QuEChERS: a new sample preparation approach for the determination of ibuprofen and its metabolites in soils

Ibuprofen is one of the most used active pharmaceutical ingredients worldwide. A new method for the analysis of ibuprofen and its metabolites, hydroxyibuprofen and carboxyibuprofen, in soils is presented. The extraction of these compounds from the soil matrices was performed by using a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. The method involves a single extraction of the investigated compounds with purified water (acidified at pH 2.5 with hydrochloric acid), and a slow and continuous addition of the QuEChERS content, followed by the addition of acidified acetonitrile (1% acetic acid), prior to the determination by liquid chromatography coupled with fluorescence detection (LC–FLD). Validation studies were carried out using soil samples with a range of organic carbon contents. Recoveries of the fortified samples ranged from 79.5% to 101%. Relative standard deviations for all matrix–compound combinations did not exceed 3%. The method quantification limits were ≤22.4 μg kg−1 in all cases. The developed method was applied to the analysis of sixteen real samples.

Voltammetric oxidation of drugs of abuse I. Morphine and metabolites

A detailed study of the electrochemical oxidative behavior of morphine in aqueous solution is reported. Through the synthesis of several metabolites and derivatives, pseudomorphine, morphine N-oxide, normorphine, dihydromorphine and 2-(N,N-dimethylaminomethyl)morphine, and their voltammetric study it was possible to identify the oxidation peaks for morphine. The anodic waves are related with the oxidation of phenolic and tertiary amine groups. It is also possible to verify that a poorly defined peak observable during morphine oxidation is not a consequence of further oxidation of pseudomorphine but due to formation of a dimer during phenolic group oxidation. The results obtained and especially those regarding the formation of a new polymer based on a C O coupling could be useful for clarifying the discoloration phenomenon occurring during storage of morphine solutions as well as leading to a better understanding of its oxidative metabolic pathways.

Voltammetric oxidation of drugs of abuse II. Codeine and metabolites

The oxidation of codeine on glassy carbon electrodes has been studied in detail using differential pulse voltammetry. The results obtained using a glassy carbon electrode clearly show a much more complex oxidation mechanism than that previously reported when platinum and gold electrodes were used. To clarify the codeine oxidative profile, several metabolites and analogues of this alkaloid, codeine N-oxide, norcodeine, dihydrocodeine, acetylcodeine and 6- chlorodesoxycodeine, were synthesized and studied. It was deduced that the anodic waves observed in codeine oxidation are related to the presence of methoxy, hydroxy and tertiary amine groups. Due to the similarity of potentials at which these oxidative processes take place, at some pHs an overlap of peaks occurs and only one anodic wave is observed.

Voltammetric oxidation of drugs of abuse III. Heroin and metabolites

The oxidative behavior of heroin in aqueous solution is reported. In order to identify its oxidation peaks, several metabolites, 6-monoacetylmorphine, 3-monoacetylmorphine and norheroin, were synthesized and their electrochemical behavior studied using differential pulse voltammetry. The anodic waves observed for heroin correspond to the oxidation of the tertiary amine group and its follow-up product (secondary amine), and to the oxidation of the phenolic group obtained from hydrolysis, at alkaline pHs, of the 3-acetyl group. The results enabled a new oxidative mechanism for heroin to be proposed in which a secondary amine, norheroin, and an aldehyde are obtained. The voltammetric behavior of 6-monoacetylmorphine and morphine was found to be similar demonstrating that the presence of an acetyl substituent on the 6-hydroxy group does not have a relevant influence on the peak potential of the wave resulting from oxidation of the 3-phenolic group.

Oxidative behaviour of apomorphine and its metabolites

The metabolism of apomorphine is quite complex due to interactions with proteins and other tissue components that affect its pharmacokinetic profile. The electrochemical oxidation mechanism of apomorphine and of some synthesised apomorphine derivatives was studied. It was found to be related to the reaction of o-diphenol and tertiary amine groups and strongly dependent on pH.

