Optimization and validation of organochlorine compounds in adipose tissue by SPE-gas chromatography

Scientific evidence has shown an association between organochlorine compounds (OCC) exposure and human health hazards. Concerning this, OCC detection in human adipose samples has to be considered a public health priority. This study evaluated the efficacy of various solid-phase extraction (SPE) and cleanup methods for OCC determination in human adipose tissue. Octadecylsilyl endcapped (C18-E), benzenesulfonic acid modified silica cation exchanger (SA), poly (styrene-divinylbenzene (EN) and EN/RP18 SPE sorbents were evaluated. The relative sample cleanup provided by these SPE columns was evaluated using gas chromatography with electron capture detection (GC–ECD). The C18-E columns with strong homogenization were found to provide the most effective cleanup, removing the greatest amount of interfering substance, and simultaneously ensuring good analyte recoveries higher than 70%. Recoveries>70% with standard deviations (SD)<15% were obtained for all compounds under the selected conditions. Method detection limits were in the 0.003–0.009 mg/kg range. The positive samples were confirmed by gas chromatography coupled with tandem mass spectrometry (GC-MS/MS). The highest percentage found of the OCC in real samples corresponded to HCB, o,p′-DDT and methoxychlor, which were detected in 80 and 95% of samples analyzed respectively. Copyright © 2012 John Wiley & Sons, Ltd.

Interlaboratory validation of an environmental monitoring method for trace analysis of endocrine disrupting compounds

Environmental pollution continues to be an emerging study field, as there are thousands of anthropogenic compounds mixed in the environment whose possible mechanisms of toxicity and physiological outcomes are of great concern. Developing methods to access and prioritize the screening of these compounds at trace levels in order to support regulatory efforts is, therefore, very important. A methodology based on solid phase extraction followed by derivatization and gas chromatography-mass spectrometry analysis was developed for the assessment of four endocrine disrupting compounds (EDCs) in water matrices: bisphenol A, estrone, 17b-estradiol and 17a-ethinylestradiol. The study was performed, simultaneously, by two different laboratories in order to evaluate the robustness of the method and to increase the quality control over its application in routine analysis. Validation was done according to the International Conference on Harmonisation recommendations and other international guidelines with specifications for the GC-MS methodology. Matrix-induced chromatographic response enhancement was avoided by using matrix-standard calibration solutions and heteroscedasticity has been overtaken by a weighted least squares linear regression model application. Consistent evaluation of key analytical parameters such as extraction efficiency, sensitivity, specificity, linearity, limits of detection and quantification, precision, accuracy and robustness was done in accordance with standards established for acceptance. Finally, the application of the optimized method in the assessment of the selected analytes in environmental samples suggested that it is an expedite methodology for routine analysis of EDC residues in water matrices.

Functional validation of LexADBDGim chimeras in saccharomyces cerevisiae

GimC/Prefoldin is a hetero-oligomeric complex involved in cytoskeleton biogenesis. In order to identify by two-hybrid system targets that directly interact with Gims and support the stress phenotypes, this work aimed the functional validation of all Gims in saccharomyces cerevisiae.

Enzymatic determination of choline in milk using a FIA system with potentiometric detection

The development of a FIA system for the determination of total choline content in several types of milk is described. The samples were submitted to hydrochloric acid digestion before injection into the system and passed through an enzymatic reactor containing choline oxidase immobilised on glass beads. This enzymatic reaction releases hydrogen peroxide which then reacts with a solution of iodide. The decrease in the concentration of iodide ion is quantified using an iodide ion selective tubular electrode based on a homogeneous crystalline membrane. Validation of the results obtained with this system was performed by comparison with results from a method described in the literature and applied to the determination of total choline in milks. The relative deviation was always < 5%. The repeatability of the method developed was assessed by calculation of the relative standard deviation (RSD) for 12 consecutive injections of one sample. The RSD obtained was < 0.6%.

Development and validation of a novel method for the analysis of chlorinated pesticides in soils using microwave-assisted extraction–headspace solid phase microextraction and gas chromatography–tandem mass spectrometry

A new procedure for determining eleven organochlorine pesticides in soils using microwave-assisted extraction (MAE) and headspace solid phase microextraction (HS-SPME) is described. The studied pesticides consisted of mirex, α- and γ-chlordane, p,p’-DDT, heptachlor, heptachlor epoxide isomer A, γ-hexachlorocyclohexane, dieldrin, endrin, aldrine and hexachlorobenzene. The HS-SPME was optimized for the most important parameters such as extraction time, sample volume and temperature. The present analytical procedure requires a reduced volume of organic solvents and avoids the need for extract clean-up steps. For optimized conditions the limits of detection for the method ranged from 0.02 to 3.6 ng/g, intermediate precision ranged from 14 to 36% (as CV%), and the recovery from 8 up to 51%. The proposed methodology can be used in the rapid screening of soil for the presence of the selected pesticides, and was applied to landfill soil samples.

