Browsing by Author "Carmo, Helena"
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- Desenvolvimento e aplicação de um método de baixo custo para avaliação de práticas de manipulação de citotóxicosPublication . Ali, Nuro; Carmo, Helena; Robalo, Raquel; Fernandes, Cristina; Rocha, Luísa; Moreira, FernandoDe acordo com a International Society of Oncology Pharmacy Practitioners (ISOPP), os hospitais devem levar a cabo programas de formação e avaliação periódica dos profissionais que manipulam citotóxicos, sendo considerado o teste de fluorescência de cloridrato de quinina como adequado para este propósito. Esta investigação pretende (i) adaptar o teste de quinina recorrendo a uma solução mais amplamente disponível e económica de água tónica e (ii) aplicar o teste qualitativo para identificar práticas e superfícies em que ocorra disseminação e deposição de citotóxicos.
- Development and validation of an analytical method to assess the importance of using gauze dressings in luer-lock connection systems during the preparation of cytotoxic drugsPublication . Silva, Clara; Carmo, Helena; Oliveira, Vânia; Carvalho, Daniel; Moreira, Fernando; Moreira, FernandoHealthcare professionals face occupational hazardous when handling cytotoxic drugs used for cancer treatment. The International Society of Oncology Pharmacy Practice (ISOPP) standards are the guidelines recommended by the European Parliament for safety procedures but, unlike others (e.g. Occupational Safety and Health Administration (OSHA)), fail to mention the need to use dressings at luer-lock connections to contain cytotoxic leaks. This study aims to develop and validate an analytical method capable of identifying and quantifying cyclophosphamide (CP) and 5-fluorouracil (5-FU) in gauze dressings. Methods: Gauze dressings (20x10 cm) containing varying concentrations of CP and 5-FU were placed in 15 mL falcons containing acetonitrile:methanol:water (19:13:68). After stirring, the samples were passed through a 0.22 µm filter. The extracted samples were then analyzed using HPLC-DAD, equipped with a C18 column (Hypersil Gold ™ 150mm x 4.6mm and 5µm particle size). The mobile phases employed were acetonitrile:methanol:water (19:13:68) (CP) and 0.5% acetic acid in water (5-FU) and detection was set at 205 nm (CP) and 260 nm (5-FU). The mean of five calibration curves generated for each drug were calculated, each exhibiting R² > 0.997, thus confirming the linearity for both drugs. Regarding sensitivity, LOD of 0.006 µg/cm2 for 5-FU and 0.11 µg/cm2 for CP and LOQs of 0.02 µg/cm2 for 5-FU and 0.32 µg/cm2 for CP, were obtained. Accuracy fell within the range of 93%-110% and precision was set between 91% and 99%. At room temperature, gauze dressings deliberately contaminated exhibited superior stability for 5-FU compared to CP. The method has been successfully validated and, once suitability is confirmed, it will be applicable for the evaluation of the importance of using gauze dressing when handling cytotoxic drugs, by testing contaminations occurring during reallife routine handling of these cytotoxic drugs.
