Biosensing with Polymerase Chain Reaction-Stable DNA-Functionalized Magnetically Susceptible Carbon-Iron Microparticles.

We demonstrate a rapid and sensitive method for DNA detection without the need for fluorescence. This is based on carbon-coated magnetic iron (Fe) microparticles with a covalent surface attachment of DNA. We show that these magnetic microparticles can capture complementary DNA. Significantly, the DNA covalent surface bonds are robust to high temperatures and can be included in a sample during polymerase chain reaction (PCR). This method is employed for the detection of targeted DNA sequences (40-50 bp). Hybridization probes on the surface of the magnetically susceptible Fe microparticle recognize the target DNA sequence-specifically. The double-stranded DNA (dsDNA) microparticles are then quickly captured with a magnet from the sample matrix. This foregoes postpurification processes, such as electrophoresis, which make our technique time- and cost-effective. Captured dsDNA can be detected with intercalating dyes such as ethidium bromide through a loss in the UV absorption signal with a limit of detection (LOD) of 24 nM within 15 min. Likewise, surface-bound DNA can act as a primer in PCR to decrease the LOD to 5 pM within 2 h. This is the first instance of a nucleotide-modified magnetically susceptible carbon substrate that is PCR-compatible. Besides DNA capture, this strategy can eventually be applied to sequence-specific nucleic acid purification and enrichment, PCR cleanup, and single-strand generation. The DNA-coated particles are stable under PCR conditions (unlike commonly used polystyrene or gold particles).

Correction: Voltammetric pH sensor based on electrochemically modified pseudo-graphite.

Correction for 'Voltammetric pH sensor based on electrochemically modified pseudo-graphite' by Haoyu Zhu et al., Analyst, 2020, 145, 7252-7259, https://doi.org/10.1039/D0AN01405B.

A Comparison of Solid Electrolyte Interphase Formation and Evolution on Highly Oriented Pyrolytic and Disordered Graphite Negative Electrodes in Lithium-Ion Batteries.

The presence and stability of solid electrolyte interphase (SEI) on graphitic electrodes is vital to the performance of lithium-ion batteries (LIBs). However, the formation and evolution of SEI remain the least understood area in LIBs due to its dynamic nature, complexity in chemical composition, heterogeneity in morphology, as well as lack of reliable in situ/operando techniques for accurate characterization. In addition, chemical composition and morphology of SEI are not only affected by the choice of electrolyte, but also by the nature of the electrode surface. While introduction of defects into graphitic electrodes has promoted their electrochemical properties, how such structural defects influence SEI formation and evolution remains an open question. Here, utilizing nondestructive operando electrochemical atomic force microscopy (EChem-AFM) the dynamic SEI formation and evolution on a pair of representative graphitic materials with and without defects, namely, highly oriented pyrolytic and disordered graphite electrodes, are systematically monitored and compared. Complementary to the characterization of SEI topographical and mechanical changes during electrochemical cycling by EChem-AFM, chemical analysis and theoretical calculations are conducted to provide mechanistic insights underlying SEI formation and evolution. The results provide guidance to engineer functional SEIs through design of carbon materials with defects for LIBs and beyond.

Voltammetric pH sensor based on electrochemically modified pseudo-graphite.

A nanocrystalline graphite-like amorphous carbon (graphite from the University of Idaho thermolyzed asphalt reaction, GUITAR) shares morphological features with classical graphites, including basal and edge planes (BP, EP). However, unlike graphites and other sp2-hybridized carbons, GUITAR has fast heterogenous electron transfer (HET) across its basal planes, and resistance to corrosion similar to sp3-C and boron-doped diamond electrodes. In this contribution, quinoid modified BP-GUITAR (q-GUITAR) is examined as a sensor for pH determination. This modification is performed by applying 2.0 V (vs. Ag/AgCl) for 150 seconds followed by 15 cyclic voltammetric scans from -0.7 to 1.0 V at 50 mV s-1 in 1.0 M H2SO4. The quinoid surface coverage of q-GUITAR is 1.35 × 10-9 mol cm-2, as measured by cyclic voltammetry. X-ray photoelectron spectroscopy analysis also confirms the high surface coverage. The quinoid surface concentration ranks highest in literature when compared with other basal plane graphitic materials. This yields a sensor that responds through a square wave voltammetric reduction peak shift of 63.3 mV per pH over a pH range from 0 to 11. The response on q-GUITAR is stable for >20 measurements and no surface re-activation is required between the measurements. The common interferents, Na+, K+ and dissolved oxygen, have no effect on the response of the q-GUITAR-based pH sensor.

