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Introduction: Stimulant medications are approved to treat attention deficit hyperactivity disorder (ADHD) in children over the age of 6 years. Fatal ingestion of stimulants by children has been reported, although most ingestions do not result in severe toxicity. Lisdexamfetamine dimesylate, a once daily long-acting stimulant, is a prodrug requiring conversion to its active form, dextroamphetamine, in the bloodstream. Based on its unique pharmacokinetics, peak levels of d-amphetamine are delayed. We describe a case of accidental ingestion of lisdexamfetamine dimesylate in an infant., Case Report: A previously healthy 10-month-old infant was admitted to the hospital with a 5-h history of tachycardia, hypertension, dyskinesia, and altered mental status of unknown etiology. Confirmatory urine testing, from a specimen collected approximately 16 h after the onset of symptoms, revealed an urine amphetamine concentration of 22,312 ng/mL (positive cutoff 200 ng/mL). The serum amphetamine concentration, from a specimen collected approximately 37 h after the onset of symptoms, was 68 ng/mL (positive cutoff 20 ng/mL). Urine and serum were both negative for methamphetamine, methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA, Ecstasy), and methylenedioxyethamphetamine (MDEA). During the hospitalization, it was discovered that the infant had access to lisdexamfetamine dimesylate prior to the onset of symptoms., Conclusion: Amphetamine ingestions in young children are uncommon but do occur. Clinicians should be aware of signs and symptoms of amphetamine toxicity and consider ingestion when a pediatric patient presents with symptoms of a sympathetic toxidrome even when ingestion is denied., Competing Interests: The authors, Kelly E. Wood and Matthew D. Krasowski, have no conflicts of information to disclose. Informed Consent Informed consent for publication of this case was obtained and provided to the journal in accordance with the Journal of Medical Toxicology policy.

Nonmedical use of prescription stimulants such as phentermine (PT) has been regulated by law enforcement authorities due to its euphorigenic and relaxing effects. Due to high potential for its abuse, reliable analytical methods were required to detect and identify PT and its metabolite in biological samples. Thus a dilute and shoot liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed and validated for simultaneous determination of PT, N-hydroxyphentermine (NHOPT) and mephentermine (MPT) in urine. A 5μL aliquot of diluted urine was injected into the LC-MS/MS system. Chromatographic separation was performed by reversed-phase C18 column with gradient elution for all analytes within 5min. Identification and quantification were based on multiple reaction monitoring (MRM) detection. Linear least-squares regression with a 1/x(2) weighting factor was used to generate a calibration curve and the assay was linear from 50 to 15000ng/mL (PT and MPT) and 5 to 750ng/mL (NHOPT). The intra- and inter-day precisions were within 8.9% while the intra- and inter-day accuracies ranged from -6.2% to 11.2%. The limits of quantification were 3.5ng/mL (PT), 1.5ng/mL (NHOPT) and 1.0ng/mL (MPT). Method validation requirements for selectivity, dilution integrity, matrix effect and stability were satisfied. The applicability of the developed method was examined by analyzing urine samples from drug abusers., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: 3-Methylmethcathinone (3-MMC) is a synthetic cathinone stimulant structurally related to the new psychoactive substance (NPS) mephedrone (4-methylmethcathinone, 4-MMC). We describe a case series of analytically confirmed intoxications involving 3-MMC presented to emergency departments in Sweden and included in the STRIDA project., Study Design: Observational case series of consecutive patients with self-reported or suspected use of NPS presenting to hospitals in Sweden between August 2012 and March 2014., Methods: NPS analysis was performed by a liquid chromatography-mass spectrometry (MS)/MS method that is updated with new substances as they appear. Data on clinical features were collected during Poisons Information Centre consultations and retrieved from medical records., Results: 3-MMC was detected in 50 (6.4%) of the 786 cases included in the STRIDA project during the 20-month study period, with the peak occurring in August 2013. The age range of patients testing positive for 3-MMC was 17-49 years (median 24) and 76% of them were men. The 3-MMC concentration in serum ranged between 0.002 and 1.49 μg/mL (median, 0.091) and between 0.007 and 290 μg/mL (median, 3.05) in urine. Co-exposure to other NPS and/or traditional drugs was very common, and 3-MMC mono-intoxication was found in only 4 (8%) cases. The most frequent clinical features were tachycardia (48% of cases) and agitation (42%). Other features included a reduced level of consciousness (32%), dilated pupils (24%), hallucinations (20%), diaphoresis (12%), seizures (8%), and hyperthermia (6%). Most patients (60%) needed hospital care for only 1 day but in 8% for 3 days or longer., Conclusion: The majority of patients with analytically confirmed 3-MMC exposure had sympathomimetic features similar to those associated with mephedrone intoxication. However, the high incidence of co-exposure to other drugs makes the clinical interpretation difficult. Nevertheless, 3-MMC was associated with a high admittance rate to intensive care (30%), and detected in two cases with a fatal outcome, suggesting that 3-MMC is a harmful drug.

