Sotiris Athanaselis

Off-line HPLC method combined to LC-MS for the determination of sildenafil and its active metabolite in post-mortem human blood according to confirmation criteria.

A simple HPLC method has been validated for the determination of sildenafil and its active metabolite (N-desmethylsildenafil) in human blood, using an octadecyl silica (ODS) hypersil column. The chromatographic run time is less than 25 min using a mobile phase of 35:65 (v/v) acetonitrile-0.015 M disodium hydrogen phosphate (Na(2)HPO(4)), triethylamine 0.1%, pH 7.4 at 1 mL/min flow rate and UV-vis detection at 230 nm. The method is linear in the concentration range of 10-500 ng/mL (r>0.999, n=5) for each analyte, with relative standard deviation (R.S.D.) less than 5.05%. Interday and intraday errors were found to be < or =11.94%. The limits of detection and quantitation for both analytes were 5.0 ng/mL (s/n>3) and 10.0ng/mL (s/n>10), respectively. The method was applied in two post-mortem human blood samples, concerning two fatal cases from sildenafil citrate use, reported for the first time in Greece, and the results were further confirmed with LC-MS. The method is proposed as supplementary to LC-MS when inadequate mass fragmentation does not provide information appropriate to meet confirmation criteria.

AH-7921: the list of new psychoactive opioids is expanded

AH-7921 is a structurally unique synthetic opioid analgesic that has recently entered the drug arena in Europe, the USA, and Japan. Although it was synthesized and patented in the mid-1970s, it was first identified in a seized sample purchased via the Internet in July 2012 and formally brought to the attention of the European Union early warning system in August 2012 by the United Kingdom. Several in vitro experiments and animal model studies established the morphine-like analgesic action of AH-7921 as a μ-opioid receptor agonist that has been found to be several times more potent than codeine and at least as potent as morphine. This novel psychoactive substance has already led to eight non-fatal intoxications and 16 deaths in Sweden, the United Kingdom, Norway, and the USA. Thus, AH-7921 is a current public health risk, and better international collaboration, effective legislation and continuous community alertness are needed to tackle this current growing problem. The aim of this review is to summarize the current knowledge about this drug concerning its chemistry, pharmacology, and toxicology, as well as its international legal status. The limited existing analytical methodologies for the determination of AH-7921 in biological samples are also presented. Published or reported AH-7921-related cases, fatalities, or intoxications, and self reports from drug users are reviewed.

The use of alcohol and other psychoactive substances by victims of traffic accidents in Greece

Alcohol is one of the main causes of traffic accidents worldwide. Its use decreases significantly the driving ability of an individual as well as the reflexes of a pedestrian increasing in this way the possibilities of their involvement in motor-vehicle accidents. The above possibilities are increased when a psychoactive substance has been taken in combination with alcohol due to their synergistic effect. The Laboratory of Forensic Medicine and Toxicology of the University of Athens is authorized to perform the toxicological investigation of traffic accidents that happen in the major part of Greece. The findings of the toxicological analyses that were performed in our laboratory during the years 1995 to 1997 are classified according to the alcohol concentration and the kind of psychoactive substance detected as well as to the gender and the age of the victims, drivers or pedestrians. The results of the above classification are evaluated and the conclusions regarding alcohol, drugs and traffic accidents in Greece are discussed thoroughly.

Forensic investigation of submersion deaths

Context:  The diagnosis of the cause and the establishment of the manner of death in submersion cases are routine challenge for forensic pathologists as it presents considerable diagnostic difficulties.

Butyrylcholinesterase: biomarker for exposure to organophosphorus insecticides.

Butyrylcholinesterase (BuChE) is routinely measured to assess exposure to or effects of organophosphorus insecticides (OP). As a biomarker, it can be used to clarify the relation between exposure to OP and health impairment. The interpretation of BuChE inhibition data, particularly of small changes in enzymatic activity, sometimes presents significant complexities. These complexities are presented in this short communication and the factors that influence the degree of BuChE inhibition are discussed. Despite the complexities of their interpretation, BuChE measurements remain a mainstay for the fast initial screening of exposure to OP; thus, they are a useful tool in the protection of humans, domestic animals and wildlife from overexposure to these toxic agents.

2C-I-NBOMe, an "N-bomb" that kills with "Smiles". Toxicological and legislative aspects.

Abstract Substituted phenethylamines are a class of designer drugs that have recently emerged in the drug abuse market. Such substances remain legal to use, possess, and supply until these compounds become classified as scheduled. 2C-I-NBOMe or 25I-NBOMe is the N-benzyl-derivative of the iodo-substituted dimethoxy-phenethylamine (2C-I) that appeared recently in the drug market under the street name “N-Bomb”. Due to its high potency, intoxications and fatal cases related to 2C-I-NBOMe use are increased worldwide. The use and trafficking of this substituted phenethylamine is banned only in some countries. A comprehensive review was performed using PubMed and Medline databases, together with additional non-peer reviewed information sources, including books and publications of state authorities in different countries, regarding chemistry, availability, pharmacology, and toxicology of 2C-I-NBOMe. Intoxications or lethal cases, published or reported, as well as the current legislation on this newly introduced drug are also reviewed.