- Development and validation of an analytical method to quantify cytotoxic drugs in gauze dressings used in luer-lock connection systems for a safer preparationPublication . Silva, Clara; Carmo, Helena; Oliveira, Vânia; Carvalho, Daniel; Moreira, FernandoDespite patient ́s benefits, healthcare professionals face occupational hazardous when handling cytotoxic drugs used for cancer treatment. Several organizations outline procedures, conditions, and equipment recommended for safe handling. The International Society of Oncology Pharmacy Practice (ISOPP) standards are the guidelines recommended by the European Parliament but, unlike others (e.g. Occupational Safety and Health Administration (OSHA)), they omit the recommendation of using dressings at luer-lock connections to contain cytotoxic leaks. This study aims to develop and validate an analytical method for cyclophosphamide (CP) and 5-fluorouracil (5-FU) identification and quantification in gauze dressings for monitoring real-life occupational situations. Gauze dressings (20x10 cm) containing varying concentrations of CP and 5-FU were placed in 15 mL falcon tubes containing acetonitrile:methanol:water (19:13:68). After stirring, the samples were filtered through a 0.22 μm filter and analyzed using HPLC-DAD, equipped with a C18 column (Hypersil Gold ™150mm x 4.6mm and 5μm particle size). The mobile phases employed were acetonitrile:methanol:water (19:13:68) (CP) and 0.5% acetic acid in water(for 5-FU), with detection set at 205 nm (CP) and 260 nm (5-FU). The mean of five calibration curves was calculated for each drug, each exhibiting R² > 0.997, confirming the linearity for both drugs. Regarding sensitivity, LOD of 0.006 μg/cm2(5-FU) and 0.11 μg/cm2(CP) and LOQs of 0.02 μg/cm2(5-FU) and 0.32 μg/cm2(CP), were obtained. Accuracy ranged between 93%-110%, while precision ranged between 91%-99%. At room temperature, gauze dressings deliberately contaminated exhibited superior stability for 5-FU compared to CP. The method has been successfully validated for analyzing CP and 5-FU residues in gauze, meeting essential validation criteria. Upon suitability verification, the method will be applicable for the evaluation of the importance of using gauze dressing when handling cytotoxic drugs, by assessing contaminations occurring during their real-life handling routine.
- Doping detection in animals: A review of analytical methodologies published from 1990 to 2019Publication . Moreira, Fernando Xavier; Carmo, Helena; Pinho, Paula Guedes de; Bastos, Maria de LourdesDespite the impressive innate physical abilities of horses, camels, greyhounds, or pigeons, doping agents might be administered to these animals to improve their performance. To control these illegal practices, anti-doping analytical methodologies have been developed. This review compiles the analytical methods that have been published for the detection of prohibited substances administered to animals involved in sports over 30 years. Relevant papers meeting the search criteria that discussed analytical methods aiming to detect and/or quantify doping substances in animal biological matrices published from 1990 to 2019 were considered. A total of 317 studies were included, of which 298 were related to horses, demonstrating significant advances toward the development of doping detection methods for equine sports. However, analytical methods for the detection of doping agents in sports involving other species are lacking. Due to enhanced accuracy and specificity, chromatographic analysis coupled to mass spectrometry detection is preferred over immunoassays. Regarding biological matrices, blood and urine remain the first choice, although alternative biological matrices, such as hair and feces, have been considered. With the increasing number and type of drugs used as doping agents, the analytes addressed in the published papers are diverse. It is very important to continue to detect and quantify these drugs, recognizing those that are most frequently used, in order to punish the abusers, protect animals' health, and ensure a healthier and genuine competition.
- In Vitro Evaluation of the Photoreactivity and Phototoxicity of Natural Polyphenol AntioxidantsPublication . Aguiar, Brandon; Carmo, Helena; Garrido, Jorge; Sousa Lobo, José M.; Almeida, Isabel F.Polyphenols are a large family of natural compounds widely used in cosmetic products due to their antioxidant and anti-inflammatory beneficial properties and their ability to prevent UV radiation-induced oxidative stress. Since these compounds present chromophores and are applied directly to the skin, they can react with sunlight and exert phototoxic effects. The available scientific information on the phototoxic potential of these natural compounds is scarce, and thus the aim of this study was to evaluate the photoreactivity and phototoxicity of five phenolic antioxidants with documented use in cosmetic products. A standard ROS assay was validated and applied to screen the photoreactivity of the natural phenolic antioxidants caffeic acid, ferulic acid, p-coumaric acid, 3,4-dihydroxyphenylacetic acid (DOPAC), and rutin. The phototoxicity potential was determined by using a human keratinocyte cell line (HaCaT), based on the 3T3 Neutral Red Uptake phototoxicity test. Although all studied phenolic antioxidants absorbed UV/Vis radiation in the range of 290 to 700 nm, only DOPAC was able to generate singlet oxygen. The generation of reactive oxygen species is an early-stage chemical reaction as part of the phototoxicity mechanism. Yet, none of the studied compounds decreased the viability of keratinocytes after irradiation, leading to the conclusion that they do not have phototoxic potential. The data obtained with this work suggests that these compounds are safe when incorporated in cosmetic products.
