RESUMEN
It is reasonable to expect the presence of multiple drugs to present a complicated picture of toxicity. We report a fatal case involving a young man who purchased illicit drugs and knowingly consumed them. After consuming these drugs and going to sleep in his friend's car, he was found unresponsive the next morning with no signs of physical violence. Drugs found in the peripheral blood at autopsy were oxymorphone, methylone and ethanol at concentrations of 0.106, 0.50 and 130 mg/dL, respectively. The levels of oxymorphone and methylone in peripheral blood were comparable to those observed in other reported fatalities. Cocaine and benzoylecgonine were detected in the urine but not in the blood. Measureable concentrations were also observed for oxymorphone and methylone in urine, liver, kidney and bile. The physical findings at autopsy included pulmonary edema. This is the only reported fatal case involving this combination of drugs encountered in our laboratory.
Asunto(s)
Sobredosis de Droga/diagnóstico , Etanol/metabolismo , Metanfetamina/análogos & derivados , Oximorfona/metabolismo , Adulto , Etanol/sangre , Etanol/orina , Resultado Fatal , Toxicología Forense , Humanos , Masculino , Metanfetamina/sangre , Metanfetamina/metabolismo , Metanfetamina/orina , Oximorfona/sangre , Oximorfona/orina , Edema Pulmonar/diagnósticoRESUMEN
Screening for emerging drugs of abuse, specifically synthetic cathinones and synthetic cannabinoids, is difficult for high-throughput laboratories as immunoassay kits are often unavailable. Consequently, most laboratories employ liquid chromatography-tandem mass spectrometry (LC-MS-MS) screening, which can be complex and time consuming as these techniques may require involved sample preparation and lengthy analysis times. The increasing demand for novel psychoactive substance testing necessitates alternative screening methods that are sensitive, fast and versatile. The RapidFire tandem mass spectrometry system (RF-MS-MS) provides a rapid and highly specific screen for these emerging drugs of abuse with minimal sample preparation and an instrumental analysis time of <14 s per sample. Presented here are two RF-MS-MS screening methods used to analyze 28 emerging drugs of abuse, 14 synthetic cannabinoids and 14 synthetic cathinones, in urine with run times of 9 and 12.6 s, respectively. Sample preparation and hydrolysis were performed in a 96-well plate with one multiple reaction monitoring transition used for the identification of each compound. Eighteen thousand urine specimens were screened by liquid-liquid extraction followed by LC-MS-MS analysis, and the results were compared with those obtained using the RF-MS-MS screening method. The analytical data illustrate the advantages of the RF-MS-MS methods.
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Alcaloides/orina , Cannabinoides/orina , Detección de Abuso de Sustancias/métodos , Humanos , Espectrometría de Masas en Tándem/métodosRESUMEN
An accidental death associated with the use of the designer drug, 2-(4-chloro-2,5-dimethoxyphenyl)-N-(2-methoxybenzyl)ethanamine (25C-NBOMe), is reported. A 23-year-old Caucasian male experienced severe respiratory distress and died after being subdued by military law enforcement. At autopsy, remarkable findings upon internal examination included mild to moderate coronary atherosclerosis, biventricular dilation, mild right ventricular hypertrophy and bilateral pulmonary edema and congestion. The decedent's blood contained no drugs, ethanol or other volatile compounds. Pseudoephedrine, nicotine and cotinine were detected in his urine. A LC-QTOF designer drug screen, employing a basic solid-phase extraction, was used to isolate 25C-NBOMe, 25C-NBOH and 2C-C from both blood and urine specimens. Quantitative analysis was performed by LC-MS-MS operating in multiple reaction monitoring mode. 25C-NBOMe and 2C-C were present in the blood (2.07 and 0.12 ng/mL) and in the urine (27.43 ng/mL and 0.38 ng/mL), respectively. 25C-NBOMe concentrations were determined by standard addition in the brain (19.10 ng/g), spleen (27.13 ng/g), lung (25.21 ng/g), liver (15.20 ng/g), kidney (25.06 ng/g) and gastric contents (30.24 µg total in 100 mL submitted). On the basis of decedent case history, autopsy and toxicological findings, the medical examiner ruled the cause of death as 25C-NBOMe toxicity temporally associated with excited delirium and forcible restraint. The manner of death was ruled accidental.
