Assessment of heavy metals bioaccumulation in Silver Birch (Betula pendula Roth) from an AMD active, abandoned gold mine waste.

Acid mine drainage (AMD) is generally outlined as one of the largest environmental concerns, characterized by very low pH value of mine waste, heavy metals and high sulphate content. This extremely hostile environment reduces plant ability to develop and grow. Present study focuses on a silver birch (Betula pendula Roth), a pioneer species that grows on an extremely hostile gold mine waste, to investigate the bioaccumulation of rare metals (thallium (Tl) and indium (In)), as well as nine other more common heavy metals (bismuth (Bi), cadmium (Cd), cobalt (Co), copper (Cu), lead (Pb), manganese (Mn), nickel (Ni), silver (Ag) and zinc (Zn)), and to asses phytoextraction and phytostabilization potential of silver birch. Additionally, parameters determining AMD process and overall contamination (pH, electrical conductivity (EC), sulphates (SO42-), arsenic (As), iron (Fe), oxidation-reduction potential (ORP), turbidity, dissolved oxygen (DO), total dissolved solids (TDS), acidity, hardness, X-ray diffraction (XRD) and radioactivity) were determined in mine waste and drainage water samples. To assess the heavy metals bioaccumulation and mine waste status, statistical geochemical indices were determined: bioaccumulation factor (BCF), pollution load index (PLI), geochemical abundance index (GAI) and exposure index (EI). The results show that silver birch bioaccumulates the essential elements Cu, Ni, Mn and Zn, and the nonessential elements Tl (average BCF = 24.99), In (average BC = 23.01) and Pb (average BCF = 0.84). Investigated mine waste was enriched by Bi, Ag and Cd according to positive values of GAI index. Present research provides a novel insight into bioaccumulation of nonessential heavy metals in silver birches who grow on the extremely hostile mine waste, and they exhibit significant phytoremediation potential.

Potentiometric Determination of Maprotiline Hydrochloride in Pharmaceutical and Biological Matrices Using a Novel Modified Carbon Paste Electrode.

Potentiometry with membrane selective electrodes is preferable for measuring the various constituents of pharmaceuticals. In this work, carbon paste electrodes (CPE) were prepared, modified, and tested for the determination of maprotiline hydrochloride, which acts as an antidepressant. The proposed CPE was based on an ionic association complex of maprotiline-tetraphenylborate, 2-nitrophenyloctyl as a binder, and sodium tetraphenylborate as an ionic lipophilic additive. The optimized composition improved potentiometric properties up to theoretical Nernst response values of -59.5 ± 0.8 mV dec-1, in the concentration range of maprotiline from 1.6 × 10-7 to 1.0 × 10-2 mol L-1, and a detection limit of 1.1 × 10-7 mol L-1. The CPE provides excellent reversibility and reproducibility, exhibits a fast response time, and is applicable over a wide pH range. No significant effect was observed in several interfering species tested. The proposed electrode was used for the precise determination of maprotiline in pure solutions, urine samples, and a real sample-the drug Ludiomil.

Highly Sensitive and Selective Graphene Nanoribbon Based Enzymatic Glucose Screen-Printed Electrochemical Sensor.

A simple, sensitive, cost effective, and reliable enzymatic glucose biosensor was developed and tested. Nitrogen-doped heat-treated graphene oxide nanoribbons (N-htGONR) were used for modification of commercially available screen-printed carbon electrodes (SPCEs), together with MnO2 and glucose oxidase. The resulting sensors were optimized and used to detect glucose in a wide linear range (0.05-5.0 mM) by a simple amperometric method, where the limit of detection was determined to be 0.008 mM. (lifetime), and reproducibility studies were also carried out and yielded favorable results. The sensor was then tested against potential interfering species present in food and beverage samples before its application to real matrix. Spiked beer samples were analyzed (with glucose recovery between 93.5 and 103.5%) to demonstrate the suitability of the developed sensor towards real food and beverage sample applications.

Highly Sensitive Amperometric Detection of Hydrogen Peroxide in Saliva Based on N-Doped Graphene Nanoribbons and MnO2 Modified Carbon Paste Electrodes.

