Horatiu Miresan, Cosmin Rosca, Nicoleta Matei, Florentina Roncea, Radu Cazacincu, Irina Iancu, Emil Stefanescu, Daniel Enache, Mihaela Bratu and Antoanela Popescu
A rapid high performance liquid chromatography method, using a monolithic column, was developed for quantitative determinations of benzodiazepines (diazepam, clonazepam, lorazepam, midazolam) in whole blood. A liquid-liquid extraction step with n-chlorobutane isolates the drugs from alkalinized blood. The separation was carried out in reversed phase conditions using a Chromolith Performance (RP-18 100x4.6 mm) column. For the mobile phase, a mixture of a phosphate buffer (pH= 2.5)/acetonitrile (65/35 v/v), in isocratic mode at 2 mL/min. An ultraviolet spectrophotometer was used as the detector at the wavelength of 220 nm. The total run time of the analytical method is less than 4-6 minutes. The calibration curves showed linearity and the correlation coefficient of each individual curve was greater than 0.995. The method was linear over a concentration range of 0.03-0.6 μg/mL for clonazepam, lorazepam and midazolam. For diazepam of linearity was over the range 0.04-5.0μg/mL. Quantification limits ranged from 0.03-0.04μg/mL and the accuracy were from 80% to 105% for the recovery test.
The results indicate that this analytical method is simple, specific, accuracy, sensitive, demonstrating from the validation data and a higher robustness. The proposed method is applied routinely in forensic toxicological analysis involving blood.
Jovanka Karadžinska-Bislimovska, Jordan Minov, Sašo Stoleski, Dragan Mijakoski, Snežana Risteska-Kuc and Snežana Milkovska
Environmental and Occupational Health Risks Among Agricultural Workers Living in a Rural Community Near Petroleum Refinery and Motorway in Skopje Region
To assess health risks in agricultural workers associated with environmental exposure to pollutants released from a petroleum refinery and from traffic, we performed a cross-sectional study that included 119 randomly selected subjects divided in two groups. Group 1 included 60 agricultural workers living in a rural community near the petroleum refinery and a motorway overpass, whereas Group 2 consisted of 59 agricultural workers performing similar activities and living in a rural community with no exposure to industrial and traffic pollutants. Risk assessment included a questionnaire, blood pressure measurement, spirometry, laboratory tests, and toxicological analysis. The groups showed a similar prevalence of health problems, with exception of muscle pain in the extremities, headache, and fatigue, which were significantly more common in Group 1. Diastolic blood pressure was higher in Group 1, but not significantly (p=0.057). The same is true for blood carbon monoxide. Significantly higher in Group 1 were blood haemoglobin (p=0.001) and blood lead (p<0.001). Serum cholinesterase activity was similar in both groups. Our findings indicate the need of regular medical exams, ambient monitoring and environmental impact assessment in agricultural population in order to detect individuals at risk and to institute adequate preventive measures.
, Am. J. Forensic Med. Pathol., 2010, 31, 156-161
 Ogden R.D., Hamilton W.K., Whitcher C., Assisted suicide by oxygen deprivation with helium at a Swiss right-to-die organisation, J. Med. Ethics, 2010, 36, 174-179
 Yoshitome K., Ishikawa T., Inagaki S., Yamamoto Y., Miyaishi S., Ishizu H., A case of suffocation by an advertising balloon filled with pure helium gas, Acta Med. Okayama, 2002, 56, 53-55
 Auwaerter V., Perdekamp M.G., Kempf J., Schmidt U., Weinmann W., Pollak S., Toxicologicalanalysis after asphyxial suicide with helium and a plastic bag
Marie Mardal, Frank Møller Aarestrup, Brian Schou Rasmussen, Christian Brinch Mollerup, Petur Weihe Dalsgaard and Kristian Linnet
, Baelum J, Schultz AC et al. Metagenomic analysis of toilet waste from long distance flights; a step towards global surveillance of infectious diseases and antimicrobial resistance. Scientific Reports 2015; 5.
 Andersen D, Rasmussen B, Linnet K. Validation of a fully automated robotic setup for preparation of whole blood samples for LC-MS toxicologyanalysis. J Anal Toxicol 2012; 36(4):280-287.
 Pedersen AJ, Dalsgaard PW, Rode AJ, Rasmussen BS, Muller IB, Johansen SS et al. Screening for illicit and medicinal drugs in whole blood using fully
Vugar Ali Turksoy, Lutfiye Tutkun, Servet Birgin Iritas, Meside Gunduzoz and Serdar Deniz
In exposure to toxic metals such as lead, determining lead and cytokine levels (IL-6, IL-10, and TNF-α) is essential for early detection of diseases. The aim of this study was to develop an model for early detection of inflammation and onset of atherosclerosis in the absence of clinical findings in young workers, which could help physicians take timely an action and start treatment. This study included 49 metal workers exposed to lead occupationally and 50 unexposed administrative workers (controls) who underwent immunological analysis for cytokines (IL-6, IL-10, and TNF-α) and atherosclerosis markers (h-FABP and VCAM-1), toxicological analysis for lead, and routine biochemical analysis (ALT, AST, creatinine) at the Ankara Occupational and Environmental Diseases Hospital in 2017. Lead levels correlated with IL-6, IL-10, and TNF-α (r=0.469, r=0.521 and r=0.279, respectively, p<0.01) but did not significantly affect h-FABP and VCAM-1 levels.
