1,721,102 research outputs found
Assunzione di alcool e abuso nei luoghi di lavoro: sviluppo di un protocollo integrato basato su questionario di screening e su nuovi metodi di campionamento e analisi biologica. applicazione in operatori sanitari secondo la legislazione italiana (Ricerca Finalizzata 2011)
No full textNonostante la sorveglianza dell’abuso di alcol sia obbligatoria nei luoghi di lavoro in ambito sanitario, non viene a tutt’oggi utilizzato uno strumento affidabile per valutare il consumo cronico di alcol e la legislazione italiana appare di difficile attuazione a causa della mancanza di indicazioni riguardo le procedure di monitoraggio. Recentemente due metaboliti dell’etanolo, l’etilglucoronide (EtG) e l’etilsolfato (EtS), sono stati considerati dei validi marcatori di assunzione sub-cronica di alcol e sono disponibili metodi per la loro determinazione nell’urina e nel sangue. Gli obiettivi di questo progetto, pertanto, sono: sviluppare e convalidare metodi analitici rapidi e poco costosi per l’analisi quantitativa di EtG e EtS in due matrici biologiche innovative (la goccia di sangue essiccata (DBS) e la goccia di urina essiccata (DUS); valutare l’affidabilità di questi marcatori per stimare l’assunzione di alcol e identificare possibili valori di soglia (cutoff); applicare le nuove procedure sviluppate su un campione di 400 lavoratori ospedalieri unitamente ai risultati del questionario AUDIT con lo scopo di verificare l’affidabilità di uno strumento integrato per controlli di routine in un grande numero di soggetti
Strategie innovative di campionamento e analisi per un efficace controllo del doping (Bando Commissione per la vigilanza ed il controllo sul doping e per la tutela della salute nelle attività sportive 2014)
No full textIl doping nello sport rappresenta attualmente una problematica concreta, in continua evoluzione, che mette gravemente a rischio la salute degli atleti, minacciandone l'integrità, e screditando la reputazione dello sport in generale. L’aggiornamento e l’introduzione di nuovi regolamenti sull’attività di controllo del doping (drug sport testing) e l’evoluzione di alcune tecniche analitiche sono riusciti solo in parte a scoraggiare l’uso di sostanze e metodi proibiti. Purtroppo esistono problematiche cruciali ancora non risolte all’interno delle procedure di prelievo e analisi, tra cui le limitazioni intrinseche dell’impiego di fluidi biologici considerati d’elezione nei controlli antidoping, primi fra tutti le urine (es. replicazione delle analisi, procedure di stoccaggio e trasporto, affidabilità dei risultati ottenuti). Il sistema sportivo riconosce come prova di avvenuto doping solo il ritrovamento nelle urine di una sostanza, o di un suo metabolita marker di assunzione, compresi nella lista delle sostanze vietate dalla World Anti-Doping Agency (WADA). Viene considerata inoltre come prova di doping una variazione fisiologica, o un livello anomalo di alcuni parametri, ascrivibili a una manipolazione fisiologica. Il sistema del controllo antidoping assume perciò una grande complessità e necessita che le singole fasi in cui si articola siano perfettamente coordinate fra loro, a garanzia della riservatezza, dell’efficienza e dell’affidabilità del sistema di testing. La disponibilità di sempre nuove strumentazioni e la progressiva automazione dei processi di laboratorio non sono tuttavia sufficienti a garantire dati analitici sicuri, perché necessitano di essere affiancati da specifiche fasi pre- e post- analitiche della massima affidabilità e che quindi non ne inficino la validità. La raccolta del campione, il trasferimento e lo stoccaggio sono infatti step cruciali per la buona riuscita dei controlli in ambito di drug sport testing. Inoltre, la scelta di utilizzare le urine come unico modello biologico, escludendone altri quali ad esempio il sangue, ha determinato alcuni dei limiti di efficacia delle analisi antidoping, facilitando tutti quegli atleti e quelle società intenzionati ad aggirare il sistema di controllo e sorveglianza. Le necessità generate dagli elevati standard e dai requisiti di qualità delle analisi antidoping moderne comportano quindi numerose sfide per i laboratori preposti a tale attività analitica.
