J. Fontecha

Major lipid classes separation of buttermilk, and cows, goats and ewes milk by high performance liquid chromatography with an evaporative light scattering detector focused on the phospholipid fraction.

An improved HPLC-ELSD method has been developed for the analysis of the lipid classes of buttermilk and milk from different species, focused in the phospholipids fraction without a prior fractionation step and in a single run. The total lipid profile analysis showed the major and minor lipid compounds as cholesterol esters, triacylglycerides, cholesterol, diacylglycerides, free fatty acids, monoacylglycerides, and also the polar compounds as glucosylceramide, lactosylceramide, phosphatidyl-ethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, sphingomyelin and lysophosphatidylcholine. The identification and quantification of the different compounds, using calibration curves made with individual standards and the low coefficients of variation obtained in the inter- and intra-assays showed the suitability of the developed method. In this study, we optimized and validated a quantitative HPLC-ELSD method at a concentration level suitable for routine analysis of the major lipid classes in milk and dairy products.

Use of β-cyclodextrin to decrease the level of cholesterol in milk fat

This study was carried out to determine optimum conditions (beta-cyclodextrin concentration, mixing time, and holding time) for cholesterol removal from pasteurized nonhomogenized milk at 4 degrees C on a commercial scale by adding beta-cyclodextrin in a specially designed bulk mixer tank. The beta-cyclodextrin (0.4, 0.6, 0.8, and 1.0%) removed from 65.42 to 95.31% of cholesterol at 4 degrees C in 20 min. Treatment of milk with 0.8 and 1.0% (wt/vol) beta-cyclodextrin was no better than treatment with 0.6% beta-cyclodextrin. Maximum cholesterol removal was seen with 6 h of treatment. The beta-cyclodextrin cholesterol complex was precipitated from milk during 20 min without stirring at 4 degrees C and removed by centrifugation. After separating the milk, approximately 0.35% of residual beta-cyclodextrin remained in the skim fraction and 0.1% in the cream from milk treated with 0.6% beta-cyclodextrin. The rest of the beta-cyclodextrin was complexed with the cholesterol and eliminated via the discharger of the separator. Individual fatty acid and triglyceride compositions did not differ between control milk and milk treated with 0.6% beta-cyclodextrin.

Total milk fat extraction and quantification of polar and neutral lipids of cow, goat, and ewe milk by using a pressurized liquid system and chromatographic techniques

Although milk polar lipids such as phospholipids and sphingolipids located in the milk fat globule membrane constitute 0.1 to 1% of the total milk fat, those lipid fractions are gaining increasing interest because of their potential beneficial effects on human health and technological properties. In this context, the accurate quantification of the milk polar lipids is crucial for comparison of different milk species, products, or dairy treatments. Although the official International Organization for Standardization-International Dairy Federation method for milk lipid extraction gives satisfactory results for neutral lipids, it has important disadvantages in terms of polar lipid losses. Other methods using mixtures of solvents such as chloroform:methanol are highly efficient for extracting polar lipids but are also associated with low sample throughput, long time, and large solvent consumption. As an alternative, we have optimized the milk fat extraction yield by using a pressurized liquid extraction (PLE) method at different temperatures and times in comparison with those traditional lipid extraction procedures using 2:1 chloroform:methanol as a mixture of solvents. Comparison of classical extraction methods with the developed PLE procedure were carried out using raw whole milk from different species (cows, ewes, and goats) and considering fat yield, fatty acid methyl ester composition, triacylglyceride species, cholesterol content, and lipid class compositions, with special attention to polar lipids such as phospholipids and sphingolipids. The developed PLE procedure was validated for milk fat extraction and the results show that this method performs a complete or close to complete extraction of all lipid classes and in less time than the official and Folch methods. In conclusion, the PLE method optimized in this study could be an alternative to carry out milk fat extraction as a routine method.

Differentiation of red wines using an electronic nose based on surface acoustic wave devices

An electronic nose, utilizing the principle of surface acoustic waves (SAW), was used to differentiate among different wines of the same variety of grapes which come from the same cellar. The electronic nose is based on eight surface acoustic wave sensors, one is a reference sensor and the others are coated by different polymers by spray coating technique. Data analysis was performed by two pattern recognition methods; principal component analysis (PCA) and probabilistic neuronal network (PNN). The results showed that electronic nose was able to identify the tested wines.

Real-Time Characterization of Electrospun PVP Nanofibers as Sensitive Layer of a Surface Acoustic Wave Device for Gas Detection

The goal of this work has been to study the polyvinylpyrrolidone (PVP) fibers deposited by means of the electrospinning technique for using as sensitive layer in surface acoustic wave (SAW) sensors to detect volatile organic compounds (VOCs). The electrospinning process of the fibers has been monitored and RF characterized in real time, and it has been shown that the diameters of the fibers depend mainly on two variables: the applied voltage and the distance between the needle and the collector, since all the electrospun fibers have been characterized by a scanning electron microscopy (SEM). Real-time measurement during the fiber coating process has shown that the depth of penetration of mechanical perturbation in the fiber layer has a limit. It has been demonstrated that once this saturation has been reached, the increase of the thickness of the fibers coating does not improve the sensitivity of the sensor. Finally, the parameters used to deposit the electrospun fibers of smaller diameters have been used to deposit fibers on a SAW device to obtain a sensor to measure different concentrations of toluene at room temperature. The present sensor exhibited excellent sensitivity, good linearity and repeatability, and high and fast response to toluene at room temperature.