Determination of permethrin and its metabolites in hearts and urines of rats by GC-ECD and GC-MS

Pyrethroids are pesticides very used in agriculture, which tend to replace organophosfate and carbamate insecticides. These pesticides have shown to exhibit cardiotoxicity. The aim of this study was to assess if cardiotoxicity is due to direct or indirect effects (metabolites) of permethrin on hearts. There were studies 8 rats; three of them were sacrificed after 24 of the end of treatment with permethrin and the other four were sacrificed after 14 days of the end of treatment. Afterwards hearts and urines were collected. The amounts of permethrin and its main metabolite (3-PBA) were evaluated on hearts and urines of female rats which were treated with permethrin and sacrificed the day after and 14 days after the treatment. Moreover has been highlighted the difference of amount of permethrin and its metabolite between rats sacrificed immediately at the end of treatment and those sacrificed after 14 days. The study of permethrin was accomplished by liquid-liquid extraction and GC-ECD. The evaluation of 3-PBA was performed by SPE procedure with 2-PBA as internal standard and gas-chromatography GC-MS. The concentration of permethrin in hearts is basically the same in 24h and 14 days. The 3-PBA concentration in urines decreased 50 times from 24h to 14 days. In hearts the 3-PBA level also decrease but only 2.24 times and a high variation of results were achieved in rats after 14 days.

Eastwards / westwards: which direction for gender studies in the 21st century?

