Browsing by Author "Pires, Bruno"
Now showing 1 - 10 of 10
Results Per Page
Sort Options
- Characterisation of Cannabis-Based Products Marketed for Medical and Non-Medical Use Purchased in PortugalPublication . Pires, Bruno; Oliveira, Patrik; Simão, Ana Y.; Reis, João; Ramos, Sofia; Duarte, Ana Paula; Margalho, Cláudia; Rosado, Tiago; Barroso, Mário; Gallardo, EugeniaCannabis-based products have gained attention in recent years for their perceived therapeutic benefits (with cannabinoids such as THC and CBD) and widespread availability. However, these products often lack accurate labelling regarding their cannabinoid content. Our study, conducted with products available in Portugal, revealed significant discrepancies between label claims and actual cannabinoid compositions. A fully validated method was developed for the characterisation of different products acquired from pharmacies and street shops (beverages, herbal samples, oils, and cosmetic products) using high-performance liquid chromatography coupled with a diode array detector. Linearity ranged from 0.4 to 100 µg/mL (0.04–10 µg/mg) (THC, 8-THC, CBD, CBG, CBDA, CBGA), 0.1–100 µg/mL (0.01–10 µg/mg) (CBN), 0.4–250 µg/mL (0.04–25 µg/mg) (THCA-A), and 0.8–100 µg/mL (0.08–10 µg/mg) (CBCA). Among sampled beverages, none contained detectable cannabinoids, despite suggestive packaging. Similarly, oils often differed from the declared cannabinoid compositions, with some containing significantly higher CBD concentrations than labelled. These inconsistencies raise serious concerns regarding consumer safety and informed decision-making. Moreover, our findings underscore the need for stringent regulation and standardised testing protocols to ensure the accuracy and safety of cannabis-based products.
- Determination of amphetamine-related drugs in hair using MEPS as sample clean-upPublication . Pires, Bruno; Simão, Ana Y.; Rosado, Tiago; Gallardo, Eugenia; Barroso, MárioIntroduction and aims: Different extraction and clean-up protocols have been used for amphetamine-related compounds determination in hair. Microextraction by packed sorbent (MEPS) is a miniaturized version of solidphase extraction (SPE) which has been applied to determine a number of drugs in hair (e.g., opiates, cocaine, ketamine, etc.) [1-4]. However, concerning amphetamine-related drugs, MEPS has only been used in hair for the determination of amphetamine (AMP) and methamphetamine (MAMP). The aim of this work was the development and validation of a method to determine AMP, MAMP, MDA, MDMA, MDE and MBDB in hair using MEPS for sample clean-up. Materials and methods: Hair (50 mg) was incubated with NaOH 1M at 45 °C overnight [5], after which HCl 10M was added for neutralization and the solution was centrifuged. The MEPS procedure for extract clean-up was optimized using the Design of Experiments (DoE) approach, and the final steps were: conditioning (1 x 250 μL of methanol and 1 x 250 μL of deionized water); loading (18 x 100 μL); and elution (7 x 100 μL of 2% NH4OH in acetonitrile). To the eluted extract, 30 μL of MBTFA was added (to avoid loss of analytes), and extracts were evaporated to dryness, following microwave-assisted derivatization with 50 μL of MBTFA. A gas chromatography coupled to mass spectrometry (GC-MS) system was used for determination. Results and discussion: The MEPS procedure resulted in recoveries of 8-14% for AMP, 14-20% for MAMP, 10-15% for MDA, 18-28% for MDMA, 25-43% for MDE and 34-52% for MBDB, in the linear range of 0.2-5.0 ng/mg. Precision and accuracy of the developed method were in accordance with the statements of international guidelines for method validation. Conclusions: This is the first analytical method using MEPS coupled to GC-MS to determine the selected amphetamines in hair samples, proving to be a great alternative to the classic procedures, being rapid, eco-friendly and less expensive.