Fentanyl is a potent, short-acting narcotic analgesic widely used as surgical anaesthetic. This article presents a case in which fentanyl was self-injected by a 41-year old nurse, an employee at the hospital emergency department, who was found dead at home. She had no known history of drug and alcohol abuse. Two syringes, one empty and one filled with a clear liquid, were found near the body, while a needle was stuck into her hand.
Toxicological analysis showed fentanyl poisoning. Fentanyl overdose was declared the cause of death and the manner of death was classified as suicide.
To our knowledge, death due to the intravenous injection of fentanyl has not previously been reported in Croatia.
Advanced oxidation processes (AOPs) are considered to be one of the most effective methods for the decomposition of a wide range of hardly-biodegradable organic compounds, including pesticides. The implementation of such processes in the water streams treatment often leads to the formation of decomposition by-products of micropollutants occurring in water. These compounds, even in concentrations of a few ng/dm3, may negatively affect the water quality. Therefore, there is a need for detailed analyses that will allow to identify intermediates found in the AOP solutions and to assess their impact on the aquatic environment. The paper presents an attempt to identify by-products of three pesticides: triclosan, triallat and oxadiazon during ozonation, chlorination and UV irradiation of their water solutions. The identification of compounds was performed based on the results of the GC-MS analysis using the NIST v17 mass spectral library. It has been shown that during all of tested advanced oxidation processes, incomplete degradation of pesticides occurs. The number of micropollutant decomposition by-products increases with the increase of the applied ozone dose and UV exposure time. During the chlorination process Cl− atoms were added to the tested compound molecules. In the case of triclosan, it led to the generation of compounds containing four or five chlorine atoms in their structure. The toxicological analysis performed by the use of the Microtox® and Lemna sp. Growth Inhibition Test showed the toxic nature of post-process solutions. The decomposition by-products of triclosan and triallate, generated during the UV irradiation process, were highly toxic against the test organisms (toxic effect > 75 %). This makes it impossible to drain these solutions into the natural environment.
Marija Definis-Gojanović, Davorka Sutlović, Dolores Britvić and Bože Kokan
Drug Analysis in Necrophagous Flies and Human Tissues
Necrophagous insects may provide useful information about the time, place and cause of death. In addition, they can serve as reliable alternative specimens for toxicological analysis in cases where human tissue and fluids, normally taken during autopsies, are not available, due to decomposition of the corpse. This paper reports the results of drug analysis of the larvae of two fly families, Calliphoridae and Sarcophagidae, collected from the body of a middle-aged man who had committed suicide approximately three weeks before his corpse was found. Multiple samples of decomposed human tissue, of the blowfly, and of the larval flesh were analysed using gas chromatography/mass spectrometry (GC/MS), and amphetamine was detected in all samples. While the screening results were beyond doubt, the quantitative analysis was less clear, and further research is needed in this area.
Hafid Belhadj-Tahar, Pierre Payoux, Mathieu Tafani, Yvon Coulais, Serge Calet and Azzedine Bousseksou
Toxicological Methods for Tracing Drug Abuse: Chromatographic, Spectroscopic and Biological Characterisation of Ecstasy Derivatives
Analysis often reveals variability in the composition of ecstasy pills from pure 3,4-methylenedioxymethamphetamine (MDMA) to mixtures of MDMA derivatives, amphetamine, and other unidentified substances. For a comprehensive toxicological analysis one needs to know all steps to MDMA synthesis which may originate impurities. The aim of this study was to synthesise and determine the chemical-physical and in vitro biological properties of a series of MDMA derivatives.
3,4-methylendioxyphenyl-2-nitropropene (MDNP) was obtained by condensation of piperonal with an excess of nitroethane in the presence of ammonium acetate. MDNP was then reduced to methylenedioxyamphetamine (MDA) by LiAlH3. All compounds were analysed using HPLC and spectroscopic technique [Raman, nuclear magnetic resonance (NMR), or infrared (IR)] at all the steps of synthesis. In addition, we assessed the biological potentials of these compounds by measuring in vitro their (i) blood cell/whole blood partition coefficient, (ii) binding to plasmatic proteins (Fbp), and (iii) membrane adsorption. Chemical structure was determined with antibody fluorescence polarisation immunoassay (FPIA). This study showed the presence of solid impurities, particularly of a neurotoxic compound of Al3+ in the final products. FPIA identified the aminoethane group close to the substituted benzene ring, but did not detect the two major precursors of MDMA: MDNP and piperonal. Raman spectroscopy is an attractive alternative technique to characterise ecstasy pills and it can identify stereoisomeric forms such as cis-MDNP and trans-MDNP, which exhibit signals at 1650 cm-1 and 1300 cm-1, respectively.
Rumyana Simeonova, Vessela Vitcheva and Mitka Mitcheva
. Biochem Pharmacol 39 : 399-405.
Etter JF, Lukas RJ, Benowitz NL, West R and Dresler CM. (2008) Cytisine for smoking cessation: a research agenda. Drug Alcohol Depend 92 : 3-8.
Fau D, Eugene A, Berson P, Letteron B, Fromenty C and Pessayre D. (1994) Toxicity of the antiandrogen Flutamide in isolated rat hepatocytes. J Pharm Exp Ther , 269 : 1-9.
Guengerich FP. (1987) Microsomal enzymes involved in toxicology. Analysis and separation. In Principals and Methods of Toxicology. (Heis AW ed