Analysis of hemoglobin-based oxygen carriers by CE-UV/Vis and CE-ESI-TOF/MS.

Blood doping involves the use of products that enhance the uptake, transport, or delivery of oxygen to the blood. One approach uses artificial oxygen carriers, known as hemoglobin-based oxygen carriers (HBOCs). This study describes an analytical strategy based on CE for detecting intact HBOCs in plasma samples collected for doping control. On-capillary detection was performed by UV/Vis at 415 nm, which offered detection selectivity for hemoproteins (such as hemoglobin and HBOCs). On-line ESI-MS detection with a TOF analyzer was further used to provide accurate masses on CE peaks and to confirm the presence of HBOCs. An immunodepletion sample preparation step was mandatory prior to analysis, in order to remove most abundant proteins that interfered with CE separation and altered the ESI process. This analytical method was successfully applied to plasma samples enriched with Oxyglobin, a commercially available HBOC used for veterinary purposes. Detection limits of 0.20 and 0.45 g/dL were achieved in plasma for CE-UV/Vis at 415 nm and CE-ESI-TOF/MS, respectively.

CE-ESI-TOF/MS for human growth hormone analysis.

CE is a powerful analytical tool used to separate intact biomolecules such as proteins. The coupling of CE with TOF/MS produces a very promising method that can be used to detect and identify proteins in different matrices. This paper describes an efficient, rapid, and simple CE-ESI-TOF/MS procedure for the analysis of endogenous human growth hormone and recombinant human growth hormone without sample preparation. Operational factors were optimized using an experimental design, and the method was successfully applied to distinguish human growth hormone and recombinant human growth hormone in unknown samples.

Development of a two-step screening ESI-TOF-MS method for rapid determination of significant stress-induced metabolome modifications in plant leaf extracts: the wound response in Arabidopsis thaliana as a case study.

To study the stress-induced effects caused by wounding under a new perspective, a metabolomic strategy based on HPLC-MS has been devised for the model plant Arabidopsis thaliana. To detect induced metabolites and precisely localise these compounds among the numerous constitutive metabolites, HPLC-MS analyses were performed in a two-step strategy. In a first step, rapid direct TOF-MS measurements of the crude leaf extract were performed with a ballistic gradient on a short LC-column. The HPLC-MS data were investigated by multivariate analysis as total mass spectra (TMS). Principal components analysis (PCA) and hierarchical cluster analysis (HCA) on principal coordinates were combined for data treatment. PCA and HCA demonstrated a clear clustering of plant specimens selecting the highest discriminating ions given by the complete data analysis, leading to the specific detection of discrete-induced ions (m/z values). Furthermore, pool constitution with plants of homogeneous behaviour was achieved for confirmatory analysis. In this second step, long high-resolution LC profilings on an UPLC-TOF-MS system were used on pooled samples. This allowed to precisely localise the putative biological marker induced by wounding and by specific extraction of accurate m/z values detected in the screening procedure with the TMS spectra.

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

A simple, robust and rapid approach to detect carbapenemases in Gram-negative isolates by MALDI-TOF mass spectrometry: validation with triple quadripole tandem mass spectrometry, microarray and PCR.

Carbapenemases should be accurately and rapidly detected, given their possible epidemiological spread and their impact on treatment options. Here, we developed a simple, easy and rapid matrix-assisted laser desorption ionization-time of flight (MALDI-TOF)-based assay to detect carbapenemases and compared this innovative test with four other diagnostic approaches on 47 clinical isolates. Tandem mass spectrometry (MS-MS) was also used to determine accurately the amount of antibiotic present in the supernatant after 1 h of incubation and both MALDI-TOF and MS-MS approaches exhibited a 100% sensitivity and a 100% specificity. By comparison, molecular genetic techniques (Check-MDR Carba PCR and Check-MDR CT103 microarray) showed a 90.5% sensitivity and a 100% specificity, as two strains of Aeromonas were not detected because their chromosomal carbapenemase is not targeted by probes used in both kits. Altogether, this innovative MALDI-TOF-based approach that uses a stable 10-μg disk of ertapenem was highly efficient in detecting carbapenemase, with a sensitivity higher than that of PCR and microarray.

Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology.

Until recently, microbial identification in clinical diagnostic laboratories has mainly relied on conventional phenotypic and gene sequencing identification techniques. The development of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) devices has revolutionized the routine identification of microorganisms in clinical microbiology laboratories by introducing an easy, rapid, high throughput, low-cost, and efficient identification technique. This technology has been adapted to the constraint of clinical diagnostic laboratories and has the potential to replace and/or complement conventional identification techniques for both bacterial and fungal strains. Using standardized procedures, the resolution of MALDI-TOF MS allows accurate identification at the species level of most Gram-positive and Gram-negative bacterial strains with the exception of a few difficult strains that require more attention and further development of the method. Similarly, the routine identification by MALDI-TOF MS of yeast isolates is reliable and much quicker than conventional techniques. Recent studies have shown that MALDI-TOF MS has also the potential to accurately identify filamentous fungi and dermatophytes, providing that specific standardized procedures are established for these microorganisms. Moreover, MALDI-TOF MS has been used successfully for microbial typing and identification at the subspecies level, demonstrating that this technology is a potential efficient tool for epidemiological studies and for taxonomical classification.

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Secondary Ion Mass Spectrometry (SIMS): principles and applications

This article outlines the basis of the technique and shows some examples of applications in order to exhibit the expectations of this technique invaried scientific fields.