Clenbuterol, a member of β-agonist family, has now been a serious threat to human health due to its illegal usage in the livestock feeding. Herein, we describe the application of melamine functionalized silver nanoparticles (M-AgNPs) as the electrochemical probe for simple, fast, highly sensitive and selective detection of clenbuterol. Generally, AgNPs are prepared and functionalized by melamine. After interacting with melamine modified gold electrode in the presence of clenbuterol, M-AgNPs can be immobilized on the surface of the electrode via the hydrogen-bonding interactions between clenbuterol and melamine. This sandwich structure permits sensitive and selective detection of clenbuterol. Since M-AgNPs can provide a couple of well-defined sharp silver stripping peaks, which stands for a highly characteristic solid-state Ag/AgCl reaction, a rather low detection limit of 10 pM can be achieved. The detection range is from 10 pM to 100 nM, which is quite wide. This developed biosensor can potentially be used for clenbuterol detection in biological fluids in the presence of various interferences.

As one of the most sensitive detection mode for CE, chemiluminescence (CL) detection is less developed compared to some other detection modes such as MS and LIF, despite its low equipment cost and simple design. The fact is partly due to the limitation of CL systems suitable for CE. In this paper, a highly sensitive CE-CL strategy was established for the determination of dopamine (DA) and cimbuterol (CM) using acridinium ester as derivatization reagent. Catalyst was not required in this CL detection system. Also, a good sensitivity was obtained due to its quite low background and strong signal. Under the optimal conditions, the presented method has been successfully applied to analyze DA and CM with LODs (S/N = 3) of 2.0 and 0.50 ng/mL, respectively. The linear ranges were 5.0–1500 ng/mL and 2.0–1000 ng/mL for DA and CM, respectively. To validate our method, the levels of DA in human urine samples were detected by this method, and the results showed acceptable accordance with those of ELISA. All the analysis procedure could be completed in 400 s, and the results showed satisfactory sensitivity and selectivity.The approach could also be further extended to rapid analysis of many other compounds including primary and second amines in clinical medicine and biopharmaceutical analysis using acridinium ester as CL derivatization reagent.

Drug-facilitated sexual assault (DFSA) has been defined as the use of a chemical agent to facilitate a sexual assault. We report two cases of the use of tetrahydrozoline for DFSA. We believe this is the first report with urinary quantification of tetrahydrozoline levels postassault. Blood and urine were obtained c. 20 h postexposure in two cases of reported DFSA. Tetrahydrozoline was not detected in blood but was identified in urine in both victims. After initial identification in the urine using the 2010 update to the AAFS mass spectrometry database library, tetrahydrozoline was quantified at 114 and 150 ng/mL, respectively, using GC/MS. Two unique clinical features reported in these cases were intermittent periods of consciousness and postexposure vomiting. Use of GC/MS was successful in identifying tetrahydrozoline in the 100 ng/mL range up to 20 h postexposure. For victims with late presentation, urine may be a better sample for evaluation for tetrahydrozoline., (© 2011 American Academy of Forensic Sciences.)

A GC method was developed for the identification and quantitation of eight sympathomimetic amines in urine, i.e., amphetamine, methamphetamine, mephentermine, ephedrine, pseudoephedrine, methylenedioxyamphetamine, methylenedioxymethamphetamine, and methylenedioxyethylamphetamine. Methoxyphenamine was used as the internal standard (IS). The assay is rapid, sensitive, and simple to perform. It involves a liquid-liquid extraction procedure with simultaneous in-solution derivatization of the organic layer with pentafluorobenzoyl chloride (PFB-CI), followed by GC/MS analysis. These derivatives and the IS were extracted from 1 mL alkaline urine into hexane before derivatization with PFB-CI. The organic layer was then removed and evaporated to dryness before dissolution with hexane for GC/MS analysis. Calibration curves for each analyte showed linearity in the range of 25-5000 ng/mL (r2 > or = 0.997). Recoveries ranged from 88 to 99%, with the precision of recoveries typically < or = 5%. The LOD values ranged from 7 to 28 ng/mL, and the LOQ values ranged from 23 to 94 ng/mL. At least four ions were available for each analyte for confirmation of identity by MS.

The urine specimens of numerous athletes were found to be positive for mephentermine both in-competition and out-of-competition in Taiwan. The donor of one specimen claimed she had only taken Mucaine (contains oxethazaine) for relieving symptomatic peptic ulcer and gastritis. Oxethazaine is not included in the prohibited list of the World Anti-Doping Agency; however, its metabolized compounds, mephentermine and phentermine, are included in that list. This study applied LC-MS-MS to analyze the excretions of three volunteers who ingested oxethazaine and presented positive results for mephentermine and/or phentermine. Thus, oxethazaine is the source of mephentermine and phentermine. Moreover, the results showed that 48 brands of gastric medicines containing oxethazaine were legally imported or locally manufactured in Taiwan, information which could be useful for limiting the misuse of oxethazaine by athletes. The data suggested that the sports associations should warn athletes about the risks of taking oxethazaine.

The sympathomimetic agent metamfepramone (2-dimethylamino-1-phenylpropan-1-one, dimethylpropion) is widely used for the treatment of the common cold or hypotonic conditions. Due to its stimulating properties and its rapid metabolism resulting in major degradation products such as methylpseudoephedrine and methcathinone, it has been considered relevant for doping controls by the World Anti-Doping Agency (WADA). The rapid degradation of the active drug complicates the detection of metamfepramone itself but the metabolites methylpseudoephedrine and methcathinone can be monitored, and the finding of the latter in particular allows the inference of a metamfepramone administration. In order to improve sports drug testing procedures, metamfepramone, methylpseudoephedrine and methcathinone were characterized using electrospray ionization-high resolution/high accuracy mass spectrometry, and a method employing liquid chromatography/tandem mass spectrometry was established that allowed the analysis of these three analytes by direct injection of 2 microL of urine specimens. The assay was validated with regard to specificity, lower limits of detection (2-10 ng mL(-1)), intraday and interday precision (3-17%) and ion suppression/enhancement effects. The developed procedure has been used to verify or falsify suspicious signals observed in routine screening procedures based on gas chromatography/mass spectrometry and yielded an adverse analytical finding concerning a metamfepramone administration in an authentic doping control sample. Although the active drug was not detected, the indicative metabolites methylpseudoephedrine and methcathinone were considered sufficient to infer the application of the prohibited drug.