Four different graphene-based nanomaterials (htGO, N-htGO, htGONR, and N-htGONR) were synthesized, characterized, and used as a modifier of carbon paste electrode (CPE) in order to produce a reliable, precise, and highly sensitive non-enzymatic amperometric hydrogen peroxide sensor for complex matrices. CPE, with their robustness, reliability, and ease of modification, present a convenient starting point for the development of new sensors. Modification of CPE was optimized by systematically changing the type and concentration of materials in the modifier and studying the prepared electrode surface by cyclic voltammetry. N-htGONR in combination with manganese dioxide (1:1 ratio) proved to be the most appropriate material for detection of hydrogen peroxide in pharmaceutical and saliva matrices. The developed sensor exhibited a wide linear range (1.0-300 µM) and an excellent limit of detection (0.08 µM) and reproducibility, as well as high sensitivity and stability. The sensor was successfully applied to real sample analysis, where the recovery values for a commercially obtained pharmaceutical product were between 94.3% and 98.0%. Saliva samples of a user of the pharmaceutical product were also successfully analyzed.

A Novel Reduced Graphene Oxide Modified Carbon Paste Electrode for Potentiometric Determination of Trihexyphenidyl Hydrochloride in Pharmaceutical and Biological Matrices.

A novel promising carbon paste electrode with excellent potentiometric properties was prepared for the analysis of trihexyphenidyl hydrochloride (THP), the acetylcholine receptor and an anticholinergic drug in real samples. It contains 10.2% trihexyphenidy-tetraphenylborate ionic pair as the electroactive material, with the addition of 3.9% reduced graphene oxide and 0.3% of anionic additive into the paste, which consists of 45.0% dibutylphthalate as the solvent mediator and 40.6% graphite. Under the optimized experimental conditions, the electrode showed a Nernstian slope of 58.9 ± 0.2 mV/decade with a regression coefficient of 0.9992. It exhibited high selectivity and reproducibility as well as a fast and linear dynamic response range from 4.0 × 10-7 to 1.0 × 10-2 M. The electrode remained usable for up to 19 days. Analytical applications showed excellent recoveries ranging from 96.8 to 101.7%, LOD was 2.5 × 10-7 M. The electrode was successfully used for THP analysis of pharmaceutical and biological samples.

Validated Stability-Indicating GC-MS Method for Characterization of Forced Degradation Products of Trans-Caffeic Acid and Trans-Ferulic Acid.

When dealing with simple phenols such as caffeic acid (CA) and ferulic acid (FA), found in a variety of plants, it is very important to have control over the most important factors that accelerate their degradation reactions. This is the first report in which the stabilities of these two compounds have been systematically tested by exposure to various different factors. Forced degradation studies were performed on pure standards (trans-CA and trans-FA), dissolved in different solvents and exposed to different oxidative, photolytic and thermal stress conditions. Additionally, a rapid, sensitive, and selective stability-indicating gas chromatographic-mass spectrometric method was developed and validated for determination of trans-CA and trans-FA in the presence of their degradation products. Cis-CA and cis-FA were confirmed as the only degradation products in all the experiments performed. All the compounds were perfectly separated by gas chromatography (GC) and identified using mass spectrometry (MS), a method that additionally elucidated their structures. In general, more protic solvents, higher temperatures, UV radiation and longer storage times led to more significant degradation (isomerization) of both trans-isomers. The most progressive isomerization of both compounds (up to 43%) was observed when the polar solutions were exposed to daylight at room temperature for 1 month. The method was validated for linearity, precision as repeatability, limit of detection (LOD) and limit of quantitation (LOQ). The method was confirmed as linear over tested concentration ranges from 1-100 mg L-1 (r2s were above 0.999). The LOD and LOQ for trans-FA were 0.15 mg L-1 and 0.50 mg L-1, respectively. The LOD and LOQ for trans-CA were 0.23 mg L-1 and 0.77 mg L-1, respectively.

Analytical Methods for Determination of Phytic Acid and Other Inositol Phosphates: A Review.