Utilizing a Single Silica Nanospring as an Insulating Support to Characterize the Electrical Transport and Morphology of Nanocrystalline Graphite.

A graphitic carbon, referred to as graphite from the University of Idaho thermolyzed asphalt reaction (GUITAR), was coated in silica nanosprings and silicon substrates via the pyrolysis of commercial roofing tar at 800 °C in an inert atmosphere. Scanning electron microscopy and transmission electron microscopy images indicate that GUITAR is an agglomeration of carbon nanospheres formed by the accretion of graphitic flakes into a ~100 nm layer. Raman spectroscopic analyses, in conjunction with scanning electron microscopy and transmission electron microscopy, indicate that GUITAR has a nanocrystalline structure consisting of ~1-5 nm graphitic flakes interconnected by amorphous sp3 bonded carbon. The electrical resistivities of 11 single GUITAR-coated nanospring devices were measured over a temperature range of 10-80 °C. The average resistivity of all 11 devices at 20 °C was 4.3 ± 1.3 × 10-3 Ω m. The GUITAR coated nanospring devices exhibited an average negative temperature coefficient of resistivity at 20 °C of -0.0017 ± 0.00044 °C-1, which is consistent with the properties of nanocrystalline graphite.

Electrochemical determination of free chlorine on pseudo-graphite electrode.

Pseudo-graphite from the University of Idaho Thermolyzed Asphalt Reaction also known as GUITAR is a new form of carbon. It shares morphological features with graphites, including basal and edge planes. Unlike graphites and other sp2-hybridized carbons, GUITAR has fast heterogeneous electron transfer across its basal planes and resistance to corrosion similar to boron-doped diamond electrodes. In this contribution GUITAR electrodes were examined as sensors for aqueous free chlorine (HOCl and OCl-) at pH 7.0 with cyclic voltammetric (CV) and chronoamperometric (CA) methods. Using CV at 50 mV s-1 GUITAR has a limit of detection of 1.0 µmol L-1, linear range of 0-5,000 µmol L-1, sensitivity of 215.8 µA L mmol-1 cm-2 and a signal stability of 4 days in constant exposure to 1 mmol L-1 free chlorine in pH 7.0, 0.1 mol L-1 phosphate buffer system. After 7 days of exposure GUITAR electrodes lost 37% of the former sensitivity, which was recovered by an in-situ regeneration procedure. The common aqueous ions, Ca2+, Na+, NO3-, SO42-, Cl-, CO32- and dissolved oxygen did not affect the response of the GUITAR-based sensor. The combination of limit of detection, linear range, sensitivity, sensor lifetime and its relative lack of interferences indicate that GUITAR is one of the best performers in free chlorine sensors.

Heavy metals removal from mine runoff using compost bioreactors.

Permeable bioreactors have gained both research and management attention as viable methods for treating mine runoff waters. We examined the operation of a field-scale bioreactor (containing mixed compost, straw and gravel) for treatment of runoff from the Mother Load (ML) mine in northern Idaho, U.S. and compared it to an experimental laboratory-scale reactor, containing a similar matrix and treating similar mine runoff water. In general both reactors were efficient in removing most of the metals assayed, Al, As, Cd, Fe, Ni, Pb and Zn, with the exception of Mn. Both systems showed evidence of bacterial-mediated sulphate reduction and concomitant metal sulphide complexes. However, the experimental laboratory bioreactor showed greater proportions of immobile metals reductions than did the ML bioreactor, presumably due to the greater action of sulphate-reducing bacteria. The major metal removal mechanism in the ML bioreactor was surmised to be adsorption. Differences in metal removal mechanisms between the reactors were hypothesized to be due to fluctuating hydraulic residence times at the ML site, in turn, due to unregulated runoff flow.

Enhanced reactivity of superoxide in water-solid matrices.

