4-hydroxy-3-methoxymethamphetamine and 3-4-methylenedioxyethamphetamine

3,4-Methylenedioxymethamphetamine (MDMA), or ecstasy, is excreted as unchanged drug, 3,4-methylenedioxyamphetamine (MDA), and free and glucuronidated/sulfated 4-hydroxy-3-methoxymethamphetamine (HMMA), and 4-hydroxy-3-methoxyamphetamine (HMA) metabolites. The aim of this paper is to describe the pattern and timeframe of excretion of MDMA and its metabolites in urine. Placebo, 1.0 mg/kg, and 1.6 mg/kg oral MDMA doses were administered double-blind to healthy adult MDMA users on a monitored research unit. All urine was collected, aliquots were hydrolyzed, and analytes quantified by gas chromatography-mass spectrometry. Median C(max), T(max), ratios, first and last detection times, and detection rates were determined. Sixteen participants provided 916 urine specimens. After 1.6 mg/kg, median C(max) were 21,470 (MDMA), 2229 (MDA), 20,793 (HMMA), and 876 ng/mL (HMA) at median T(max) of 13.9, 23.0, 9.2 and 23.3 h. In the first 24 h, 30.2-34.3% total urinary excretion occurred. HMMA last detection exceeded MDMA's by more than 33 h after both doses. Identification of HMMA as well as MDMA increased the ability to identify positive specimens but required hydrolysis. These MDMA, MDA, HMMA, and HMA pharmacokinetic data may be useful for interpreting workplace, drug treatment, criminal justice, and military urine drug tests. Measurement of urinary HMMA provides the longest detection of MDMA exposure yet is not included in routine monitoring procedures.

Topics: 3,4-Methylenedioxyamphetamine; Adolescent; Adult; Female; Gas Chromatography-Mass Spectrometry; Hallucinogens; Humans; Male; Methamphetamine; N-Methyl-3,4-methylenedioxyamphetamine; Substance Abuse Detection; Young Adult

3,4-Methylenedioxymethamphetamine (MDMA, or Ecstasy) is a popular recreational drug. Analysis of MDMA and metabolites in human plasma, particularly in pharmacokinetic studies, requires low limits of quantification. Two-dimensional GC/MS with cryofocusing is a chromatographic technique recognized for its increased selectivity and resolution.. This method simultaneously quantifies 3,4-methylenedioxyethylamphetamine (MDEA), MDMA, and its metabolites, 3,4-methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxymethamphetamine (HMMA), and 4-hydroxy-3-methoxyamphetamine (HMA) in human plasma. With hydrochloric acid, we hydrolyzed 1 mL plasma, fortified with internal standard. Analytes were subjected to solid-phase extraction, derivatized with heptafluorobutyric acid anhydride, and quantified using cryofocused 2-dimensional GC/MS operated in electron-impact selected ion-monitoring mode.. Limits of quantification were 1.0 microg/L for MDA and 2.5 microg/L for MDEA, MDMA, HMMA, and HMA. Calibration curves were linear to 100 microg/L for MDA and HMA and to 400 microg/L for MDEA, MDMA, and HMMA, with r(2) > 0.997. At 3 concentrations spanning the linear dynamic range of the assay, mean overall extraction efficiencies from plasma were > or =85% for all compounds of interest. Recoveries were 85.6% to 107.2% of target, and intra- and interassay imprecision (CV) was <8.5% for all drugs at 3 concentrations within the range of the assay. None of the 66 exogenous compounds tested interfered with analyte quantification.. This GC/MS assay provides low limits of quantification for simultaneous determination of MDEA, MDMA, and metabolites MDA, HMMA, and HMA in human plasma. The 2D chromatographic system should be suitable for application to other analytes and to other complex matrices.

