Liquid Chromatography-Tandem Mass Spectrometry Analysis of Perfluorooctane Sulfonate and Perfluorooctanoic Acid in Fish Fillet Samples

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic (PFOA) acid are persistent contaminants which can be found in environmental and biological samples. A new and fast analytical method is described here for the analysis of these compounds in the edible part of fish samples. The method uses a simple liquid extraction by sonication, followed by a direct determination using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The linearity of the instrumental response was good, with average regression coefficients of 0.9971 and 0.9979 for PFOS and PFOA, respectively, and the coefficients of variation (CV) of the method ranged from 8% to 20%. Limits of detection (LOD) were 0.04 ng/g for both the analytes and recoveries were 90% for PFOS and 76% for PFOA. The method was applied to samples of homogenized fillets of wild and farmed fish from the Mediterranean Sea. Most of the samples showed little or no contamination by perfluorooctane sulfonate and perfluorooctanoic acid, and the highest concentrations detected among the fish species analyzed were, respectively, 5.96 ng/g and 1.89 ng/g. The developed analytical methodology can be used as a tool to monitor and to assess human exposure to perfluorinated compounds through sea food consumption

CHEMICAL COMPOSITION, ANTIMICROBIAL, ANTIOXIDANT AND CYTOTOXIC ACTIVITIES OF EUCALYPTUS CHAPMANIANA GROWN IN IRAQ

The chemical composition of the essential oils extracted from immature flowers, leaves and seeds of Eucalyptus chapmaniana grown in Iraq were analyzed for the first time by gas chromatography/mass spectrometry. Twenty-four different compounds were identified and the predominant compound is eucalyptol, which accounted for 59.9, 55.6 and 8.6% of total compounds, respectively. To asses the possible therapeutic uses of the extracts, their antioxidant properties were assessed via DPPH free radical scavenging. The extracts showed significant antioxidant and antimicrobial activities against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Proteus volgaris, Staphylococcus aureus and Candida albicans. The cytoxicity of flower extract against the Human Leukemia (HL-60) cells was evaluated and the extracts significantly reduced the viability of HL-60 cells in a dose- and time-dependent response relationship. The results indicated that essential oils from immature flowers are highly cytotoxic to HL-60 cells and that their antitumor potential was confirmed

Flexible high resolution-mass spectrometry approach for screening new psychoactive substances in urban wastewater

The number of new psychoactive substances (NPS) on the recreational drug market has increased rapidly in the last years, creating serious challenges for public health agencies and law enforcement authorities. Epidemiological surveys and forensic analyses to monitor the consumption of these substances face some limitations for investigating their use on a large scale in a shifting market. The aim of this work was to develop a comprehensive and flexible screening approach for assessing the presence of NPS in urban wastewater by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Almost 200 substances were selected as “priority NPS” among those most frequently and recently reported by the Early Warning Systems (EWS) of different agencies and were included in the screening. Wastewater samples were collected from several cities all over Europe in 2016 and 2017, extracted using different solid-phase cartridges and analysed by LC-HRMS. The screening workflow comprised two successive analytical steps and compounds were identified and confirmed following specific criteria from the current guidelines. Thirteen NPS were identified at different confidence levels by using analytical standards or information from libraries and literature, and about half of them were phenethylamines. As far as we know, this is the first time that four of them (i.e. 3,4-dimethoxy-α-pyrrolidinovalerophenone, para-methoxyamphetamine, 2-phenethylamine and α – methyltryptamine) have been found in urban wastewater. The proposed screening approach was successfully applied in the largest NPS European wastewater monitoring, providing an innovative and easily adapted procedure for investigating NPS. In the light of current challenges and specific future research issues, this approach may complement epidemiological information and help in establishing measures for public health protection

Identification of metabolites from type III F2-isoprostane diastereoisomers by mass spectrometry.