From the early precipitation-based techniques, introduced more than a century ago, to the latest development of enzymatic bio- and nano-sensor applications, the analysis of phytic acid and/or other inositol phosphates has never been a straightforward analytical task. Due to the biomedical importance, such as antinutritional, antioxidant and anticancer effects, several types of methodologies were investigated over the years to develop a reliable determination of these intriguing analytes in many types of biological samples; from various foodstuffs to living cell organisms. The main aim of the present work was to critically overview the development of the most relevant analytical principles, separation and detection methods that have been applied in order to overcome the difficulties with specific chemical properties of inositol phosphates, their interferences, absence of characteristic signal (e.g., absorbance), and strong binding interactions with (multivalent) metals and other biological molecules present in the sample matrix. A systematical and chronological review of the applied methodology and the detection system is given, ranging from the very beginnings of the classical gravimetric and titrimetric analysis, through the potentiometric titrations, chromatographic and electrophoretic separation techniques, to the use of spectroscopic methods and of the recently reported fluorescence and voltammetric bio- and nano-sensors.

Development of the New Fluoride Ion-Selective Electrode Modified with FexOy Nanoparticles.

A new modified ion-selective electrode with membranes of LaF3 single crystals with different internal contacts (solid steel or electrolyte) and with FexOy nanoparticles as loading was developed. The best response characteristic with linear potential change was found in the fluoride concentration range from 10-1 to 3.98 × 10-7 M. The detection limit for the electrolyte contact was determined at 7.41 × 10-8 M with a regression coefficient of 0.9932, while the regression coefficient for the solid contact was 0.9969. The potential change per concentration decade ranged from 50.3 to 62.4 mV, depending on whether the contact was solid or electrolytic. The prepared modified electrode has a long lifetime, as well as the possibility of application in different positions (solid contact), and it can also be used for the determination of iron ions. The electrode characterization was performed with scanning electron microscopy and elemental analysis with the technique of electron-dispersive X-ray spectroscopy.

Choline Chloride Based Natural Deep Eutectic Solvents as Extraction Media for Extracting Phenolic Compounds from Chokeberry (Aronia Melanocarpa).

For the isolation of selected phenolic compounds from dried chokeberries, natural deep eutectic solvents (NADESs) were investigated as a green alternative to conventionally used extraction solvents. Four types of NADESs were synthesised, with choline chloride as the hydrogen bond acceptor in combination with different hydrogen bond donors (sugars, organic acid and urea). Ultrasound-assisted extraction was used to improve the extractability of the phenolic compounds and the results were compared to those obtained with 80% methanol as the extraction media. The highest values of total phenols and total flavonoids were found in the extract obtained with choline chloride-fructose NADES (36.15 ± 3.39 mg gallic acid g-1 dry weight (DW) and 4.71 ± 0.33 mg rutin g-1 DW, respectively). The extraction recoveries for the individual phenolic compounds depended strongly on the phenolic compound's structure, with relative mean values between 70% and 97%.

Measurement of breath ammonia for detection of patients with chronic kidney disease.

BACKGROUND: In a healthy individual, ammonia is converted to urea in the liver. Urea is then transported through the bloodstream and then excreted into the urine by the kidneys. In patients with chronic kidney disease (CKD), the accumulated urea is degraded by salivary urease into ammonia, which is then excreted by breathing. Breath ammonia can therefore be used for detecting the increased nitrogen-bearing wastes. In our pilot study, an electrochemical sensor was used to measure and analyze breath ammonia in healthy volunteers and patients with CKD. PATIENTS AND METHODS: In our study, 8 patients with CKD (stages 4 and 5) and 6 healthy volunteers were enrolled. All participants were nonsmokers and without pulmonary or liver disease. One controlled breath sample was collected from each participant. Immediately after the sample was collected, a gas analyzer was used for measuring breath ammonia in our participants. RESULTS: Mean creatinine value of CKD patients was 455.2 ± 294.1 µmol/L and 62.1 ± 7.5 µmol/L for healthy volunteers. Breath ammonia levels (3.32 ± 2.19 ppm vs. 0.49 ± 0.08 ppm; p = 0.003) and measured electric current (4.33 ± 0.25 mA vs. 4.01 ± 0.01 mA; p = 0.003) were significantly higher in the CKD group. CONCLUSIONS: The results of our pilot study show that breath monitoring of ammonia can be a simple, useful, fast, and noninvasive tool for detection of advanced kidney impairment.
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Chemometric Characterization of Slovenian Red Wines.