Liquid-liquid-liquid microextraction (LLLME) in combination with HPLC and UV detection has been used as a sensitive method for the determination of ephedrine in urine samples. Extraction process was performed in a homemade total glass vial without using a Teflon ring, usually employed. Ephedrine was first extracted from 3.5 mL of urine sample (pH 12) into a microfilm of toluene/benzene (50:50). The analyte was subsequently back extracted into an acidic microdrop solution (pH 2) suspended in the organic phase. The extract was then injected into the HPLC system directly. An enrichment factor of 137 along with a good sample clean-up was obtained under the optimized conditions. The calibration curve showed linearity in the range of 0.01-50 mg/L with regression coefficient corresponding to 0.998. The LODs and LOQs, based on a S/N of 3 and 10, were 5 and 10 microg/L, respectively. The method was eventually applied for the determination of ephedrine in urine sample after oral administration of 5 mg single dose of drug.

Background: Immunoassay-based screening for amphetamines has a variable positive predictive value (PPV) for detecting amphetamine abuse. The lack of immunoassay specificity necessitates confirmatory testing by gas chromatography-mass spectrometry (GC/MS), but the technical complexity and expense of GC/MS limit its availability. Physicians may make decisions regarding patient disposition based on unverified results. In this study we assessed the utility of using dose-response properties to distinguish urine samples containing amphetamines from samples containing cross-immunoreactive species., Methods: Urine was supplemented with known concentrations of amphetamine, methamphetamine, methylenedioxymethamphetamine (MDMA), or pseudoephedrine. Using a series of dilutions, we determined the maximum change in rate over the fractional change in concentration for each compound in the Emit II amphetamine/methamphetamine immunoassay. Patient urine samples that screened positive for amphetamines were diluted 1:1, 1:10, and 1:20, and maximum slope estimates within the dynamic assay range were determined. An optimal slope cutoff that differentiated samples containing (meth)amphetamine from those containing cross-reacting species was determined by ROC analysis., Results: The slope of the dose response was largest for amphetamine and methamphetamine, followed by MDMA and pseudoephedrine. The optimum slope cutoff for identifying patient specimens containing (meth)amphetamine was 320 (sensitivity, 96%; specificity, 90%; PPV, 92%). High concentrations of less reactive compounds may mask low concentrations of amphetamines., Conclusions: Use of the slope of the dose-response relationship in patient urine specimens can enhance the PPV of presumptive positive immunoassay results but does not exclude the presence of low amphetamine concentrations in samples containing high concentrations of cross-reactive species.

Aims: To study the elimination of ephedrines with reference to the International Olympic Committee (IOC) doping control cut-off levels, following multiple dosing of over-the-counter decongestant preparations., Methods: A double-blind study was performed in which 16 healthy male volunteers were administered either pseudoephedrine or phenylpropanolamine in maximal recommended therapeutic doses over a 36-h period. Urine was collected every two hours between 08:00 and 24:00 h and at 04:00 h throughout the testing period of three days. Urine drug levels were quantified using high performance liquid chromatography. Side-effects were assessed, including heart rate and blood pressure, every four hours between 08:00 and 20:00 h., Results: Mean (95% CI) total phenylpropanolamine and pseudoephedrine eliminated unchanged was 75 (88, 61) and 81 (92, 71)%, respectively. Maximum urine concentrations of phenylpropanolamine and pseudoephedrine were 112.1 (164.2, 59.9) and 148.5 (215.0, 82.1) mg.l(-1), respectively. A peak in drug urine concentration occurred four hours following the final dose. There were no adverse cardiovascular effects and only mild CNS stimulation was evident., Conclusions: Following therapeutic, multiple dosing, drug levels remain above the IOC cut-off levels for a minimum of 6 h and 16 h following final doses of phenylpropanolamine and pseudoephedrine, respectively. Athletes require informed advice on this from their healthcare professionals.

The purpose of this study was to evaluate the ability of two amphetamine class screening reagents to exclude ephedrine (EPH), pseudoephedrine (PSEPH), and phenylpropanolamine (PPA) from falsely producing positive immunoassay screening results. The study also sought to characterize the prevalence and concentration distributions of EPH, PSEPH, and PPA in samples that produced positive amphetamine screening results. Approximately 27,400 randomly collected human urine samples from Navy and Marine Corps members were simultaneously screened for amphetamines using the DRI and Abuscreen online immunoassays at a cutoff concentration of 500 ng/mL. All samples that screened positive were confirmed for amphetamine (AMP), methamphetamine (MTH), 3,4-Methylenedioxyamphetamine (MDA), 3,4-Methylenedioxymethamphetamine (MDMA), EPH, PSEPH, and PPA by gas chromatography/mass spectrometry (GC/MS). The DRI AMP immunoassay identified 1,104 presumptive amphetamine positive samples, of which only 1.99% confirmed positive for the presence of AMP, MTH, MDA, or MDMA. In contrast, the online AMP reagent identified 317 presumptive amphetamine positives with a confirmation rate for AMP, MTH, MDA, or MDMA of 7.94%. The presence of EPH, PSEPH, or PPA was confirmed in 833 of the 1,104 samples that failed to confirm positive for AMP, MTH, MDA, or MDMA; all of the 833 samples contained PSEPH. When compared to the entire screened sample set, PSEPH was present in approximately 3%, EPH in 0.9%, and PPA in 0.8% of the samples. The results indicate that cross reactivities for EPH, PSEPH, and PPA are greater than reported by the manufacturer of these reagents. The distribution of concentrations indicates that very large concentrations of EPH, PSEPH, and PPA are common.