Asunto(s)
Bencilaminas/sangre , Fenetilaminas/sangre , Psicotrópicos/sangre , Adulto , Cromatografía Liquida , Resultado Fatal , Toxicología Forense , Humanos , Masculino , Espectrometría de Masas en Tándem , Adulto JovenRESUMEN
Synthetic cannabinoids emerged on the designer drug market in recent years due to their ability to produce cannabis-like effects without the risk of detection by traditional drug testing techniques such as immunoassay and gas chromatography-mass spectrometry. As government agencies work to schedule existing synthetic cannabinoids, new, unregulated and structurally diverse compounds continue to be developed and sold. Synthetic cannabinoids undergo extensive metabolic conversion. Consequently, both blood and urine specimens may play an important role in the forensic analysis of synthetic cannabinoids. It has been observed that structurally similar synthetic cannabinoids follow common metabolic pathways, which often produce metabolites with similar metabolic transformations. Presented are two validated quantitative methods for extracting and identifying 15 parent synthetic cannabinoids in blood, 17 synthetic cannabinoid metabolites in urine and the qualitative identification of 2 additional parent compounds. The linear range for most synthetic cannabinoid compounds monitored was 0.1-10 ng/mL with the limit of detection between 0.01 and 0.5 ng/mL. Selectivity, specificity, accuracy, precision, recovery and matrix effect were also examined and determined to be acceptable for each compound. The validated methods were used to analyze a compilation of synthetic cannabinoid investigative cases where both blood and urine specimens were submitted. The study suggests a strong correlation between the metabolites detected in urine and the parent compounds found in blood.
Asunto(s)
Cannabinoides/metabolismo , Cromatografía Liquida/métodos , Espectrometría de Masas en Tándem/métodos , Drogas de Diseño/metabolismo , Humanos , Redes y Vías MetabólicasRESUMEN
Synthetic cannabinoids, including JWH-018 and JWH-073, belong to a class of aminoalkylindoles (AAIs) that are smoked to produce an effect similar to tetrahydrocannabinol. Compounds in this class are often collectively known as 'Spice'. After ingestion, these compounds are extensively metabolized to their hydroxy and carboxylic acid metabolites. During forensic analysis, detection of these metabolites in urine is an indication of past exposure to the parent compounds. The analytical process involved hydrolysis of conjugated metabolites by glucuronidase, solvent extraction, derivatization by trifluoroacetic anhydride and hexafluoroisopropanol and GC-EIMS detection. Identification of the unknown was based on the criteria of GC retention time within ±2% and mass spectral ion ratio within ±20% of that of a standard. Deuterated internal standards of the carboxylic acid metabolites were used for quantification. The acid (JWH-018-COOH, JWH-073-COOH) and hydroxy (JWH-018-OH, JWH-073-OH) metabolites were linear over the concentration range of 0.1-10 and 0.2-10 ng/mL, respectively, with a correlation coefficient-square, R(2) > 0.999 (N = 5). Extraction recoveries of the metabolites were within 79 and 87%. The method was applied to 17 urine specimens collected as part of a military law enforcement investigation. Nine of the specimens tested positive for one or more of the metabolites. When the procedure was extended to screen other AAI compounds, two of the specimens were found to contain JWH-210, JWH-250 (JWH-302 or JWH-201) and JWH-250 (C4 isomers). The GC-EIMS method presented here was found to be suitable for detecting JWH-018 and JWH-073 metabolites and other AAI compounds in urine.
Asunto(s)
Ácidos Carboxílicos/orina , Toxicología Forense/métodos , Cromatografía de Gases y Espectrometría de Masas , Drogas Ilícitas/orina , Indoles/orina , Naftalenos/orina , Detección de Abuso de Sustancias/métodos , Trastornos Relacionados con Sustancias/orina , Biotransformación , Calibración , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas/normas , Humanos , Hidroxilación , Límite de Detección , Modelos Lineales , Personal Militar , Valor Predictivo de las Pruebas , Estándares de Referencia , Reproducibilidad de los Resultados , Fumar , Detección de Abuso de Sustancias/normas , Trastornos Relacionados con Sustancias/diagnóstico , Espectrometría de Masas en TándemRESUMEN
A multidrug fatality involving sumatriptan is reported. Sumatriptan is a tryptamine derivative that acts at 5-HT(1B/1D) receptors and is used for the treatment of migraines. The decedent was a 21-year-old white female found dead in bed by her spouse. No signs of physical trauma were observed and a large number of prescription medications were discovered at the scene. Toxicological analysis of the central blood revealed sumatriptan at a concentration of 1.03 mg/L. Following therapeutic dosing guidelines, sumatriptan concentrations do not exceed 0.095 mg/L. Sumatriptan was isolated by solid-phase extraction and analyzed using liquid chromatography-tandem mass spectrometry in multiple reaction monitoring mode. A tissue distribution study was completed with the following concentrations measured: 0.61 mg/L in femoral blood, 0.56 mg/L in iliac blood, 5.01 mg/L in urine, 0.51 mg/kg in liver, 3.66 mg/kg in kidney, 0.09 mg/kg in heart, 0.32 mg/kg in spleen, 0.01 mg/kg in brain, 15.99 mg/kg in lung and 78.54 mg/45 mL in the stomach contents. Carisoprodol, meprobamate, fluoxetine, doxylamine, orphenadrine, dextromethorphan and hydroxyzine were also present in the blood at the following concentrations: 3.35, 2.36, 0.63, 0.19, 0.06, 0.55 and 0.16 mg/L. The medical examiner ruled the cause of death as acute mixed drug toxicity and the manner of death as accident.