Total phenolic (TPC), flavonoid (TFC) and tannin (TTC) contents, total SO2, total acids, pH, and reducing sugars were measured in twenty five Slovenian red wines from three key wine producing regions, Podravje, Posavje and Primorska. The results were chemometrically analysed and the wines were classified according to wine growing region and vine variety. Principal component analysis proved that TPC, TFC and TTC contents were primarily responsible for variation in the wines. Additionally, linear discriminant analysis (LDA) was performed and resulted in the satisfactory classification of samples by both vine variety and region.

Simultaneous GC-MS Determination of Free and Bound Phenolic Acids in Slovenian Red Wines and Chemometric Characterization.

Several phenolic acids (PAs), caffeic, vanillic, syringic, p-coumaric and ferulic acid, found in Slovenian red wines were studied using gas chromatography and mass spectrometry. For isolation of the PAs from wine samples, solid phase extraction using hydrophilic modified styrene - HLB cartridges was used. The bound PAs were extracted after basic hydrolysis and o-coumaric acid was used as the internal standard. The developed method was validated and the linear concentration range for all analytes was from 1 to 100 mg L-1 with correlation coefficients above 0.999. We show that the method is repeatable (RSD<2%), recoveries were above 96%, and LOD and LOQ values were acceptable. In all of the wine samples tested, caffeic and p-coumaric acid were determined to be the predominant PAs (17-72 mg L-1), while other compounds were found in lower concentrations. Principal Component Analysis and Cluster Analysis were used to study differences between wines related towards varieties and Slovenian wine regions. The results demonstrate that variety has more influence on PAs content than wine regions in Slovenian red wines.

In-situ enzyme-initiated production of hexanal from sunflower oil and its release from double emulsion electrospun bio-active membranes.

The aim of the presented study was to evaluate the release of the enzymatically initiated production of hexanal from double emulsion electrospun bio-active membranes at a temperature of fruit storage. Among different formulations of water-in-oil (W1/O) primary emulsions, the emulsion composed of 12% w/v Tween20 and 0.1 M NaCl in water (W1) and 6% of poly(glycerol) poly(ricinoleate) dissolved in sunflower oil (O) using W1/O ratio of 80/20 (w/w) (Tween20-NaCl/6% PGPR) was selected, for further incorporation of enzymes, based on the lowest average droplet size (391.0 ± 15.6 nm), low polydispersity index (0.255 ± 0.07), and good gravitational stability also after 14 days. Both enzymes, lipase and lipoxygenase are needed to produce hexanal (up to 58 mg/L). Additionally, double emulsions were prepared with sufficient conductivity and viscosity using different W1/O to W2 ratios for electrospinning. From the selected electrospun membrane, up to 4.5 mg/L of hexanal was released even after 92 days.

Introduction of a spectrophotometric method for salivary iodine determination on microplate based on Sandell-Kolthoff reaction.

BACKGROUND: Iodine is an essential element for the synthesis of thyroid hormones. Therefore, a reliable marker of iodine supply is important. Iodine is predominantly excreted via kidneys, but also via salivary glands. Our aim was to introduce a new and simple method for determination of salivary iodine concentration (SLIC). MATERIALS AND METHODS: Self-prepared chemicals and standards for Sandell-Kolthoff reaction on microplate with ammonium peroxydisulfate (AP) in the range 0-400 µg/L were used. Suitability of water-based standards (WBS) and artificial saliva-based standards (ASS) for standard curve were tested. We followed standards for method validation, defined concentration of used AP and compared our results with Inductively Coupled Plasma Mass Spectrometry (ICP-MS). RESULTS: WBS gave more reliable results than ASS as an underestimation of iodine concentration was found for ASS. LoB was 6.5 µg/L, LoD 12.0 µg/L, therefore analytical range was 12-400 µg/L. Intra- and inter-assay imprecisions at iodine concentrations, namely 20, 100, 165, and 350 µg/L were 18.4, 5.1, 5.7, and 2.8%, respectively, and 20.7, 6.7, 5.1, and 4.3%, respectively. Suitable molarity of AP was 1.0 mol/L and showed no difference to 1.5 mol/L (P values for samples with concentration 40, 100, and 150 µg/L, were 0.761, 0.085, and 0.275, respectively), whereas there was a significant change using 0.5 mol/L (P<0.001). Saliva samples could be diluted up to 1:8. There was no interference of thiocyanate and caffeine up to 193.5 mg/L. Our original method was comparable to ICP-MS. Spaerman coefficient was 0.989 (95% CI: 0.984-0.993). CONCLUSIONS: The new method for SLIC determination is in excellent agreement with ICP-MS and easy-to-use.