Objectives: It is generally accepted that the aetiology of craniomandibular dysfunction (CMD) is multifactorial. Different types of malocclusion, oral parafunctions especially bruxism, trauma of the mandible or temporomandibular joint (TMJ) and emotional stress are known aetiologic factors. Research has been conducted into the relationship between each of these aetiologic factors and the signs and symptoms of CMD. However, such an approach does not control for the simultaneous effect of other factors responsible for the development of the dysfunction. The purpose of this study was to investigate the effect of each aetiologic factor on the signs and symptoms of CMD in children, controlling for the effect of all other known factors by means of a multifactorial analysis., Methods: A sample of 314 children, aged 6-8 years, was examined clinically for signs of CMD and morphologic and functional malocclusion. Symptoms of CMD and oral parafunctions were recorded by the same investigator in an interview. Emotional stress was measured through urinary catecholamines including epinephrine, norepinephrine and dopamine, detected in a 24-h urine sample, using high performance liquid chromatography. A questionnaire was distributed to the parents to collect information regarding socioeconomic factors and the history of dentofacial injuries. A logistic multiple regression was carried out to estimate the partial effect of each aetiologic factor. A 95% probability level was used., Results: Posterior crossbite with lateral shift significantly affected the probability of child developing deviation of the mandible on opening. Similarly, posterior crossbite and epinephrine had a significant impact on TMJ tenderness, overjet had an effect on clicking, clenching and biting of objects had an effect on muscle tenderness, and lip/cheek biting influenced dysfunctional opening. Of the symptoms reported, pain on wide opening was affected significantly by lip/cheek biting., Conclusion: On the basis of these results, it can be suggested that parafunctional and some structural and psychological factors may increase the probability of the child developing the signs and symptoms of CMD.

Application of citric acid/acetic anhydride reagent (CAR), a colour reagent selective for tertiary amines in solution, improves detection of abused tertiary amino drugs on the TLC plate. The plate is pretreated by a brief immersion in phosphoric acid/acetone solution to suppress colouration. After suppressing, the plate is sprayed with CAR and heated at 100 degrees C, causing tertiary amines to turn red purple within 3 minutes. The sensitivity of this new CAR method is 2.5 to 15-times greater than that of conventional detection with Dragendorff reagent for some of the tertiary amines dimethylamphetamine, methylephedrine, levomepromazine, chlorpromazine, caffeine, theophylline, theobromine and nicotine. This present method provides rapid TLC detection of abused tertiary amino drugs such as phenethylamine, phenothiazine, xanthine derivative, nicotine and narcotics.

A rapid and highly sensitive flow-injection chemiluminometric method was developed for determination of 3 sympathomimetic drugs, namely, etilefrine hydrochloride, isoxsuprine hydrochloride, and prenalterol hydrochloride. The method is based on chemiluminescence induced by oxidation of drugs with acidified potassium permanganate in the presence of formic acid as a carrier. The calibration graphs were linear over the concentration ranges 0.2-9, 0.2-12.5, and 0.025-1.25 microg/mL for the 3 compounds, respectively. The method was applied successfully in determining the drugs in dosage forms and in biological fluids. A proposal for the reaction pathway is suggested.

A simple and highly sensitive voltammetric method was developed for the determination of isoxsuprine HCl (I) and fenoterol HBr (II) in dosage forms and biological fluids. The method is based on treatment of the two compounds with nitrous acid followed by measuring the cathodic current produced by the resulting nitroso derivatives. The voltammetric behavior was studied adopting Direct Current (DCt), Differential Pulse (DPP) and Alternating Current (ACt) polarography. Both compounds produced well-defined, diffusion-controlled cathodic waves over the whole pH range in Britton-Robinson buffers (BRb). At pH 11 and pH 9, the values of diffusion-current constants (Id), were 9.4 +/- 0.3 and 7.7 +/- 0.4 for I and II, respectively. The current-concentration plots for I were rectilinear over the range of 0.6-12 microg/ml and 0.1-12 microg/ml in the DCt and DPP modes, respectively. As for II, the range was 1-20 microg/ml and 0.1-20 microg/ml in the DCt and DPP modes, respectively. The minimum detectability (S/N = 2) were 0.02 microg/ml (approximately 6 x 10(-8) M) and 0.01 microg/ml (approximately 2.6 x 10(-8) M) for I and II, respectively, adopting the DPP mode. The proposed method was applied to the determination of both compounds in dosage forms and the results obtained were in good agreement with those obtained using reference methods. The proposed method was further applied to the determination of isoxsuprine in spiked human urine and plasma. The percentage recoveries adopting the DPP mode were 98.84 +/- 1.18 and 99.26 +/- 0.97, respectively.