Asunto(s)
Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/sangre , Sumatriptán/envenenamiento , Autopsia , Carisoprodol/sangre , Cromatografía Liquida , Dextrometorfano/sangre , Doxilamina/sangre , Estudios de Evaluación como Asunto , Resultado Fatal , Femenino , Fluoxetina/sangre , Toxicología Forense , Humanos , Hidroxizina/sangre , Hígado/efectos de los fármacos , Hígado/metabolismo , Meprobamato/sangre , Orfenadrina/sangre , Reproducibilidad de los Resultados , Manejo de Especímenes , Sumatriptán/farmacocinética , Espectrometría de Masas en Tándem , Distribución Tisular , Adulto JovenRESUMEN
The detection of new designer drugs is often a difficult issue in forensic urine drug testing as immunoassays are the primary screening methodology for drugs of abuse in many of these laboratories. Cross-reactivity of compounds with immunoassay kits can either aid or complicate the detection of a variety of drug and drug metabolites. For instance, emerging designer drugs that share structural similarities to amphetamines and phencyclidine (PCP) have the potential to cross-react with assays designed to detect these compounds. This study evaluates the cross-reactivity of five commercially available immunoassay reagent kits for 94 designer drugs on a Roche/Hitachi Modular P automated screening instrument. The compounds used in this study are grouped by structural class as follows: 2,5-dimethoxyamphetamines, 2C (2,5-dimethoxyphenethylamines), ß-keto amphetamines, substituted amphetamines, piperazines, α-pyrrolidinopropiophenones, tryptamines and PCP analogs. A drug concentration of 100 µg/mL was used to determine cross-reactivity for each assay and resulted in the following positive rates: Microgenics DRI(®) Ecstasy enzyme assay (19%), Microgenics DRI(®) Phencyclidine enzyme assay (20%), Lin-Zhi Methamphetamine enzyme immunoassay (39%), Siemens/Syva(®) EMIT(®)II Plus Amphetamines assay (43%) and CEDIA(®) DAU Amphetamine/Ecstasy assay (57%). Of the 94 designer drugs tested, 14% produced a negative response for all five kits. No designer drug used in this study generated a positive result for all five immunoassay kits.
Asunto(s)
Drogas de Diseño/análisis , Inmunoensayo/métodos , Juego de Reactivos para Diagnóstico , Reacciones CruzadasRESUMEN
In recent years, a class of new designer drugs commonly referred to as 'bath salts' have made their way to the illicit drug market. The most common drugs encountered are designer amphetamines and cathinones. Many analytical methods for analysis and identification of bath salts have been published, but there has been little reported on their impact on existing gas chromatography-mass spectrometry (GC-MS) amine confirmation methods. Due to structural similarities, the potential exists that designer amphetamines may interfere with methods used for analysis of sympathomimetic amines. Methiopropamine, 4-fluoroamphetamine, 4-fluoromethamphetamine (4-FMA) and 4-methylamphetamine were examined for potential interference with immunoassays and GC-MS confirmation analysis utilizing three derivatization procedures: R(-)-α-methoxy-α-trifluoromethylphenylacetyl chloride (R-MTPAC), heptafluorobutyric anhydride (HFBA) and chlorodifluoroacetic anhydride (ClF(2)AA). Significant cross-reactivity was observed with all the four compounds on the Syva Emit(®) II Plus Amphetamines and Roche KIMS Amphetamines II immunoassays. Laboratories utilizing GC-MS selected-ion-monitoring confirmation methods with R-MTPAC, HFBA or ClF(2)AA derivatives could experience potential chromatographic and mass spectral interferences from 4-fluroamphetamine, 4-FMA and methiopropamine in the form of ion ratio and quantitative failures. Careful ion selection, proper selectivity and specificity studies during method validation and rigid chromatographic and spectral acceptance criteria are required to assure the robustness and accuracy of GC-MS methods.
