3,4-methylenedioxymethamphetamine (MDMA) intoxication in an infant chronically exposed to cocaine

Accidental ingestion of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) was detected in an infant admitted at the Pediatric Emergency Department by drug testing in urine. Concentrations of MDMA and its principal metabolite 4-hydroxy-3-methoxymethamphetamine (HMMA) in the infant's hydrolyzed urine were 11.7 mg/L and 34.4 mg/L, respectively. Apparent febrile convulsions and cardiovascular side effects resolved within 1 day after treatment with benzodiazepines. Chronic exposure to cocaine was evidenced by segmental hair analysis. Continuous maternal denial of the presence of any drug in the household made diagnosis of accidental ingestion of MDMA and chronic exposure to cocaine problematic. Periodic clinical and laboratory follow-ups were requested to check eventual long-term effects of exposure to illicit drugs and discontinuation of the child from exposure to dangerous environments.     

Distribution study of 3,4-methylenedioxymethamphetamine and 3,4-methylenedioxyamphetamine in a fatal overdose

In this study, regional tissue distributions of the amphetamine analogue 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") and its metabolite 3,4-methylenedioxyamphetamine (MDA) in a fatal overdose are presented. Quantitation of MDMA and MDA levels occurred in blood samples taken centrally (right and left heart and main adjacent great vessels) and peripherally (subclavian and femoral blood). In addition, MDMA and MDA concentrations were determined in cardiac and iliopsoas muscle, both lungs, liver, both kidneys, spleen, the four brain lobes, cerebellum and brainstem, and adipose tissue. Finally, MDMA and MDA levels were determined in serum, vitreous humor, urine, and bile. For all samples, a fully validated high-pressure liquid chromatography procedure with fluorescence detection was used. The found substances were also identified with liquid chromatography-tandem mass spectrometry. Our data confirm that blood sampling from an isolated peripheral vein is recommended for MDMA and MDA. In addition, the vitreous humor MDMA level indicates that this fluid can be an interesting alternative when a suitable blood sample is missing. Considering the substantial differences in concentrations in blood samples taken from various sites in the body and the high levels in some tissues (e.g., in liver), we concluded that the influence of postmortem redistribution should be taken into account in the interpretation of toxicological data when an appropriate peripheral sample cannot be obtained or when blood samples are not available because of putrefaction.     

MDMA and MDA concentrations in antemortem and postmortem specimens in fatalities following hospital admission

Over the last 15 years, numerous deaths involving "Ecstasy" (3,4-methylenedioxymethamphetamine, MDMA) have been reported and described in the literature. In most cases, either antemortem or postmortem concentration data are available. Because of the wide range of results and potential idiosyncratic nature of MDMA toxicity, interpretation of both antemortem and postmortem concentrations is difficult. The possible influence of postmortem redistribution may be an overlooked factor, but existing data involve postmortem concentrations from varying anatomical sites. However, this paper describes for the first time an evaluation of the concentrations of MDMA and 3,4-methylenedioxyamphetamine (MDA) found in five fatalities admitted to hospital where both antemortem and postmortem blood samples were available. Admission MDMA and MDA concentrations ranged between 0.55 and 4.33 mg/L and 0 and 0.10 mg/L, respectively, in antemortem serum/plasma. Postmortem blood MDMA and MDA concentrations ranged between 0.47 and 28.39 mg/L and 0.02 and 1.33 mg/L, respectively. Postmortem concentrations were higher than corresponding antemortem concentrations in all 5 cases with postmortem/antemortem ratios between 1.1 and 6.6 for MDMA and 1.5 and 13.3 for MDA. Differences in concentrations were also observed between anatomical sites, with central sites (e.g., heart) having much higher concentrations than peripheral sites (e.g., femoral). Overall, MDMA and MDA appear to exhibit postmortem redistribution and concentrations measured in postmortem specimens (even from peripheral sites) are not directly comparable with antemortem findings close to or prior to death.     