A method was developed that permitted rapid identification in urine of the following sympathomimetic amines: amphetamine, benzphetamine, cathinone, desmethylsegiline, diethylpropion, ephedrine, fenfluramine, mazindol, methylenedioxyamphetamine, methylenedioxyethylamphetamine, methylenedioxymethamphetamine, mescaline, methamphetamine, methcathinone, methylaminorex, methylphenidate, pemoline, phendimetrazine, phenylepherine, phentermine, phenylpropanolamine, pseudoephedrine, and selegiline. In addition, two alpha-phenylethylamine-like monoamine oxidase inhibitors, phenelizine and tranylcypromine, were studied. Those sympathomimetic amines containing a primary or secondary amine, a hydrazine, and/or hydroxyl (except mazindol) functional groups were derivatized effectively using an on-column derivatization technique that used a reagent consisting of 10% fluoroanhydride in hexane, whereas the other sympathomimetic amines, including mazindol, were analyzed underivatized. Three different fluoroanhydrides, trifluoroacetic (TFAA), pentafluoropropionic (PFPA), and heptafluorobutyric (HFBA), and three different injection-port temperatures (160, 200, and 260 degrees C) were investigated. Both TFAA and PFPA gave sympathomimetic amine derivatives with essentially identical retention times, whereas HFBA gave longer retention times and better separation of individual compounds. The base fragmentation ion was noted to increase 50 amu (CF2) for each derivatized sympathomimetic amine as the length of the carbon-fluorine chain increased. Fragmentation ion abundance was maximized at an injection-port temperature of 260 degrees C, and this enhanced sensitivity coupled with the better chromatographic resolution of the individual sympathomimetic amines prompted the selection of HFBA as the derivatizing agent of choice. Assignments were made for the fragmentation ions produced by each derivatized drug. The developed method was adapted to analyze urine specimens that might be encountered in emergency toxicology testing. For identification of sympathomimetic amines requiring derivatization, 0.1 mL of the patient specimen had amphetamine-d5 and methamphetamine-d5 added as internal standard followed by adjustment of pH to 9.3 with borate buffer, extraction with 9:1 chloroform/isopropanol, centrifugation and separation of the organic phase, addition of 10% methanolic HCI and evaporation under nitrogen, reconstitution with HFBA reagent, and on-column derivatization during gas chromatographic-mass spectrometric (GC-MS) analysis. For those sympathomimetic amines not requiring derivatization, 1.0 mL of urine specimen had diazepam-d5 added as internal standard followed by the same extraction procedure and reconstitution accomplished with ethyl acetate. Because precolumn derivatization was eliminated and only 8 min was required for GC-MS analysis, complete analysis time was approximately 30 min, making the method suitable for clinical emergency toxicology purposes.

A rapid, sensitive, and solvent-free procedure for the simultaneous determination of amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA) in urine was developed using solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) in the selected ion monitoring mode. A headspace vial containing the urine sample, NaOH, NaCl, and amphetamine-d3 as the internal standard was heated at 100 degrees C for 20 min. A polydimethylsiloxane fiber was maintained in the vial headspace for 10 min in order to adsorb the amphetaminic compounds, which were subsequently derivatized by exposing the fiber to trifluoroacetic anhydride for 20 min in the headspace of another vial maintained at 60 degrees C for 20 min. The trifluoroacetyl derivatives were desorbed in the GC injection port for 5 min. Several parameters were considered during the method optimization process. These included a comparison of SPME with or without headspace, the required derivatization procedure, and the influence of temperature on the headspace extraction and derivatization methods. The optimized method was validated for the four compounds tested. Calibration curves showed linearity in the range 50-1000 ng/mL (r = 0.9946-0.9999). Recovery data were 71.89-103.24%. The quantitation limits were 10 ng/mL for amphetamine and methamphetamine and 20 ng/mL for MDA and MDMA. All of these data recommend the applicability of the method for use in the analytical routine of a forensic laboratory.

A rapid and sensitive fluorimetric method for the determination of prenalterol hydrochloride is presented, based on the oxidation of the analyte with potassium hexacyanoferrate(III) in a slightly alkaline medium (pH 9.23). The different experimental parameters were carefully studied and incorporated into the procedure. The oxidation product exhibits a blue fluorescence with its emission maximum at 427 nm, and excitation maximum at 314 nm. Fluorescence intensity is a linear function of prenalterol hydrochloride concentration over the range of 0.2-3.6 microg/ml(-1) in the solution finally measured. The method was successfully applied to the determination of prenalterol hydrochloride in pharmaceutical formulations and biological fluids. A proposal for the reaction pathway is suggested.

The effectiveness of capillary electrophoresis (CE) in the field of stereoselective determination of drugs in biological matrices is demonstrated by analyzing clenbuterol in human urine. Due to the very low therapeutical doses of 20-40 microg per day the total concentrations in urine are 1-10 ng/ml. The sample was extracted with hexane-tert.-butyl methyl ether (99.5:0.5). The reconstituted sample was injected electrokinetically (50 s, 10 kV). Using phosphate buffer, pH 3.3 and hydroxyethyl-beta-cyclodextrin as chiral selector the total analysis time was below 15 min. The limit of determination was estimated as 0.5 ng/ml per enantiomer. S-(-)-Bupranolol was used as internal standard. Both precision and accuracy of the method were within the limits for biological samples. The application to human urine from patients having received therapeutical doses showed a slightly predominant excretion of the (+)-enantiomer to the (-)-enantiomer.

