A hydrodynamic injection approach for capillary electrophoresis using rotor–stator valves

Sample injection is a critical step in a capillary electrophoresis (CE) analysis. Electrokinetic injection is the simplest approach and is often selected for implementation in portable CE instruments. However, in order to minimize the effect of sample matrix upon the results of a CE analysis, hydrodynamic injection is preferred. Although portable CE instruments with hydrodynamic injection have been reported, injection has always been performed at the grounded end of the capillary. This simplifies fluidic handling but limits coupling with electrochemical detectors and electrospray ionization–mass spectrometry (ESI–MS). We demonstrated previously that injection at the high-voltage (HV) end of the capillary could be performed using an HV-compatible rotary injection valve (fixed-volume injection). However, the mismatch between the bore sizes of the channels on the rotor–stator valve and the separation capillary caused peak tailing and undesired mixing, impairing analytical performance. In this work, we present an HV-compatible hydrodynamic injection approach that overcomes the issues associated with the fixed-volume injection approach reported previously. The performance of the CE instrument was demonstrated by analyzing a mixture of 13 amino acids by CE coupled to laser-induced fluorescence, which showed relative standard deviations for peak area and migration time below 5% and 1%, respectively, for triplicate analysis. Additionally, replicate measurements of a mixture of amino acids, peptides, nucleobases, and nucleosides by CE coupled to electrospray ionization–mass spectrometry (CE–ESI–MS) were performed to evaluate peak tailing, and results were similar to those obtained with a commercial CE–ESI–MS setup.     

A voltage trade study for the design of capillary electrophoresis instruments for spaceflight

Capillary electrophoresis (CE) systems have undergone extensive development for spaceflight applications. A flight-compatible high voltage power supply and the necessary voltage isolation for other energized components can be large contributors to both the volume and mass of a CE system, especially if typical high voltage levels of 25–30 kV are used. Here, we took advantage of our custom CE hardware to perform a trade study for simultaneous optimization of capillary length, high voltage level, and separation time, without sacrificing method performance. A capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C⁴D) method recently developed by our group to target inorganic cations and amino acids relevant to astrobiology was used as a test case. The results indicate that a 50 cm long capillary with 15 kV applied voltage (half of that used in the original method) can be used to achieve measurement goals while minimizing instrument size.     

Carbon-isotopic composition of soil-respired carbon dioxide in static closed chambers at equilibrium

The carbon-isotopic composition (delta13C) of soil-respired CO2 has been employed to evaluate soil carbon-cycling processes and the contribution of soil CO2 emissions to canopy and tropospheric air. These evaluations can be successful only when accurate isotope values of soil-respired CO2 are available. Here, we tested the robustness of delta13C values of soil-respired CO2 obtained after long incubations in static closed chambers that were initially flushed with soil air. The rationale of this approach is that the equilibrium carbon-isotope values of chamber-headspace CO2 are theoretically equal to those of CO2 produced within the soil. Static closed chambers were installed in replicated grass monocultures, and measurements of headspace CO2 concentrations and delta13C values were performed at regular time intervals for 24 h in July 2005. The results revealed no significant effects of grass species on headspace CO2 concentrations or delta13C values (repeated measures analysis of variance (ANOVA), P>0.1). As predicted by theory, isotope values asymptotically approached equilibrium conditions, which in our experimental setting occurred after 10 h. This good match between model predictions and our results suggests that an accurate determination of delta13C values of CO2 produced within soils is obtained through the isotopic measurement of chamber-headspace CO2 once equilibrium conditions have been reached with the underlying soils. An additional advantage of this approach is that only one sample per chamber is required, which, combined with the low uncertainties of these measurements, facilitates the investigation of the spatial (landscape) variability of soil-respired CO2.     

The effects of alkyl sulfates on the analysis of phenolic compounds by microchip capillary electrophoresis with pulsed amperometric detection

The effects of different surfactants (sodium 2-ethylhexyl sulfate, sodium decyl sulfate, sodium dodecyl sulfate and sodium tetradecyl sulfate) on the analysis of phenolic compounds by microchip-CE with pulsed amperometric detection were investigated. Using sodium decyl sulfate as a model surfactant, the effects of concentration and pH were examined. Under the optimized conditions, the analysis of six phenolic compounds was performed and compared with control runs performed without surfactant. When these surfactants were present in the run buffer, decreases in the migration time and increases in the run-to-run reproducibility were observed. Systematic improvements in the electrochemical response for the phenolic compounds were also obtained. According to the results presented, surfactants enhance the analyte-electrode interaction and facilitate the electron transfer process. These results should allow a more rational selection of the surfactants based on their electrophoretic and electrochemical effects.     