Superoxide is unreactive in deionized water, but aqueous systems containing added solvents, including H2O2 at >100 mM, show significantly increased reactivity of superoxide with oxidized organic compounds such as highly chlorinated aliphatics. The potential for solid surfaces to similarly increase the reactivity of superoxide in water was investigated. Heterogeneous birnessite (gamma-MnO2)-catalyzed decomposition of H2O2 promoted the degradation of the superoxide probe hexachloroethane (HCA) at H202 concentrations as low as 7.5 mM, while no measurable HCA degradation was found in parallel homogeneous iron(III)-EDTA-H2O2 systems at H2O2 concentrations <100 mM. Electron spin resonance spectroscopy confirmed that superoxide was the dominant reactive species generated in the birnessite-catalyzed decomposition of H2O2. Increased superoxide reactivity was also found in aqueous superoxide-glass bead heterogeneous systems, and the rates of HCA degradation increased as a function of the surface area of the glass beads. The results of this research show that, similar to the addition of solvents, the presence of surfaces also enhances the reactivity of superoxide in water, possibly by altering the superoxide solvation shell. On the basis of these findings, superoxide generated in catalyzed H2O2 propagations (CHP; modified Fenton's reagent) used for in situ chemical oxidation (ISCO) may have greater reactivity with highly oxidized contaminants than previously thought.

Electrochemical detection of triacetone triperoxide employing the electrocatalytic reaction of iron(II/III)-ethylenediaminetetraacetate and hydrogen peroxide.

An electrochemical method for the detection of triacetone triperoxide (TATP) is proposed and examined. In this method, TATP solutions were treated with 1.08 M HCl for 10min releasing H2O2 and/or hydroperoxides. Subsequently, these peroxides undergo an electrocatalytic reduction through the Fe(II/III)ethylenediaminetetraacetate (EDTA) complex at a glassy carbon electrode. Cyclic voltammetric results indicate that no redox reaction was observed between Fe(II)EDTA and TATP. Acid treated TATP yielded voltammograms indicative of electrocatalysis of ROOH/HOOH reduction via Fe(II/III)EDTA redox cycling. Chronoamperometric results yielded a detection limit of 0.89 microM for TATP and a sensitivity of 0.025 mAmM(-1). The influence of pH and O2 interference on the analytical signal is briefly discussed.

EDTA degradation induced by oxygen activation in a zerovalent iron/air/water system.

A method for the removal of ethylenediaminetetraacetic acid (EDTA) at room temperature and 1 atm is demonstrated. EDTA (1 mM, 50 mL) containing 2.5 g of granular zerovalent iron (ZVI) (20-40 mesh) was degraded in 2.5 h. Using a recently developed form of O2 activation, reactive oxygen species are generated in situ, resulting in the degradation of EDTA when complexed with FeII. ESI-MS measurements indicate that degradation of EDTA yields low-molecular carboxylic acids. The presence of oxygen is crucial: the observed pseudo-first-order rate constants for EDTA removal are kobs = 1.02 h(-1) (kSA = 1.85 +/- 0.046 L h(-1) m(-2)) and kobs = 0.04 h(-1) (kSA = 0.00724 +/- 0.002 L h(-1) m(-2)) under air and under N2 purge, respectively. kSA represents surface area normalized rate constants. Large excesses of EDTA in the reaction mixture slowthe rate of degradation. Increasing the concentration of EDTA from 1.0 to 10.0 mM while holding all other parameters constant gave observed rates of kobs = 1.02 +/- 0.26 h(-1) (kSA = 1.85 +/- 0.046 L h(-1) m(-2)) and kobs = 0.044 +/- 0.01 h(-1) (kSA = 0.00796 +/- 0.002 L h(-1) m(-2)), respectively. The rate-limiting step is determined to be homogeneous oxygen activation.

Voltammetric studies of Zn and Fe complexes of EDTA: evidence for the push mechanism.