A simple, rapid and highly sensitive high-performance liquid chromatographic method for the determination of methamphetamine and its metabolites in human urine was developed. Without tedious pretreatment procedures, aliquots of methamphetamine spiked or abusers' urine samples were simply acidified, dried under a stream of N2 gas, and then derivatized with 4-(4,5-diphenyl-1H-imidazol-2-yl) benzoyl chloride in acetonitrile in the presence of traces of triethylamine. Five derivatives were isocratically separated within 33 min by an ODS column, and the effluent was monitored at 440 nm (lambda(ex), 330 nm). Using 1-phenylethylamine as an internal standard calibration curves were confirmed to be linear up to at least 2x10(-5) M in urine with correlation coefficients of 0.998-1.000, while the detection limits at S/N=3 were 0.6-5.2 fmol per 5-microl injection. The relative standard deviations (n=5) of inter- and intra-day variations were less than 8.9%. The correlation between the concentrations of methamphetamine and amphetamine in urine determined by the proposed method and another currently accepted one was satisfactory. The method was successfully applied to urine samples collected from methamphetamine addicts.

The pharmacokinetics of norephedrine enantiomers were determined after the independent i.v. administration of (+/-)-norephedrine (20 mg/kg), (-)-norephedrine (10 mg/kg), and (+)-norephedrine (10 mg/kg) to rats. Significant differences were observed in the pharmacokinetic parameters of each enantiomer when the enantiomers were administered singly and as a racemate. For example, the values of total body clearance (Cltot) and urinary excretion clearance (Clr) of (-)-norephedrine administered as a racemate were higher than those of the norephedrine enantiomer administered singly. The areas under the curve of concentration versus time (AUC) of (-)-norephedrine administered as a racemate had a tendency to increase. While Cltot of (+)-norephedrine administered as a racemate showed a lower value and AUC showed a higher value. The value of Clr of (+)-norephedrine administered as a racemate showed a tendency to decrease. There was no difference in the in vitro serum protein binding of (-)- and (+)-norephedrine. The data from this study reveal that pharmacokinetic interactions exist between the norephedrine enantiomers and also reveal that the serum protein binding is not concerned with those interactions. The differences in the pharmacological effects after the individual administration of (-)-norephedrine or (+/-)-norephedrine may be coused by the differences in their concentrations in the plasma.

Background: Drug use has been associated with chest pain in adults presenting for emergency care. The association of drug use and chest pain in adolescents presenting to a pediatric emergency department has not been evaluated., Methods: Urine drug testing was conducted in a convenience sample of healthy adolescents with chest pain (cases) and compared to a control group of adolescents presenting with other complaints to a pediatric emergency department. Exclusion criteria were known diagnoses associated with chest pain (e.g., cardiac disease, sickle cell disease) and major trauma (due to its association with drug use). Urine drug testing consisted of 2 screening tests and gas chromatography-mass spectrometry confirmation of all positive or indeterminate results. All patients completed a questionnaire regarding recently prescribed, over-the-counter, and illicit drug use., Results: Twenty-eight cases and 26 controls completed the study over an 11-month study period. No cases or controls were positive for cocaine whereas marijuana was detected in 7 (25.0%) cases and in 7 (26.7%) controls. Five (17.8%) of the cases but none of the controls were positive for ephedrine (p = 0.05)., Conclusions: Ephedrine exposure appeared to be associated with chest pain in adolescents presenting for emergency care and marijuana was the most common drug of abuse, regardless of presenting complaint.

A young man (22 years old) died of a cardiorespiratory arrest a few hours following admission to the emergency department of a hospital. He was found lying seriously ill in the parking lot of a dance club. Screening of postmortem blood and urine with enzyme multiplied immunoassay (EMIT) detected only amphetamines, caffeine, and cotinine. Further screening of blood, urine, and stomach contents with thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) was negative for all three matrices. Specific conditions for amphetamines were used for the gas chromatographic (GC) screening (GC-mass spectrometric [MS] and GC-nitrogen-phosphorus detection). This resulted in the preliminary identification of amphetamine in both blood and urine. Confirmation of the presence of amphetamine in all available postmortem specimens was provided by mass and infrared spectral data (GC-MS and GC-Fourier transform infrared spectrometry) after derivatization. Quantitative results and differentiation between the enantiomers of amphetamine were obtained after chiral derivatization. The calculated concentrations disclosed amphetamine ingestion as the cause of this fatality.

A toxicological method of sensitive and specific confirmation of methamphetamine and other primary and secondary amines in biological samples after extractive perfluorobenzoylation is described. The method is based on the principle of gas chromatography with parallel specific nitrogen detection and electron capture detection. The other alternative is the gas chromatography combined with mass spectrometry in chemical ionization mode. The method described allows the detection of methamphetamine and amphetamine in urine in concentrations below 10 ng/ml.

Objectives: To evaluate lorry drivers' work stress by measurement of adrenaline and noradrenaline excreted in the urine, and to find out which factors in their working situation are related to the excretion rates of these catecholamines., Methods: The urinary excretion of adrenaline and noradrenaline of 32 lorry drivers, who also had loading and unloading activities to perform, was studied for one working day and one rest day. Each driver was asked to provide six urine samples on both days., Results: For all samples, except the first (overnight) sample, the excretion rates of both catecholamines on the working day were higher than those on the rest day. Hierarchical multiple regression analyses were carried out to find out which factors in the drivers' working situation were related to the excretion rate of the working day. The excretion rate of adrenaline on the rest day, age, and psychosomatic complaints were positively related to the excretion rate on the working day (all P < 0.05). Body mass index and physical workload during loading and unloading were positively related to noradrenaline excretion rate (both P < 0.01). Psychosocial job strain did not significantly contribute to the proportion of variance explained in the excretion rates of both catecholamines., Conclusions: The excretion rates of adrenaline and, especially, noradrenaline on the working day were higher than those found in earlier studies among professional drivers and insufficient recovery took place after the work was ended. The only association between excretion rate on the working day and work stressors was found for noradrenaline and physical workload. The drivers' sympathoadrenal medullary reactivity to everyday work demands shows the characteristics of sustained activation.