Eastwards / Westwards: Which Direction for Gender Studies in the XXIst Century? is a collection of essays which focus on themes and methods that characterize current research into gender in Asian countries in general. In this collection, ideas derived from Gender Studies elsewhere in the world have been subjected to scrutiny for their utility in helping to describe and understand regional phenomena. But the concepts of Local and Global – with their discoursive productions – have not functioned as a binary opposition: localism and globalism are mutually constitutive and researchers have interrogated those spaces of interaction between the ‘self’ and the ‘other’, bearing in mind their own embeddedness in social and cultural structures and their own historical memory. Contributors to this collection provided a critical transnational perspective on some of the complex effects of the dynamics of cultural globalization, by exploring the relation between gender and development, language, historiography, education and culture. We have also given attention to the ideological and rhetorical processes through which gender identity is constructed, by comparing textual grids and patterns of expectation. Likewise, we have discussed the role of ethnography, anthropology, historiography, sociology, fiction, popular culture and colonial and post-colonial sources in (re)inventing old/new male/female identities, their conversion into concepts and circulation through time and space. This multicultural and trans-disciplinary selection of essays is totally written in English, fully edited and revised, therefore, it has a good potential for an immediate international circulation. This project may trace new paths and issues for discussion on what concerns the life, practices and narratives by and about women in Asia, as well as elsewhere in the present day global experience. Academic readership: Researchers, scholars, educators, graduate and post-graduate students, doctoral students and general non-fiction readers, with a special interest in Gender Studies, Asia, Colonial and Post-Colonial Literature, Anthropology, Cultural Studies, History, Historiography, Politics, Race, Feminism, Language, Linguistics, Power, Political and Feminist Agendas, Popular Culture, Education, Women’s Writing, Religion, Multiculturalism, Globalisation, Migration. Chapter summary: 1. “Social Gender Stereotypes and their Implication in Hindi”, Anjali Pande, Jawaharlal Nehru University, New Delhi, India. This essay looks at the subtle ways in which gender identities are constructed and reinforced in India through social norms of language use. Language itself becomes a medium for perpetuating gender stereotypes, forcing its speakers to confirm to socially defined gender roles. Using examples from a classroom discussion about a film, this essay will highlight the underlying rigid male-female stereotypes in Indian society with their more obvious expressions in language. For the urban woman in India globalisation meant increased economic equality and exposure to changed lifestyles. On an individual level it also meant redefining gender relations and changing the hierarchy in man-­woman relationships. With the economic independence there is a heightened sense of liberation in all spheres of social life, a confidence to fuzz the rigid boundaries of gender roles. With the new films and media celebrating this liberated woman, who is ready to assert her sexual needs, who is ready to explode those long held notions of morality, one would expect that the changes are not just superficial. But as it soon became obvious in the course of a classroom discussion about relationships and stereotypes related to age, the surface changes can not become part of the common vocabulary, for the obvious reason that there is still a vast gap between the screen image of this new woman and the ground reality. Social considerations define the limits of this assertiveness of women, whereas men are happy to be liberal within the larger frame of social sanctions. The educated urban woman in India speaks in favour of change and the educated urban male supports her, but one just needs to scratch the surface to see the time tested formulae of gender roles firmly in place. The way the urban woman happily balances this emerging promise of independence with her gendered social identity, makes it necessary to rethink some aspects of looking at gender in a gradually changing, traditional society like India. 2. “The Linguistic Dimension of Gender Equality”, Alissa Tolstokorova, Kiev Centre for Gender Information and Education, Ukraine. The subject-matter of this essay is gender justice in language which, as I argue, may be achieved through the development of a gender-related approach to linguistic human rights. The last decades of the 20th century, globally marked by a “gender shift” in attitudes to language policy, gave impetus to the social movement for promoting linguistic gender equality. It was initiated in Western Europe and nowadays is moving eastwards, as ideas of gender democracy progress into developing countries. But, while in western societies gender discrimination through language, or linguistic sexism, was an issue of concern for over three decades, in developing countries efforts to promote gender justice in language are only in their infancy. My argument is that to promote gender justice in language internationally it is necessary to acknowledge the rights of women and men to equal representation of their gender in language and speech and, therefore, raise a question of linguistic rights of the sexes. My understanding is that the adoption of the Universal Declaration of Linguistic Rights in 1996 provided this opportunity to address the problem of gender justice in language as a human rights issue, specifically as a gender dimension of linguistic human rights. 