Topics: 3,4-Methylenedioxyamphetamine; Calibration; Central Nervous System Stimulants; Dopamine; Gas Chromatography-Mass Spectrometry; Humans; Hydrolysis; Methamphetamine; N-Methyl-3,4-methylenedioxyamphetamine; Reproducibility of Results; Substance Abuse Detection

3,4-Methylenedioxymethamphetamine (MDMA), a commonly encountered drug of abuse, has been shown in a variety of studies to cause neurotoxic effects. Because MDMA itself is not neurotoxic, identifying the potential neurotoxic metabolite(s) was of significant importance. Evaluation of urine and plasma concentrations of MDMA and three of its main metabolites, 3,4-methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxyamphetamine (HMA), and 4-hydroxy-3-methoxymethamphetamine (HMMA), following administration of a neurotoxic dose (20 mg/kg) to male Dark Agouti rats was accomplished. Currently there are no data available describing urine and plasma concentrations of MDMA and these metabolites over a period of 7 days. The rats received a single 20 mg/kg i.p. dose of MDMA. Blood and urine samples were collected prior to administration and at 2, 4, 8, 12, 16, 20, 24, 48, 96, and 168 h following drug administration. Plasma and urine samples were extracted using solid-phase extraction, derivatized with N-methyl-bis(trifluoroacetamide), then analyzed using gas chromatography-mass spectrometry. Urine samples showed peak concentrations of MDMA at 4 h, MDA at 8 h, HMMA at 12 h, and HMA at 16 h post dose. MDMA and its metabolites were detectable (limit of detection 25 ng/mL) in the urine for up to 168 h post dose. Plasma samples showed mean peak concentrations of MDMA and MDA at 2 h post dose and HMMA at 4 h. Although the highest mean concentration of HMA was seen at 24 h post dose, variability between sample results for this time point was significant. No detectable levels of MDMA, MDA, HMA, and HMMA (LOD 10 ng/mL) were found in plasma at 96 and 168 h post dose.

Topics: 3,4-Methylenedioxyamphetamine; Animals; Biotransformation; Calibration; Dopamine; Gas Chromatography-Mass Spectrometry; Male; Methamphetamine; N-Methyl-3,4-methylenedioxyamphetamine; Psychotropic Drugs; Rats; Reproducibility of Results; Toxicology

3,4-Methylenedioxymethamphetamine (MDMA) is a recreational drug with neurotoxic potential. Pharmacokinetic data of MDMA and its metabolites may shed light on the mechanism of MDMA neurotoxicity. An LC-MS assay with electrospray ionization (ESI) is presented for quantifying MDMA and its metabolites 3,4-methylenedioxyamphetamine (MDA), 3,4-dihydroxymethamphetamine (HHMA), and 4-hydroxy-3-methoxymethamphetamine (HMMA) in squirrel monkey plasma. The method involved enzymatic conjugate cleavage and protein precipitation. Separation was achieved within 14min. The method was validated according to international guidelines with respect to selectivity, linearity, accuracy, precision, recovery, and matrix effect. The present method should prove useful for acquiring pharmacokinetic and toxicokinetic data in squirrel monkeys.

Topics: 3,4-Methylenedioxyamphetamine; Animals; Calibration; Chromatography, Liquid; Deoxyepinephrine; Hallucinogens; Methamphetamine; N-Methyl-3,4-methylenedioxyamphetamine; Saimiri; Spectrometry, Mass, Electrospray Ionization

A sensitive gas chromatography-mass spectrometry method was developed and validated for the simultaneous measurement of MDEA, MDMA, and its metabolites, 3,4-methylenedioxy-N-ethylamphetamine (MDEA), 3,4-methylenedioxymethamphetamine (MDMA or Ecstasy), and its metabolites, 4-hydroxy-3-methoxyamphetamine (HMA), 3,4-methylenedioxyamphetamine (MDA), and 4-hydroxy-3-methoxyamphetamine (HMMA) in human urine.. We hydrolyzed 1 mL urine, fortified with MDMA-d5, MDA-d5, and MDEA-d6, with 100 microL of concentrated hydrochloric acid at 120 degrees C for 40 min, then added 100 microL 10 N sodium hydroxide and 3 mL phosphate buffer 0.1 N (pH 6.0) were added to hydrolyzed urine specimens before solid-phase extraction. After elution and evaporation, we derivatized extracts with heptafluorobutyric acid anhydride and analyzed with gas chromatography-mass spectrometry operated in EI-selected ion-monitoring mode.. Limits of quantification were 25 microg/L for MDEA, MDMA, and its metabolites. Calibration curves were linear to 5000 microg/L for MDEA, MDMA, HMA, MDA, and HMMA, with a minimum r2 > 0.99. At 3 concentrations spanning the linear dynamic range of the assay, mean overall extraction efficiencies from urine were >85.5% for all compounds of interest. Intra- and interassay imprecisions, produced as CV, were <15% for all drugs at 30, 300, and 3000 microg/L.. This gas chromatography-mass spectrometry assay provides adequate sensitivity and performance characteristics for the simultaneous quantification of MDEA, MDMA, and its metabolites HMMA, MDA, and HMA in human urine. The method meets and exceeds the requirements of the proposed Substance Abuse and Mental Health Services Administration's guidelines for federal workplace drug testing of MDEA and MDMA in urine.