F 2 -isoprostanes (F 2 -iPs) are prostaglandin (PG)-like products of non-enzymatic free radical-catalyzed peroxida- tion of arachidonic acid that are now widely used as indices of lipid peroxidation in vivo. Knowledge of the metabolic fate of F 2 -iPs in vivo is still scant, despite its importance for defining their overall formation and biological effects in vivo. Type III F 2 -iPs, which are diastereoisomers of cyclo- oxygenase-derived PGF 2 a , may be metabolized through the pathways of PG metabolism. We therefore studied the in vitro metabolism of eight synthetic Type III F 2 -iP diastereo- isomers in comparison with PGF 2 a . We used gas chroma- tography-mass spectrometry and high performance liquid chromatography-electrospray-tandem mass spectrometry for structural identification of metabolites formed after in- cubation of the various compounds with isolated rat hepato- cytes. PGF 2 a was metabolized to several known products, resulting from a combination of b -oxidation, reduction of D 5 and/or D 13 double bonds, and 15-OH oxidation, plus other novel products deriving from conjugation with tau- rine of PGF 2 a and its metabolites. Of the eight F 2 -iP diaste- reoisomers, some were processed similarly to PGF 2 a , whereas others showed peculiar metabolic profiles according to spe- cific stereochemical configurations. These data represent the first evidence of biodegradation of selected Type III F 2 -iP isomers other than 8- epi- PGF 2 a , through known and novel pathways of PGF 2 a metabolism. The analytical characterization of these products may serve as a basis for identifying the most significant products formed in vivo. — Chiabrando, C., C. Rivalta, R. Bagnati, A. Valagussa, T. Durand, A. Guy, P. Villa, J-C. Rossi, and R. Fanelli. Identifi- cation of metabolites from Type III F 2 -isoprostane diastereo- isomers by mass spectrometry. J. Lipid Res. 2002. 43: 495-509

Chronic cholesterol administration to the brain supports complete and long-lasting cognitive and motor amelioration in Huntington's disease

: Evidence that Huntington's disease (HD) is characterized by impaired cholesterol biosynthesis in the brain has led to strategies to increase its level in the brain of the rapidly progressing R6/2 mouse model, with a positive therapeutic outcome. Here we tested the long-term efficacy of chronic administration of cholesterol to the brain of the slowly progressing zQ175DN knock-in HD mice in preventing ("early treatment") or reversing ("late treatment") HD symptoms. To do this we used the most advanced formulation of cholesterol loaded brain-permeable nanoparticles (NPs), termed hybrid-g7-NPs-chol, which were injected intraperitoneally. We show that one cycle of treatment with hybrid-g7-NPs-chol, administered in the presymptomatic ("early treatment") or symptomatic ("late treatment") stages is sufficient to normalize cognitive defects up to 5 months, as well as to improve other behavioral and neuropathological parameters. A multiple cycle treatment combining both early and late treatments ("2 cycle treatment") lasting 6 months generates therapeutic effects for more than 11 months, without severe adverse reactions. Sustained cholesterol delivery to the brain of zQ175DN mice also reduces mutant Huntingtin aggregates in both the striatum and cortex and completely normalizes synaptic communication in the striatal medium spiny neurons compared to saline-treated HD mice. Furthermore, through a meta-analysis of published and current data, we demonstrated the power of hybrid-g7-NPs-chol and other strategies able to increase brain cholesterol biosynthesis, to reverse cognitive decline and counteract the formation of mutant Huntingtin aggregates. These results demonstrate that cholesterol delivery via brain-permeable NPs is a therapeutic option to sustainably reverse HD-related behavioral decline and neuropathological signs over time, highlighting the therapeutic potential of cholesterol-based strategies in HD patients. DATA AVAILABILITY: This study does not include data deposited in public repositories. Data are available on request to the corresponding authors

Cocaine in surface waters: a new evidence-based tool to monitor community drug abuse

BACKGROUND: Cocaine use seems to be increasing in some urban areas worldwide, but it is not straightforward to determine the real extent of this phenomenon. Trends in drug abuse are currently estimated indirectly, mainly by large-scale social, medical, and crime statistics that may be biased or too generic. We thus tested a more direct approach based on 'field' evidence of cocaine use by the general population. METHODS: Cocaine and its main urinary metabolite (benzoylecgonine, BE) were measured by mass spectrometry in water samples collected from the River Po and urban waste water treatment plants of medium-size Italian cities. Drug concentration, water flow rate, and population at each site were used to estimate local cocaine consumption. RESULTS: We showed that cocaine and BE are present, and measurable, in surface waters of populated areas. The largest Italian river, the Po, with a five-million people catchment basin, steadily carried the equivalent of about 4 kg cocaine per day. This would imply an average daily use of at least 27 ± 5 doses (100 mg each) for every 1000 young adults, an estimate that greatly exceeds official national figures. Data from waste water treatment plants serving medium-size Italian cities were consistent with this figure. CONCLUSION: This paper shows for the first time that an illicit drug, cocaine, is present in the aquatic environment, namely untreated urban waste water and a major river. We used environmental cocaine levels for estimating collective consumption of the drug, an approach with the unique potential ability to monitor local drug abuse trends in real time, while preserving the anonymity of individuals. The method tested here – in principle extendable to other drugs of abuse – might be further refined to become a standardized, objective tool for monitoring drug abuse