Asunto(s)
Anfetamina/análisis , Drogas de Diseño/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , InmunoensayoRESUMEN
This study examined the potential of abalone ß-glucuronidase as a viable and cost effective alternative to current hydrolysis procedures using acid, Helix pomatia ß-glucuronidase and Escherichia coli ß-glucuronidase. Abalone ß-glucuronidase successfully hydrolyzed oxazepam-glucuronide and lorazepam-glucuronide within 5% of the spiked control concentration. Benzodiazepines present in authentic urine specimens were within 20% of the concentrations obtained with the current hydrolysis procedure using H. pomatia ß-glucuronidase. JWH 018 N-(5-hydroxypentyl) ß-d-glucuronide was hydrolyzed within 10% of the control concentration. Authentic urine specimens showed improved glucuronide cleavage using abalone ß-glucuronidase with up to an 85% increase of drug concentration, compared with the results obtained using E. coli ß-glucuronidase. The JWH 018 and JWH 073 carboxylic acid metabolites also showed increased drug concentrations of up to 24%. Abalone ß-glucuronidase was able to completely hydrolyze a morphine-3-glucuronide control, but only 82% of total morphine was hydrolyzed in authentic urine specimens compared with acid hydrolysis results. Hydrolysis of codeine and hydromorphone varied between specimens, suggesting that abalone ß-glucuronidase may not be as efficient in hydrolyzing the glucuronide linkages in opioid compounds compared with acid hydrolysis. Abalone ß-glucuronidase demonstrates effectiveness as a low cost option for enzyme hydrolysis of benzodiazepines and synthetic cannabinoids.
Asunto(s)
Gastrópodos/enzimología , Glucuronidasa/metabolismo , Urinálisis/métodos , Analgésicos Opioides/orina , Animales , Benzodiazepinas/orina , Cannabinoides/orina , Codeína/metabolismo , Análisis Costo-Beneficio , Escherichia coli/enzimología , Caracoles Helix/enzimología , Hidrólisis , Lorazepam/análogos & derivados , Lorazepam/metabolismo , Derivados de la Morfina/metabolismo , Oxazepam/análogos & derivados , Oxazepam/metabolismo , Manejo de EspecímenesRESUMEN
A case is presented of a 19-year-old white male who was found dead in bed by a friend. While no anatomic cause of death was observed at autopsy, toxicological analysis of his blood identified AH-7921, a synthetic opioid. AH-7921 was isolated by liquid-liquid extraction into n-butyl chloride from alkalinized samples. Extracts were analyzed and quantified by gas chromatography mass spectrometry in selected ion monitoring mode. The heart blood had an AH-7921 concentration of 3.9 mg/L and the peripheral blood concentration was 9.1 mg/L. In addition to the blood, all submitted postmortem specimens including urine, liver, kidney, spleen, heart, lung, brain, bile and stomach content were quantified. The following concentrations of AH-7921 were reported: 6.0 mg/L in urine, 26 mg/kg in liver, 7.2 mg/kg in kidney, 8.0 mg/kg in spleen, 5.1 mg/kg in heart, 21 mg/kg in lung, 7.7 mg/kg in brain, 17 mg/L in bile and 120 mg/125 mL in the stomach content. The medical examiner reported that the cause of death was opioid intoxication and the manner of death was accident.
