Preparation and evaluation of carriers for detection of cholinesterase inhibitors.

OBJECTIVES: The aim of the study was to use methods of pharmaceutical technology, and prepare carriers in the form of pellets suitable as a filling of detection tubes for enzymatic detection of cholinesterase inhibitors. The enzymatic detection was based on enzymatic hydrolysis of acetylthiocholine iodide and the subsequent colour reaction of its hydrolysis product with Ellman's reagent. The suitable carriers should be in the form of white, regular and sufficiently mechanically resistant particles of about 1 mm allowing it to capture the enzyme during the impregnation process and ensuring its high activity for enzymatic detection. METHODS: Carriers consisting of microcrystalline cellulose, lactose, povidone, and sodium carboxymethyl cellulose were prepared using extrusion-spheronization method under three different drying conditions in either a hot air oven or a microwave oven. Subsequently, the carriers were impregnated with acetylcholinesterase and their size, shape, mechanical resistance, bulk, tapped and pycnometric density, Hausner ratio, intraparticular and total tapped porosity, and activity were measured and recorded. RESULTS: In this procedure, carriers with different physical parameters and different acetylcholinesterase activity were evaluated. It was found that higher acetylcholinesterase activity was associated not only with a higher intraparticular porosity but also with more regular particles characterized by high sphericity and low total tapped porosity. CONCLUSION: This unique finding is important for the preparation of detection tubes based on enzymatic detection which is still irreplaceable especially in the field of detection and analysis of super-toxic cholinesterase inhibitors.

Electrochemical Corrosion Tests in Low-Conductivity Ethanol-Gasoline Blends: Application of Supporting Electrolyte for Contaminated E5 and E10 Fuels.

Ethanol-based E5 and E10 fuels have extensively been used as automotive fuels in gasoline engines. However, especially when contaminated, these fuels can exhibit corrosion effects on some engine construction parts such as mild steel. Thus, the study of mild steel corrosion resistance has become of the utmost importance. Electrochemical methods such as electrochemical impedance spectroscopy (EIS) and polarization characteristics measurements (Tafel scan-TS) were proven to be very valuable in studying the mild steel corrosion behavior in ethanol-gasoline blends (EGBs). However, the use of these methods was, so far, very limited for low-ethanol-content EGBs such as E5 and E10 due to their low conductivity. In this study, we present modified EIS and TS corrosion measurements based on the use of tetrabutylammonium tetrafluoroborate (TBATFB) at 500 mg/L as a supporting electrolyte. This modification led to an increase in the contaminated E5 and E10 fuels' conductivity, which allowed us to successfully perform the electrochemical corrosion tests. The corrosion current densities were determined to be 1.5 × 10-3 and 1.5 × 10-2 µA/cm2 for the tested E5 and E10 fuels, respectively. These modified methods present a significant extension of an electrochemical testing apparatus for steel corrosion studies in EGBs. They can allow one to obtain instantaneous information about the occurring corrosion process and, thus, estimate the materials' lifetime in corrosive environments and potentially help to prevent corrosion.

Utilization of Pharmaceutical Technology Methods for the Development of Innovative Porous Metasilicate Pellets with a Very High Specific Surface Area for Chemical Warfare Agents Detection.

Pharmaceutical technology offers various dosage forms that can be applied interdisciplinary. One of them are spherical pellets which could be utilized as a carrier in emerging second-generation detection tubes. This detection system requires carriers with high specific surface area (SSA), which should allow better adsorption of toxic substances and detection reagents. In this study, a magnesium aluminometasilicate with high SSA was utilized along with various concentrations of volatile substances (menthol, camphor and ammonium bicarbonate) to increase further the carrier SSA after their sublimation. The samples were evaluated in terms of physicochemical parameters, their morphology was assessed by scanning electron microscopy, and the Brunauer-Emmett-Teller (BET) method was utilized to measure SSA. The samples were then impregnated with a detection reagent o-phenylenediamine-pyronine and tested with diphosgene. Only samples prepared using menthol or camphor were found to show red fluorescence under the UV light in addition to the eye-visible red-violet color. This allowed the detection of diphosgene/phosgene at a concentration of only 0.1 mg/m3 in the air for samples M20.0 and C20.0 with their SSA higher than 115 m2/g, thus exceeding the sensitivity of the first-generation DT-12 detection tube.

Properties and Composition of Products from Hydrotreating of Straight-Run Gas Oil and Its Mixtures with Light Cycle Oil Over Sulfidic Ni-Mo/Al2O3 Catalyst.

