Evaluation of an Improved Indirect Method for the Analysis of 3-MCPD Esters Based on Acid Transesterification

The quantification of 3-chloropropane-1,2-diol (3-MCPD) esters in fat-based matrices is currently carried out according to a number of methods that involve either the conversion of all the esters to the free form that is then quantified (indirect methods), or the separation and quantification of the individual esters (direct methods). Indirect methods of analysis show a better sensitivity, however, the series of chemical reactions that take place during sample preparation may affect the reliability of the results because of the potential ex-novo formation of 3-MCPD from precursors present in the sample. This study is focused on the evaluation of the selectivity and robustness of an indirect acid-catalysed method of 3-MCPD esters analysis. The interference of chloride ions and glycidyl esters was evaluated. 3-MCPD esters are overestimated only when high levels of chloride ions (>1.7 mmol/kg oil) were added to the samples. The interference by chloride ions can be easily eliminated by a single extraction step of the samples before analysis. In contrast, glycidyl esters did not interfere with the determination of 3-MCPD esters. Further investigation on the robustness of the method showed that the time allowed for the transesterification, a major drawback of the previous version of the method, can be reduced from 16 to 4 h without any significant reduction of the accuracy and repeatability (RSD = 0.7%).     

Polymer anion-selective membranes for electrolytic splitting of water. Part II: Enhancement of ionic conductivity and performance under conditions of alkaline water electrolysis

An attempt was made to increase the ionic conductivity of novel, heterogeneous, anion-selective membranes by increasing the porosity of their surface skin. This was based on the addition of a water-soluble component, namely poly(ethylene-ran-propylene glycol), to an inert polymer matrix, based on low-density polyethylene, while mixing it with the ion-exchange particles. A series of membranes was prepared, consisting of 66 wt% of anion-exchange phase represented by a styrene-divinyl benzene copolymer matrix with quaternary ammonium functional groups and an inert polymer matrix in a mixture with variable amounts of water-soluble component added. The membranes were subsequently tested with respect to their morphology, mechanical properties, apparent ion-exchange capacity, ionic conductivity, and performance under conditions of alkaline water electrolysis. When added in the appropriate amount, the addition of a water-soluble component was found to improve the electrochemical properties of the resulting membrane efficiently, while at the same time not reducing its mechanical properties to below a critical level.     

Determination of Total Carbohydrate Content in Beer Using Its Pre-column Enzymatic Cleavage and HPLC-RI

Beside ethanol, carbohydrates are the main source of total energy in beer. While analyses of fermentable carbohydrates are important from the technological point of view, the total content of carbohydrates is relevant in terms of nutrition. A high-performance liquid chromatography (HPLC) method with refractometric (RI) detection was developed for determination of total carbohydrate content in beer. Using enzymatic reaction with amyloglucosidase, the carbohydrates were cleaved to yield glucose and short glucose oligomers of less than 10 units, and separated on HPLC ion exchanger Rezex RSO-Oligosaccharide column in Ag⁺ mode. Optimum parameters were established for the enzymatic sample treatment and for the HPLC separation of reaction products. Calibration curves of glucose, fructose, maltose and simultaneously analyzed glycerol ranged from 0.001 to 0.5 g/100 ml, correlation coefficients of all calibration curves were 0.9999. The instrumental limits of quantification were 0.001 g/100 ml and they were verified using repetitive injections, with coefficient of variation (CV) less than 10 % in five replicates. The method limit of quantification was 0.01 g/100 ml since it was necessary to dilute the beer samples before chromatographic analysis. Recovery of the method in non-alcoholic and alcoholic beer was 98.5 %, and 92.3 %, respectively. Finally, ten non-alcoholic and 15 alcoholic beers from Czech market were analyzed using the method, the average content of total carbohydrates in non-alcoholic and alcoholic beers being 4.21 and 3.70 g/100 ml, respectively. These results are in a good correlation with the real extract of beer, which is on average 4.58 and 4.27 g/100 ml.     

Design and control of a demand flow system assuring spontaneous breathing of a patient connected to an HFO ventilator

Lung protective ventilation is intended to minimize the risk of ventilator induced lung injury and currently aimed at preservation of spontaneous breathing during mechanical ventilation. High-frequency oscillatory ventilation (HFOV) is a lung protective ventilation strategy. Commonly used high-frequency oscillatory (HFO) ventilators, SensorMedics 3100, were not designed to tolerate spontaneous breathing. Respiratory efforts in large pediatric and adult patients impose a high workload to the patient and may cause pressure swings that impede ventilator function. A Demand Flow System (DFS) was designed to facilitate spontaneous breathing during HFOV. Using a linear quadratic Gaussian state feedback controller, the DFS alters the inflow of gas into the ventilator circuit, so that it instantaneously compensates for the changes in mean airway pressure (MAP) in the ventilator circuit caused by spontaneous breathing. The undesired swings in MAP are thus eliminated. The DFS significantly reduces the imposed work of breathing and improves ventilator function. In a bench test the performance of the DFS was evaluated using a simulator ASL 5000. With the gas inflow controlled, MAP was returned to its preset value within 115 ms after the beginning of inspiration. The DFS might help to spread the use of HFOV in clinical practice.     