Asunto(s)
Analgésicos Opioides/farmacocinética , Analgésicos Opioides/envenenamiento , Benzamidas/farmacocinética , Benzamidas/envenenamiento , Accidentes , Adulto , Analgésicos Opioides/sangre , Analgésicos Opioides/orina , Benzamidas/sangre , Benzamidas/orina , Resultado Fatal , Cromatografía de Gases y Espectrometría de Masas , Humanos , Extracción Líquido-Líquido , Masculino , Intoxicación/sangre , Intoxicación/etiología , Intoxicación/orina , Cambios Post Mortem , Distribución Tisular , Adulto JovenRESUMEN
Salvia divinorum, a member of the mint plant family, has hallucinogenic properties that have become increasingly sought after by recreational drug users. The main psychoactive component, salvinorin A, has potency comparable to lysergic acid diethylamide. Though still legal to possess in most of the United States and much of Europe, little is known regarding the compound's long-term health effects, addiction liability, and pharmacokinetics. Limited data are available in the scientific literature, and few analytical methods are published for the detection in human biological fluids. These factors contribute to the unfamiliarity of the compound and complicate the method development process necessary to accommodate special requested testing for salvinorin A. A sensitive analytical method for the detection and quantitation of salvinorin A in human biological fluids was developed and validated to resolve analytical shortcomings. The method utilizes a solid-phase extraction technique coupled with liquid chromatography-electrospray ionization mass spectrometry operated in selected ion monitoring mode. The assay has a linear range of 5.0-100 ng/mL with a correlation coefficient of 0.997. The limit of detection and limit of quantitation were experimentally determined as 2.5 and 5.0 ng/mL, respectively. The method has been applied to blood and urine samples successfully and can be used to detect the presence of salvinorin A in forensic testing.
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Diterpenos de Tipo Clerodano/análisis , Psicotrópicos/análisis , Salvia/química , Cromatografía Liquida , Diterpenos de Tipo Clerodano/sangre , Diterpenos de Tipo Clerodano/orina , Humanos , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Delta9-Tetrahydrocannabinol (THC) is the main psychoactive compound present in marijuana. THC can also be found, as a contaminant, in some commercially available hemp products marketed in health food stores and on the internet as a good source of essential fatty acids. The products range from oil to alcoholic beverages to nutritional bars to candies, with oil being the most popular and commonly available. The analytical results are separated into two groups, products tested prior to and after publication of 21 CFR Part 1308, "clarification of listing of tetrahydrocannabinols." The data presented are a summary of 79 different hemp products tested for THC. THC was separated by a liquid-liquid or solid-liquid extraction, depending upon the product matrix. THC concentrations range from none detected to 117.5 microg THC/g material. Typical limits of detection for the assay (depending on matrix) are 1.0-2.5 microg THC/g material. Products that were of aqueous base (beer, tea) had much lower limits of detection (2.5 ng/mL). No THC was detected in 58% of the products from group 1 and 86% of the products from group 2. The amounts indicate that THC levels in currently marketed hemp products are significantly lower than in those products available before 2003 and reported in previous studies. The results reported here may be used as a general guideline for the THC content of hemp products recently found in the marketplace today.
Asunto(s)
Cannabis/química , Dronabinol/análisisRESUMEN
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/mL except for ecgonine, which was 16 ng/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/mL in LV and 0-75 ng/mL in PM specimens. Ethyl ecgonidine was found only in PM urine at concentrations 0-39 ng/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 nor-cocaethylene were consistently the lowest of all cocaine metabolites. At benzoylecgonine concentrations below 100 ng/mL, ecgonine was present at the highest concentrations. In 20 urine specimens, benzoylecgonine and ecgonine median concentrations (range) were 54 (0-47) and 418 ng/mL (95-684), respectively. Therefore, detection of ecgonine is advantageous when benzoylecgonine concentrations are below 100 ng/mL.
Asunto(s)
Cocaína/análogos & derivados , Cocaína/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Cocaína/sangre , Cocaína/orina , HumanosRESUMEN
Forensic Urine Drug Testing Laboratories often requires two confirmatory methods for a methamphetamine positive screen. First, methamphetamine is identified and quantitated using gas chromatography-mass spectrometry. If the total methamphetamine concentration is above the administrative cutoff level, the isomeric composition must be determined. This eliminates a possible contribution by over-the-counter cold medications that contain l-methamphetamine (Vick's inhalers). Products that contain only the l-isomer of methamphetamine must be distinguishable from prescription or illicitly manufactured methamphetamine, which consists mainly of the d-isomer. Optically impure derivatizing reagents will produce an impure mixture from a pure isomeric compound. Therefore, methods utilizing impure reagents can prove problematic when interpreting results. Use of an optically pure chiral derivatizing reagent, such as R(-)-alpha-methoxy-alpha-trifluoromethylphenylacetic acid chloride, allows for the creation and measurement of chromatographically separable isomeric compounds. The novel method described here utilizes a polymer-based solid-phase column adapted to a positive pressure manifold extraction system and a one-step derivatization process that occurs directly in the elution solvent. This methodology eliminates an elution solvent dry-down step that may adversely affect recovery of volatile amphetamine compounds. Although the method was designed for the quantitative analysis of the isomers of amphetamine and methamphetamine, it can be adapted for use with a wide range of phenethylamines including methylenedioxyamphetamine, N-methylenedioxymethamphetamine, and possibly N-methylenedioxyethylamphetamine. The linear range for quantitation was 25-10,000 ng/mL for d,l-methamphetamine and d,l-amphetamine, and correlation coefficients were 0.997 or better. The coefficient of variation for all four analytes did not exceed 2.8%. Concentrations analyzed ranged from 500 to 4000 ng/mL (n=40). The method allows for a simple and accurate quantitation and isomeric determination of amphetamine and methamphetamine using a process that eliminates extraction and derivatization complications common in current methods.
