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- Novel synthetic opioids – toxicological aspects and analysisPublication . Tabarra, Inês; Soares, Sofia; Rosado, Tiago; Gonçalves, Joana; Luís, Ângelo; Malaca, Sara; Barroso, Mário; Keller, Thomas; Restolho, José; Gallardo, EugeniaOver the past few years, there has been an emerging number of new psychoactive drugs. These drugs are frequently mentioned as “legal highs”, “herbal highs”, “bath salts” and “research chemicals”. They are mostly sold and advertised on online forums and on the dark web. The emerging new psychoactive substances are designed to mimic the effects of psy- choactive groups, which are often abused drugs. Novel synthetic opioids are a new trend in this context and represent an alarming threat to public health. Given the wide number of fatalities related to these compounds reported within the last few years, it is an important task to accurately identify these compounds in biologic matrices in order to administer an effective treatment and reverse the respiratory depression caused by opioid related substan- ces. Clinicians dealing with fentanyl intoxication cases should consider that it could, in fact, be a fentanyl analogue. For this reason, it is a helpful recommendation to include synthetic opioids in the routine toxicological screening procedures, including analysis in alternative matrices, if available, to investigate poly-drug use and possible tolerance to opioids. To address this public health problem, better international collaboration, effective legislation, effective investigation, control of suspicious “research chemicals” online forums and continu- ous community alertness are required. This article aims to review diverse reported fatalities associated with new synthetic opioids describing them in terms of pharmacology, metabol- ism, posology, available forms, as well as their toxic effects, highlighting the sample proce- dures and analytical techniques available for their detection and quantification in biological matrices
- Editorial: Current Analytical Trends in Drug Testing in Clinical and Forensic ToxicologyPublication . Gallardo, Eugenia; Barroso, Mário; Concheiro-Guisan, Marta; de-Castro-Ríos, AnaEditorial on the Research Topic: Current Analytical Trends in Drug Testing in Clinical and Forensic Toxicology The articles included in this collection cover novel analytical approaches, including chromatographic and spectrometric methods, and sample preparation techniques for the investigation and analysis of several classes of compounds. These compounds include novel psychoactive substances (NPS) as well as other drugs and substances within the scope of clinical and forensic toxicology, and other fields, such as doping control. Current trends in bioanalysis require the constant development of novel analytical tools, which includes efficient sample collection procedures and adequate sample preparation protocols in order to maximize compound detection, even at trace levels. Taking into account that the number of substances possibly present in a sample are increasing, efficient multi-analyte methods are usually necessary.
- Evaluation of the Cytotoxicity of Ayahuasca BeveragesPublication . Simão, Ana Y.; Gonçalves, Joana; Gradillas, Ana; García, Antonia; Restolho, José; Fernández, Nicolás; Rodilla, Jesus M.; Barroso, Mário; Duarte, Ana Paula; Cristóvão, Ana C.; Gallardo, EugeniaAyahuasca is a beverage consumed at shamanic ceremonies and currently has gained popularity on recreational scenarios. It contains beta-carboline alkaloids and N,N-dimethyltryptamine, which possesses hallucinogenic effects. Only a few studies have elicited the psychoactive effects and the dose of such compounds on neurological dopaminergic cells or animals. In this work, we aimed to study the cytotoxic effects of these compounds present in ayahuasca beverages and on five different teas (Banisteriopsis caapi, Psychotria viridis, Peganum harmala, Mimosa tenuiflora and Dc Ab (commercial name)) preparations on dopaminergic immortalized cell lines. Moreover, a characterization of the derivative alkaloids was also performed. All the extracts were characterized by chromatographic systems and the effect of those compounds in cell viability and total protein levels were analyzed in N27 dopaminergic neurons cell line. This is the first article where cytotoxicity of ayahuasca tea is studied on neurological dopaminergic cells. Overall, results showed that both cell viability and protein contents decreased when cells were exposed to the individual compounds, as well as to the teas and to the two mixtures based on the traditional ayahuasca beverages. View Full-Tex