Il progetto prevede l’utilizzo di matrici urinarie essiccate su appositi supporti in forma di “spot” anziché “in provetta”, come i Dried Urine Spot (DUS), come approccio alternativo e altamente innovativo che permette di eseguire monitoraggi della massima affidabilità, garantendo la stabilità dei campioni biologici. In particolare, la presenza di batteri nelle urine fresche dovuta a comuni infezioni delle vie urinarie o a contaminazioni durante il campionamento in provetta, può causare degradazione dei composti in esame, con conseguenti risultati falsi-negativi. Anche se le tecniche di congelamento e/o refrigerazione dei campioni urinari in provetta e l’utilizzo di preservanti tentano di ridurre in parte tali fenomeni di metabolismo, non sono efficaci tanto quanto una strategia di deposito ed essiccamento su speciali supporti, che consente di arrestare immediatamente qualsiasi attività enzimatica e batteriologia. In aggiunta a questi evidenti vantaggi offerti dal microcampionamento con DUS, va sottolineata la facilità e la rapidità di prelievo, senza specifiche esigenze di trattamento, conservazione e trasporto. L’approccio tramite DUS rappresenta quindi un’innovativa strategia in grado di processare un numero maggiore di campioni in tempi brevi (high throughput screening); tutti gli step preanalitici (dalla raccolta degli spot al momento dell’analisi) risultano semplificati; la stabilità dei composti è garantita; il controllo antidoping è potenziato al massimo in tutte le sue fasi, dal prelievo del campione alla consegna del risultato delle analisi.
La strategia di campionamento tramite spot può spaziare anche ad altre matrici biologiche, come il sangue, con la possibilità di ottenere preziose informazioni complementari sulla positività al doping degli atleti al momento della competizione. La sola analisi urinaria infatti difficilmente fornisce informazioni su tale problematica, che può invece essere affrontata con l’impiego di campioni aggiuntivi come i Dried Blood Spot (DBS), la saliva e il sudore.
Il progetto rappresenta un significativo avanzamento della conoscenza scientifica in materia di metodologia analitica, consentendo un notevole rafforzamento di entrambi gli aspetti fondamentali delle strategie di controllo antidoping, cioè la dissuasione dall’utilizzo di pratiche illecite nello sport e la tutela degli atleti
Metabolic Syndrome in Schizophrenia: Focus on the Role of Antipsychotic Medications and Indications for Therapeutic Drug Monitoring (TDM) Methods
No full textMetabolic syndrome is a complex pathology characterized by imbalances in lipid and glucose metabolism and weight gain, and consequently by an increase in the incidence of type II diabetes and cardiovascular disease. Metabolic syndrome is rapidly becoming one of the most important side effects of treatment with modern “atypical” antipsychotic agents, probably due to their specific mechanisms of action. Although the most recent members of this class (aripiprazole, asenapine, ziprasidone) seem to produce a reduce incidence of metabolic syndrome, the problem is far from being resolved. In this chapter, the three most important classical neuroleptics (chlorpromazine, haloperidol and loxapine) and the most important atypical antipsychotics will be individually analyzed in relation to their propensity to cause metabolic syndrome. The most reliable, current data will be presented, also in the perspective of possible interventions to mitigate metabolic imbalances, comparative studies, switching studies and augmentation strategies. An important strategy for metabolic syndrome prevention could also be the performing of an accurate therapeutic drug monitoring (TDM). Thus, an up-to-date overview will also be presented of recent and significant analytical methods for the determination of the drugs of interest and their main metabolites in human biological fluids
Mass spectrometry investigation for drugs of abuse control in unknown specimens
No full textThe detection and identification of abuse substances in unknown specimens poses unusual analytical problems in the field of forensic toxicology. A wide variety of compounds may be encountered, from natural and synthetic opioids, cocaine, amphetamine derivatives, ketamine, to the newest and less widely used classes of drugs such as synthetic cannabinoids, cathinones and smart drugs.
The Pharmaco-Toxicological Analysis Laboratory of the University of Bologna received a finding from a subject who claimed it could contain traces of illicit drugs; the finding (a steel spoon) appeared almost completely turned in on itself. The outer convex portion showed obvious signs of blackening of the metal, presumably due to a prolonged heating to direct flame. The inner concave portion showed the presence of a greyish white solid, probably the last remnants of a dried solution.
Thus, an extensive screening assessment was carried out in order to identify and quantify possible drugs of abuse or psychotropic substances: the powder residues obtained from the finding were dissolved in different organic solvents and screened via infusion in an advanced electrospray ionisation (ESI+) triple quadrupole mass spectrometer. The following chromatographic analysis was carried out by means of a LC-MS/MS methodology, developed for the detection of the major classes of abuse substaces. The screening suggested the presence of drugs of abuse, namely heroin and cocaine, in each sample collected from the specimen under examination.