Forty-one stimulant drugs banned by the International Olympic Committee were studied after they were administered to human volunteers. The parent drugs and their metabolites in free or conjugated forms in human urine collected within 24 hours after administration of the drugs were extracted, separated and identified. The separation was performed on a capillary gas chromatograph with nitrogen-phosphorous detector, while the identification was achieved on a capillary gas chromatograph with mass-selective detector. The extract was injected into the gas chromatograph both directly and after derivatization with trifluoroacetic anhydride (TFAA) or N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) as well as TFAA and MSTFA combined. The conjugated metabolites were studied after acid hydrolysis of the extract and then selectively derivatized as above. The results are summarized in 4 tables in the text.

Hordenine cross-reacted with various enzyme linked immunosorbent assay (ELISA) or radioimmunoassay (RIA) kits used for the screening of urine samples. Morphine-ELISA kit was most sensitive, whereas etorphine- and buprenorphine-ELISA kits were least sensitive to hordenine cross-reactivity. Hordenine also interfered with the thin layer chromatography of oxymorphone, hydromorphone and apomorphine. The major source of hordenine in humans is beer brewed from barley, whereas the major source of hordenine in horses is canary grass or barley. Therefore, the presence of hordenine in the urine of humans consuming beer or in the urine of horses consuming canary grass may give false positive values when the immunoassay and TLC methods are used for the screening of the urine sample. In order to distinguish hordenine from the opiate drugs, simple and sensitive gas chromatographic/mass spectrometric (GC/MS) and high performance liquid chromatographic (HPLC) methods have been developed in this study.

Derivatives of seven commonly used sympathomimetic amines and two "designer amines" were isolated from urine, separated chromatographically from amphetamine and methamphetamine, and determined by mass spectrometry with selected ion monitoring. The drugs included ephedrine, propylhexedrine, pseudoephedrine, phenylpropanolamine, hydroxynorephedrine, phenylephrine, phentermine, methylenedioxyamphetamine (MDA), and methylenedioxy methamphetamine (MDMA). The drugs were liquid extracted from urine and derivatized by either heptafluorobutyric anhydride (HFBA) or 4-carbethoxyhexafluorobutyryl chloride (4-CB). Because the base peak ions for ephedrine, pseudoephedrine, propylhexedrine, MDMA, and phentermine are identical to methamphetamine (e.g. 254 amu for HFBA) and those for phenylephrine, hydroxynorephedrine, phenylpropanolamine, and MDA are identical to amphetamine (e.g. 240 amu for HFBA), a table of selected ions was developed for all 11 drugs that distinguished amphetamine and methamphetamine from the sympathomimetic amines with either HFBA or 4-CB. The distinguishing ions rely on the ring structure of the different drugs and fragmentation associated with that structure. The 4-CB reagent partially derivatized the hydroxy-containing sympathomimetic amines, while the HFBA completely derivatized all the sympathomimetic amines. Furthermore, false positive results for the 4-CB reagent were found only for methamphetamine (20-2250 ng/mL of methamphetamine) when high concentrations (greater than 5 micrograms) of ephedrine or pseudoephedrine were present in the specimen. These results are related to a combination of injection port temperature and cleanliness of the injection port sleeve.

A specific, sensitive and reliable gas chromatography-mass spectrometry (GC-MS) technique for detection of sympathomimetic amines following urinary extraction is proposed. Amphetamine, phentermine, ephedrine, mephenorex, methylphenidate, benzphetamine, clobenzorex and internal standard (fenfluramine) are extracted from urines at pH 7.0 using elution by chloroform-isopropanol on C18 cartridges. Derivatization followed by GC-MS analysis allows identification of these drugs founded on relative retention times and mass spectra. The quantitation limit for derivatizable drugs was found to be 200 ng/ml and 500 ng/ml for underivatizable drugs.

An analytical procedure for determination of phenolalkylamines, narcotic analgesics, and beta-blockers in urine by gas chromatography/mass spectrometry (GC/MS) is described. The detection of phenolalkylamines, narcotic analgesics, and beta-blockers is based on acid hydrolysis, liquid-liquid extraction, and selective derivatization. For screening of phenolalkylamines the m/e 179 and 267 ions were monitored by GC/MS. With narcotic analgesics, the extracted ion corresponded to the molecular ion (M+) of the drug and two additional characteristic ions. Beta-blockers were analyzed as the selectively derivatized forms of the parent molecule and its metabolites by GC/MS with selected ion monitoring. The ions monitored for screening of beta-blockers containing an isopropylamine group were m/e 284 and 129 ions. The ion at m/e 86 was monitored to characterize the tert-butylamine group of beta-blockers.