Prevalence of use study for amphetamine (AMP), methamphetamine (MAMP), 3,4-methylenedioxy-amphetamine (MDA), 3,4-methylenedioxy-methamphetamine (MDMA), and 3,4-methylenedioxy-ethylamphetamine (MDEA) in military entrance processing stations (MEPS) specimens

The Roche Abuscreen Onlinetrade mark Amphetamine immunoassay (IA), modified to include sodium periodate, and the Microgenics DRI Ecstasy IA were used to determine the prevalence of amphetamine (AMP), methamphetamine (MAMP), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA) in urine specimens from applicants seeking to join the United States Armed Forces. Over a 4-month period, a total of 85,658 specimens were IA screened using the Department of Defense 500 ng/mL administrative cutoff level for AMP and MDMA. All presumptively positive specimens were confirmed using a solid-phase extraction procedure coupled with simultaneous analysis of AMP, MAMP, MDA, MDMA, and MDEA by fast gas chromatography-mass spectrometry using the same cutoff levels as the IA. The Roche Online Amphetamine IA identified 216 specimens as presumptively positive; of these, 70 specimens confirmed positive for AMP and 87 specimens confirmed positive for AMP and/or MAMP, resulting in a confirmation rate of 73%. The Microgenics DRI Ecstasy IA identified eight specimens as presumptively positive; of these, five specimens confirmed positive for MDMA and/or MDA, resulting in a confirmation rate of 63%. The total use prevalence for AMP, MAMP, MDA, MDMA, and/or MDEA in military entrance processing stations specimens over the testing period was determined to be 0.19%.     

An unusual multiple drug intoxication case involving citalopram

A 47-year-old male with a history of drug abuse and suicide attempts was found dead at home. The death scene investigation showed evidence of cocaine abuse and multiple drug ingestion. Citralopram, a new selective serotonin reuptake inhibitor, cocaine, oxycodone, promethazine, propoxyphene, and norpropoxyphene were identified and quantitated in the postmortem samples by gas chromatography-mass spectrometry. The concentration of citalopram in the femoral blood was 0.88 mg/L. The heart blood concentration was 1.16 mg/L. Femoral blood concentrations of the other drugs were as follows: cocaine, 0.03 mg/L; oxycodone, 0.06 mg/L; promethazine, 0.02 mg/L; propoxyphene, 0.02 mg/L; and norpropoxyphene, 0.07 mg/L. Other tissue samples were also analyzed. The concentrations of cocaine, oxycodone, promethazine, and propoxyphene in the blood, liver, brain, and gastric contents did not suggest an intentional overdose. However, the possibility of multiple drug interactions including citalopram was evident. In this case, the citalopram concentrations were consistent with those reported in fatal cases involving multiple drug use. Citalopram was present in urine at a concentration of 0.9 mg/L.     

Detection of 1-benzylpiperazine, 1-(3-trifluoromethylphenyl)-piperazine, and 1-(3-chlorophenyl)-piperazine in 3,4-methylenedioxymethamphetamine-positive urine samples

Historically, ecstasy tablets contained 3,4-methylenedioxymethamphetamine (MDMA) as the psychoactive component. In recent years, the Drug Enforcement Administration (DEA) and other law enforcement agencies have seized ecstasy tablets that are comprised of psychoactive drugs or drug mixtures other than MDMA. Many jurisdictions have reported the presence of piperazine derivatives including 1-benzylpiperazine (BZP), 1-(3-trifluoromethylphenyl)-piperazine (TFMPP), and 1-(3-chlorophenyl)-piperazine (mCPP) in ecstasy tablets. These piperazine derivatives produce stimulant and psychoactive effects similar to those produced by MDMA, amphetamine, and methamphetamine. In many countries, their use is not controlled, and therefore they have become a legal alternative to MDMA. For this study, a targeted population of 251 MDMA-positive urine samples were analyzed for designer drugs, including the piperazine derivatives. A basic liquid-liquid extraction followed by pentafluoropropionic anhydride (PFPA) derivatization and a full scan (m/z 42-550) gas chromatography-mass spectrometry analysis was used to screen the urine samples for 33 designer drugs. Overall, in 36% of the specimens analyzed, a stimulant or psychoactive compound other than MDMA and 3,4-methylenedioxyamphetamine (MDA) was detected. BZP, TFMPP, and mCPP were detected in 15%, 7%, and 1% of the samples, respectively.     

