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To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.12.006 Byline: Jochen Beyer (a), Olaf H. Drummer (a), Hans H. Maurer (b) Keywords: Alkaloids; Natural substances; Detection; Blood; Urine; Toxicology Abstract: Many plants contain toxic alkaloids which may be dangerous to humans. Despite the large number of poisonous plants, cases of fatal plant poisonings are relatively rare. The frequencies of poisonings and the plants involved are often regionally specific. Plant poisonings can be aggregated into three categories: unintended ingestions, intended ingestions, and poisoning due to abuse of plant material. Unintended ingestions often occur in children or from a mix-up of plants and mushrooms in adults. Intended ingestions are common in homicides and suicides. Increasingly common is the abuse of plants for hallucinogenic reasons. Toxicological analysis of such alkaloids may help in diagnosis of poisoning or abuse cases. This review describes the toxic alkaloids aconitine, atropine, coniine, colchicine, cytisine, dimethyltryptamine, harmine, harmaline, ibogaine, kawain, mescaline, scopolamine, and taxine, which are often involved in fatal and non-fatal poisonings. The paper summarizes the symptoms of the intoxications and reviews the methods of detection of their toxic constituents in biological fluids. Author Affiliation: (a) Victorian Institute of Forensic Medicine and Monash University, Department of Forensic Medicine, 57-83 Kavanagh Street, Southbank, 3006, Australia (b) Department of Experimental & Clinical Toxicology, Saarland University, D-66421 Homburg, Saarland, Germany Article History: Received 6 August 2008; Revised 25 September 2008; Accepted 3 December 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.12.001 Byline: Frank T. Peters (a), Calin-A. Dragan (b), Andrea E. Schwaninger (a), Christoph Sauer (a), Josef Zapp (c), Matthias Bureik (b), Hans H. Maurer (a) Keywords: Drugs; Fission yeast; Cytochrome P450; Synthesis; Biotechnology; PCEEA Abstract: Standards of drug metabolites are required for drug metabolism studies as a basis for toxicological risk assessment with respect to drug interactions and pharmacogenetic polymorphisms. They are further needed as reference compounds in analytical toxicology. However, metabolite standards are often not commercially available, particularly in the case of new designer drugs. As an alternative to often cumbersome chemical synthesis, human cytochrome P450 (CYP) isoenzymes heterologously expressed in the fission yeast Schizosaccharomyces pombe can be used for the biotechnological synthesis of drug metabolites. In the present study this concept was applied to the synthesis of N-(1-phenylcyclohexyl)-2-hydroxyethanamine (PCHEA), the common O-dealkyl metabolite of the designer drugs N-(1-phenylcyclohexyl)-2-methoxyethanamine (PCMEA) and N-(1-phenylcyclohexyl)-2-ethoxyethanamine (PCEEA). After adding 250[mu]mol PCEEA*HCl (62mg), a 1l culture of CAD65 (S. pombe strain co-expressing human CYP reductase and CYP2B6) was fermented over 65h (pH 8, 30[degrees]C) and centrifuged. PCHEA and remaining parent drug were isolated from the supernatant by solid-phase extraction (SPE). The eluate was evaporated to dryness and reconstituted in HPLC solvent. Aliquots were separated by semi-preparative HPLC. From the respective fraction, PCHEA was extracted by liquid-liquid extraction and precipitated as hydrochloric salt. Approximately 80% of PCEEA was converted to PCHEA. The final yield of PCHEA*HCl was 9mg (35[mu]mol). Its identity was confirmed by GC-MS,.sup.1H NMR and.sup.13C NMR. The product purity, as determined by HPLC-UV, was 95%. Author Affiliation: (a) Department of Experimental and Clinical Toxicology, Saarland University, Building 46, D-66421 Homburg (Saar), Germany (b) PomBioTech GmbH, Building A1-1, D-66123 Saarbrucken, Germany (c) Department of Pharmacy, Pharmaceutical Biology, Saarland University, Building C2-2, D-66123 Saarbrucken, Germany Article History: Received 6 October 2008; Revised 22 November 2008; Accepted 1 December 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.12.002 Byline: Melanie Mueller (a)(b), Frank T. Peters (a), Marilyn A. Huestis (c), George A. Ricaurte (b), Hans H. Maurer (a) Keywords: MDMA; Metabolites; LC-ESI-MS; Human plasma; Validation; Acidic hydrolysis Abstract: 3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) is a psychoactive drug with abuse liability and neurotoxic potential. Specimen preparation of a recently presented LC-MS assay with electrospray ionization for quantifying MDMA and its main metabolites in squirrel monkey plasma was modified to include acidic hydrolysis to obtain total 3,4-dihydroxymethamphetamine and 4-hydroxy-3-methoxy-methamphetamine. Method re-validation for squirrel monkey plasma and full validation for human plasma showed selectivity for all analytes. Recoveries were [greater than or equal to]71.0%. Changed specimen preparation or matrix did not affect accuracy or precision. No instability was observed after repeated freezing or in processed samples. Plasma MDMA and metabolites quantification, derived pharmacokinetic and toxicokinetic data and neurotoxicity research will benefit from this validated method. Author Affiliation: (a) Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Saarland University, D-66421 Homburg (Saar), Germany (b) Department of Neurology, The Johns Hopkins Bayview Medical Center, Baltimore, MD, USA (c) Chemistry and Drug Metabolism, Intramural Research Program, National Institute on Drug Abuse, National Institute of Health, Baltimore, MD, USA Article History: Received 22 September 2008; Revised 27 November 2008; Accepted 1 December 2008

To link to full-text access for this article, visit this link: http://dx.doi.org/10.1016/j.forsciint.2008.09.001 Byline: Christoph Sauer (a), Frank T. Peters (a), Roland F. Staack (a), Giselher Fritschi (b), Hans H. Maurer (a) Keywords: N-(1-phenylcyclohexyl)-3-methoxypropanamine; PCMPA; Metabolism; GC-MS; Urinalysis Abstract: Studies on the metabolism and the toxicological detection of the phencyclidine-derived designer drug N-(1-phenylcyclohexyl)-3-methoxypropanamine (PCMPA) in rat urine are described using gas chromatographic-mass spectrometric (GC-MS) techniques. Based on the identified metabolites, the following metabolic pathways could be postulated: N-dealkylation, O-demethylation partially followed by oxidation of the resulting alcohol to the corresponding carboxylic acid, hydroxylation of the cyclohexyl ring at different positions, and aromatic hydroxylation. The formed metabolites were identical to those of the homologue N-(1-phenylcyclohexyl)-3-ethoxypropanamine (PCEPA) with exception of the mono hydroxyl metabolites of PCEPA. All PCMPA metabolites were partially excreted in conjugated form. An intake of a common drug users' dose of PCMPA could be detected in rat urine by the authors' systematic toxicological analysis (STA) procedure using full-scan GC-MS after acid hydrolysis, liquid-liquid extraction and microwave-assisted acetylation. The STA should be suitable for proof of an intake of PCMPA also in human urine assuming similar metabolism. Author Affiliation: (a) Department of Experimental and Clinical Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, Saarland University, D-66421 Homburg (Saar), Germany (b) Hessisches Landeskriminalamt, D-65187 Wiesbaden, Germany Article History: Received 28 May 2008; Revised 5 August 2008; Accepted 2 September 2008