Estratègies d'immbolització orientada d'anticossos per a la detecció directa en biosensors òptics: influència en la sensibilitat i especificitat

Els assajos de diagnòstic clínic i biomèdic requereixen tècniques de detecció sensibles, selectives i ràpides que permeten mesurar concentracions de substàncies directament de mostres biològiques com sang, orina o sèrum. Els dispositius biosensors compleixen en gran mesura aquests requisits i en particular, molts esforços van dirigits actualment al disseny de versions compactes de mida reduïda tipus "point of care" (POC) que permeten, per exemple, deslocalitzar les anàlisis dels hospitals i que proporcionen un seguiment del pacient més proper i eficient. Tot i així, la transferència d'aquestes tecnologies a dispositius miniaturitzats i portàtils amb aplicacions definides encara no està totalment assolida. Apart de les dificultats inherents a la integració de components electrònics, òptics, de distribució de líquids, etc. en un sistema menut i portàtil, un important repte en la miniaturització de biosensors és la biofuncionalització de la superfície sensora i la seua estabilitat una vegada s'han immobilitzat els reactius biològics. La detecció de biomarcadors proteics es duu a terme principalment mitjançant immunoassajos, basats en el reconeixement específic de la proteïna per part d'anticossos específics. En el cas de biosensors, el format directe, en el qual un anticòs s'immobilitza sobre la superfície sensora i la proteïna diana es detecta directament de la mostra, hauria de ser el format preferible. Tanmateix, és un dels menys eficients en termes d'estabilitat i sensibilitat, degut en part a la necessitat d'immobilitzar l'anticòs en una orientació adequada sense perjudicar l'afinitat per la molècula diana. En aquest treball s'ha fet un estudi comparatiu entre diferents metodologies d'immobilització d'anticossos sobre un transductor òptic i s'han desenvolupat i optimitzat diferents estratègies de bloqueig de superfície a fi d'obtindre un procediment robust, estable, aplicable a superfícies sensores de diferent naturalesa i que a més, permeta la detecció sensible i selectiva en mostres complexes.

Use of NIR spectroscopy and multivariate process spectra calibration methodology for pharmaceutical solid samples analysis

Accomplish high quality of final products in pharmaceutical industry is a challenge that requires the control and supervision of all the manufacturing steps. This request created the necessity of developing fast and accurate analytical methods. Near infrared spectroscopy together with chemometrics, fulfill this growing demand. The high speed providing relevant information and the versatility of its application to different types of samples lead these combined techniques as one of the most appropriated. This study is focused on the development of a calibration model able to determine amounts of API from industrial granulates using NIR, chemometrics and process spectra methodology.

X-ray photoelectron spectroscopy analysis of ion¿beam¿induced oxidation of GaAs and AlGaAs

The oxidation of GaAs and AlGaAs targets subjected to O2+ bombardment has been analyzed, using in situ x¿ray photoelectron spectroscopy, as a function of time until steady state is reached. The oxides formed by the O2+ bombardment have been characterized in terms of composition and binding energy. A strong energy and angular dependence for the oxidation of As relative to Ga is found. Low energies as well as near normal angles of incidence favor the oxidation of As. The difference between Ga and As can be explained in terms of the formation enthalpy for the oxide and the excess supply of oxygen. In an AlGaAs target the Al is very quickly completely oxidized irrespective of the experimental conditions. The steady state composition of the altered layers show in all cases a preferential removal of As.

Comparative study of the LC-MS/MS and UPLC-MS/MS for the multi-residue analysis of quinolones, penicillins and cephalosporins in cow milk, and validation according to the regulation 2002/657/EC

The aim of this study was to develop and validate an analytical method to simultaneously determine European Union-regulated beta-lactams (penicillins and cephalosporins) and quinolones in cow milk. The procedure involves a new solid phase extraction (SPE) to clean-up and pre-concentrate the three series of antibiotics before analysis by liquid chromatography¿tandem mass spectrometry (LC-MS/MS) and ultra-high-performance liquid chromatography¿tandem mass spectrometry (UPLC-MS/MS). LC-MS/MS and UPLC-MS/MS techniques were also compared. The method was validated according to the Directive 2002/657/EC and subsequently applied to 56 samples of raw cow milk supplied by the Laboratori Interprofessional Lleter de Catalunya (ALLIC) (Laboratori Interprofessional Lleter de Catalunya, Control Laboratory Interprofessional of Milk of Catalunya).