Honey Origin Authentication via Mineral Profiling Combined with Chemometric Approaches.

In the present study, the potential of elemental analysis combined with statistical tools to identify honey origin was evaluated by mineral characterization of 173 honeys of 13 floral types (acacia, fir, spruce, linden, chestnut, lavender, coriander, thistle, honeydew, rosemary, sage, euphorbia and ziziphus plant species) collected from five geographical regions (Slovenia, Croatia, Bulgaria, Turkey, and Morocco). The objective of the study was to accurately and reliably differentiate the mineral composition among honey varieties. The aim was to establish traceability, to ensure product authenticity and to improve quality control measures within the honey industry. For this purpose, 18 major, minor and trace elements were quantified using microwave digestion, followed by ICP-MS measurement. Statistical evaluation of elemental concentration was undertaken using principal component analysis (PCA) to distinguish honey floral types. The research give light on the specific elements that can serve as indicators for determining the geographical and botanical source of honey. Our findings indicate that certain elements, such as Mn, K, and Ca, are primarily influenced by the type of pollen present in the honey, making them indicative of the floral source. On the other hand, levels of Na, Mg, and Fe were found to be more strongly influenced by environmental factors and can be considered as markers of geographical origin. One novel aspect of this research is the exploration of the relationship between honey minerals and honey botanical source. This was achieved through the analysis of chestnut tree samples and a subsequent comparison with the composition of chestnut honey.

Extraction of Polyphenols from Slovenian Hop (Humulus lupulus L.) Aurora Variety Using Deep Eutectic Solvents: Choice of the Extraction Method vs. Structure of the Solvent.

Polyphenols from Slovenian hops (Humulus lupulus L.) of the Aurora variety were extracted by different methods and using classical solvents and several deep eutectic solvents (DES) based on choline chloride as the hydrogen bond acceptor component. The obtained extract solutions were analyzed by HPLC for the content of extracted α- and ß-acids and extracted xanthohumol. It was found that choline chloride:phenol DES concentrated aqueous solution had an extraction efficiency close to that of diethyl ether, which is considered one of the best classical extraction solvents for polyphenols from hops. The comparison of the extraction efficiency with other choline chloride-based DESs showed that the chemical similarity of the phenol ring in the solvent DES with the polyphenols in hops may be crucial for a highly efficient extraction with choline chloride:phenol DES. On the other hand, the choice of extraction method and the viscosity of the solvents tested seem to play only a minor role in this respect. As far as we know, this is the first study to attempt to relate extraction efficiency in the extraction of hydrophobic solutes to the compressibility of the DES extractants, the latter of which may be correlated with the extent of hydrophobic hydration around the DES components. In addition, using the heating and stirring method for the preparation of choline chloride-based DES concentrated aqueous solutions we found no support for the occurrence of water in two different roles (in the structural and in the dilution role) in these solvents.

Trace metals and macrominerals in common seaweeds in the Marchica (a restored lagoon, Mediterranean Sea): Nutritional value and health risk assessment.

This study investigated the contents of macrominerals (Na, K, Ca, Mg and P) and essential trace metals (Fe, Mn, Cu, Zn and Se) in four species of seaweeds (Gracilaria sp., Alsidium corallinum, Caulerpa prolifera, and Chaetomorpha sp.) from Marchica Lagoon. The contents of macrominerals with mean values (% dw) can be sequenced in this descending order, [Ca > Mg > Na > K > P] for Caulerpa prolifera, and the decreasing sequence [K > Ca > Mg > P > Na] for Chaetomorpha sp. In red seaweeds, Gracilaria sp. and Alsidiumcorallinum followed these orders: [K > Ca > Na > Mg > P] and [K > Na > Ca > Mg > P] respectively. The essential trace metals mean values (mg kg-1) followed the decreasing order [Fe > Mn > Zn > Cu > Se] for Alsidiumcorallinum, Chaetomorpha sp. and Gracilaria sp., and the following order [Fe > Mn > Zn > Se > Cu] for Caulerpa prolifera. Based on the calculated recommended dietary allowance (RDA), targeted hazard quotient (THQ) and the hazard index (HI), the studied seaweeds did not pose any health risk for human consumption.

