Elsa Ramalhosa

Microwave treatment of biological samples for methylmercury determination by high performance liquid chromatography–cold vapour atomic fluorescence spectrometry

A simple and rapid microwave-assisted alkaline digestion procedure was developed in combination with high performance liquid chromatography-ultraviolet post-column oxidation-cold vapour atomic fluorescence spectrometric detection for methylmercury determination in biological tissues. Since the stability of methylmercury in methanolic potassium hydroxide solution under microwave irradiation was verified, the microwave-assisted extraction procedure was optimized in terms of quantitative recovery of methylmercury and minimum time required. The alkaline extracts were subjected to clean-up steps with dichloromethane and hydrochloric acid in order to reduce matrix interferences in methylmercury determination. The effects of matrix interferences were checked by comparison of the slopes corresponding to calibration and standard addition curves. The accuracy of the method was evaluated by the analysis of two biological certified reference materials, NRC TORT-2 and BCR 463. The results obtained by the proposed method were in good agreement with the certified values of methylmercury concentration in both materials. The detection limit was 10 microg kg(-1) and the relative standard deviation was < 8% for methylmercury concentrations ranging from 0.15 to 3.0 mg kg(-1).

Storage and export of mercury from a contaminated bay (Ria de Aveiro, Portugal)

Mercury has been determined in water, suspended particulate matter (SPM)and sediments (solid fraction and pore water) collected in a heavilycontaminated confined area (Largo do Laranjo) of the Ria de Aveiro.Mercury has been analysed in seabass (Dicentrarchus labrax) capturedin several locations of the lagoon. The levels of mercury in water and SPMin the contaminated sampled places fluctuated daily with the tide (from 10to 53 ng.dm-3 and 3.5 to 26 μg.g-1, respectively),indicating the export at the tidal rhythm. Bottom sediments exhibitedhigher concentrations, the maximum (48 μg.g-1) occurringnear the anthropogenic source at depths corresponding to the periods ofhigher industrial discharges. Mercury concentrations in pore waters (max.80 ng.dm-3) exceed the values found in the water column and werehigher at depths where Acid Volatile Sulphides (AVS) presented broadmaximums. This implies a remobilization at anoxic conditions. However theexport across the sediment-water interface predicted through moleculardiffusion is low (between 1 to 4 × 10-3 ng.cm-2.h-1) incomparison to the industrial input. The anthropogenic mercury appears tobe rapidly accumulated in seabass, a predator fish that enter seasonally inLargo do Laranjo.

The progression from a lower to a higher invasive stage of bladder cancer is associated with severe alterations in glucose and pyruvate metabolism

Cancer cells present a particular metabolic behavior. We hypothesized that the progression of bladder cancer could be accompanied by changes in cells glycolytic profile. We studied two human bladder cancer cells, RT4 and TCCSUP, in which the latter represents a more invasive stage. The levels of glucose, pyruvate, alanine and lactate in the extracellular media were measured by Proton Nuclear Magnetic Resonance. The protein expression levels of glucose transporters 1 (GLUT1) and 3 (GLUT3), monocarboxylate transporter 4 (MCT4), phosphofructokinase-1 (PFK1), glutamic-pyruvate transaminase (GPT) and lactate dehydrogenase (LDH) were determined. Our data showed that glucose consumption and GLUT3 levels were similar in both cell lines, but TCCSUP cells displayed lower levels of GLUT1 and PFK expression. An increase in pyruvate consumption, concordant with the higher levels of lactate and alanine production, was also detected in TCCSUP cells. Moreover, TCCSUP cells presented lower protein expression levels of GPT and LDH. These results illustrate that bladder cancer progression is associated with alterations in cells glycolytic profile, namely the switch from glucose to pyruvate consumption in the more aggressive stage. This may be useful to develop new therapies and to identify biomarkers for cancer progression.