Asunto(s)
Trastornos Relacionados con Anfetaminas/orina , Anfetamina/orina , Estimulantes del Sistema Nervioso Central/orina , Metanfetamina/orina , Fenilacetatos/química , Detección de Abuso de Sustancias/métodos , Anfetamina/química , Trastornos Relacionados con Anfetaminas/diagnóstico , Estimulantes del Sistema Nervioso Central/química , Medicina Legal/métodos , Humanos , Metanfetamina/química , SolventesRESUMEN
Current Department of Defense (DoD) and Department of Health and Human Services (HHS) procedures for the detection of heroin abuse by testing urine utilize an initial opiate (codeine/morphine) immunoassay (IA) screen followed by gas chromatography-mass spectrometry (GC-MS) confirmation of 6-acetylmorphine (6-AM), if the morphine concentration is above established cutoff. An alternative to the current opiates screen for heroin abuse is the direct IA for the metabolite of heroin, 6-acetylmorphine. In this regard, the performance of the Microgenics CEDIA heroin metabolite (6-AM) screening reagent was assessed. This evaluation was conducted on the P module of a Hitachi Modular automated IA analyzer calibrated using 6-AM at 10 ng/mL. Reproducibility, linearity, accuracy, sensitivity, and interferences associated with use of the 6-AM IA reagent were evaluated. The IA reagent precision (percent coefficient of variation (%CV)) around each of seven standards was less than 0.63%, with a linearity (r(2)) value of 0.9951. A total of 37,713 active duty service members' urine samples were analyzed simultaneously using the CEDIA heroin metabolite (6-AM) reagent and the Roche Abuscreen ONLINE opiate reagent to evaluate both the prevalence rate of 6-AM in the demographic group and the sensitivity and specificity of the reagents for the detection of heroin use. Of the 37,713 samples tested using the CEDIA heroin metabolite (6-AM) reagent, three samples screened positive at the DoD and HHS cutoff of 10 ng/mL. One of the three samples confirmed positive for 6-AM by GC-MS above the cutoff of 10 ng/mL, the two remaining samples confirmed negative for 6-AM at a GC-MS limit of detection (LOD) of 2.1 ng/mL. In contrast, the Roche Abuscreen ONLINE opiate IA produced 74 opiate-positive results for codeine/morphine, with 6 of the 74 specimens confirming positive for morphine above the DoD cutoff concentration of 4000 ng/mL (8% DoD morphine confirmation rate), only one of the 74 opiate-positive screen specimens confirmed positive for 6-AM above the 10 ng/mL GC-MS cutoff concentration. As a further check of the sensitivity and specificity of the Microgenics 6-AM IA reagent, human urine samples (n = 87) known to contain 6-AM by GC-MS, were re-analyzed using both IA reagents. All 87 of the samples screened positive using the CEDIA heroin metabolite (6-AM) assay. However, using the Roche ONLINE opiate reagent, 12 of the known 6-AM positives screened negative at the DoD and HHS screening cutoff of 2000 ng/mL (morphine). Of the remaining 75 samples that screened positive by the ONLINE opiate reagent, five of the samples did not contain morphine above the DoD GC-MS cutoff concentration of 4000 ng/mL and would not have required 6-AM analysis. However, under the HHS GC-MS morphine cutoff concentration of 2000 ng/mL all 75 samples would have required 6-AM analysis. Furthermore, using the current DoD opiate screen, 17 out of 87 samples known to contain 6-AM would have gone undetected (19.5% false-negative rate); additionally, even under the more stringent HHS opiate screening standards 12 out of the 87 samples known to contain 6-AM would also have gone undetected (13.8% false-negative rate). The Microgenics CEDIA heroin metabolite (6-AM) reagent assay appears well adapted for the rapid and specific detection of heroin abuse as an alternative for, or an adjunct test to, the current opiates (codeine/morphine) IA screening procedure.