Comparison of urine and hair testing for drugs of abuse in the control of abstinence in driver's license re-granting

The purpose of the study was to compare the detection rate of illicit drugs in urine and hair specimens. The samples were taken from subjects trying to regain their revoked driver's license after a drug- or alcohol-related traffic offence. In 2010, we screened 14 000 urine and 3900 hair samples for amphetamines, methamphetamines, cannabinoids, cocaine, opiates, methadone, and benzodiazepines as well as for ethylglucuronide. We used the low threshold values of the new German guidelines for Medical Psychological Assessment (MPA). Positive screening tests were confirmed with gas chromatography-mass spectrometry (GC-MS), gas chromatography-tandem mass spectrometry (GC-MS/MS) or liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results show that positivity rates for methamphetamines, MDMA, cocaine, and monoacetylmorphine were 1.7-, 5.7-, 3.8- and 9.3-fold higher in hair than in urine. In contrast, the detection rate for benzodiazepines was higher in urine than in hair (oxazepam, 0.21% versus 0%, nordiazepam 0.10% versus 0.03%). The positivity rate in hair for ethylglucuronide was 6-fold (12.7%) that for urine testing (2.1%). The study reveals that in the control of abstinence in the context of driving license re-granting there are in part large differences of positivity rates for some drugs or metabolites between hair and urine samples. These differences should be kept in mind by physicians and psychologists in traffic medicine who are ordering the drug testing.     

Usefulness of roadside urine drug screening in drivers suspected of driving under the influence of drugs (DUID)

In Belgium, the driving under the influence of drugs (DUID) procedure consists of three steps: observation of external signs of drug consumption by a police officer; an on-site urine test for amphetamines, cannabinoids, cocaine, and opiates; and blood sampling by a physician for gas chromatography-mass spectrometry analysis. The driver is sanctioned if THC is greater than 2 ng/mL, morphine is greater than 20 ng/mL, or amphetamine, methylenedioxymethamphetamine (MDMA), methylenedioxyethylamphetamine, N-methyl-1-(3.4-methylenedioxyphenyl)-2-butanamine, cocaine, or benzoylecgonine are greater than 50 ng/mL in plasma. We analyzed the results of 450 blood samples taken from May 2000 to February 2005. Cannabis was most often found above the cut-off (73.5% of the cases), followed by MDMA (20.4%), amphetamine (19.8%), benzoylecgonine (17.9%), cocaine (6.9%), and morphine (2.7%). One drug was found in 72.0% of the cases, two drugs in 22.6%, three drugs in 5.2%, and four drugs in 0.25%. In 10.7% of the plasma samples no target drugs were found above the legal cut-off. This percentage was 8.4% when urine was obtained and tested on-site and 21.2% when no urine was obtained (chi2 = 8.574, P = 0.0034). In 64.6% of these samples, a target drug (THC in 74.2%) was found under the legal cut-off. These data indicate that roadside urine testing significantly decreases the number of unnecessary blood analyses in DUID.     

A novel procedure for the analysis of drugs in whole blood by homogeneous enzyme immunoassay (EMIT)

This paper describes a procedure for the analysis of whole blood by six commonly used EMIT d.a.u. assays: barbiturates, benzodiazepines, cocaine metabolite, opiates, amphetamines, and the cannabinoid 20 assays using the Syva Autocarousel and the COBAS MIRA. The sensitivity of the method rivals that of other immunoassay methods, such as RIA, for the detection of drugs in blood. The procedure has been successfully applied to the analysis of several hundred postmortem forensic blood specimens.     