- Stability of Cocaine, Opiates, and Metabolites in Dried Saliva SpotsPublication . Almeida, Ema; Soares, Sofia; Gonçalves, Joana; Rosado, Tiago; Fernández, Nicolás; Rodilla, Jesus M.; Passarinha, Luís A.; Barroso, Mário; Gallardo, EugeniaDrug abuse still represents a global problem, and it is associated with an increased risk of diseases, injuries, and deaths. Cocaine (COC) and opiates are the most abused drugs and account for a significant number of fatalities. Therefore, it is important to develop methods capable of effectively identifying and quantifying these substances. The present study aims to evaluate the long- term stability of COC, ecgonine methylester (EME), benzoylecgonine (BEG), cocaethylene (COET), norcocaine (NCOC), morphine (MOR), codeine (COD) and 6-monoacetylmorphine (6-MAM) in oral fluid samples. The analytes of interest were isolated from the matrix (50 μL) using the dried saliva spots (DSS) sampling approach and were subsequently analyzed by gas chromatography coupled with tandem mass spectrometry (GC–MS/MS). The parameters that could influence the stability of the target compounds were studied, and these were storage temperature, light, use of preservatives (and respective concentrations), and time. The effects of each parameter were evaluated using the design of experiments (DOE) approach. The stability of the target analytes was improved when the DSS were stored at room temperature, in the presence of light and using 1% sodium fluoride. The best conditions were then adopted for the DSS storage and long-term stability was assessed. COD was only stable for 1 day, EME was stable for 3 days, COC, COET, NCOC and 6-MAM were stable for 7 days, MOR for 14 days and BEG remained stable throughout the study (136 days). This is the first study that evaluates the stability of these compounds in oral fluid samples after application in DSS cards, and optimizes the conditions in order to improve their stability
- An Update on the Implications of New Psychoactive Substances in Public HealthPublication . Simão, Ana Y.; Antunes, Mónica; Cabral, Emanuel; Oliveira, Patrik; Rosendo, Luana M.; Brinca, Ana Teresa; Alves, Estefânia; Marques, Hernâni; Rosado, Tiago; Passarinha, Luís A.; Andraus, Maristela; Barroso, Mário; Gallardo, EugeniaThe emergence of new psychoactive substances has earned a great deal of attention, and several reports of acute poisoning and deaths have been issued involving, for instance, synthetic opiates. In recent years, there have been profound alterations in the legislation concerning consumption, marketing, and synthesis of these compounds; rapid alert systems have also been subject to changes, and new substances and new markets, mainly through the internet, have appeared. Their effects and how they originate in consumers are still mostly unknown, primarily in what concerns chronic toxicity. This review intends to provide a detailed description of these substances from the point of view of consumption, toxicokinetics, and health consequences, including case reports on intoxications in order to help researchers and public health agents working daily in this area.
- Advances on Bioanalysis: Recent Approaches in the Determination of Biomarkers, Drugs of Abuse and Medicines (editorial)Publication . Barroso, Mário; Gallardo, Eugenia; Passarinha, Luís A.
- Analysis of opiates in urine using microextraction by packed sorbent and gas Chromatography- Tandem mass spectrometryPublication . Simão, Ana Y.; Monteiro, Catarina; Marques, Hernâni; Rosado, Tiago; Margalho, Cláudia; Barroso, Mário; Andraus, Maristela; Gallardo, EugeniaOpiates recreational consumption has always been a concern in society, public health, and in clinical toxicology analysis. The aim of this study was to develop and fully validate an analytical method, which was simple and rapid for the determination of tramadol, codeine, morphine, 6- acetylcodeine, 6-monoacetylmorphine and fentanyl using gas chromatography coupled to tandem mass spectrometry. The procedure includes the use of microextraction by packed sorbent for sample clean-up. A mixed mode sorbent was used, allowing the minimal use of solvents. The method was validated in urine samples, with the ability to detect and quantify all analytes with satisfactory linearity (in the range of 1 – 1000 ng/mL for all analytes, except for fentanyl (10–1000 ng/mL)). Extraction efficiency varied from 17 to 107%, which did not impair sensitivity, taking into account the low LLOQs obtained (1 ng/ mL for all analytes; and 10 ng/mL for fentanyl). The developed procedure proved to be fast, selective, and accurate for use in routine analysis, with a low volume of sample (250 µL).
