the fate of 6-MAM after exposure to nitrite and to identify any formed oxidation products that may potentially be used for monitoring heroin abuse despite nitrite adulteration. Potassium nitrite (0.05 M and 0.6 M) was reacted with 6-MAM (5-10,000 ng/mL) in both water and blank urine with pH adjusted to range from 3 to 8. Following reaction at room temperature for varying periods, the reaction mixtures were monitored by both the CEDIA® Heroin Metabolite (6-AM) immunoassay and liquid chromatography-mass spectrometry (LC-MS) methods. Structural elucidation of the isolated oxidation products was based on mass spectrometry and nuclear magnetic resonance spectroscopic evidence. Nitrite, under acidic environment (pH<7), was shown to be effective in masking the detection of 6-MAM by both the CEDIA® immunoassay and the LC-MS methods. 2-Nitro-6-monoacetylmorphine (2-nitro-MAM) was identified as the sole oxidation product, which remained detectable in urine for at least 11 days under the experimental conditions investigated. 2-Nitro-MAM was detectable in a urine sample of a heroin user after nitrite exposure. 2-Nitro-MAM has shown potential to serve as a marker for monitoring heroin abuse when urine is adulterated with nitrite. Certification of 2-nitro-MAM reference standard for further development of its quantitative testing methods is thus warranted.

Topics: Biomarkers; Chromatography, Liquid; Humans; Immunoassay; Magnetic Resonance Spectroscopy; Mass Spectrometry; Morphine Derivatives; Nitrites; Opioid-Related Disorders

In the last ten years advances in analytical methods have enabled the determination of xenobiotics in alternative material such as sweat, saliva, and hair. The aim of this study was to develop an analytical method and measure the concentration of the main opiates in serum saliva and hair of subjects from a detoxification and methadone treatment programme. The analytical strategy in the presented study, based on enzymoimmunoassay screening of opiates in urine and GC/MS confirmation, meets the needs of forensic and clinical toxicology. Blood and saliva samples from thirty seven patients and hair from twenty three with a history of intravenous opiate use were collected for analysis. The ranges of morphine in serum and saliva were 0-2081 and 0-208 ng/ml respectively; corresponding concentrations of codeine were 0-580 and 0-428 ng/ml respectively. The concentration of morphine, codeine and 6-MAM in hair of addicts ranged respectively from 0-32.4, 0-12.5 and 0-2.8 ng/mg. From the clinical toxicology point of view, hair analysis is supplementary to urine, serum or saliva determination, but in drug testing at the workplace it can play a crucial role.

Topics: Adult; Codeine; Female; Gas Chromatography-Mass Spectrometry; Hair; Humans; Hydrogen-Ion Concentration; Male; Methadone; Morphine; Morphine Derivatives; Narcotics; Opioid-Related Disorders; Saliva

Hair samples taken from 850 individuals with presumed drug abuse were tested simultaneously for delta 9-tetrahydrocannabinol (THC), cocaine, heroin, the primary heroin metabolite 6-monoacetylmorphine (6-MAM) and morphine. The drugs were extracted with methanol under sonication. Compared to other extraction procedures this solvent extraction technique provides high extraction yields and less experimental effort. The analyses were carried out using gas chromatography-mass spectrometry (GCMS) in selected ion monitoring (SIM) mode. This procedure allows the simultaneous detection of amphetamine, methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA) and methylenedioxylamphetamine (MDE). THC was found in 104 (12.2%), cocaine in 230 (27%) and 6-MAM in 141 (16.6%) samples. In addition to 6-MAM, morphine was detected in 87 (10.2%) and heroin in 38 samples (4.5%). The concentrations found were in a range 0.009-16.7 ng/mg for THC, 0.037-129.68 ng/mg for cocaine, 0.028-79.82 ng/mg for 6-MAM, 0.045-53.14 ng/mg for heroin and 0.011-7.800 ng/mg for morphine. The statistical distribution of the drug concentrations compared with the self-reported consumption behaviour of the users may possibly lead to a better understanding of the relationship between drug dosage and corresponding concentrations in hair.

Topics: 3,4-Methylenedioxyamphetamine; Accidents, Traffic; Amphetamine; Cocaine; Crime; Designer Drugs; Dronabinol; Expert Testimony; Gas Chromatography-Mass Spectrometry; Hair; Heroin Dependence; Humans; Illicit Drugs; Marijuana Abuse; Morphine Derivatives; N-Methyl-3,4-methylenedioxyamphetamine; Opioid-Related Disorders; Psychotropic Drugs; Sensitivity and Specificity

Opiates were extracted from sixteen hair samples of drug addicts using a supercritical fluid extraction method with supercritical carbon dioxide and a modifier solution of methanol-triethylamine-water (2:2:1, v/v). the concentrations, as determined by GC-MS, ranged from 1.22 to 21.73 (mean 7.60 ng/mg), 0.17 to 1.54 (mean 0.69 ng/mg) and 0.15 to 14.009 ng/mg hair (mean 3.78 ng/mg) for codeine, morphine and 6-monoacetylmorphine, respectively. The reproducibility of the total procedure had a relative standard deviation of 13%, 17% and 14% for codeine, morphine and 6-monoacetylmorphine, respectively. But this method, concentrations of 0.3, 0.2 and 0.1 ng/mg hair for codeine, morphine and 6-monoacetylmorphine, respectively, could be detected. Relative extraction recoveries were 61%, 53% and 96% for codeine, morphine and 6-monoacetylmorphine, respectively.

Topics: Cannabinoids; Carbon Dioxide; Cocaine; Codeine; Ethylamines; Gas Chromatography-Mass Spectrometry; Hair; Humans; Methanol; Morphine; Morphine Derivatives; Narcotics; Opioid-Related Disorders; Reproducibility of Results; Solutions; Temperature; Water

A solid-phase extraction and gas chromatographic-mass spectrometric method for the simultaneous determination of codeine, dihydrocodeine, morphine, and 6-monoacetylmorphine in serum, blood or postmortem blood is described. The extraction technique allows the determination of free or total morphine (morphine plus morphine glucuronide). Experiments with spiked blood samples resulted in recoveries of 96.4% +/- 4.2% for codeine, 95.8% +/- 5.1% for dihydrocodeine, 90.3% +/- 7.8% for 6-monoacetylmorphine and 92.5% +/- 8.1% for morphine. Excellent linearity was obtained over the range 1-1500 ng/mL. The detection limit for all analytes is less than 1 ng/mL.

Topics: Codeine; Gas Chromatography-Mass Spectrometry; Humans; Morphine; Morphine Derivatives; Opioid-Related Disorders; Postmortem Changes