Postmortem drug analysis: analytical and toxicological aspects

Publications focusing on the analysis of postmortem specimens for the presence of drugs were reviewed with particular reference to systematic toxicological analysis. Specimens included blood, liver, other solid specimens, and fly larvae. Extraction techniques published during the past 10 years most commonly used traditional solvent extraction techniques. High-performance liquid chromatography coupled to multichannel wavelength detection was most commonly used, which would easily lend itself to liquid chromatography-mass spectrometry. There were few practical differences in the assays validated for a range of postmortem specimens to those in other forms of forensic toxicology, unless substantially decomposed tissue was used. When putrefied specimens were analyzed, a back-extraction or other form of specimen cleanup was recommended to reduce interfering substances. Many immunoassays designed for urine have been adapted for use in blood and tissue homogenates. Immunoassays designed for blood analysis, however, are likely to have more useful cutoff values than immunoassays optimized for urine testing. Postmortem specimens provide less stability for a number of drugs than other types of specimens. This is particularly a problem for cocaine, heroin, and some antidepressants, antipsychotics, and benzodiazepines. A number of artifacts occur postmortem, which affects the concentration of drug in specimens. This includes postmortem redistribution for drugs with a high tissue concentration relative to blood. Consequently, the likely extent of any change in concentration is relevant to the interpretation of doses and drug effects.     

Children victims of drug abuser parents: Hair testing as a forensic tool to assess exposure—A cohort of 37 cases from Spain

Hair testing is a useful tool to investigate suspected pediatric exposure to drugs of abuse. Newborns and young children are at high risk of exposure to drugs of abuse from parents or caregivers who consumed these substances, a fact prosecuted by Spanish authorities as child abuse. A retrospective study based on a cohort of 37 cases classified using several parameters, which involve children under 12 years old, were analyzed at the Drugs Laboratory of the National Institute of Toxicology and Forensic Sciences (Madrid, Spain) between 2009 and 2021. Hair samples were tested for the presence of opiates, cocaine, ketamine, amphetamines, methadone, and cannabis using a gas chromatography–mass spectrometry (GC–MS) method. A 59% of the studied children had ages in the range of 1–3 years old, and in 81% of cases, victims required hospitalization. In 81% of cases (n = 30), hair was submitted only or in combination with other samples, and these were classified in four categories according to analyzed samples: A (only hair), B (hair and blood), C (hair and urine), and D (hair, blood, and urine). The 93.3% of these cases (n = 28) showed a positive result of cannabinoids (THC and CBN in hair and THC-COOH in urine; 71.4% n = 20), cocaine and metabolites (benzoylecgonine and cocaethylene; 46.4% n = 13), opiates (morphine and 6-acetylmorphine), and amphetamines (MDMA and MDMA; 3.10% n = 1). Hair analysis matched positive results in cases where urine screening test was carried out previously (n = 24) and in those cases where blood and/or urine were also submitted (35.6% n = 11). As a conclusion, hair analysis was confirmed as a useful tool to detect previous exposure to acute poisoning events in children.     

Three fatal cases of PMA and PMMA poisoning in Denmark

Paramethoxyamphetamine (PMA) and paramethoxymetamphetamine (PMMA) are methoxylated phenylethylamine derivatives with effects similar to methylenedioxymetamphetamine (MDMA) and sold as such. However, PMA and PMMA are more potent than MDMA, but have a slower onset of action, which encourages users to take more. Three fatal cases involving PMA and PMMA in Denmark in year 2000 are investigated including history, pathological, and toxicological findings. The methods used for extraction, identification, and quantitation of PMA and PMMA are described. In two of the cases, lethal postmortem blood concentrations of PMA and PMMA were determined at 3.4 and 3.3 mg/kg (case 1) and 0.78 and 0.68 mg/kg (case 3), respectively. In addition, other drugs such as MDMA, tetrahydrocannabinol, cocaine, and alcohol were involved in these cases. In the third case, death occurred four days after the ingestion of tablets containing PMA and PMMA, and therefore only low postmortem concentrations of PMA and amphetamine were detected. However, in a serum sample taken at admission to the hospital, PMA and PMMA were found, but not quantitated. It is believed that the cause of death in case 2, multiple-organ failure, was caused by overdoses of PMA and PMMA.     