Antioxidant activity and bioactive compounds of lettuce improved by espresso coffee residues

The antioxidant activity and individual bioactive compounds of lettuce, cultivated with 2.5-30% (v/v) of fresh or composted espresso spent coffee grounds, were assessed. A progressive enhancement of lettuces antioxidant capacity, evaluated by radical scavenging effect and reducing power, was exhibited with the increment of fresh spent coffee amounts, while this pattern was not so clear with composted treatments. Total reducing capacity also improved, particularly for low spent coffee concentrations. Additionally, very significant positive correlations were observed for all carotenoids in plants from fresh spent coffee treatments, particularly for violaxanthin, evaluated by HPLC. Furthermore, chlorophyll a was a good discriminating factor between control group and all spent coffee treated samples, while vitamin E was not significantly affected. Espresso spent coffee grounds are a recognised and valuable source of bioactive compounds, proving herein, for the first time, to potentiate the antioxidant pool and quality of the vegetables produced.

Optimization of mead production using Response Surface Methodology

The main aim of the present work was to optimize mead production using Response Surface Methodology. The effects of temperature (x₁: 20-30°C) and nutrients concentration (x₂: 60-120g /hL) on mead quality, concerning the final concentrations of glucose (Y₁), fructose (Y₂), ethanol (Y₃), glycerol (Y₄) and acetic acid (Y₅), were studied. Twelve operational conditions were tested. No delays and moods were observed during fermentations. The second order polynomial models determined produced satisfactory fittings of the experimental data with regard to glucose (R²=0.646, p=0.001), ethanol (R²=0.741, p=0.049), glycerol (R²=0.899, p=0.002), fructose (R²=0.902, p=0.033) and acetic acid (R²=0.913, p=0.001). The optimum extraction conditions determined in order to maximize the combined responses were 24°C and a nutrients concentration of 0.88g/L. The mead produced under these conditions had the following characteristics: ethanol concentration of 10.2%, acetic acid 0.54 g/L, glycerol 7.8 g/L, glucose 1.8 g/L and fructose 2.5 g/L. These values were in agreement with the predicted and were within the safe limit established for acetic acid and the recommended range for glycerol. Furthermore, the residual sugars concentration was also low, decreasing the possibility of occurring undesirable refermentations.

YEAST dynamics during the natural fermentation process of table olives (Negrinha de Freixo cv.).

Yeast population and dynamics associated to spontaneous fermentation of green table olives Negrinha de Freixo cv. were evaluated. Olives and brine samples were taken at different fermentation times, and yeast were enumerated by standard plate count and identified by sequencing of the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (rDNA). Saccharomyces cerevisiae was the most frequent, followed by Candida tropicalis, Pichia membranifaciens and Candida boidini, representing together 94.8% of the total isolates. Galactomyces reessii was also identified for the first time in table olives. The highest species diversity was found between 44 and 54 days of fermentation, both in brine and olive pulp. Furthermore, high similarity was observed between brine and olive pulp microbiotas. In conclusion, these results give valuable information to table olive industrials in order to achieve more knowledge on the fermentation process of this important Protected Designation of Origin product.

Microwave Heating: A Time Saving Technology or a Way to Induce Vegetable Oils Oxidation?

The use of microwave radiation for food heating was discovered unintentionally in 1945 by Dr. Percy Spencer (Osepchuk, 1984). In the following years several experiments were conducted in order to improve the technology and food application fields, being exclusively used at the industrial level, mostly for drying, baking, and thawing. The popularization of domestic microwave ovens started in the 70’s (Osepchuk, 1984) once the improvements allowed price and size reduction. Microwaves are now indispensible equipments in westerns modern kitchens. Although there is no formal definition of the frequency range for microwave radiation, these electromagnetic waves occur in the 300MHz 300GHz region. Nevertheless, and in accordance with the industrial, scientific and medical (ISM) frequency bands for noncommunication purposes, only 915 MHz and 2.45GHz are used for food applications, especially the second due to its worldwide availability. In domestic equipments, these microwaves (high frequency oscillating electric and magnetic fields) are produced inside the oven, when electrons resonate at high frequencies in an electron tube called magnetron. The electric field is created through an inner cathode and an outer anode presenting a large potential difference between both. Permanent magnets, which compose the magnetron, are responsible for the magnetic field. Once heated, the cathode releases the electrons, traveling from the cathode in an outward spiraling path, eventually making their way to the anode. As the electrons go by resonance chambers, energy is released and received as microwaves by the output antenna. The microwaves are applied to the cavity oven, where the food is heated, via waveguides (Mutyala et al., 2010). The microwaves are reflected and distributed by the stirrer fan and then reflected again by the metallic walls of the container, being absorbed by food and lead to its temperature rise (Fig. 1). This capability to absorb microwave energy is governed by food dielectric properties, and involves primarily two mechanisms: dipolar relaxation and ionic conduction. Food water is often the primary component responsible for dielectric heating. Because of its dipolar nature, water molecules tend to re-orientate with the high frequencies