Amperometric and spectrophotometric determination of carbaryl in natural waters and commercial formulations

The work presented describes the development and evaluation of two flow-injection analysis (FIA) systems for the automated determination of carbaryl in spiked natural waters and commercial formulations. Samples are injected directly into the system where they are subjected to alkaline hydrolysis thus forming 1-naphthol. This product is readily oxidised at a glassy carbon electrode. The electrochemical behaviour of 1-naphthol allows the development of an FIA system with an amperometric detector in which 1-naphthol determination, and thus measurement of carbaryl concentration, can be performed. Linear response over the range 1.0×10–7 to 1.0×10–5 mol L–1, with a sampling rate of 80 samples h–1, was recorded. The detection limit was 1.0×10–8 mol L–1. Another FIA manifold was constructed but this used a colorimetric detector. The methodology was based on the coupling of 1-naphthol with phenylhydrazine hydrochloride to produce a red complex which has maximum absorbance at 495 nm. The response was linear from 1.0×10–5 to 1.5×10–3 mol L–1 with a detection limit of 1.0×10–6 mol L–1. Sample-throughput was about 60 samples h–1. Validation of the results provided by the two FIA methodologies was performed by comparing them with results from a standard HPLC–UV technique. The relative deviation was <5%. Recovery trials were also carried out and the values obtained ranged from 97.0 to 102.0% for both methods. The repeatability (RSD, %) of 12 consecutive injections of one sample was 0.8% and 1.6% for the amperometric and colorimetric systems, respectively.

Epidemiologic transition in northern Angola: discussion of cumulative results from CISA (Health Research Centre of Angola)

The 41 years of armed conflict (1961 to 2002) resulted in a poor development of the health care and education infrastructures, and forced the relocation of people to safer places, namely major urban cities like Luanda. This phase was characterized by typical demographic, nutritional and epidemiological profiles. With the end of this period Angola has been repeatedly ranked as one of the three fastest growing economies in the world, and along with the social stabilization and globalization, the country is facing the introduction of new medical technologies, improvement of health sys-tems and services, better access to them, and overall better quality of life. These changes could also be translating into socio-cultural, demographic and nutritional changes which in turn may leading to changes in the epidemiological profile of the country. Thus, the emergence of non-communicable diseases are likely to become an increasingly im-portant public health problem in Angola. Also, considering that several infectious diseases persist, our weakened health system will have to face a double burden. Thus, disease surveillance data on non-communicable diseases to determine their prevalence and impact, along with the major behavioural risk factors like consumption of tobacco, alcohol, diets and physical inactivity are urgently needed.

Electroanalytical study of the pesticide asulam

The electrochemical behaviour of the herbicide Asulam was studied by cyclic and square wave voltammetry. Asulam may be irreversibly oxidised at a glassy carbon electrode. Maximum currents were obtained at pH=1.9 in aqueous electrolyte solution. Based on the electrochemical behaviour of Asulam, two analytical methodologies were developed for its determination in water samples, using square wave voltammetry (SWV) and flow injection analysis (FIA) coupled with an amperometric detector. Limits of detection of 7.1x10-6 mol L-1 and 1.2x10-8 mol L-1 for SWV and FIA respectively, were achieved. Repeatability was calculated by assessing the relative standard deviation (%) for 10 consecutive determinations of one sample. The found values were 2.1% for SWV and 5.0% for FIA. Validation of the results provided by SWV and FIA methodologies was performed by comparison with results from an HPLC-DAD technique. Good relative deviations were found (<5%). Recovery trials were performed to assess the accuracy of the results and the obtained values were between 84% and 107% for both methods.

Validation of a single-extraction procedure for sequential analysis of vitamin E, cholesterol, fatty acids, and total fat in seafood

Food lipid major components are usually analyzed by individual methodologies using diverse extractive procedures for each class. A simple and fast extractive procedure was devised for the sequential analysis of vitamin E, cholesterol, fatty acids, and total fat estimation in seafood, reducing analyses time and organic solvent consumption. Several liquid/liquid-based extractive methodologies using chlorinated and non-chlorinated organic solvents were tested. The extract obtained is used for vitamin E quantification (normal-phase HPLC with fluorescence detection), total cholesterol (normal-phase HPLC with UV detection), fatty acid profile, and total fat estimation (GC-FID), all accomplished in <40 min. The final methodology presents an adequate linearity range and sensitivity for tocopherol and cholesterol, with intra- and inter-day precisions (RSD) from 3 to 11 % for all the components. The developed methodology was applied to diverse seafood samples with positive outcomes, making it a very attractive technique for routine analyses in standard equipped laboratories in the food quality control field.