Noves metodologies d’anàlisi de compostos volàtils mitjançant trampes d’agulla. Aplicació a l’anàlisi d’alè, atmosfèrica i d’aigües

New methodologies for the analysis of volatile compoundsusing needle traps. Applications to breath, atmospheric andwater analysis. A new preconcentration technique has been developed for the analysis of volatile compounds based on the use of needle traps. These traps are based on stainless steel needles filled with one or more adsorbents, which allows the preconcentration of the anilities inside the trap by passing a gas flow through the needle. The parameters affecting the sorption/desorption process have been assessed (e.g. needle and liner dimensions, injector temperature, split less time, memory effects, and stability inside the needle). For liquid samples, four different sampling methodologies were studied, including passive and active sampling methods. The best results, considering the simplicity and sensitivity, are obtained by sampling the headspace volume using various cycles of a small and fix volume. Once the best conditions of analysis have been found, the method has been validated for gas and liquid samples. The results obtained show that needle traps are a good analytical methodology for the analysis of breath, environmental and liquid samples

Studies on the characterization of dental whitening process by using both hyperspectral imaging and colorimeter techniques

A beautiful smile is directly related with white teeth. Nowadays oral care has increased and developed processes for beautiful smiles. Dental bleaching is frequently used in odontology, not just for health care also for aesthetic treatment. With the possibility of teeth bleaching, now the importance is in, how white the tooth is? Because color is relate to an individual perception. In order to assets teeth correct color identification has been developed many color guides, models, spaces and analytical methods. Spite all of these useful tools the color interpretation depends on environmental factors, position of the sample in the data acquisition and most importantly the instrument sensitivity. The commons methods have proved to be useful. They are easy to handle, some are portable but they do not have a high sensitivity. The present work is based on the integration of a new analytical technique for color acquisition. High spectral Image (HSI) is able to performed image analysis with high quality and efficiency. HSI is used in many fields and we used it for color image analysis within the bleaching process. The main comparison was done with the HSI and the colorimeter through the processes of two different bleaching protocols. The results showed that HSI has higher sensitivity than the colorimeter. During the analysis the dental surface with the HSI we were able to notice surface changes. These changes were analyzed by roughness studies.

Acylation of chiral alcohols: a simple procedure for chiral GC analysis

The use of iodine as a catalyst and either acetic or trifluoroacetic acid as a derivatizing reagent for determining the enantiomeric composition of acyclic and cyclic aliphatic chiral alcohols was investigated. Optimal conditions were selected according to the molar ratio of alcohol to acid, the reaction time, and the reaction temperature. Afterwards, chiral stability of chiral carbons was studied. Although no isomerization was observed when acetic acid was used, partial isomerization was detected with the trifluoroacetic acid. A series of chiral alcohols of a widely varying structural type were then derivatized with acetic acid using the optimal conditions. The resolution of the enantiomeric esters and the free chiral alcohols was measured using a capillary gas chromatograph equipped with a CP Chirasil-DEX CB column. The best resolutions were obtained with 2-pentyl acetates (α = 3.00) and 2-hexyl acetates (α = 1.95). This method provides a very simple and efficient experimental workup procedure for analyzing chiral alcohols by chiral-phase GC.

Migration of antimony from polyethylene terephthalate used in mineral water bottles.

The influence of storage time and temperature on Sb migration from PET bottles into mineral water was studied in short-term tests lasting up to 15 days and long-term studies lasting up to 220 days. Samples purchased were stored in three different coloured bottles: clear (CL), light blue (LB) and dark blue (DB). Sb migration was assayed by HG-AFS for total determination and HPLC-ICP-MS for speciation analysis. Migration studies showed that waters stored at 4 and 20 oC were not subject to Sb migration. At 40 oC there was a significant increase in Sb concentration, although the maximum limit established by the European Union (5.0 ug/L) was not exceeded, whereas at 60 oC samples were subject to considerable Sb migration after 30 days of storage. In this case, the maximum limit established by the European Union was exceeded and both Sb (V) and Sb (III) were detected.