Drug-impaired driving and traffic collisions: Study on a cross section of the Italian population

The present study focuses on the association between road accidents and the presence of drugs of abuse markers in the biological fluids of the drivers. Biological fluids collected from 1236 drivers involved in road accidents (54 fatal and 1182 non-fatal crashes) in the Rome area were analyzed for alcohol and psychotropic drugs, as required by judicial authorities. The substance most frequently detected was alcohol (in 19% of non-fatal and 32% of fatal crashes), followed by cannabinoids (12% of non-fatal crashes) and cocaine (9% of non-fatal and 20% of fatal crashes). The results obtained for cocaine and cannabinoids in blood and urine were compared. We observed the absence or low concentrations of the active drug in blood (cocaine was often below 5 ng/ml and THC below 1 ng/ml), whereas urinary concentrations of metabolites were generally high (benzoylecgonine 250–above 5000 ng/ml, THCCOOH 15–270 ng/ml). The risk of being involved in a road accident if cocaine or cannabis markers were present in the urine specimens was evaluated compared to a control population. The odds ratios calculated, being 8.13 for cannabis and 5.32 for cocaine, suggest a strong association between the presence of these drugs in the urine of drivers and traffic accidents, regardless of their presence in blood samples. The present data suggest that the chance of being involved in a road accident is higher than in the control population even if the subject is no longer “under the influence” of cannabis or cocaine at the time of the accident.     

Highly sensitive micro-plate enzyme immunoassay screening and NCI-GC-MS confirmation of flunitrazepam and its major metabolite 7-aminoflunitrazepam in hair.

Flunitrazepam (Rohypnol) is a benzodiazepine used in the treatment of insomnia as a sedative hypnotic and as preanesthetic medication in European countries and Mexico. Although it has no medicinal purpose in the United States, the occurrence of its abuse is increasing. Sexual abuse of both men and women while under the influence of so-called "date-rape" drugs has been the focus of many investigations. Reported date-rape drugs include flunitrazepam (FN), clonazepam, diazepam, oxazepam, gamma-hydroxybutyrate, and many others. FN has been banned in the United States because of its alleged use in such situations. Unfortunately, the detection of FN or its metabolites 7-aminoflunitrazepam (7-AFN) and desmethylflunitrazepam in a single specimen such as urine or blood is difficult in criminal situations because of the likelihood of single-dose ingestion and the length of time since the alleged incident. Hair provides a solution to the second of these problems in that drugs tend to incorporate into hair and remain there for longer periods of time than either urine or blood. There are various techniques for the detection of FN in plasma, blood, and urine, but little work has been done with hair. Hair collection is a virtually noninvasive procedure that can supply information on drug use for several months preceding collection. The objective of this paper was to determine if a commercially available micro-plate enzyme immunoassay system was sufficiently sensitive for the routine screening of 7-AFN in hair by the development of extraction procedures and optimization of the immunoassay kit. Further, this study used the same solid-phase extraction to isolate FN and its major metabolite, 7-AFN, and gas chromatography-mass spectrometry with negative ion chemical ionization for confirmation. Two seven-point standard curves were established ranging from 0.5 pg/mg to 100 pg/mg for 7-AFN and 2.5 pg/mg to 200 pg/mg for FN with respective deuterated internal standards. A replicate analysis of controls was performed to establish inter- and intraday variabilities. Two suicide cases along with one alleged date-rape case and one case of an emergency room patient whose blood screened positive for benzodiazepines were analyzed. All the hair specimens screened positive for benzodiazepines using micro-plate enzyme immunoassay. Two cases, including the date-rape case, were negative for FN and 7-AFN, and two postmortem hair samples were confirmed positive for FN and its metabolite.     