- Preventing exposure to cytotoxic drugs by analysing cyclophosphamide residues in the manipulation areas of a northern portuguese hospitalPublication . Nuro, Ali; Carmo, Helena; Correia, Patrícia; Leitão, Mariana; Rocha, Luísa; Fernando, Cristina; Moreira, FernandoThe manipulation of cytotoxic agents has been increasing as chemotherapy remains the first line treatment for several malignant diseases of growing incidence. However, cytotoxic agents such as cyclophosphamide do not specifically target cancer cells, being therefore able to harm the professionals that prepare them. To prevent occupational exposure, an adequate monitoring of the working environment is warranted. The present study aimed at validating an analytical method for the evaluation of the presence of cyclophosphamide in the surface of three different rooms of a northern Portuguese hospital, in which cytotoxic drugs are received or manipulated. The extraction procedure consisted of wiping with a 100cm2 gauze moistened with ethyl-acetate followed by mixing with 15mL methanol: acetonitrile: water (25:10:65) and filtration with a PTFE 0.2µm filter. The samples were then analysed by HPLC-DAD (205nm), with the separation being obtained with a C18 column. The analytical method was validated regarding linearity, intra-day (92-98%) and inter-day (91-98%) precision, accuracy (80-106%), recovery (97-114%) and sensibility (LOD = 0.089 µg/cm2; LOQ = 0.269 µg/cm2). A total of 50 real samples were analysed and all of them presented values below the LOD. Despite using analytical methods with lower sensibility than the one herein presented, previous published studies about the presence of cytotoxic drugs on surfaces of other Portuguese hospitals revealed severe contamination in the manipulation areas. These negative results likely indicate that in this hospital the preparation protocols efficiently limit exposure, further evidencing the importance of complying with the guidelines. By adopting these procedures, it is possible to eliminate or at least greatly limit the occupational exposure to cytotoxic drugs, by nurses, pharmacy technicians and pharmaceutics. Analytical tools that efficiently monitor contamination are a helpful tool to monitor and prevent hazardous exposure.
- A simple and inexpensive method to monitor and minimize exposure from manipulation of cytotoxic drugsPublication . Ali, Nuro; Carmo, Helena; Robalo, Raquel; Rocha, Luísa; Fernandes, Cristina; Moreira, FernandoPharmacy personnel that manipulate cytotoxic drugs are under continuous exposure risk. Therefore, training and strict adherence to recommended practices should always be promoted. The main objective of this study was to develop and apply a safe, effective and low-cost method for the training and assessment of the safe handling of cytotoxic drugs, using commercially available tonic water. To evaluate the potential of tonic water as a replacement marker for quinine hydrochloride, deliberate spills of 1 mL of four different tonic waters (one coloured and three non-coloured) were analysed under ultraviolet light (300–400 nm). The pigmented sample did not produce fluorescence under ultraviolet (UV) light. The three commercially available tonic waters that exhibited fluorescence were further analysed by UV/Vis spectrophotometry (300–500 nm). Afterwards, a protocol of simulated manipulation of cytotoxic drugs was developed and applied to 12 pharmacy technicians, that prepared 24 intravenous bags according to recommended routine procedures using tonic water. Participants responded to a brief questionnaire to evaluate the adequacy and applicability of the activity. Seven of the participants had spillages during manipulation, the majority of which recorded during manipulation with needles. All participants scored the tonic water manipulation simulation with 4 or 5 points for simplicity, efficiency and feasibility. The obtained results suggest that tonic water can be used to simulate the manipulation of cytotoxic drugs in training and assessment programs. By using this replacement marker for quinine hydrochloride, it is possible to perform a more cost-effective, yet equally effective, assessment.