Liquid chromatographic determination of caffeine and adrenergic stimulants in food supplements sold in Brazilian e-commerce for weight loss and physical fitness

Methyl-xanthines and adrenergic stimulants, such as caffeine and synephrine, are commonly added to food supplements due to their stimulating and thermogenic effects. In addition, the abusive consumption of food supplements with ergogenic and aesthetic purposes has been observed worldwide. This work describes the study of caffeine, p-synephrine, hordenine, octopamine, tyramine, ephedrine and salicin as stimulants in dietary supplements marketed in Brazil for weight loss and physical fitness claims. A total of 94 different products were acquired from 30 Brazilian websites. Thus, the sampling of marketed supplements was performed in virtual commerce (e-commerce) with claims of weight loss, appetite reduction, fat burning and metabolism acceleration. The developed analytical method involved the separation of the stimulants by HPLC with diode array detection (HPLC-DAD) by using a gradient elution of flow rate (0.7–2.5 ml min^?1) and mobile phase composition (0.1% H₃PO₄/methanol). The validated method was applied to the study of 46 dietary supplements. Caffeine, p-synephrine and ephedrine were found to be present as stimulants in 52% of the studied samples marketed as encapsulated or bulk forms. Caffeine was found to be present in concentrations that represent doses from 25.0 to 1476.7 mg day^–1. Synephrine was found in concentrations that represent doses from 59.1 to 127.0 mg day^–1. Ephedrine was found to be associated with caffeine in one formulation at a concentration representing a 26.1 mg day^–1 dosage.     

Chemo-Enzymatic Fluorescence Labeling Of Genomic DNA For Simultaneous Detection Of Global 5-Methylcytosine And 5-Hydroxymethylcytosine**

5-Methylcytosine and 5-hydroxymethylcytosine are epigenetic modifications involved in gene regulation and cancer. We present a new, simple, and high-throughput platform for multi-color epigenetic analysis. The novelty of our approach is the ability to multiplex methylation and de-methylation signals in the same assay. We utilize an engineered methyltransferase enzyme that recognizes and labels all unmodified CpG sites with a fluorescent cofactor. In combination with the already established labeling of the de-methylation mark 5-hydroxymethylcytosine via enzymatic glycosylation, we obtained a robust platform for simultaneous epigenetic analysis of these marks. We assessed the global epigenetic levels in multiple samples of colorectal cancer and observed a 3.5-fold reduction in 5hmC levels but no change in DNA methylation levels between sick and healthy individuals. We also measured epigenetic modifications in chronic lymphocytic leukemia and observed a decrease in both modification levels (5-hydroxymethylcytosine: whole blood 30 %; peripheral blood mononuclear cells (PBMCs) 40 %. 5-methylcytosine: whole blood 53 %; PBMCs 48 %). Our findings propose using a simple blood test as a viable method for analysis, simplifying sample handling in diagnostics. Importantly, our results highlight the assay‘s potential for epigenetic evaluation of clinical samples, benefiting research and patient management.     

Field evaluation of a Portable Fine Particle Concentrator (PFPC) for ice nucleating particle measurements

The custom-built Portable Fine Particle Concentrator (PFPC) is evaluated for the measurement of ice nucleating particles (INPs) in the atmosphere. The concentrations of INPs in remote regions of the atmosphere are very low, often close to instrumental detection limits. The PFPC is a dual slit-nozzle virtual impactor where particles are concentrated from an input flow of 250 LPM (litres per minute) into an output flow of 10 LPM. The enrichment factors (EFs) for ambient particles with diameters between 0.4 and 2.5?µm were found to be 21?±?5 at sea level and 18?±?2 at a field station 3580 meters above sea level for the PFPC operated in horizontal configuration. Similar enhancement factors (16?±?5) in the concentrations of INPs measured by the Horizontal Ice Nucleation Chamber at the high-altitude station were observed when the air mass was characterized by high numbers of particles larger than 0.5?µm. When the number size distribution was dominated by particles smaller than 0.5?µm, the INP EF was considerably lower. Corroborating short-term measurements were provided by additional INP-measuring instruments, the Fast Ice Nucleus CHamber and the Frankfurt Ice Deposition Freezing Experiment. Results from two aerosol mass spectrometers also indicate significant particle enhancement using the PFPC. These results indicate that the PFPC can be usefully deployed to improve the detection efficiency of ambient INP measurements.

Copyright © 2019 American Association for Aerosol Research     

Crosslinking kinetics of SBR composites containing vulcanized ground scraps as filler

The aim of this study was to characterize the cure reaction of styrene–butadiene rubber (SBR) composites containing industrial rubber scraps. Different proportions of SBR ground scraps (SBR-r), varying from 10 to 80 parts per hundred of rubber, were incorporated into a base formulation of identical composition. Crosslink formation and the kinetics of the cure reaction were evaluated through oscillatory disk rheometry, differential scanning calorimetry, and crosslink density. Cure characteristics, such as scorch time and cure time, decreased with increasing SBR-r content. Minimum torque indicated only a small variation in the viscosity with the incorporation of SBR-r. The maximum torque decreased with the addition of scrap rubber, as a consequence of the reduction in virgin rubber content where crosslinks had been formed. Crosslink density values corroborated these findings, presenting a slight decrease with the increase in the SBR-r content. The kinetic study indicated lower enthalpy values for SBR-r composites compared to those of the control sample. The kinetic parameters, such as activation energy and reaction order, indicated a change in the mechanism of reaction, related to the increased complexity of the systems.     