3. “The Rebirth of an Old Language: Issues of Gender Equality in Kazakhstan”, Maria Helena Guimarães, Polytechnic Institute of Porto, Portugal. The existing language situation in Kazakhstan, while peaceful, is not without some tension. We propose to analyze here some questions we consider relevant in the frame of cultural globalization and gender equality, such as: free from Russian imperialism, could Kazakhstan become an easy prey of Turkey’s “imperialist dream”? Could these traditionally Muslim people be soon facing the end of religious tolerance and gender equality, becoming this new old language an easy instrument for the infiltration in the country of fundamentalism (it has already crossed the boarders of Uzbekistan), leading to a gradual deterioration of its rich multicultural relations? The present structure of the language is still very fragile: there are three main dialects and many academics defend the re-introduction of the Latin alphabet, thus enlarging the possibility of cultural “contamination” by making the transmission of fundamentalist ideas still easier through neighbour countries like Azerbaijan, Uzbekistan and Turkmenistan (their languages belong to the same sub-group of Common Turkic), where the Latin alphabet is already in use, and where the ground for such ideas shown itself very fruitful. 4. “Construction of Womanhood in the Bengali Language of Bangladesh”, Raasheed Mahmood; University of New South Wales, Sydney. The present essay attempts to explore the role of gender-based language differences and of certain markers that reveal the status accorded to women in Bangladesh. Discrimination against women, in its various forms, is endemic in communities and countries around the world, cutting across class, race, age, and religious and national boundaries. One cannot understand the problems of gender discrimination solely by referring to the relationship of power or authority between men and women. Rather one needs to consider the problem by relating it to the specific social formation in which the image of masculinity and femininity is constructed and reconstructed. Following such line of reasoning this essay will examine the nature of gender bias in the Bengali language of Bangladesh, holding the conviction that as a product of social reality language reflects the socio-cultural behaviour of the community who speaks it. This essay will also attempt to shed some light on the processes through which gender based language differences produce actual consequences for women, who become exposed to low self-esteem, depression and systematic exclusion from public discourse. 5. “Marriage in China as an expression of a changing society”, Elisabetta Rosado David, University of Porto, Portugal, and Università Ca’Foscari, Venezia, Italy. In 29 April 2001, the new Marriage Law was promulgated in China. The first law on marriage was proclaimed in 1950 with the objective of freeing women from the feudal matrimonial system. With the second law, in 1981, values and conditions that had been distorted by the Cultural Revolution were recovered. Twenty years later, a new reform was started, intending to update marriage in the view of the social and cultural changes that occurred with Deng Xiaoping’s “open policy”. But the legal reform is only the starting point for this case-study. The rituals that are followed in the wedding ceremony are often hard to understand and very difficult to standardize, especially because China is a vast country, densely populated and characterized by several ethnic minorities. Two key words emerge from this issue: syncretism and continuity. On this basis, we can understand tradition in a better way, and analyse whether or not marriage, as every social manifestation, has evolved in harmony with Chinese culture. 6. “The Other Woman in the Portuguese Colonial Empire: The Case of Portuguese India”, Maria de Deus Manso, University of Évora, Portugal. This essay researches the social, cultural and symbolic history of local women in the Portuguese Indian colonial enclaves. The normative Portuguese overseas history has not paid any attention to the “indigenous” female populations in colonial Portuguese territories, albeit the large social importance of these social segments largely used in matrimonial and even catholic missionary strategies. The first attempt to open fresh windows in the history of this new field was the publication of Charles Boxer’s referential study about Women in lberian Overseas Expansion, edited in Portugal only after the Revolution of 1975. After this research we can only quote some other fragmentary efforts. In fact, research about the social, cultural, religious, political and symbolic situation of women in the Portuguese colonial territories, from the XVI to the XX century, is still a minor historiographic field. In this essay we discuss this problem and we study colonial representations of women in the Portuguese Indian enclaves, mainly in the territory of Goa, using case studies methodologies. 7. “Heading East this Time: Critical Readings on Gender in Southeast Asia”, Clara Sarmento, Polytechnic Institute of Porto, Portugal. This essay intends to discuss some critical readings of fictional and theoretical texts on gender condition in Southeast Asian countries. Nowadays, many texts about women in Southeast Asia apply concepts of power in unusual areas. Traditional forms of gender hegemony have been replaced by other powerful, if somewhat more covert, forms. We will discuss some universal values concerning conventional female roles as well as the strategies used to recognize women in political fields traditionally characterized by male dominance. Female empowerment will mean different things at different times in history, as a result of culture, local geography and individual circumstances. Empowerment needs to be perceived as an individual attitude, but it also has to be facilitated at the macro­level by society and the State. Gender is very much at the heart of all these dynamics, strongly related to specificities of historical, cultural, ethnic and class situatedness, requiring an interdisciplinary transnational approach.