Comparison of HPLC and GLC methodologies for determination of glucosinolates using reference material

 The reference material rapeseed of known glucosinolate composition, was analysed by HPLC and GLC and two commercially available glucosinolates (glucotropaeolin and sinigrin) were used as internal standards. The HPLC method enabled determination of 11 different glucosinolates (as desulphoglucosinolates) that occur in the rapeseed. Only seven glucosinolates (as trimethylsilylated desulphoglucosinolates) were separated by GLC. The latter method did not allow the determination of methylsulphinylalkyl glucosinolates (glucoiberin, glucoraphanin and glucoallysin) and did not separate optical isomers of 2-hydroxy-3-butenyl glucosinolate (progoitrin and epi-progoitrin). Statistical evaluation of data (t-test, F-test) revealed no significant differences between the tested methods at the 95% confidence level. The advantages and disadvantages of both widely used chromatographic methods are discussed. Received: 26 May 1997 / Revised version: 11 July 1997     

Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis

Sucrose phosphorylase is a promising biocatalyst for the glycosylation of a wide variety of acceptor molecules, but its low thermostability is a serious drawback for industrial applications. In this work, the stability of the enzyme from Bifidobacterium adolescentis has been significantly improved by a combination of smart and rational mutagenesis. The former consists of substituting the most flexible residues with amino acids that occur more frequently at the corresponding positions in related sequences, while the latter is based on a careful inspection of the enzyme's crystal structure to promote electrostatic interactions. In this way, a variant enzyme could be created that contains six mutations and whose half-life at the industrially relevant temperature of 60 °C has more than doubled compared with the wild-type enzyme. An increased stability in the presence of organic co-solvents could also be observed, although these effects were most noticeable at low temperatures.     

<Emphasis Type="Italic">N</Emphasis>-Acylated Bacteriohopanehexol-Mannosamides from the Thermophilic Bacterium <Emphasis Type="Italic">Alicyclobacillus acidoterrestris</Emphasis>

Identification of molecular species of various N-acylated bacteriohopanehexol-mannosamides from the thermophilic bacterium Alicyclobacillus acidoterrestris by semipreparative HPLC and by RP-HPLC with ESI is described. We used triple-quadrupole type mass spectrometer, ¹H and ¹³C NMR for analyzing this complex lipid. CD spectra of two compounds (model compound—7-deoxy-d-glycero-d-allo-heptitol obtained by stereospecific synthesis, and an isolated derivative of hopane) were also measured and the absolute configuration of both compounds was determined. On the basis of all the above methods, we identified the full structure of a new class of bacteriohopanes, represented by various N-acylated bacteriohopanehexol-mannosamides.     

Comparative impact assessment for flax fibre versus conventional glass fibre reinforced composites: Are bio-based reinforcement materials the way to go?

In many applications the use of composite materials can offer significant weight reduction opportunities, which can have a positive influence on the life cycle impact of a component or system primarily through energy saving effects in the use phase. The impact associated with the production and end-of-life (EOL) phases, however, forms a possible counter indication for systematic replacement of conventional structures by composite solutions.     

The thermal protection of a specific experimental instrument for monitoring of combustion conditions on the grate of municipal solid waste incinerators

The thermal protection of the specific experimental instrument for monitoring of combustion conditions on the grate of municipal solid waste incinerators (MSWI) represents a very important part of the assembled measuring system. The inner part of the instrument with control electronics and diverse sensors (temperature and flue gas concentration measurements) requires sufficient thermal protection against the high temperature environment of the combustion process with a surrounding temperature higher than 1200 °C. The influence of the high temperature environment is limited, but not fully eliminated, by thermal protection. The thermal protection is applied in two different ways. The standard thermal insulation with low thermal conductivity represents passive protection. The optimal thickness of the insulation itself partially defines the heat flux to the central part of the measuring system and its impacts upon increase of the inner temperature. The second type of protection, in an active mode, in addition to low conductivity, also provides heat storage. The full-scale thermal protection is designed by means of computer calculations with boundary and initial conditions or laboratory measurements within a similar environment to that on the grate of the MSWI. An additional aim of the sufficient thermal protection development is also to minimize the final size of the experimental instrument. The experimental trials have approved the functionality of the measuring instrument in high temperatures by keeping the inner temperature at the very low level.     

Silica Nanoparticles Inhibit Responses to ATP in Human Airway Epithelial 16HBE Cells

Because of their low cost and easy production, silica nanoparticles (SiNPs) are widely used in multiple manufacturing applications as anti-caking, densifying and hydrophobic agents. However, this has increased the exposure levels of the general population and has raised concerns about the toxicity of this nanomaterial. SiNPs affect the function of the airway epithelium, but the biochemical pathways targeted by these particles remain largely unknown. Here we investigated the effects of SiNPs on the responses of 16HBE14o- cultured human bronchial epithelial (16HBE) cells to the damage-associated molecular pattern ATP, using fluorometric measurements of intracellular Ca²⁺ concentration. Upon stimulation with extracellular ATP, these cells displayed a concentration-dependent increase in intracellular Ca²⁺, which was mediated by release from intracellular stores. SiNPs inhibited the Ca²⁺ responses to ATP within minutes of application and at low micromolar concentrations, which are significantly faster and more potent than those previously reported for the induction of cellular toxicity and pro-inflammatory responses. SiNPs-induced inhibition is independent from the increase in intracellular Ca²⁺ they produce, is largely irreversible and occurs via a non-competitive mechanism. These findings suggest that SiNPs reduce the ability of airway epithelial cells to mount ATP-dependent protective responses.