The Singular Molecular Conformation of Humic Acids in Solution Influences Their Ability to Enhance Root Hydraulic Conductivity and Plant Growth

Some studies have reported that the capacity of humic substances to improve plant growth is dependent on their ability to increase root hydraulic conductivity. It was proposed that this effect is directly related to the structural conformation in solution of these substances. To study this hypothesis, the effects on root hydraulic conductivity and growth of cucumber plants of a sedimentary humic acid and two polymers—polyacrylic acid and polyethylene glycol—presenting a molecular conformation in water solution different from that of the humic acid have been studied. The results show that whereas the humic acid caused an increase in root hydraulic conductivity and plant growth, both the polyacrylic acid and the polyethylene glycol did not modify plant growth and caused a decrease in root hydraulic conductivity. These results can be explained by the different molecular conformation in water solution of the three molecular systems. The relationships between these biological effects and the molecular conformation of the three molecular systems in water solution are discussed.     

Effect of the Addition of Alginate and/or Tetracycline on Brushite Cement Properties

Calcium phosphate cements have the advantage that they can be prepared as a paste that sets in a few minutes and can be easily adapted to the shape of the bone defect, which facilitates its clinical application. In this research, six formulations of brushite (dicalcium phosphate dihydrated) cement were obtained and the effect of the addition of sodium alginate was analyzed, such as its capacity as a tetracycline release system. The samples that contain sodium alginate set in 4 or 5 min and showed a high percentage of injectability (93%). The cements exhibit compression resistance values between 1.6 and 2.6 MPa. The drug was released in a range between 12.6 and 13.2% after 7 days. The antimicrobial activity of all the cements containing antibiotics was proven. All samples reached values of cell viability above 70 percent. We also observed that the addition of the sodium alginate and tetracycline improved the cell viability.     

ACE-Inhibitory and Antioxidant Activities of Peptide Fragments Obtained from Tomato Processing By-Products Fermented Using <Emphasis Type="Italic">Bacillus subtilis</Emphasis>: Effect of Amino Acid Composition and Peptides Molecular Mass Distribution

The effects of amino acid composition and peptide molecular mass on ACE-inhibitory and antioxidant activities of protein fragments obtained from tomato waste fermented using Bacillus subtilis were evaluated. The addition of B. subtilis increased the relative amounts of aromatic and positively-charged amino acids which have been described to influence the biological activities of peptide fragments. IC₅₀ values of hydrolysates for ACE-inhibitory and 2, 2′-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities were found to be 1.5 and 8.2 mg/mL, respectively. Size-exclusion chromatography (SEC) pattern of the hydrolysate indicated the breakdown of parent proteins to smaller peptides with molecular weights mainly below 1400 Da. MALDI-TOF mass spectrometry analysis revealed that the highest ACE-inhibitory activity was due to peptides showing molecular mass range 500–800 Da, while the most active antioxidant peptides were found to be mainly at the two different peptide weight ranges 500–800 Da and 1200–1500 Da.     

Visualization and Quantification of Transmembrane Ion Transport into Giant Unilamellar Vesicles

Transmembrane ion transporters (ionophores) are widely investigated as supramolecular agents with potential for biological activity. Tests are usually performed in synthetic membranes that are assembled into large unilamellar vesicles (LUVs). However transport must be followed through bulk properties of the vesicle suspension, because LUVs are too small for individual study. An alternative approach is described whereby ion transport can be revealed and quantified through direct observation. The method employs giant unilamellar vesicles (GUVs), which are 20–60 μm in diameter and readily imaged by light microscopy. This allows characterization of individual GUVs containing transporter molecules, followed by studies of transport through fluorescence emission from encapsulated indicators. The method provides new levels of certainty and relevance, given that the GUVs are similar in size to living cells. It has been demonstrated using a highly active anion carrier, and should aid the development of compounds for treating channelopathies such as cystic fibrosis.     

Order Types of Convex Bodies

We prove a Hadwiger transversal-type result, characterizing convex position on a family of non-crossing convex bodies in the plane. This theorem suggests a definition for the order type of a family of convex bodies, generalizing the usual definition of order type for point sets. This order type turns out to be an oriented matroid. We also give new upper bounds on the Erdős–Szekeres theorem in the context of convex bodies.