Marine cyanobacteria compounds with anticancer properties: a review on the implication of apoptosis

Marine cyanobacteria have been considered a rich source of secondary metabolites with potential biotechnological applications, namely in the pharmacological field. Chemically diverse compounds were found to induce cytoxicity, anti-inflammatory and antibacterial activities. The potential of marine cyanobacteria as anticancer agents has however been the most explored and, besides cytotoxicity in tumor cell lines, several compounds have emerged as templates for the development of new anticancer drugs. The mechanisms implicated in the cytotoxicity of marine cyanobacteria compounds in tumor cell lines are still largely overlooked but several studies point to an implication in apoptosis. This association has been related to several apoptotic indicators such as cell cycle arrest, mitochondrial dysfunctions and oxidative damage, alterations in caspase cascade, alterations in specific proteins levels and alterations in the membrane sodium dynamics. In the present paper a compilation of the described marine cyanobacterial compounds with potential anticancer properties is presented and a review on the implication of apoptosis as the mechanism of cell death is discussed.

Effect of oxidative stress upon absorption of glucose by the human placenta: in vitro studies with BeWo cells

Pregnancy is a dynamic state and the placenta is a temporary organ that, among other important functions, plays a crucial role in the transport of nutrients and metabolites between the mother and the fetus, which is essential for a successful pregnancy. Among these nutrients, glucose is considered a primary source of energy and, therefore, fundamental to insure proper fetus development. Several studies have shown that glucose uptake is dependent on several morphological and biochemical placental conditions. Oxidative stress results from the unbalance between reactive oxygen species (ROS) and antioxidants, in favor of the first. During pregnancy, ROS, and therefore oxidative stress, increase, due to increased tissue oxygenation. Moreover, the relation between ROS and some pathological conditions during pregnancy has been well established. For these reasons, it becomes essential to understand if oxidative stress can compromise the uptake of glucose by the placenta. To make this study possible, a trophoblastic cell line, the BeWo cell line, was used. Experiments regarding glucose uptake, either under normal or oxidative stress conditions, were conducted using tert-butylhydroperoxide (tBOOH) as an oxidative stress inducer, and 3H-2-deoxy-D-glucose (3H-DG) as a glucose analogue. Afterwards, studies regarding the involvement of glucose facilitative transporters (GLUT) and the phosphatidylinositol 3-kinases (PI3K) and protein kinase C (PKC) pathways were conducted, also under normal and oxidative stress conditions. A few antioxidants, endogenous and from diet, were also tested in order to study their possible reversible effect of the oxidative effect of tBOOH upon apical 3H-DG uptake. Finally, transepithelial studies gave interesting insights regarding the apical-to-basolateral transport of 3H-DG. Results showed that 3H-DG uptake, in BeWo cells, is roughly 50% GLUT-mediated and that tBOOH (100 μM; 24h) decreases apical 3H-DG uptake in BeWo cells by about 33%, by reducing both GLUT- (by 28%) and non-GLUT-mediated (by 40%) 3H-DG uptake. Uptake of 3H-DG and the effect of tBOOH upon 3H-DG uptake are not dependent on PKC and PI3K. Moreover, the effect of tBOOH is not associated with a reduction in GLUT1 mRNA levels. Resveratrol, quercetin and epigallocatechin-3-gallate, at 50 μM, reversed, by at least 45%, the effect of tBOOH upon 3H-DG uptake. Transwell studies show that the apical-to-basolateral transepithelial transport of 3H-DG is increased by tBOOH.In conclusion, our results show that tBOOH caused a marked decrease in both GLUT and non-GLUT-mediated apical uptake of 3H-DG by BeWo cells. Given the association of increased oxidative stress levels with several important pregnancy pathologies, and the important role of glucose for fetal development, the results of this study appear very interesting.

Monitorização de pesticidas em águas de Esposende

De forma a proteger o ambiente e a saúde humana, é imperativo evitar, prevenir ou reduzir as concentrações prejudiciais de poluentes nocivos na água subterrânea. A necessidade da obtenção de níveis de protecção da água subterrânea, encontra-se estabelecida em normas de qualidade e devem ser desenvolvidas metodologias que permitam a avaliação do estado químico da água subterrânea. Este trabalho experimental centrou-se no desenvolvimento de uma metodologia analítica de detecção e quantificação por cromatografia gasosa com detector de captura de electrões dos pesticidas atrazina e respectivos metabolitos (desetilatrazina e deisopropilatrazina), simazina, terbutilazina e o metabolito desetiterbutilazina, folpete, dimetoato, diazinão, malatião, cloropirifos e o azinfos-metilo em águas de poços. O estudo progressivo baseou-se na colheita de água a 20 poços agrícolas na zona de Esposende, área considerada pelo Ministério da Agricultura do Desenvolvimento Rural e Pescas como sendo uma zona vulnerável. O método utilizado para a validação da técnica cromatográfica baseou-se na norma ISO 8466-1:1990. Os parâmetros de validação considerados foram: especificidade/selectividade, capacidade de identificação, limites de detecção e quantificação, relação sinal/ruído, linearidade e curva de calibração, precisão (repetibilidade, precisão intermédia e reprodutibilidade), eficiência de extracção e arrastamento. O método demonstrou ser capaz de identificar e quantificar os analitos, sem interferência de outros compostos. Obteve-se um valor para os parâmetros da precisão inferior a 10%, enquanto os mais baixos limites de detecção e de quantificação foram, respectivamente, 0,014 e 0,047 μg L-1. Na preparação de amostras optou-se pelo método de extracção em fase sólida, tendo sido testadas cinco diferentes tipos de colunas extractivas; Lichrolut® EN/RP-18; Strata SDB-L e C18-E; Chromabond HR-P e HR-X, sendo que as colunas Lichrolut® EN/RP- 18 apresentaram melhores resultados para a globalidade dos pesticidas. Da análise efectuada aos 20 poços agrícolas verificou-se que apenas 3 não apresentavam qualquer vestígio dos pesticidas monitorizados, sendo que as restantes apresentavam valores entre 0,05 e 53,2 μg L-1, valores superiores aos impostos pela legislação em vigor (Decreto-Lei n.º 208/2008 de 28 de Outubro para água subterrânea e Decreto-Lei nº306/2007 referente a água para consumo). Verificou-se que os proprietários dos poços agrícolas, dos quais se procedeu à amostragem de água para análise não têm a consciência da falta de qualidade dessa água, nem dos malefícios que possam advir do seu consumo.