Surveillance of drug abuse in Hong Kong by hair analysis using LC‐MS/MS

The aim of this study is to reveal the habits of drug abusers in hair samples from drug rehabilitation units in Hong Kong. With the application of liquid chromatography–tandem mass spectrometry (LC–MS/MS) technology, a total of 1771 hair samples were analyzed during the period of hair testing service (January 2012 to March 2016) provided to 14 drug rehabilitation units including non‐governmental organizations (NGOs), rehabilitation centers, and medical clinics. Hair samples were analyzed for abused drugs and their metabolites simultaneously, including ketamine, norketamine, cocaine, benzoylecgonine, cocaethylene, norcocaine, codeine, MDMA, MDA, MDEA, amphetamine, methamphetamine, morphine, 6‐acetylmorphine, phencyclidine, and methadone. The results showed that ketamine (77.2%), cocaine (21.3%), and methamphetamine (16.5%) were the frequently detected drugs among those drug abusers, which is consistent with the reported data. In addition, the usage of multiple drugs was also observed in the hair samples. About 29% of drug‐positive samples were detected with multiple drug use. Our studies prove that our locally developed hair drug‐testing method and service can be a valid tool to monitor the use of abused drugs, and which could facilitate rehabilitation program management.     

Simultaneous determination of amphetamine, methamphetamine, methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA), and methylenedioxyethylamphetamine (MDEA) enantiomers by GC-MS.

A method is described for the simultaneous determination of the ratio of l- and d-enantiomers of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA) in urine. The assay uses liquid-liquid extraction followed by derivatization with trifluoroacetyl-l-prolyl chloride (l-TPC) and analysis by gas chromatography-mass spectrometry. The assay was developed using prepared samples containing varying concentrations of each of the analytes over a range of percentages of each enantiomer. Results showed the method to provide accurate and reliable results in samples containing > or = 10 ng/mL amphetamine and methamphetamine and > or = 25 ng/mL MDA, MDMA, and MDEA. The assay was used to analyze urine samples from subjects of a controlled MDMA study. Results for each of the eight subjects showed a greater percentage of the l-enantiomer of MDMA initially, and the percentage increased with time postdose. Analysis of the metabolite MDA revealed that the proportion of d-enantiomer was initially greater than the l-enantiomer followed by a gradual increase in the proportion of l-enantiomer until it exceeded the amount of the d-enantiomer. In all cases, the l-MDA exceeded the d-MDA within the first 36 h postdose.     

S(+)- and R(-)N-methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane (MDMA) as discriminative stimuli: effect of cocaine

Racemic N-methyl-1-(3,4-methylenedioxyphenyl)-2-aminopropane (methylenedioxymethamphetamine, MDMA), a central stimulant and empathogenic agent, and cocaine are drugs of abuse that function as training drugs in drug discrimination studies. In tests of stimulus generalization (substitution), asymmetric generalization occurs between the two agents: a (+/-)MDMA stimulus generalized to cocaine, but a cocaine stimulus did not generalize to (+/-)MDMA. A possible explanation may be found, at least in part, in the stimulus effects of the optical isomers of MDMA. In the present study, groups of male Sprague-Dawley rats were trained to discriminate either S(+)MDMA (training dose=1.5 mg/kg, i.p.; n=10; ED50=0.6 mg/kg) or R(-)MDMA (training dose=1.75 mg/kg, i.p.; n=7; ED50=0.4 mg/kg) from saline vehicle using a VI-15s schedule of reinforcement. Tests of stimulus generalization with cocaine were conducted in each of the two groups. Cocaine only partially substituted for the S(+)MDMA stimulus (maximum=39% drug-appropriate responding), and various doses of cocaine did not enhance the percent drug-appropriate responding produced by a low dose (0.5 mg/kg) of S(+)MDMA. In contrast, the R(-)MDMA stimulus generalized completely to cocaine (ED50=1.3 mg/kg). Taken together with an earlier report that a (+/-)MDMA stimulus generalizes to cocaine, it would seem that the stimulus actions of cocaine might share greater similarity with R(-)MDMA than with S(+)MDMA.     