Determination of Oxygen by Means of a Biogas and Gas - Interference Study Using an Optical Tris (4,7-Diphenyl-1,10-Phenanthroline) Ruthenium(II) Dichloride Complex Sensor.

Biogas is a mixture of gases produced by anaerobic fermentation where biomass or animal waste is decomposed and methane and carbon dioxide are mainly released. Biogas also has a very high moisture content (up to 80%), temperatures of around 60 °C, high pressure, and can contain other gases (N2, H2S, NH3 and H2). We searched for an appropriate measuring system for the determining of oxygen in biogas, since the production process of biogas must be run under anaerobic conditions; as the presence of oxygen decreases the quality of the biogas. Ruthenium (II) complexes are by far the most widely-used oxygen dyes within optical oxygen sensors. In general, they have efficient luminescences, relatively long-life metal-ligand charge-transfer excited states, fast response times, strong visible absorptions, large Stokes shifts, and high-photochemical stability. The purpose of this work was to characterise and optimize an optical oxygen sensor using tris (4,7-diphenyl-1,10-phenanthroline) ruthenium(II) dichloride complex for measuring oxygen. Different sensor properties were additionally studied, focusing on the interference of external light, temperature, and various gases. A special gas-mixing chamber was developed for gas interference study, and on-line experiments are presented for oxygen determination within the pilot biogas reactor.

Citrate-induced local ionized calcium decrease in pediatric hemodialysis settings: An in-vitro study.

BACKGROUND: Citrate is instilled into the dialysis catheter to prevent clotting and to maintain patency between dialysis sessions. A significant amount of citrate leaks from the catheter at the injection time, which decreases the blood ionized calcium concentration (Ca2+) due to chelation. We aimed to evaluate the local impact of concentrated (i.e. 30%) citrate spilling on Ca2+ at the catheter tip in real-time pediatric conditions. METHODS: An in-vitro model was constructed, involving an ion-selective electrode (Ca-ISE). A pre-curved catheter and the Ca-ISE were submerged in a glass vessel with the tips positioned adjacent to each other. The vessel was filled with 30 and 80 ml of normal saline with added calcium to simulate normal right atrium size in children and adults, respectively, and normal blood Ca2+. The amount of instilled citrate matched the filling volume of the catheter. Measurements were performed with 4% and 30% citrate solutions. RESULTS: The mean Ca2+ measured at the tip of the catheter was 0.457 and 0.058 mmol/l when using 4% and 30% citrate, respectively (p < 0.001). The mean Ca2+ recorded in 30 and 80 ml after instilling 30 % citrate was 0.058 and 0.055 mmol/l, respectively (p = 0.878). CONCLUSIONS: The spilling of 30% citrate caused a strikingly greater decrease of Ca2+ at the catheter tip than the standard 4% citrate. The atrial volume did not influence the test results implying similar safety concerns for pediatric and adult patients. The used static experimental setting might have overestimated the spilling effect.

Improving Electroactivity of N-Doped Graphene Derivatives with Electrical Induction Heating.

Graphene derivatives doped with nitrogen have already been identified as active non-noble metal materials for oxygen reduction reaction (ORR) in PEM and alkaline fuel cells. However, an efficient and scalable method to prepare active, stable, and high-surface-area non-noble metal catalysts remains a challenge. Therefore, an efficient, potentially scalable strategy to improve the specific surface area of N-doped graphene derivatives needs to be developed. Here, we report a novel, rapid, and scalable electrical induction heating method for the preparation of N-doped heat-treated graphene oxide derivatives (N-htGOD) with a high specific surface area. The application of the induction heating method has been shown to shorten the reaction time and improve the energy efficiency of the process. The materials synthesized by induction heating exhibited very high specific surface area and showed improved ORR activity compared to the conventional synthesis method. Moreover, we demonstrated that the temperature program of induction heating could fine-tune the concentration of nitrogen functionalities. In particular, the graphitic-N configuration increases with increasing final temperature, in parallel with the increasing ORR activity. The presented results will contribute to the understanding and development of nonmetal N-htGOD for energy storage and conversion applications.