Monomethylmercury behaviour in sediments collected from a mercury-contaminated lagoon

Surface sediments and sediment cores were collected in a mercury (Hg)-contaminated lagoon, namely Largo do Laranjo – Ria de Aveiro, Portugal and analysed in order to establish the monomethylmercury (MeHg) behaviour in this kind of environment. In surface sediments, this compound was only detected in one place (13.2 ng g−1 (dry weight)). In this site, it was determined one of the lowest redox potentials (22 mV), indicative of oxic/anoxic conditions, which favours Hg methylation by enhancing the sulphate-reducing bacteria activity. However, the MeHg percentage obtained was low, namely less than 0.1% of the total Hg. This is probably due to Hg deposition with organic matter and iron oxyhydroxides, decreasing Hg availability to methylation. At the deeper layers, MeHg was also determined, reaching 46.4 ng g−1 (dry weight) and representing less than 0.1% of the total Hg. The higher MeHg percentages were observed near the surface, where Hg seems to be faster methylated as a result of the lower sulphide concencentrations that render bioavailable the inorganic Hg. At depth the low MeHg percentages obtained are due to the formation of HgS and to the adsorption of Hg to iron monosulphides.

Effect of High Hydrostatic Pressure (HHP) Treatment on Edible Flowers’ Properties

Edible flowers are increasing worldwide because they can improve the appearance, taste, and aesthetic value of food, aspects that the consumer appreciates. However, some of these are highly perishable and have a short shelf-life. To overcome these problems, high hydrostatic pressure (HHP) food processing might be applied, allowing producing high-quality food with enhanced safety and increased shelf-life. The application of HHP to vegetables has been extensively discussed and is already an industrial reality, but information on edible flowers is scarce and incomplete. Thus, in order to summarize the current knowledge on potential applications of HHP treatment in edible flowers and to determine the effect of this treatment on physical (e.g., color and texture) and nutritional characteristics as well as on microbial and enzymatic inactivation, a literature review was performed. It was stated that broccoli and cauliflower (inflorescences, usually not considered by consumers as flowers) have been the most studied, existing few information for other edible flowers. Thus, much more works are needed to better understand the effect and mechanisms behind HHP, and to define the adequate technological conditions for each flower.

Influence of sweetness and ethanol content on mead acceptability.

Abstract Mead is a traditional alcoholic beverage obtained by fermenting mead wort; however, its production still remains frequently an empirical exercise. Different meads can be produced, depending on fermentation conditions. Nevertheless, to date few studies have been developed on factors that may influence mead quality. The main objective of this work was to study the influence of sweetness and ethanol content on mead acceptability. Different meads were produced with two sweetness levels (sweet and dry meads) and three ethanol contents (18, 20, 22% (v/v)), adjusted by brandy addition. Afterwards, meads acceptability was evaluated by sensory analysis through a consumers’ panel (n=108) along with chemical analysis by HPLC-RID of glucose, fructose, ethanol, glycerol and acetic acid. The sweet (75 gglucose+fructose/L) and dry (23 gglucose+fructose/L) meads presented glycerol contents equal to 5.10±0.54 and 5.96±0.95 g/L, respectively, that were desirable since glycerol improves mead quality. Low concentrations of acetic acid were determined (0.46±0.08 and 0.57±0.09 g/L), avoiding the vinegar off-character. Concerning sensory analysis, the alcohol content of mead had no effect on the sensory attributes studied, namely, aroma, sweetness, flavour, alcohol feeling and general appreciation. Regarding sweetness, the “sweet meads” were the most appreciated by the consumers (score of 5.4±2.56), whereas the “dry meads” (score of 2.7±2.23) showed low acceptability. In conclusion, this work revealed that sweetness is a sensory key attribute for mead acceptance by the consumers, whereas ethanol content (18 to 22% (v/v)) is not.