Organochlorine pesticide residues in strawberries from integrated pest management and organic farming

A rapid, specific, and sensitive method based on theQuick Easy Cheap Effective Rugged and Safe (QuEChERS) method and a cleanup using dispersive solid-phase extraction with MgSO4, PSA, and C18 sorbents has been developed for the routine analysis of 14 pesticides in strawberries. The analyses were performed by three different analytical methodologies: gas chromatography (GC) with electron capture detection (ECD), mass spectrometry (MS), and tandem mass spectrometry (MS/MS). The recoveries for all the pesticides studied were from 46 to 128%, with relative standard deviation of <15% in the concentration range of 0.005-0.250 mg/kg. The limit of detection (LOD) for all compoundsmetmaximumresidue limits (MRL) accepted in Portugal for organochlorine pesticides (OCP). A survey study of strawberries produced in Portugal in the years 2009-2010 obtained from organic farming (OF) and integrated pest management (IPM) was developed. Lindane and β-endosulfan were detected above the MRL in OF and IPM. Other OCP (aldrin, o,p0-DDT and their metabolites, and methoxychlor) were found below the MRL. The OCP residues detected decreased from 2009 to 2010. The QuEChERS method was successfully applied to the analysis of strawberry samples.

Antitumor activity of hierridin B, a cyanobacterial secondary metabolite found in both filamentous and unicellular marine strains

Cyanobacteria are widely recognized as a valuable source of bioactive metabolites. The majority of such compounds have been isolated from so-called complex cyanobacteria, such as filamentous or colonial forms, which usually display a larger number of biosynthetic gene clusters in their genomes, when compared to free-living unicellular forms. Nevertheless, picocyanobacteria are also known to have potential to produce bioactive natural products. Here, we report the isolation of hierridin B from the marine picocyanobacterium Cyanobium sp. LEGE 06113. This compound had previously been isolated from the filamentous epiphytic cyanobacterium Phormidium ectocarpi SAG 60.90, and had been shown to possess antiplasmodial activity. A phylogenetic analysis of the 16S rRNA gene from both strains confirmed that these cyanobacteria derive from different evolutionary lineages. We further investigated the biological activity of hierridin B, and tested its cytotoxicity towards a panel of human cancer cell lines; it showed selective cytotoxicity towards HT-29 colon adenocarcinoma cells.

Deteção eletroquímica da norfloxacina e trimetoprim em amostras de urina utilizando polímero molecularmente impresso modificado e grafeno