The analytical approach described in this case report shows how mass spectrometry can be a very powerful tool for the investigation of drugs of abuse in specimens of unknown composition
Threat-or-treat: Chemical risk assessment of confectionery products in various age groups of European population
No full textTwo studies on chemical contaminants in food matrices were carried out. The first study addressed health concerns about the dietary exposure to bisphenol A (BPA) contamination due to consumption of soft drink by Polish population. BPA is an organic additive used in the production of epoxy resins and polycarbonate plastics and because of this it is used in the internal coating of cans and in plastic bottle production. Depending on several factors, BPA can migrate from these materials to the soft drink and so, it can be ingested by consumers causing hormonal and reproductive disorders. To estimate the Polish population exposure to BPA, several soft drinks belonging to different brands were purchased from a supermarket in the city of Warsaw and analysed. The result of the analysis highlight that mean BPA exposure in the Polish population exceeds the tolerable daily intake proposed by the EFSA scientific opinion, raising health concerns. On the other hand, the second study, focused on cadmium exposure due to chocolate consumption by Polish population, did not raise any health concern. Cadmium is a heavy metal that naturally occurs in its inorganic form in the environment and its presence in chocolate derives only from the cocoa beans and not from contamination during processing. Its accumulation in the human body can create several adverse effects, including renal dysfunction and failure. To estimate the Polish population exposure to cadmium, several chocolate bars were purchased from a supermarket in the city of Warsaw and analysed. The results of the analysis show that cadmium exposure in the Polish population does not exceed the tolerable weekly intake proposed by the EFSA scientific opinion
An effective DBS-LC-MS/MS strategy for the monitoring of alcohol biomarkers
No full textWorkplace monitoring of alcohol consumption habits is highly advisable especially in the healthcare framework, due to the serious consequences alcohol use and abuse could bring.
Within this project, two highly selective alcohol consumption biomarkers were selected, namely ethyl glucuronide (EtG) and ethyl sulfate (EtS), to perform a reliable quali-quantitative analysis as a tool to investigate alcohol consumption behaviours. The two analytes, as medium-term biomarkers, are suitable for workplace monitoring. To this aim, a minimally invasive biosampling strategy has been proposed and developed within this research, also allowing simplified sample storage and shipping of numerous samples: a dried blood spots (DBS) microsampling approach. It is based on haematic collection through a finger prick, avoiding an invasive procedure (i.e. phlebotomy). An accurate blood volume (10 μL) was collected from a fingertip and spotted on special cellulose DBS cards; after spot drying, the cards were stored at room temperature without the need for cryopreservation. The use of a miniaturised, dried biological matrix also ensures enhanced EtG and EtS stability in blood, when compared to the same analytes in classic fluid blood samples.
On the other hand, the reduced volumes and the low expected analyte levels required the development and validation of a highly sensitive and selective instrumental analytical method: the microsampling approach was coupled to an original LC-MS/MS method. In the first phase of the project, an in-depth optimisation of mass spectrometry parameters was carried out and led to the definition of electrospray ionization (ESI) polarity, acquisition mode, fragmentation parameters, selection of mass ion transitions leading to the highest selectivity and sensitivity. Chromatographic conditions were also optimised for the simultaneous determination of both analytes in a short time (3.5 minutes) and validated according to the main International Guidelines. In the second phase of the research, the developed method was applied to evaluate the analyte levels in DBS from volunteers, after alcohol controlled dosing and aiming at discriminating between abstinence, accidental alcohol intake, acute and chronic consumption.
At the same time, an attempt was made to define EtG and EtS baseline levels, as well as effective cut-off reference values. The original high-throughput analytical methodology proposed herein combines the advantages of microsampling for sample collection, transportation and storage with the high sensitivity and selectivity of LC-MS/MS. It has granted the successful analysis of samples from healthcare professionals, recruited in the last phase of the study, for the monitoring of workplace alcohol consumption. This study was funded by the Italian Ministry of Health within the Finalized Research Project 2011-2012 (RF-2011-02352096) and performed at the Pharmaco-Toxicological Analysis Laboratory (PTA Lab) of the Department of Pharmacy and Biotechnology (FaBiT), Alma Mater Studiorum - University of Bologna
Ion-Channel Antiepileptic Drugs: An Analytical Perspective on the Therapeutic Drug Monitoring (TDM) of Ezogabine, Lacosamide, and Zonisamide
Full text linkThe term seizures includes a wide array of different disorders with variable etiology, which cur-rently represent one of the most important classes of neurological illnesses. As a consequence, many different antiepileptic drugs (AEDs) are currently available, exploiting different activity mechanisms and providing different levels of performance in terms of selectivity, safety, and ef-ficacy. AEDs are currently among the psychoactive drugs most frequently involved in therapeutic drug monitoring (TDM) practices. Thus, the plasma levels of AEDs and their metabolites are monitored and correlated to administered doses, therapeutic efficacy, side effects, and toxic ef-fects. As for any analytical endeavour, the quality of plasma concentration data is only as good as the analytical method allows. In this review, the main techniques and methods are described, suitable for the TDM of three AEDs belonging to the class of ion channel agents: ezogabine (or re-tigabine), lacosamide, and zonisamide. In addition to this analytical overview, data are provid-ed, pertaining to two of the most important use cases for the TDM of antiepileptics: drug–drug interactions and neuroprotection activity studies. This review contains 146 references
Enhanced urinary stability and detection window of peptide hormones and growth factors by dried urine microsampling (WADA 2017)
No full textUrine samples collected worldwide for anti-doping testing are not always shipped in refrigerated conditions from collection sites to WADA accredited laboratories, in particular when several transport days are required. Commensal urethral microbiota, urinary pathogens and environmental bacteria may contaminate urine. Enzymatic activity by microbial contamination can cause severe modifications to the excreted compounds, and in particular to doping-relevant peptides such as hormones and growth factors. In recent years, the anti-doping scientific community has also raised strong concerns about the possible use of proteolytic enzymes (proteases) during collection to alter analysis results. Since delays are inevitable and sample refrigeration is not always possible and effective, sound methods for preserving and stabilizing urine samples are desirable.