Validation of a new reference values’s database for semiquantification of 123I‐FP‐CIT SPECT scans in one nuclear medicine department

The DatScanTM and its Semiquantification (SQ) can provide advantages in the diagnosis of Parkinsonian Syndromes (PS). To improve the SQ is recommended the creation of adapted database (DB) with reference values for the Nuclear Medicine Departments. Previously to this work was created a adapted database (DBRV) to Nuclear Medicine Department's protocol and population of Infanta Cristina's Hospital located in Badajoz, for patients between the ages of 60 and 75, and reference values of the SQ were calculated. Aim: To evaluate the discrimination capacity of a department's adapted DB reference's values of healthy controls for DatScanTM.

Development and validation of a psychometric scale for assessing PA chest image quality

Purpose - To develop and validate a psychometric scale for assessing image quality for chest radiographs.

Exploring the experience of stigma in severe mental illness : a Portuguese contribution to the validation of a psychometric instrument

RESUMO: Fizemos uma análise da evolução do conceito de estigma, das suas correlações e das suas consequências e analisámos os instrumentos psicométricos utilizados para estudar experiências pessoais de estigma. Revimos os principais estudos de investigação sobre estigma em Portugal. Revimos, igualmente, os estudos relevantes utilizando o “Consumer Experiences of Stigma Questionnaire” (CESQ) e as propriedades psicométricas já documentadas. O nosso estudo teve como objetivos: explorar as experiências de estigma numa amostra portuguesa de pessoas com perturbação mental grave e contribuir para a documentação das propriedades psicométricas do “Consumer Experiences of Stigma Questionnaire” e para a validação da sua versão portuguesa. Fizemos um estudo transversal, descritivo e analítico, recolhemos dados sociodemográficos e clínicos e medimos as experiências de estigma e o funcionamento global. A frequência das respostas da secção de estigma foi semelhante à dos restantes estudos utilizando a CESQ. A frequência das respostas na secção de discriminação foi ligeiramente inferior à reportada noutros estudos. Verificámos a existência de uma associação entre a pontuação da subescala de discriminação, o sexo masculino e o facto de se viver na comunidade. A pontuação da subescala de discriminação está também correlacionada de forma positiva com o funcionamento global. Os alfas de Cronbach para a CESQ e para as suas subescalas foram considerados bons. Os coeficientes de correlação intraclasse foram igualmente considerados igualmente bons. Utilizando técnicas de análise fatorial, verificámos que a maior parte dos itens da CESQ se enquadrava em dois fatores, correspondendo sensivelmente às subescalas definidas previamente. Concluímos que o presente estudo explorou com sucesso a questão do estigma em Portugal, contribuindo em simultâneo para a validação do “Consumer Experiences of Stigma Questionnaire.--------------ABSTRACT: We reviewed the evolution of concept of stigma, its correlates and consequences, and analysed psychometric instruments that were used to study personal experiences of stigma. We provided an insight over research of stigma in Portugal. We reviewed relevant studies that use Consumer Experiences of Stigma Questionnaire and documented psychometric properties of this instrument. Our study aimed both to explore experiences of stigma in a Portuguese sample of people with severe mental illness and to contribute to the assessment of the psychometric properties of Consumer Experiences of Stigma Questionnaire and to the validation of its Portuguese translation. We performed a cross sectional descriptive and analytic study, collected socio-demographic data and measured experiences of stigma and global functioning. Frequency of responses regarding stigma section of CESQ matched previous studies using that scale. Frequency of responses in discrimination section was slightly lower than previously reported studies. We found an association between the discrimination score of CESQ and both male gender and living in the community. The discrimination score also positively correlated with global functioning. Cronbach alphas for CESQ and its subscales were good. Intraclass correlation coefficients for CESQ and stigma subscale were also good. Using factor analysis we found most of the items in CESQ would fit 2 factors, grossly corresponding to the previously defined subscales. We conclude that this study successfully explored stigma in Portugal, contributing in simultaneous to the validation of Consumer Experiences Questionnaire.

Insurance segmentation as unfair discrimination: what to expect next in the wake of Test-Achats

Proceedings of the 16th Annual Conference organized by the Insurance Law Association of Serbia and German Foundation for International Legal Co-Operation (IRZ), entitled "Insurance law, governance and transparency: basics of the legal certainty" Palic Serbia, 17-19 April 2015.