Topics: 3,4-Methylenedioxyamphetamine; Amphetamines; Calibration; Central Nervous System Stimulants; Dopamine; Gas Chromatography-Mass Spectrometry; Humans; Hydrolysis; Methamphetamine; N-Methyl-3,4-methylenedioxyamphetamine; Sensitivity and Specificity; Substance Abuse Detection

In order to clarify the mechanism of drug incorporation into hair, disposition of 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), 3-methoxy-4,5-methylenedioxyamphetamine (MMDA) and metabolites of MDMA, 4-hydroxy-3-methoxyamphetamine (HMAP) and 4-hydroxy-3-methoxymethamphetamine (HMMA), into hair was investigated with an animal model. After the intraperitoneal administration of those six drugs to pigmented hairy rats (5 mg/kg/day, 10 days, n = 3), the parent compounds and their metabolites in the rat plasma (5, 15, 30, 60, 120, 360 min after administration) and in the newly grown rat hair for 4 weeks were determined by GC/MS-SIM. When the ratio of hair concentration to area under the concentration versus time curves (AUCs) in plasma was represented as an index of incorporation rate (ICR) of drugs into hair, the order of ICRs was HMAP < MDA < HMMA < MDMA < MDEA < MMDA. In the comparison between MDA, MDMA and MDEA, their ICRs increased according to the length of carbon branches from proton to ethyl at the N position. From the point of view that the ICRs of MMDA was 2.3 times as much as that of MDA, the methoxy group on the benzene ring seemed to serve as a positive factor for the ICR. However, the ICRs of 4-hydroxy-3-methoxy compounds, HMAP and HMMA, were lower in comparison with those of MDA and MDMA, respectively. On the other hand, the ICRs of MDA, MDMA and MDEA were 5.5-6.1 times larger than those of amphetamine, methamphetamine and ethylamphetamine, suggesting that the methylenedioxy group on the benzene ring raises their ICRs very positively. Moreover, in order to apply the results from the animal experiments to human cases, the scalp hair samples of seven MDMA abusers were analyzed. MDMA and its metabolites, MDA; were simultaneously detected in all the samples by GC/MS. In the two samples, MDEA was found in addition to MDMA and MDA. It was shown that a hair sample is a good specimen for the confirmation of retrospective use of methylenedioxyamphetamines.

Topics: 3,4-Methylenedioxyamphetamine; Amphetamines; Animals; Gas Chromatography-Mass Spectrometry; Hair; Hallucinogens; Humans; Lactates; Male; Methamphetamine; N-Methyl-3,4-methylenedioxyamphetamine; Rats; Substance Abuse Detection; Substance-Related Disorders

A gas chromatography-mass spectrometry assay based on perfluorotributylamine-enhanced ammonia positive ion chemical ionization has been developed for MDMA and three of its primary metabolites in biological specimens; the assay is linear from 2 to 1000 ng ml-1. Quantitatively, more of an administered dose of 10 mg kg-1 MDMA was excreted by the mouse (72%) than by the rat (35%); most in both species was excreted in urine and within 24 h. The difference in per cent excretion is entirely due to proportionally greater excretion of the parent drug by the mouse. 4-Hydroxy-3-methoxymethamphetamine (HMM) is the major urinary metabolite in both species. HMM and another primary metabolite, 4-hydroxy-3-methoxyamphetamine (HMA), were excreted mainly as glucuronide and sulphate conjugates (> 85%).

Topics: 3,4-Methylenedioxyamphetamine; Ammonia; Animals; Dopamine; Fluorocarbons; Gas Chromatography-Mass Spectrometry; Male; Methamphetamine; Mice; Mice, Inbred Strains; N-Methyl-3,4-methylenedioxyamphetamine; Rats; Rats, Sprague-Dawley; Species Specificity