Array of Love-wave sensors based on quartz/Novolac to detect CWA simulants.

An array of Love-wave sensors based on quartz and Novolac has been developed to detect chemical warfare agents (CWAs). These weapons are a risk for human health due to their efficiency and high lethality; therefore an early and clear detection is of enormous importance for the people safety. Love-wave devices realized on quartz as piezoelectric substrate and Novolac as guiding layer have been used to make up an array of six sensors, which have been coated with specific polymers by spin coating. The CWAs are very dangerous and for safety reasons their well known simulants have been used: dimethylmethyl phosphonate (DMMP), dipropyleneglycol methyl ether (DPGME), dimethylmethyl acetamide (DMA), dichloroethane (DCE), dichloromethane (DCM) and dichloropentane (DCP). The array has been exposed to these CWA simulants detecting very low concentrations, such as 25 ppb of DMMP, a simulant of nerve agent sarin. Finally, principal component analysis (PCA) as data pre-processing and discrimination technique, and probabilistic neural networks (PNN) as patterns classification technique have been applied. The performance of the sensor array has shown stability, accuracy, high sensitivity and good selectivity to these simulants.

Graphene oxide as sensitive layer in Love-wave surface acoustic wave sensors for the detection of chemical warfare agent simulants

A Love-wave device with graphene oxide (GO) as sensitive layer has been developed for the detection of chemical warfare agent (CWA) simulants. Sensitive films were fabricated by airbrushing GO dispersions onto Love-wave devices. The resulting Love-wave sensors detected very low CWA simulant concentrations in synthetic air at room temperature (as low as 0.2 ppm for dimethyl-methylphosphonate, DMMP, a simulant of sarin nerve gas, and 0.75 ppm for dipropylene glycol monomethyl ether, DPGME, a simulant of nitrogen mustard). High responses to DMMP and DPGME were obtained with sensitivities of 3087 and 760 Hz/ppm respectively. Very low limit of detection (LOD) values (9 and 40 ppb for DMMP and DPGME, respectively) were calculated from the achieved experimental data. The sensor exhibited outstanding sensitivity, good linearity and repeatability to all simulants tested. The detection mechanism is here explained in terms of hydrogen bonding formation between the tested CWA simulants and GO.

Characterization of an array of Love-wave gas sensors developed using electrospinning technique to deposit nanofibers as sensitive layers.

The electrospinning technique has allowed that very different materials are deposited as sensitive layers on Love-wave devices forming a low cost and successful sensor array. Their excellent sensitivity, good linearity and short response time are reported in this paper. Several materials have been used to produce the nanofibers: polymers as Polyvinyl alcohol (PVA), Polyvinylpyrrolidone (PVP) and Polystirene (PS); composites with polymers as PVA+SnCl4; combined polymers as PS+Poly(styrene-alt-maleic anhydride) (PS+PSMA) and metal oxides (SnO2). In order to test the array, well-known chemical warfare agent simulants (CWAs) have been chosen among the volatile organic compounds due to their importance in the security field. Very low concentrations of these compounds have been detected by the array, such as 0.2 ppm of DMMP, a simulant of sarin nerve gas, and 1 ppm of DPGME, a simulant of nitrogen mustard. Additionally, the CWA simulants used in the experiment have been discriminated and classified using pattern recognition techniques, such as principal component analysis and artificial neural networks.

Love-Wave Sensors Combined with Microfluidics for Fast Detection of Biological Warfare Agents

The following paper examines a time-efficient method for detecting biological warfare agents (BWAs). The method is based on a system of a Love-wave immunosensor combined with a microfluidic chip which detects BWA samples in a dynamic mode. In this way a continuous flow-through of the sample is created, promoting the reaction between antigen and antibody and allowing a fast detection of the BWAs. In order to prove this method, static and dynamic modes have been simulated and different concentrations of BWA simulants have been tested with two immunoreactions: phage M13 has been detected using the mouse monoclonal antibody anti-M13 (AM13), and the rabbit immunoglobulin (Rabbit IgG) has been detected using the polyclonal antibody goat anti-rabbit (GAR). Finally, different concentrations of each BWA simulants have been detected with a fast response time and a desirable level of discrimination among them has been achieved.

Nanocrystalline Tin Oxide Nanofibers Deposited by a Novel Focused Electrospinning Method. Application to the Detection of TATP Precursors

A new method of depositing tin dioxide nanofibers in order to develop chemical sensors is presented. It involves an electrospinning process with in-plane electrostatic focusing over micromechanized substrates. It is a fast and reproducible method. After an annealing process, which can be performed by the substrate heaters, it is observed that the fibers are intertwined forming porous networks that are randomly distributed on the substrate. The fiber diameters oscillate from 100 nm to 200 nm and fiber lengths reach several tens of microns. Each fiber has a polycrystalline structure with multiple nano-grains. The sensors have been tested for the detection of acetone and hydrogen peroxide (precursors of the explosive triacetone triperoxide, TATP) in air in the ppm range. High and fast responses to these gases have been obtained.