Recent Advances in Intracellular and In Vivo ROS Sensing: Focus on Nanoparticle and Nanotube Applications

Reactive oxygen species (ROS) are increasingly being implicated in the regulation of cellular signaling cascades. Intracellular ROS fluxes are associated with cellular function ranging from proliferation to cell death. Moreover, the importance of subtle, spatio-temporal shifts in ROS during localized cellular signaling events is being realized. Understanding the biochemical nature of the ROS involved will enhance our knowledge of redox-signaling. An ideal intracellular sensor should therefore resolve real-time, localized ROS changes, be highly sensitive to physiologically relevant shifts in ROS and provide specificity towards a particular molecule. For in vivo applications issues such as bioavailability of the probe, tissue penetrance of the signal and signal-to-noise ratio also need to be considered. In the past researchers have heavily relied on the use of ROS-sensitive fluorescent probes and, more recently, genetically engineered ROS sensors. However, there is a great need to improve on current methods to address the above issues. Recently, the field of molecular sensing and imaging has begun to take advantage of the unique physico-chemical properties of nanoparticles and nanotubes. Here we discuss the recent advances in the use of these nanostructures as alternative platforms for ROS sensing, with particular emphasis on intracellular and in vivo ROS detection and quantification.     

Influence of the chemical structure of additives poly(ethylene-co-vinyl acetate)-based on the pour point of crude oils

One of the methods to prevent wax precipitation, during petroleum production, transport, and refining, is the use of polymer additives that can reduce the oil pour point. However, no single additive work for all types of crude oil and this relation is not yet well known. In this study, a family of polymers based on poly(ethylene-co-vinyl acetate), containing hydroxyl groups and long pendant hydrocarbon chains (from C6 to C18), were synthesized and characterized by H¹ nuclear magnetic resonance and solubility test. Four crude oil samples containing different amounts and size distribution of the wax were used. The additive's action is favored by higher contents of iso + cycloalkanes and lower contents of n-paraffins with larger chain sizes. The presence of the CH₃COO group in the copolymers promoted the lowering of the pour point, supported by a low OH concentration and the presence of a long pendant hydrocarbon chain: the best results were obtained with C10 and C12 chain lengths. © 2020 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48969.     

Prediction and comparison of textural properties of magnetic copolymer supports for enzyme immobilization

Magnetic polymers supports have proven to be valuable materials for enzyme immobilization, as they allow recovering the catalyst by magnetic separation, precluding the need for costly and time-consuming separation steps. In this study, magnetic copolymer supports were synthesized using styrene (STY) and different crosslinking agents (divinylbenzene, ethylene glycol dimethacrylate, or triethylene glycol dimethacrylate) and initiators (azobisisobutyronitrile or benzoyl peroxide) and used to immobilize Candida antarctica lipase B (CALB). The aim was to obtain biocatalysts with high enzymatic activity and satisfactory morphological properties for use in biotransformation reactions. Two morphological properties known to influence the immobilization yield were taken into consideration, specific surface area, and swelling index. Experimental data were compared to the predictions of a model based on molar balance, method of moments, numerical fractionation, and elementary gel structures. The high correlation (R² = 0.9974) between experimental and predicted values demonstrated the suitability of the model for estimating the textural properties of enzyme supports. CALB was successfully immobilized, showing high hydrolytic activity (500–700 U g⁻¹) and good thermal stability at 50°C. CALB/STY-EGDMA-M was 14 times more stable than free CALB. The results confirm the efficiency of the immobilization method and the suitability of the copolymers for enzyme immobilization.     

Synthesis of Isopropyl Palmitate by Lipase Immobilized on a Magnetized Polymer Matrix

Penicillium camemberti lipase immobilized on a magnetized poly(styrene-co-divinylbenzene) was used as a biocatalyst for isopropyl palmitate synthesis. The reaction conditions were determined by 2² factorial central composite design. A mathematical model based on a simplified kinetic approach was developed to describe the system and validated with the experimental data. An assay carried out in a stirred-tank reactor confirmed the proposed model. The ester was purified and the properties such as density and water content were similar to those found in commercially available isopropyl palmitate.     

Direct ink writing of ceramic matrix composite structures

We present the development of an ink containing chopped fibers that is suitable for direct ink writing (DIW), enabling to obtain ceramic matrix composite (CMC) structures with complex shape. We take advantage of the unique formability opportunities provided by the use of a preceramic polymer as both polymeric binder and ceramic source. Inks suitable for the extrusion of fine filaments (<1 mm diameter) and containing a relatively high amount of fibers (>30 vol% for a nozzle diameter of 840 μm) were formulated. Despite some optimization of ink rheology still being needed, complex CMC structures with porosity of ~75% and compressive strength of ~4 MPa were successfully printed. The process is of particular interest for its ability to orient the fibers in the extrusion direction due to the shear stresses generated at the nozzle tip. This phenomenon was observed in the production of polymer matrix composites, but it is here employed for the first time for the production of ceramic matrix ones. The possibility to align high aspect ratio fillers using DIW opens the path to layer‐by‐layer design for optimizing the mechanical and microstructural properties within a printed object, and could potentially be extended to other types of fillers.     

Biodegradation of PHBV/GNS nanocomposites by Penicillium funiculosum

Biodegradable polymer nanocomposites are an essential alternative to minimize the generation of polymeric solid waste that shows short shelf life and difficult degradation. In this study, nanocomposites based on poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (PHBV) were prepared by the incorporation of different contents, 0.25, 0.50, 0.75, and 1.00 wt % of graphite nanosheets (GNS), using a solution casting method. The investigation of the PHBV samples biodegradation was made using filamentous fungi (Penicillium funiculosum) in solid medium. Characterization of the material was performed by weight loss, differential scanning calorimetry, carbonyl index determined by Fourier transformed infrared spectroscopy, contact angle, roughness, and scanning electron microscopy. Results revealed that PHBV/GNS nanocomposites can be totally degraded in the presence of Penicillium funiculosum; however, it will be necessary high incubation period. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44234.