A norfloxacina e o trimetoprim são dois antibióticos antibacterianos usados para o tratamento de infeções urinárias, intestinais e respiratórias. A maioria dos fármacos exige uma dosagem que garanta os níveis de segurança e eficácia de atuação. A necessidade de dosear os medicamentos e os seus metabolitos é assim um controlo imperioso e em muitos casos regular no tratamento de um paciente. Neste trabalho desenvolveram-se dois sensores eletroquímicos para a deteção da norfloxacina (NFX) e do trimetoprim (TMP), usando como superfície de suporte o carbono vítreo. A busca de novos materiais que conferiram maior seletividade e sensibilidade aos sistemas de deteção e por outro lado apresentem menores riscos para o paciente quando usados em dispositivos que permitam uma análise point-of-care, é especialmente importante e pode ser uma parte crucial do processo de decisão clínica. Assim, os polímeros molecularmente impresos enquadram-se nesse perfil e o seu uso tem vindo a ser cada vez mais avaliado. A impressão molecular é uma tecnologia capaz de produzir polímeros que incorporam as moléculas do analito e que após remoção por solventes específicos, permitem dotá-los de locais específicos de reconhecimento estereoquímico. A seleção do pirrol como polímero molecularmente impresso (MIP) permitiu construir com sucesso os sensores para doseamento dos antibióticos. A fim de aumentar a sensibilidade do método incorporou-se grafeno na superfície do elétrodo. Este material tem vindo a ser largamente utilizado devido às suas propriedades: estrutura molecular, condutividade elétrica e aumento da superfície são algumas das características que mais despertam o interesse para a sua aplicação neste projeto. Os sensores desenvolvidos foram incorporados em sistemas eletroquímicos. Os métodos voltamétricos aplicados foram a voltametria cíclica, a voltametria de onda quadrada e ainda a impedância. As condições de análise foram otimizadas no que respeita à polimerização do pirrol (concentração do polímero, número de ciclos de eletropolimerização e respetivos potenciais aplicados, tempo de incubação, solvente de remoção do analito), ao pH da solução do fármaco, à gama de concentrações dos antibióticos e aos parâmetros voltamétricos dos métodos de análise. Para cada um dos antibióticos um elétrodo não-impresso foi também preparado, usando o procedimento de polimerização mas sem a presença da molécula do analito, e foi usado como controlo. O sensor desenvolvido para o trimetoprim foi usado no doseamento do fármaco em amostras de urina. As amostras foram analisadas sem qualquer diluição, apenas foram centrifugadas para remoção de proteínas e algum interferente. Os sensores construídos apresentaram comportamento linear na gama de concentrações entre 102 e 107 mol/L. Os resultados mostram boa precisão (desvios padrão inferiores a 11%) e os limites de deteção foram de 8,317 e 1,307 mol/L para a norfloxacina e o trimetoprim, respetivamente. Para validação do método foram ainda efetuados ensaios de recuperação tendo obtido valores superiores a 94%.

Antitumor activity of hierridin B, a cyanobacterial secondary metabolite found in both filamentous and unicellular marine strains

Cyanobacteria are widely recognized as a valuable source of bioactive metabolites. The majority of such compounds have been isolated from so-called complex cyanobacteria, such as filamentous or colonial forms, which usually display a larger number of biosynthetic gene clusters in their genomes, when compared to free-living unicellular forms. Nevertheless, picocyanobacteria are also known to have potential to produce bioactive natural products. Here, we report the isolation of hierridin B from the marine picocyanobacterium Cyanobium sp. LEGE 06113. This compound had previously been isolated from the filamentous epiphytic cyanobacterium Phormidium ectocarpi SAG 60.90, and had been shown to possess antiplasmodial activity. A phylogenetic analysis of the 16S rRNA gene from both strains confirmed that these cyanobacteria derive from different evolutionary lineages. We further investigated the biological activity of hierridin B, and tested its cytotoxicity towards a panel of human cancer cell lines; it showed selective cytotoxicity towards HT-29 colon adenocarcinoma cells.

Development of a simple analytical method for the simultaneousdetermination of paracetamol, paracetamol-glucuronide andp-aminophenol in river water

Paracetamol is among the most worldwide consumed pharmaceuticals. Although its occurrence in the environment is well documented, data about the presence of its metabolites and transformation products is very scarce. The present work describes the development of an analytical method for the simultaneous determination of paracetamol, its principal metabolite (paracetamol-glucuronide) and its main transformation product (p-aminophenol) based on solid phase extraction (SPE) and high performance liquid chromatography coupled to diode array detection (HPLC-DAD). The method was applied to analysis of river waters, showing to be suitable to be used in routine analysis. Different SPE sorbents were compared and the use of two Oasis WAX cartridges in tandem proved to be the most adequate approach for sample clean up and pre-concentration. Under optimized conditions, limits of detection in the range 40–67 ng/L were obtained, as well as mean recoveries between 60 and 110% with relative standard deviations (RSD) below 6%. Finally, the developed SPE-HPLC/DAD method was successfully applied to the analysis of the selected compounds in samples from seven rivers located in the north of Portugal. Nevertheless all the compounds were detected, it was the first time that paracetamol-glucuronide was found in river water at concentrations up to 3.57 μg/L.