Straight-run gas oil (SRGO) and its mixtures with 5, 10, 15, and 20 wt % light cycle oil (LCO) from fluid catalytic cracking (FCC) were hydrotreated on a commercial NiMo/Al2O3 catalyst in a laboratory tubular reactor with the cocurrent flow of the raw material and hydrogen. The hydrotreating of the raw material was undertaken at a temperature of 350 °C, a pressure of 4 MPa, a weight hourly space velocity of ca 1.0 h-1, and a hydrogen-to-raw-material ratio of 240 m3·m-3. The LCO had a high density due to the high content of bicyclic aromatics and the high content of sulfur species, which are difficult to desulfurize. Therefore, increasing the content of the LCO in the raw material resulted in increasing the density and increasing the content of the sulfur and polycyclic aromatics in the hydrotreated products. Only the products prepared from the raw material with LCO content up to 10 wt % fulfilled the density requirement of EN 590. To improve the product density, the products prepared from the raw material containing 15 wt % LCO were blended with refined kerosene. The addition of the kerosene decreased the density of the mixtures prepared, but the cold filter plugging point (CFPP) of the mixtures was only lowered by about 1-2 °C. It was necessary to add a depressant in an amount of 600 mg·kg-1 to achieve a cold filter plugging point of -20 °C. Some refined products were blended with desulfurized heavy naphtha from the FCC. The addition of the heavy naphtha was mainly limited by its high density. Up to 10 wt % heavy naphtha could be added to the product obtained by hydrotreating the raw material containing 10 wt % LCO. More than 15 wt % heavy naphtha could be added to the mixture of the hydrotreated product and 20 wt % kerosene.

A Strip Biosensor with Guinea Green B and Fuchsin Basic Color Indicators on a Glass Nanofiber Carrier for the Cholinesterase Detection of Nerve Agents.

This paper deals with the innovation of the Czech colorimetric biosensor Detehit designed for the simple, fast, and sensitive detection of nerve agents. The innovation is based on the use of an indicator consisting of a mixture of two triphenylmethane dyes, Guinea green B and a basic fuchsin, on a glass nanofiber filter paper carrier. The advantage of this solution is the blue-red color transition, which is much more visible than the white-yellow transition of other Detehit biosensors. The newly designed biosensor allows the users to visually detect (with the naked eye) the presence of the most significant paralytic substances (sarin, soman, cyclosarin, tabun, VX) in water at concentrations of at least 0.001 µg/mL. This biosensor design also enables one to detect these substances in air or on contaminated surfaces.

Cyclic Potentiometric Polarization and Resistance of Mild Steel in an Environment of Alcohols and Their Blends with Gasoline.

Nowadays, there is an effort to increase the more widespread use of biofuels that are a renewable energy source in transportation and an alternative to conventional, petroleum-based fuels. These biofuels include alcohols such as biomethanol, bioethanol, and biobutanol that have a high octane number, but generally different physical and chemical properties than petroleum fuels. The different properties of alcohols may cause low material compatibility with carbon steel. Here, we used cyclic potentiodynamic polarization (CPP) to study the behavior of carbon steel in an environment of alcohols and alcohol-gasoline blends (AGBs). Using CPP, we proved that the corrosion of mild steel can be significantly influenced by alcohol properties, such as the chain length, pK a, and solubility of oxygen and water. In the environment of pure alcohols (not blended by gasoline), a very good passivation ability of steel was proven, especially for n-butanol. In AGBs, steel corrosion can also be influenced by the gasoline amount. When these pure alcohols or their gasoline blends are contaminated by water-containing chlorides and organic acids, the corrosion rate of carbon steel can increase by up to 4 orders of magnitude. In an anhydrous environment of alcohols, the CPP can give results with a very good informative value.

Efficiency of Steel Corrosion Inhibitors in an Environment of Ethanol-Gasoline Blends.

Ethanol produced from renewable sources (i.e., bioethanol) is a first-generation biofuel that is currently being added as a biocomponent into gasolines. Mixtures of ethanol and gasoline are designated as ethanol-gasoline blends (EGBs). Ethanol has high polarity and moisture affinity, which considerably influence the properties of the resulting EGBs including their aggressiveness to many metallic and nonmetallic materials. The corrosion aggressiveness of EGBs can be minimized by suitable corrosion inhibitors. In this study, we tested three different corrosion inhibitors on mild steel in the environment of aggressive E10, E25, E60, and E85 fuels. The inhibitors tested were diethylene triamine (DETA) and two mixed inhibitors containing propargyl alcohol, dibenzyl sulfoxide, and octadecyl amine. To study the efficiency of the corrosion inhibitors, we used static and dynamic corrosion tests and electrochemical measurements including impedance spectroscopy and potentiodynamic polarization. The highest corrosion aggressiveness on mild steel was observed for the E60 fuel. The highest inhibitory efficiency was, for all the fuels tested, observed for the DETA inhibitor. For the DETA concentration of 100 mg·L-1, the inhibitory efficiency in the E60 fuel was determined to be around 98%.