This Project involves the use of Volumetric Absorptive Microsampling (VAMSTM) and Dried Urine Spot (DUS) strategies for the collection of dried microsamples processed by means of streamlined workflows to be developed ad hoc, for high-throughput LC-MS/MS analysis of several peptide hormones and growth factors. DUS and VAMSTM approaches represent promising alternatives to the procedures currently performed by anti-doping laboratories, allowing analyte stabilization by water loss and the consequent broadening of their detection window. This innovative sampling produces logistics savings due to the small transported volume (by air shipments and through customs) and to the possibility to keep the specimens at room temperature, with significant implications on overall analysis cost.
The ultimate goal of this project is to establish and validate feasible but reliable protocols for the collection of urine microvolumes, unlikely to be tampered, stably storable and shippable with no particular precautions. The use of innovative microsampling media offers perspectives towards the development of engineered, highly reliable devices, as well as the expansion of their use for the sampling of complementary and promising biological matrices, such as oral fluid
Recent trends in the analysis of amino acids in fruits and derived foodstuffs
No full textThe amino acid content of fruit and fruit-derived foods is intensely studied, because of their contribution to nutritional value, aroma, taste and health-promoting effects and their possible use as markers of origin and authenticity. In this review, based on 101 references, the most recent trends in the analysis of amino acids are presented: the most important techniques, the different sample treatment procedures (including derivatisation) and the most frequent applications are described and compared.
Pertinent publications were retrieved from Scopus and Web of Knowledge database searches lastly performed in February 2012 with the keywords "amino acid", "analysis", "liquid chromatography", "gas chromatography", "electrophoresis", "fruit", "vegetables"; the time limit was set from the year 2000 onwards.
Although amino acids have been analysed in foods for decades, new technical possibilities and advancements have allowed ever increasing accuracy and targeting of the methods, in order to overcome the challenges posed by the complex plant matrices and by their high intrinsic variability
Evaluation of Mediterranean diet health benefits: lycopene analysis in tomato-based products
No full textCardiovascular diseases (CVD) are the leading cause of death in Western countries and represent about one third of all deaths worldwide. Compared to Northern Europe or other Western countries, the Mediterranean area has lower rates of mortality from CVD and cancer, and this is attributed, at least partially, to the so-called Mediterranean diet, rich in bioactive phytochemicals. Identification of the active constituents of the Mediterranean diet is therefore essential for the formulation of appropriate dietary guidelines. Lycopene is a natural carotenoid found in tomatoes, an essential component of the Mediterranean diet. Although lycopene belongs to the carotenoid family, it has no pro-vitamin A functions, but many other biochemical functions as a scavenger antioxidant, hypolipidemic agent, inhibitor of pro-inflammatory and pro-thrombotic factors, and is therefore potentially beneficial in the prevention of CVD. Although many aspects of lycopene in vivo metabolism, functions, and clinical indications remain to be clarified, the integration of low doses of lycopene has already been proposed as a preventive measure to counter CVD and improve many aspects of their therapy.
The purpose of this study is the quantitative determination of lycopene in tomatoes by means of an HPLC system coupled to Diode Array Detection (DAD), to investigate its content in some varieties (such as globe, plum and cherry tomatoes) coming from different areas, in order to define nutrient profiles and to identify species with high contents of lycopene. Moreover, the analytical method is being applied to the evaluation of lycopene levels in tomato-based products (tomato paste, canned tomato, tomato sauce). Matrix purification was obtained by means of a liquid-liquid extraction, followed by a chromatographic separation with a C18 stationary phase and a methanolic mobile phase. The method is being validated and seems promising for the quantitative determination of lycopene in complex vegetal matrices such as tomatoes and tomato-based products
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