The pharmacokinetic behaviour of doxaminol (N-methyl-N-(2-hydroxy-3-phenoxy-propyl)-11-(2-amino-ethyl)-6, 11-dihydrodibenz[b,e]oxepine, neutral fumarate; BM 10.188) was examined in dogs using peroral and intravenous application of the 14C-labelled drug. The maximum plasma concentration was reached 1 h after application, indicating a relatively quick absorption of doxaminol. Decrease of total radioactivity after intravenous and peroral application is characterized by two phases, the elimination half-lives being 1.33 and 1.55 h, respectively, and 24.05 and 21.05 h, respectively. The biological availability of doxaminol was ca. 60%. The plasma levels of the unchanged drug showed that doxaminol was very rapidly eliminated and metabolized. Within the examined period of 96 h, the elimination of doxaminol and its metabolites via urine and faeces amounted to 76.5% after intravenous application, and 44.1% of the applied dose after peroral application. The major amount of radioactivity is eliminated via faeces (61.5% and 31.2% of dose, respectively) while the elimination through urine is found to be 15.0 and 12.9% of the dose, respectively.

Recoveries of a series of sympathomimetic central nervous stimulants in human urine are measured using either adsorption chromatography on self-filled columns (method A) or with a special resin method suitable for racehorse urine (method B). The Amberlite resins used are XAD-2, XAD-4, XAD-7 and XAD-8 and elution is performed using chloroform. The reported comparative drug extractabilities indicate that in most instances the recoveries follow the sequence XAD-4 greater than XAD-2 approximately XAD-8 greater than XAD-7 using method A. Based on the recovery and purity of the extracts obtained, XAD-8 is preferred for gas chromatographic analysis while XAD-4 is very suitable for thin-layer chromatographic screening work. Comparing the two methods, equally good or better results were obtained with method A for all of the resins studied except XAD-7. Finally, it was found that the effect of refrigerated storage of the resins on the drug extractabilities for central nervous stimulants could be neglected.

The equipment, methods, logistics, and results of doping-control analyses for the 1984 Los Angeles Olympic Games are discussed in this article. Within 15 days, 1510 different urine specimens underwent 9440 screening analyses by a combination of gas chromatography, gas chromatography-mass spectrometry, "high-performance" liquid chromatography, and radioimmunoassay. These tests covered more than 200 different drugs and metabolites, including psychomotor stimulants, sympathomimetic amines, central nervous system stimulants, narcotic analgesics, and anabolic steroids. The results are summarized by class of drug. Less than 2% of the samples were found to contain a banned drug.

A sensitive and rapid method for the gas chromatographic (with electron-capture detection) confirmation of derivable sympathomimetic amines is described. Extractive derivatization with pentafluorobenzoyl chloride is performed on 2-ml urine or plasma samples. Especially for primary amines, the method appears to be very sensitive. Mass spectral data allowed confirmation of the monobenzoylation of all congeners.

Single oral doses of 16, 32 and 64 mg oxilofrine as dragées as well as 16mg as drops were given to 12 healthy male volunteers in an open Latin-square design with one week interval between dosing days. Concentrations of unchanged drug were monitored in plasma over 24 hours, in urine concentrations of free and total oxilofrine were monitored over 36 hours. Drug concentrations were measured by a specific high pressure liquid chromatography method with electrochemical detection. Medians of maximum plasma concentrations (Cmax) of oxilofrine were 9.1 ng/ml, 11.4 ng/ml, 31.4 ng/ml and 122.9 ng/ml for 16 mg drops, 16, 32, and 64 mg dragées, respectively. The times to maximum plasma concentration (tmax) were between 0.7 and 1.7 h, respectively. The values for areas under the plasma concentration-time curve (AUC0-24) were 12.8, 17.7, 61.0 and 268.2 ng/ml.h for the four treatments. The results show that both Cmax- and AUC0-24-values increase faster with increasing dosing than a linear first-pass would suggest and give evidence for a saturable first-pass metabolism. There is also evidence for an enterohepatic circulation. About 50% of the administered dose is found in the urine, urinary recovery shows a dose-linear dependency. General tolerability was good; no side-effects were reported.

A high-performance liquid chromatographic (HPLC) method with electrochemical detection for the determination of oxilofrine [1-(4-hydroxyphenyl)-2-methylaminopropanol] in human plasma and urine (before and after cleavage of the metabolic conjugates) is described. Isolation from biological fluids is performed batchwise by weak acid cation exchange. Separation of plasma and urine components is achieved on a reversed-phase C18 column as an ion pair with heptanesulphonic acid. For amperometric detection the potential of the electrode was set at 0.95 V versus an Ag/AgCl reference electrode. The detection limit for oxilofrine in plasma is 1 ng/ml and in urine 12.5 ng/ml at a signal-to-noise ratio of 2.0 using 1.0 ml of plasma and 0.02 ml of urine. The method was compared with a gas chromatographic-mass spectrometric method and showed a good concordance for plasma (r = 0.996) and urine (r = 0.994). With the HPLC method it is also possible to determine related sympathomimetic drugs, e.g., etilefrine, norefenefrine or octopamine, after a slight modification of the mobile phase.

(+/-)-[(14)C]Amphetamine sulphate (20mg) was administered orally to each of two human male subjects, and 80-85% of the (14)C was excreted in the urine in 2 days. The metabolites in the first day's urine were examined quantitatively. Apart from the metabolites previously described (Dring et al., 1970), norephedrine (2.2 and 2.6% of dose in the two subjects respectively) and 4-hydroxynorephedrine (0.3 and 0.4% of dose in the two subjects respectively) were also found.