- Determination of amphetamine-type psychostimulants in hair samples using MEPS as sample clean-up and gas-chromatography coupled with mass spectrometryPublication . Pires, Bruno; Simão, Ana Y.; Rosado, Tiago; Gallardo, Eugenia; Barroso, MárioNumerous protocols for the analysis of amphetamine-type psychostimulants (ATS) in hair have been employed over the years. Microextraction by packed sorbent (MEPS), a downsized version of solid-phase extraction (SPE), has been effectively applied in hair drug analysis, including substances like opiates, cocaine, and ketamine. However, concerning ATS, MEPS has primarily been used to the determination of amphetamine (AMP) and methamphetamine (MAMP) in hair1-4. The main objective of this study was to develop and validate a method using MEPS as a sample clean-up, for the determination of AMP, MAMP, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 1-(1,3-benzodioxol-5-yl)propan-2-yl(ethyl)amine (MDE), and N-methyl-1-(1,3-benzodioxol-5-yl)-2-aminobutane (MBDB) in hair. The extraction procedure involved incubating 50 mg of hair with 1M NaOH at 45 ºC overnight, followed by neutralization with 10M HCl and centrifugation5. A Design of Experiments (DoE) approach was employed to optimize MEPS clean-up process, encompassing conditioning, loading, and elution steps. The eluted extract underwent derivatization and was analysed using gas chromatography coupled to mass spectrometry (GC-MS). The developed MEPS method yielded recoveries ranging from 8 to 52% for the different analytes in hair samples and linearity was obtained between 0.2 (cut-off proposed by SoHT) and 5.0 ng/mg. The precision and accuracy were in accordance to international method validation standards. This study introduces the first analytical method integrating MEPS with GC-MS for the detection of these specific amphetamines in hair samples. Notably, this method offers a viable alternative to conventional procedures, characterized by its speed, eco-friendliness, and cost-effectiveness.
- Evaluation of hair hydrolysis and MEPS columns for the detection of amphetamines derivatives in hairPublication . Pires, Bruno; Simão, Ana Y.; Rosado, Tiago; Gallardo, Eugenia; Barroso, MárioIntroduction: Different extraction and cleanup procedures have been used to identify amphetamines in hair samples. The microextraction by packed sorbent (MEPS), a miniaturized approach of the classic solid-phase extraction (SPE), has only been used so far to determine the presence of amphetamine (AMP) and methamphetamine (MAMP) in hair. The aim of this work was to evaluate different hair sample extraction conditions, namely hydrolysis, to maximize the detection of AMP, MAMP, MDA, MDMA, MDE, and MBDB in combination with MEPS and GC-MS. Methodology: Pulverized authentic hair (50mg) was incubated with 500μL of NaOH 1M at 80ºC for 1H, 60ºC overnight and 45ºC overnight1. After incubation, the extracts were neutralized with 50μL of 10M HCl and subsequently centrifuged. The supernatant was tested with several conditions to access the most suitable when paired with MEPS clean-up. The studied conditions were as follows: the addition of 500μL of H2O; supernatant filtration followed by addition of H2O; and precipitation with cold ACN followed by centrifugation. After clean-up, 30μL of MBTFA were added for evaporation under nitrogen gas. After being fully dried, 50μL of MBTFA was added and derivatization was done with a microwave oven. Finally, the extract was injected into a GC-MS and analysed. Results: Results obtained from these tests indicated that the best hydrolysis for this purpose was obtained with 500μL NaOH 1M at 45ºC overnight, with the addition of H2O before MEPS clean-up. The clean-up procedure was previously optimized by design of experiments (not being the scope of this study). Furthermore, M1 and C18 MEPS sorbents were tested to check which one would provide cleaner extracts and greater analyte signals. A concentration of 2.00 ng/mg of AMP, MAMP, MDA, MDMA, MDE, and MBDB was used for this test. Discussion: Considering the results, both columns presented similar recoveries for each amphetamine, when using equal hydrolysis and MEPS conditions.
- Microextraction action by packed sorbent in forensic drug analysisPublication . Rosado, Tiago; Pelixo, Rodrigo; Pires, Bruno; Catarro, João Miguel; Rosendo, Luana M.; Brinca, Ana T.; Antunes, Mónica; Soares, Sofia; Simão, Ana Y.; Barroso, Mário; Gallardo, EugeniaMicroextraction by packed sorbent (MEPS) efficiently combines extraction, pre-concentration, and cleanup in a single device comprising two parts: the MEPS syringe and the packed sorbent bed1. MEPS has been used for several bioanalytical applications, including extraction of endogenous metabolites, biomarkers, and drugs from biological samples. It is particularly useful in metabolomics and pharmacokinetic studies2,3. Regarding MEPS applicability to forensic toxicology, urine is the most used specimen, followed by oral fluid (despite of its relatively high viscosity). Protein precipitation followed by centrifugation and with, or without dilution of the supernatant is the most commonly reported approach. The most detected compounds in forensic settings using MEPS are drugs of abuse [opiates and opioids (26%), cocaine (13%) cathinones (11%), dissociative hallucinogens (11%), cannabinoids and amphetamines (9% each) and other drugs (10%)] and medicinal drugs [antidepressants (9%), benzodiazepines/Z drugs (4%)]. MEPS was also applied to a beverage for forensic purposes e.g. to evaluate its composition in drug-facilitated crimes. An important feature in MEPS is the miniaturization of the sorbent. A careful selection of the sorbent will allow working with complex matrices, separating the target analytes from interferences and improve recoveries. The most widely selected sorbent was the silica based C18 that is a popular reversed-phase material (41%). Starting in the 2000s, new modifications of sorbents appeared. Overall, what the future holds for MEPS applications in forensic toxicology is promising, and ongoing research and technological advancements are likely to enhance the capabilities of MEPS approaches, making this technique an increasingly valuable tool for toxicological investigations.