Analysis of raw hams using SELDI-TOF-MS to predict the final quality of dry-cured hams

The relationship between protein profiles of Gluteus medius (GM) muscles of raw hams obtained from 4 pure breed pigs (Duroc, Large White, Landrace, and Piétrain) with the final quality of the Semimembranosus and Biceps femoris muscles of dry-cured hams was investigated. As expected, Duroc hams showed higher levels of marbling and intramuscular fat content than the other breeds. Piétrain hams were the leanest and most conformed, and presented the lowest salt content in dry-cured hams. Even if differences in the quality traits (colour, water activity, texture, composition, intramuscular fat, and marbling) of dry-cured hams were observed among the studied breeds, only small differences in the sensory attributes were detected. Surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS) was used to obtain the soluble protein profiles of GM muscles. Some associations between protein peaks obtained with SELDI-TOF-MS and quality traits, mainly colour (b*) and texture (F0, Y2, Y90) were observed. Candidate protein markers for the quality of processed dry-cured hams were identified

UPLC-TOF-MS for plant metabolomics: a sequential approach for wound marker analysis in Arabidopsis thaliana.

The model plant Arabidopsis thaliana was studied for the search of new metabolites involved in wound signalling. Diverse LC approaches were considered in terms of efficiency and analysis time and a 7-min gradient on a UPLC-TOF-MS system with a short column was chosen for metabolite fingerprinting. This screening step was designed to allow the comparison of a high number of samples over a wide range of time points after stress induction in positive and negative ionisation modes. Thanks to data treatment, clear discrimination was obtained, providing lists of potential stress-induced ions. In a second step, the fingerprinting conditions were transferred to longer column, providing a higher peak capacity able to demonstrate the presence of isomers among the highlighted compounds.

INFLUÊNCIA DE DIVERSOS SUBSTRATOS NO DESENVOLVIMENTO DE MUDAS DE MARACUJAZEIRO AZEDO (Passiflora edulis Sims f. flavicarpa DEG)

Objetivando avaliar a influência de diversos substratos no desenvolvimento de mudas de maracujazeiro-azedo, conduziu-se um experimento em casa de vegetação da Emprapa Cerrados. Utilizou-se o delineamento experimental de blocos casualizados, em esquema fatorial 2 x 4 x 3 x 2, totalizando 48 tratamentos, 18 plantas úteis por parcela e 4 repetições. Os tratamentos constituíram-se das combinações de: dois substratos comerciais (PlantmaxR -- à base de vermiculita mais casca de Pinus sp e VermiculitaR); três fontes orgânicas (f.o.) (húmus, esterco de curral e NutriplantaR (produto à base de bactérias) e ausência de f.o., na proporção de 3:1 do substrato básico para a f.o.); duas formulações de adubo [OsmocoteR na fórmula 14-14-14 (produto de lenta liberação de nutrientes) e 4-14-8 (de liberação normal)], além da ausência de adubo; e Glomus etunicatum, ausência e presença. O substrato comercial PlantmaxR foi superior à VermiculitaR em todas as características analisadas. Dentre as f.o., o NutriplantaR junto com o esterco proporcionaram o melhor desempenho. O OsmocoteR promoveu o maior desenvolvimento das mudas, seguido pelo 4-14-8. A presença ou ausência de f.o. combinada com PlantmaxR praticamente não influenciou nas características analisadas. Não se deve utilizar o Glomus etunicatum associado a PlantmaxR, devido ao alto teor de fósforo presente neste substrato.

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Los adhesivos empleados en el entelado de carteles modernos. Uso de la Espectrometría de Masas de Ionización/Desorción por Láser asistida por Matriz y analizador de Tiempo de Vuelo (MALDI-TOF-MS) para la identificación de éteres de celulosa

"XVIII Congreso Internacional de Conservación y Restauración de Bienes Culturales - 18th International Meeting on Heritage Conservation", Granada 9 al 11 novembre de 2011

Efeito dos substratos artificiais no enraizamento e no desenvolvimento de estacas de maracujazeiro-azedo (Passiflora edulis Sims f. flavicarpa Deg)

Os ensaios foram instalados em casa de vegetação na Embrapa Cerrados, Planaltina-DF, em agosto de 1999. Nos primeiros 30 dias, estacas da cultivar MSC (Marília Seleção Cerrados) foram mantidas sob nebulização com duas regas diárias. Avaliaram-se os seguintes parâmetros: percentagens de estacas brotadas, enraizadas, com fungos, peso de matéria fresca e seca da parte aérea e das raízes, aos 90 dias após o plantio. O estudo foi composto de dois experimentos. No primeiro, utilizaram-se estacas coletadas em agosto de 1999, e oito tipos de substratos. O Plantmax Florestal EstacaÒ proporcionou maior brotação e enraizamento, e ainda menorincidência de estacas contaminadas por Colletotrichum gloeosporioides, comparativamente ao substrato à base de Plantmax Hortaliças.Ò O aditivo NutriplantaÒ quando combinado com o fertilizante OsmocoteÒ, no substrato à base de Plantmax Florestal EstacaÒ, proporcionou maior brotação e enraizamento e também menor incidência de estacas contaminadas com C. gloeosporioides. No segundo ensaio, utilizaram-se estacas do híbrido RC1 ((F1: Marília x Roxo Australiano) x Marília), coletadas em agosto de 2000, tratadas com 2.000 ppm de AIB, plantadas em bandejas de poliestireno expandido com doze tipos de substratos. Para enraizamento das estacas do Híbrido RC1, não houve diferença entre os substratos testados. A adição de um g de Osmocote® por estaca, aos 30 dias após o plantio no substrato Plantmax Florestal Estaca® , proporcionou o melhor desenvolvimento da parte aérea das estacas do Híbrido RC1.