- The Determination of Cannabinoids in Urine Samples Using Microextraction by Packed Sorbent and Gas Chromatography-Mass SpectrometryPublication . Rosendo, Luana M.; Rosado, Tiago; Oliveira, Patrik; Simão, Ana Y.; Margalho, Cláudia; Costa, Suzel; Passarinha, Luís A.; Barroso, Mário; Gallardo, EugeniaCannabis is the most consumed illicit drug worldwide, and its legal status is a source of concern. This study proposes a rapid procedure for the simultaneous quantification of Δ9-tetrahydrocannabinol (THC), 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC), 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH), cannabidiol (CBD), and cannabinol (CBN) in urine samples. Microextraction by packed sorbent (MEPS) was used to pre-concentrate the analytes, which were detected by gas chromatography–mass spectrometry. The procedure was previously optimized, and the final conditions were: conditioning with 50 µL methanol and 50 µL of water, sample load with two draw–eject cycles, and washing with 310 µL of 0.1% formic acid in water with 5% isopropanol; the elution was made with 35 µL of 0.1% ammonium hydroxide in methanol. This fast extraction procedure allowed quantification in the ranges of 1–400 ng/mL for THC and CBD, 5–400 ng/mL for CBN and 11-OH-THC, and 10–400 ng/mL for THC-COOH with coefficients of determination higher than 0.99. The limits of quantification and detection were between 1 and 10 ng/mL using 0.25 mL of sample. The extraction efficiencies varied between 26 and 85%. This analytical method is the first allowing the for determination of cannabinoids in urine samples using MEPS, a fast, simple, and low-cost alternative to conventional techniques
- New Method for the Monitoring of Antidepressants in Oral Fluid Using Dried Spot SamplingPublication . Soares, Sofia; Rosado, Tiago; Barroso, Mário; Gallardo, EugeniaThe increase in the consumption of antidepressants is a public health problem worldwide, as these are a class of compounds widely used in the treatment of several illnesses, such as depression and anxiety. This work aimed to develop and optimize a method for the quantification of a number of antidepressants and their metabolites (fluoxetine, venlafaxine, O-desmethylvenlafaxine, citalopram, sertraline, and paroxetine) in 100 μL of oral fluid using the dried saliva spots (DSS) sampling approach and gas chromatography coupled with tandem mass spectrometry (GC–MS/MS). The method was validated, presenting linearity within the studied range, with detection and quantification limits ranging between 10 and 100 ng/mL, and coefficients of determination (R2) of at least 0.99 for all analytes. Recoveries were between approximately 13 and 46%. The analysis of precision and accuracy presented acceptable coefficients of variation and relative errors, considering the criteria usually accepted in the validation of bioanalytical procedures. The method herein described is the first to be reported using DSS for the extraction of antidepressants, proving to be a sensitive, simple, and fast alternative to conventional techniques, and capable of being routinely applied in clinical and forensic toxicology scenarios
- A review of current bioanalytical approaches in sample pretreatment techniques for the determination of antidepressants in biological specimensPublication . Soares, Sofia; Barroso, Mário; Gallardo, EugeniaAntidepressants are a class of compounds widely used in clinical settings for the treatment of several diseases. In the last years there has been a considerable increase in their consumption, representing an important public health issue in several countries. Because they are substances with narrow therapeutic windows, and since they are capable of interacting with other classes of compounds, monitoring of these compounds is of relevance, minimizing the risk of medical interactions as well as side and toxic effects. In addition, understanding the extent of their use, their detection through routine toxicology tests and development of new methods for detection and monitoring is of extreme importance concerning public health, patient well-being, and implications in clinical and forensic situations. The main objective of this work is to perform a critical review on the biological samples used in the detection and quantification of antidepressants with special focus on the techniques for sample preparation.