- Otimização de um método para a determinação de anfetaminas e derivados em cabelo por microextração em seringa empacotadaPublication . Pires, Bruno; Simão, Ana Y.; Rosado, Tiago; Gallardo, Eugenia; Barroso, MárioTêm sido utilizados diferentes protocolos de extração para a determinação de compostos relacionados com anfetaminas em cabelo. A microextração em seringa empacotada (MEPS) é uma versão miniaturizada da extração em fase sólida que já tem sido aplicada para determinar diversas drogas em cabelo (por exemplo, opiáceos, cocaína, ketamina, etc.). No entanto, no que diz respeito às anfetaminas, a MEPS só foi ainda utilizada em cabelo para a determinação de anfetamina (AMP) e metanfetamina (MAMP). O objetivo deste trabalho foi o desenvolvimento e otimização de um método para determinar não só a AMP e a MAMP, como também MDA, MDMA, MDE e MBDB em amostras cabelo, utilizando a MEPS como técnica de extração. O cabelo pulverizado (50 mg) foi incubado com NaOH 1M a 45 ºC durante a noite, após o que se adicionou HCl 10M para neutralizar. De seguida, a solução foi centrifugada, recolhendo cuidadosamente o sobrenadante. Após se realizar a extração recorrendo ao MEPS, adicionou-se 30 μL de MBTFA ao eluato obtido, e as amostras foram evaporadas à secura. Posteriormente foram derivatizadas com recurso a microondas e utilizando 50 μL de MBTFA. O produto derivatizado foi então analisado por GC-MS em modo SIM. O procedimento de MEPS foi optimizado usando os softwares MiniTab e SPSS, mediante dois testes de Desenho Experimental (DOE) e uma análise não paramétrica, respetivamente. Inicialmente, foi conduzido um DOE fatorial, com o estudo dos seguintes parâmetros: número de aspirações de amostra (entre 5 e 15 aspirações de 100 μL); número de aspirações para a lavagem [entre 1 e 3 aspirações com 50 μL de H2O ultrafiltrada e 50 μL de H2O:MeOH (95:5)]; e número de aspirações para a eluição (entre 2 e 6 eluições com 100 μL de 2% NH4OH em ACN). Foi necessário implementar um Desenho Experimental de Superfície de Resposta (SR), estendendo os limites dos parâmetros mencionados para uma análise mais detalhada. Os intervalos ampliados foram os seguintes: 0 a 26,1 aspirações de 100 μL de amostra; 0 a 7,7 aspirações de lavagem; e 0 a 10,3 aspirações de eluição. Após esta optimização, o método final de MEPS foi o seguinte: 18 aspirações de 100 μL de amostra; 0 aspirações para a lavagem; 7 aspirações com 100 μL de 2% NH4OH em ACN para a eluição. Este método permite a análise sensível de compostos derivados das anfetaminas em amostras de cabelo, podendo ser utilizado em situações onde estejam envolvidos.
- A review of publications in hair testing: 2020-2023Publication . Simão, Ana Y.; Antunes, Mónica; Pires, Bruno; Catarro, Gonçalo; Soares, Sofia; Matos, Francisco; Rosado, Tiago; Gallardo, Eugenia; Barroso, MárioIntroduction and Aims: Since its first use until now, the relevance of hair in the toxicological expertise has evolved, mainly due to its advantages over traditional samples. Despite the fact that it is one of the most used alternative specimens, hair still presents important challenges. The objective of this study was to perform a bibliographic review of scientific publications on toxicological applications of hair testing between the years 2020 and 2023. Materials & Methods: The Scopus database was used for literature search, for papers published from 2020 to 2023 (10th April), and only articles written in English were considered. The search strings for terms included in the title, abstract or keywords were “hair testing” and “hair samples”. The papers were independently reviewed by four of the authors to determine their relevance in the context of this study. Results & Discussion: Taking into consideration these criteria and search strings, 218 articles were found, from which 18 were excluded since animal hair specimens had been used. The papers were classified according to type (e.g. review, research paper), analytes, extraction and clean-up approaches. Each paper could be placed under more than one category, except review papers (32 articles). Concerning analytes, the highest number of papers involved drugs of abuse (175, from which 45 involving new psychoactive substances), drugs (44) and alcohol biomarkers (17). Other biomarkers included hormones, endocannabinoids, etc. (44 papers, 25 of which involved cortisol). Most studies involve protein precipitation and direct injection (91), and only 12 studies address the issues of cosmetic treatments or drug adulterants. Conclusions: Hair testing is still mainly directed to the determination of drugs of abuse, and new psychoactive substances are still seldom included. Additionally, other applications are becoming more relevant, for instance the use of hair to determine biomarkers in some diseases.