Toxicological investigation of drug‐facilitated sexual assaults

Drug‐facilitated sexual assaults have drawn a lot of attention during the last years. The role of the non‐forensic doctors in the early recognition and management of such cases is crucial. Diagnosis assisted by samplings must be ascertained without delay, in victims initially brought to the emergency room for a sleepy state, a suspected abuse of drug or a pseudo‐alcoholic clinical state. Further, where amnesia is present, it may be prudent to assume that sexual activity has occurred and offer pregnancy and sexually transmitted disease prophylaxis. Forensic doctors should entertain the possibility of poisoning in any presentation involving amnesia. Forensic toxicological laboratories should develop sensitive and specific methods to assure a low threshold of detection of the drugs. The awareness of a delinquent or criminal use of alcohol and/or drugs could permit a better medical recognition, so that proper and timely forensic examination is achieved, and adequate evidence is collected for the establishment of an effective investigation and prosecution.

The cannabinoid content of marihuana samples seized in Greece and its forensic application

Abstract The three major cannabinoids, Δ 9 -tetrahydrocannabinol (Δ 9 -THC), cannabidiol (CBD) and cannabinol (CBN) were identified and determined quantitatively using a GCD (GC–EI) instrument, in samples of illicit herbal cannabis, seized by Customs and Police authorities in two areas of Greece (Ipiros and Lakonia) during 1996. These samples were sent by the above authorities to the Department of Forensic Medicine and Toxicology, University of Athens, for forensic chemical analysis. The cannabinoid content of these samples led to the classification of cannabis into two chemical phenotypes and to the differentiation of resinous and textile plants by using three different classification indexes. The cannabinoid content of cannabis plants is of forensic value in determining the geographical origin of cannabis samples, since it can be used for their classification, allocating this way the area of cultivation of the relative plants. The forensic aspects of cannabis classification are discussed.

Validated GC/MS method for the simultaneous determination of clozapine and norclozapine in human plasma. Application in psychiatric patients under clozapine treatment

A sensitive and specific GC/MS method for the determination of clozapine (CLZ) and its major metabolite norclozapine (NCLZ), in plasma has been developed, optimized and validated. Specimen preparation includes solid-phase extraction of both analytes using Bond-Elut Certify cartridge and further derivatization with TFAA. Clozapine-d8 was used as internal standard for the determination of CLZ and NCLZ. Limits of detection were 0.45 ng/mL for CLZ and 1.59 ng/mL for NCLZ, while limits of quantification were 1.37 ng/mL for CLZ and 4.8 ng/mL for NCLZ, as calculated by the calibration curves. The calibration curves were linear up to 600 ng/mL for CLZ and NCLZ. Absolute recovery ranged from 82.22% to 95.35% for both analytes. Intra- and interday accuracy was less than 7.13% and --12.52%, respectively, while intra- and interday precision was between 9.47% and 12.07%, respectively, for CLZ and NCLZ. The method covers all therapeutic range and proved suitable for the determination of CLZ and NCLZ not only in psychiatric patients but also in forensic cases with clozapine implication.

Validated method for the simultaneous determination of methadone and its main metabolites (EDDP and EMDP) in plasma of umbilical cord blood by gas chromatography-mass spectrometry

A sensitive and specific GC/MS method for the determination of methadone (MDN) and its two main metabolites, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and 2-ethyl-5-methyl-3,3-diphenylpyrroline (EMDP), in plasma samples obtained from venous and arterial umbilical cord blood and maternal blood has been developed, optimized and validated. Specimen preparation includes protein precipitation with acetonitrile and simultaneous solid-phase extraction of the three analytes. Methadone-d9 was used as internal standard for the determination of MDN and EMDP, while EDDP-d3 for EDDP. Limits of detection were 0.6 microg/L for MDN and 0.3 microg/L for EDDP and EMDP, while limits of quantification were 2.0 microg/L for MDN and 1.0 microg/L for EDDP and EMDP. The calibration curves were linear up to 2000 microg/L for MDN and up to 1000 microg/L for EDDP and EMDP. Absolute recovery ranged from 94.8 to 99.7% for all three analytes. Intra- and interday accuracy was less than 5.3 and 5.5%, respectively, while intra- and interday precision was less than 3.5 and 5.0%, correspondingly, for all analytes. The method proved suitable for the determination of MDN and its two main metabolites in plasma samples obtained from umbilical cord and maternal blood of a woman participating in a MDN maintenance program, during the prenatal and postpartum period.