New Carrier Made from Glass Nanofibres for the Colorimetric Biosensor of Cholinesterase Inhibitors.

Cholinesterase inhibitors are widely used as pesticides in agriculture, but also form a group of organophosphates known as nerve chemical warfare agents. This calls for close attention regarding their detection, including the use of various biosensors. One such biosensor made in the Czech Republic is the Detehit, which is based on a cholinesterase reaction that is assessed using a colour indicator-the Ellman's reagent-which is anchored on cellulose filter paper together with the substrate. With the use of this biosensor, detection is simple, quick, and sensitive. However, its disadvantage is that a less pronounced yellow discoloration occurs, especially under difficult light conditions. As a possible solution, a new indicator/substrate carrier has been designed. It is made of glass nanofibres, so the physical characteristics of the carrier positively influence reaction conditions, and as a result improve the colour response of the biosensor. The authors present and discuss some of the results of the study of this carrier under various experimental conditions. These findings have been used for the development of a modified Detehit biosensor.

Metal Corrosion and the Efficiency of Corrosion Inhibitors in Less Conductive Media.

Material corrosion can be a limiting factor for different materials in many applications. Thus, it is necessary to better understand corrosion processes, prevent them and minimize the damages associated with them. One of the most important characteristics of corrosion processes is the corrosion rate. The measurement of corrosion rates is often very difficult or even impossible especially in less conductive, non-aqueous environments such as biofuels. Here, we present five different methods for the determination of corrosion rates and the efficiency of anti-corrosion protection in biofuels: (i) a static test, (ii) a dynamic test, (iii) a static test with a reflux cooler and electrochemical measurements (iv) in a two-electrode arrangement and (v) in a three-electrode arrangement. The static test is advantageous due to its low demands on material and instrumental equipment. The dynamic test allows for the testing of corrosion rates of metallic materials at more severe conditions. The static test with a reflux cooler allows for the testing in environments with higher viscosity (e.g., engine oils) at higher temperatures in the presence of oxidation or an inert atmosphere. The electrochemical measurements provide a more comprehensive view on corrosion processes. The presented cell geometries and arrangements (the two-electrode and three-electrode systems) make it possible to perform measurements in biofuel environments without base electrolytes that could have a negative impact on the results and load them with measurement errors. The presented methods make it possible to study the corrosion aggressiveness of an environment, the corrosion resistance of metallic materials, and the efficiency of corrosion inhibitors with representative and reproducible results. The results obtained using these methods can help to understand corrosion processes in more detail to minimize the damages caused by corrosion.

Modified Biosensor for Cholinesterase Inhibitors with Guinea Green B as the Color Indicator.

Colorimetric biosensors of cholinesterase inhibitors are ideal for fast, reliable, and very simple detection of agents in air, in water, and on surfaces. This paper describes an innovation of the Czech Detehit biosensor, which is based on a biochemical enzymatic reaction visualized by using Ellman's reagent as a chromogenic indicator. The modification basically consists of a much more distinct color response of the biosensor, attained through optimization of the reaction system by using Guinea Green B as the indicator. The performance of the modified biosensor was verified on the chemical warfare agents (sarin, soman, cyclosarin, and VX) in water. The detection limits ascertained visually (with the naked eye) were about 0.001 µg/mL in water (exposure time 60 s, inhibition efficiency 25%).

The development of a butyrylcholinesterase porous pellet for innovative detection of cholinesterase inhibitors.

The aim of the presented research was the preparation of an innovative carrier with significantly improved properties for the fast and sensitive detection of cholinesterase inhibitors such as nerve agents. This innovative carrier was in the form of spherical pellets containing different amounts of Neusilin. Neusilin is a synthetic and amorphous form of magnesium aluminometasilicate with a high specific surface area, and the immobilized enzyme butyrylcholinesterase with an activity of 50nkat·g-1. Pellets were prepared by the extrusion-spheronization method and dried in a hot air oven under two conditions - at 30°C for 72h and at 60°C for 24h. Dried pellets were consequently impregnated with a solution containing butyrylcholinesterase. Impregnated pellets were evaluated for their quality parameters, enzymatic activity and inhibition. Activity and inhibition were tested according to the standard Ellman's method. It was observed that the addition of Neusilin significantly increased the hardness, intraparticular porosity, sphericity and activity of the carriers as well as intensity of the color transition. Therefore it is shown that these carriers have unquestionable advantages over common carriers of their kind. Drying temperatures have been shown to have no effect on properties of pellets except for a change in their size. Results were confirmed by statistical evaluation using ANOVA and PCA.