- Sensors in the Detection of Abused Substances in Forensic Contexts: A Comprehensive ReviewPublication . Rosendo, Luana M.; Antunes, Mónica; Simão, Ana Y.; Brinca, Ana Teresa; Catarro, Gonçalo; Pelixo, Rodrigo; Martinho, João; Pires, Bruno; Soares, Sofia; Cascalheira, José Francisco; Passarinha, Luís; Rosado, Tiago; Barroso, Mário; Gallardo, Eugeniaorensic toxicology plays a pivotal role in elucidating the presence of drugs of abuse in both biological and solid samples, thereby aiding criminal investigations and public health initiatives. This review article explores the significance of sensor technologies in this field, focusing on diverse applications and their impact on the determination of drug abuse markers. This manuscript intends to review the transformative role of portable sensor technologies in detecting drugs of abuse in various samples. They offer precise, efficient, and real-time detection capabilities in both biological samples and solid substances. These sensors have become indispensable tools, with particular applications in various scenarios, including traffic stops, crime scenes, and workplace drug testing. The integration of portable sensor technologies in forensic toxicology is a remarkable advancement in the field. It has not only improved the speed and accuracy of drug abuse detection but has also extended the reach of forensic toxicology, making it more accessible and versatile. These advancements continue to shape forensic toxicology, ensuring swift, precise, and reliable results in criminal investigations and public health endeavours.
- The Therapeutic Potential of Amphetamine-like PsychostimulantsPublication . Pires, Bruno; Rosendo, Luana M.; Brinca, Ana Teresa; Simão, Ana Y.; Barroso, Mário; Rosado, Tiago; Gallardo, EugeniaThis review delves into the therapeutic applications of amphetamine-type stimulants such as lisdexamphetamine dimesylate, mixed amphetamine salts, 3,4-methylenedioxymethamphetamine (MDMA), dextroamphetamine, and phentermine. These compounds have been investigated for their potential in treating a range of psychiatric disorders, including attention deficit hyperactivity disorder (ADHD), drug dependence, post-traumatic stress disorder (PTSD), and obesity. Lisdexamphetamine dimesylate has shown promise in effectively treating ADHD symptoms in both children and adults. Additionally, it has been explored as a potential treatment for drug dependency and withdrawal, demonstrating encouraging results. Mixed amphetamine salts have also exhibited efficacy in reducing ADHD symptoms in adults. Future research should explore their potential use in treating bipolar disorder and cocaine dependence, considering the associated risks and benefits. MDMA-assisted psychotherapy has emerged as an innovative approach to treating PTSD, leading to sustained reductions in symptoms and even promoting post-traumatic growth. Furthermore, it has shown promise in managing anxiety related to life-threatening illnesses. Dextroamphetamine and phentermine have demonstrated efficacy in treating cocaine and opioid dependence, ADHD, and obesity. However, careful consideration and monitoring by medical professionals are essential due to the potential risks and benefits associated with them. In conclusion, amphetamine-type stimulants present a promising avenue for therapeutic interventions in various psychiatric conditions. Nevertheless, further research is necessary to comprehensively understand their mechanisms of action, dosage requirements, and long-term effects in different patient populations.
- Volumetric Absorptive Microsampling in ToxicologyPublication . Pires, Bruno; Catarro, Gonçalo; Soares, Sofia; Gonçalves, Joana; Rosado, Tiago; Barroso, Mário; Araujo, André R. T. S.; Gallardo, EugeniaVolumetric absorptive microsampling (VAMS) is an emerging technique in clinical and forensic toxicology. It is recognized as a promising alternative to traditional sampling methods, offering an accurate and minimally invasive means of collecting small volumes of biological samples, such as blood, urine, and saliva. Unlike conventional methods, VAMS provides advantages in terms of sample stability, storage, and transportation, as it enables samples to be collected outside laboratory environments without requiring refrigeration. This review explores several VAMS methodologies, with a particular focus on its application for the quantification of drugs and other substances in clinical and forensic toxicology. It compares VAMS to other microsampling techniques, such as dried blood spots (DBSs), highlighting VAMS's superiority in addressing issues related to sample volume consistency and environmental impact. Despite its advantages, VAMS also presents certain limitations, including higher costs and difficulties in detecting underfilled samples. Overall, VAMS stands out as a microsampling technique with the potential to enhance patient compliance and operational efficiency, positioning itself as a viable tool for toxicological analysis in both clinical and forensic contexts.