Method development for the simultaneous determination of methylmercury and inorganic mercury in seafood

This paper reports the method development for the simultaneous determination of methylmercury MeHgþ) and inorganic mercury (iHg) species in seafood samples. The study focused on the extraction and quantification of MeHgþ (the most toxic species) by liquid chromatography coupled to on-line UV irradiation and cold vapour atomic fluorescence spectroscopy (LC-UV-CV-AFS), using HCl 4 mol/L as the extractant agent. Accuracy of the method has been verified by analysing three certified reference materials and different spiked samples. The values found for total Hg and MeHgþ for the CRMs did not differ significantly from certified values at a 95% confidence level, and recoveries between 85% and 97% for MeHgþ, based on spikes, were achieved. The detection limits (LODs) obtained were 0.001 mg Hg/kg for total mercury, 0.0003 mg Hg/kg for MeHgþ and 0.0004 mg Hg/kg for iHg. The quantification limits (LOQs) established were 0.003 mg Hg/kg for total mercury, 0.0010 mg Hg/kg for MeHgþ and 0.0012 mg Hg/kg for iHg. Precision for each mercury species was established, being 12% in terms of RSD in all cases. Finally, the developed method was applied to 24 seafood samples from different origins and total mercury contents. The concentrations for Total Hg, MeHg and iHg ranged from 0.07 to 2.33, 0.003-2.23 and 0.006-0.085 mg Hg/kg, respectively. The established analytical method allows to obtain results for mercury speciation in less than 1 one hour including both, sample pretreatment and measuring step.

Analytical challenges in breath analysis and its application to exposure monitoring

There is an increasing interest in the use of breath analysis for monitoring human physiology and exposure to toxic substances or environmental pollutants. This review focuses on the current status of the sampling procedures, collection devices and sample-enrichment methodologies used for exhaled breath-vapor analysis. We discuss the different parameters affecting each of the above steps, taking into account the requirements for breath analysis in exposure assessments and the need to analyze target compounds at sub-ppbv levels. Finally, we summarize the practical applications of exposure analysis in the past two decades

Looking beneath Dalí's paint: non-destructive canvas analysis

A new analytical method was developed to non-destructively determine pH and degree of polymerisation (DP) of cellulose in fibres in 19th 20th century painting canvases, and to identify the fibre type: cotton, linen, hemp, ramie or jute. The method is based on NIR spectroscopy and multivariate data analysis, while for calibration and validation a reference collection of 199 historical canvas samples was used. The reference collection was analysed destructively using microscopy and chemical analytical methods. Partial least squares regression was used to build quantitative methods to determine pH and DP, and linear discriminant analysis was used to determine the fibre type. To interpret the obtained chemical information, an expert assessment panel developed a categorisation system to discriminate between canvases that may not be fit to withstand excessive mechanical stress, e.g. transportation. The limiting DP for this category was found to be 600. With the new method and categorisation system, canvases of 12 Dalí paintings from the Fundació Gala-Salvador Dalí (Figueres, Spain) were non-destructively analysed for pH, DP and fibre type, and their fitness determined, which informs conservation recommendations. The study demonstrates that collection-wide canvas condition surveys can be performed efficiently and non-destructively, which could significantly improve collection management.

Potencialidades e aplicações do ligante dietilditiofosfato de amônio em análises químicas

Este trabalho descreve uma revisão a respeito do uso do ligante dietilditiofosfato de amônio em química analítica e suas potencialidades no campo da análise química espectrofotométrica (UV/VIS) e espectrométrica (AAS, ICP-AES, ICP-MS) em diferentes matrizes e meios. A discriminação de espécies químicas por meio de complexações seletivas de elementos químicos com dietilditiofosfato e a hifenação de técnicas espectroscópicas a etapas de separação como extração em fase sólida, extração líquido-líquido, extração por "cloud point", são outros assuntos abordados neste trabalho.

Sensibilidad de Mycrobacterium tuberculosis a estreptomicina e isoniacida determinada por cromatografía de líquidos de alta resolución y por dilusión y siembra en placa

Tesis (Maestría en Ciencias con Especialidad en Química Analítica) UANL

Estudio químico biodirigido de la raíz de Karwinskia parvifolia

Tesis (Maestría en Ciencias con Especialidad en Química Analítica Biomédica) UANL

Desarrollo de un método analítico para la determinación de aflatoxinas M1 y B1 en fluidos biológicos empleando cromatografía de líquidos de alta resolución

Tesis (Maestría en Ciencias con Especialidad en Química Analítica Biomédica) UANL