Analysis of 3,4-methylenedioxymetamphetamine: whole blood versus dried blood spots

Analysis of dried blood spots is an increasingly accepted method in therapeutic drug monitoring, whereas its application by analogy to forensic samples has not been studied in detail. Therefore, we investigated whether determination of 3,4-methylenedioxymethamphetamine (MDMA) and its main metabolite 3,4-methylendioxyamphetamine (MDA) from dried blood spots (DBS) is as reliable as that from whole blood specimens. Analysis was performed by liquid chromatography-tandem mass spectrometry following liquid-liquid extraction of blood and corresponding DBS samples from 20 volunteers participating in a controlled driving experiment under the influence of MDMA. The assay was checked for carryover, ion suppression/enhancement, linearity of response, lower limits of detection (LLOD) and quantitation, extraction efficiency and the within-run and between-run assay imprecision for both whole blood and DBS. The LLODs were 2.0 and 1.6 ng/mL for MDMA in whole blood and DBS, respectively, using a volume of 100 μL. LLODs of MDA were determined to be 0.25 ng/mL in whole blood specimens and 0.12 ng/mL in DBS. Extraction efficiency and imprecision did not differ significantly between the two methods for both MDMA and MDA. The mean concentration ratio of corresponding whole blood and DBS samples, t-test, and the Bland-Altman difference plot were used to test hypothesis of equality. Statistical analyses revealed that methods did not significantly differ for MDMA or MDA. Thus, DBS analysis has potential as a precise and inexpensive alternative to whole blood analysis of MDMA.     

Effects of MDMA (ecstasy) and two of its metabolites on rat embryos in vitro

MDMA consumers are young people of childbearing age. Consequently, developmental exposure to this drug is a potential public health concern. Several studies have addressed MDMA neurotoxicity in adults; however, knowledge of the effects of MDMA on developing embryos is limited. After administration, MDMA is metabolized species specifically via two main pathways. One leads to the formation of MDA and the other to the formation of HHMA. Here we evaluated the embryotoxic effects of MDMA, and also those of MDA, a main metabolite of MDMA in rats, and HHMA, a main metabolite in humans. For this purpose, we used the whole embryo culture (WEC). Our results show a concentration-dependent embryotoxic effect of MDMA, MDA and HHMA at a concentration range of 25-50μg/ml. The embryotoxic potential of the parent compound and the two metabolites was comparable in vitro.     

Rapid detection of benzoylecgonine in vitreous humor by enzyme immunoassay

The usual specimens submitted by a medical examiner for toxicological analysis include blood, urine, bile, vitreous humor, stomach contents, and solid-organ tissue. The detection of drugs in these specimens typically involves a combination of techniques including colorimetry, immunoassay, and gas chromatography. Although many laboratories rely principally on urine for the detection of drugs of abuse by immunoassay, these assays may be applied to other specimen types. An evaluation of Microgenics Corporation's cloned enzyme donor immunoassay (CEDIA) was conducted in order to evaluate its use in the detection of cocaine/cocaine metabolites in vitreous humor specimens. During a 14-month period, 392 vitreous humor specimens were analyzed by the CEDIA DAU Cocaine assay. Instrument parameters were set according to published manufacturer's guidelines. All presumptive positive immunoassay results prompted confirmatory testing and quantitation by gas chromatography-mass spectrometry (GC-MS) of other specimens including blood. Vitreous humor specimens were not tested by GC-MS. Using a approximately 100-ng/mL cutoff, the CEDIA assay produced 23 presumptive positive results, 22 of which were confirmed by GC-MS. The only specimen which could not be confirmed, elicited an immunoassay screen value near the cutoff limit. Routine analysis of blood, urine, bile, and/or bladder wash specimens by gas chromatography-nitrogen phosphorus detection revealed the presence of cocaine/cocaine metabolites in only 7 (31.8%) of the 22 confirmed cases. The concentration ranges of cocaine and benzoylecgonine in the blood specimens were none detected to 337 ng/mL and 17 to 8598 ng/mL, respectively. Cocaethylene was not detected in these cases. Analysis of vitreous humor specimens by CEDIA improved the detection rate of cocaine/cocaine metabolites by 0.7% in the cases submitted to our laboratory during the 14-month period.