The 'push' hypothesis for the antioxidant action of Zn2+ is based on its displacement of iron from a low molecular weight pro-oxidant complex. In this study, the chemical plausibility of that proposed function is investigated by cyclic voltammetry. As a model for a pro-oxidative low molecular weight iron complex the Fe(II/III)/EDTA couple was examined. This complex was selected for its well-defined electrochemical, iron stability constants, and similarity to other low molecular weight chelates in physiological fluids in terms of logical binding sites, i.e. amino, and carboxylate groups. Also investigated were iron complexes of nitrilotriacetic acid and DL-glutamic acid. Results demonstrate that approximately 90% of the cyclic voltammetric peak current for Fe(III)EDTA reduction and the EC' current for the mediated reduction of H2O2 by Fe(II/III)EDTA (Fenton Reaction) are lost when Zn2+ is introduced to a 1:1 molar ratio relative to iron. All experiments were conducted in HEPES buffered solutions at pH 7.4. Iron (II/III) complexes of nitrilotriacetic acid and DL-glutamic acid followed the same trends. Cyclic voltammetric experiments indicate that Zn2+ displaces Fe(III) from EDTA despite the much larger stability constant for the iron complex (10(25.1)) versus zinc (10(16.50)). The hydrolysis aided displacement of Fe(III) from EDTA by Zn2+ is considered by the equilibria modeling program, HySS. With Fe(III) hydrolysis products included, Zn2+ is able to achieve 90% displacement of iron from EDTA, a result consistent with cyclic voltammetric observations.

Stability of ferric complexes with 3-hydroxyflavone (flavonol), 5,7-dihydroxyflavone (chrysin), and 3',4'-dihydroxyflavone.

The acid dissociation and ferric stability constants for complexation by the flavonoids 3-hydroxyflavone (flavonol), 5,7-dihydroxyflavone (chrysin), and 3',4'-dihydroxyflavone in 50:50 (v/v) ethanol/water are determined by pH potentiometric and spectrophotometric titrations and the linear least-squares curve-fitting program Hyperquad. Over the entire range of pH and reagent concentrations spanning the titration experiments, the stoichiometry for iron-flavonoid complex formation was 1:1 for all three flavonoids examined. The three flavonoids were chosen for their hydroxy substitution pattern, with each possessing one of the three most commonly suggested sites for metal binding by the flavonoids. On the basis of the calculated stability constants, the intraflavonoid-binding site competition is illustrated as a function of pH via speciation curves. The curves indicate that the binding site comprised of the 3',4'-hydroxy substitutions, the catecholic site, is most influential for ferric complexation at the physiological pH of 7.4. The possibility for antioxidant activity by flavonoid chelation of ferric iron in the presence of other competitive physiological complexing agents is demonstrated through additional speciation calculations.

A hypothesis for the basis of the pro-oxidant nature of calcium ions.

A new hypothesis describing the role of the redox inactive Ca2+ ion in the expression of physiological oxidative damage is described. The hypothesis is based on the optimization of the chelation characteristics of iron complexes for pro-oxidant activity. In a previous investigation it was found that an excess of ligand kinetically hindered the Fenton reaction activity of the FeII/III EDTA complex (Bobier et al. 2003). EDTA, citrate, NTA, and glutamate were selected as models for the coordination sites likely encountered by mobile iron, i.e. proteins. The optimal [EDTA]:[FeIII] ratio for Fenton reaction activity as measured by electrocatalytic voltammetry in a solution was found to be 1:1. An excess of EDTA in the amount of 10:1 [ligand]: [metal] suppresses the Fenton reaction activity to nearly the control. It is expected that the physiological coordination characteristics of mobile Fe would have a very large excess of [ligand]:[metal] and thus not be optimized for the Fenton reaction. Introduction of Ca2+ in to a ratio of 10:10:1 [Ca2+]:[EDTA]:[FeIII] to the system reinvigorated the Fenton reaction activity to nearly the value of the optimal 1:1 [EDTA] :[FeIII] complex. The pH distribution diagrams of Ca2+ in the presence of EDTA and FeII/III indicate that Ca2+ has the ability to uptake excess EDTA without displacing either FeII of FeIII from their respective complexed forms. The similarity in the presence for hard ligand sites albeit with a lower binding constant for Ca2+ accounts for this action.

Variability of the Fenton reaction characteristics of the EDTA, DTPA, and citrate complexes of iron.

The common metal chelation agents, DTPA and EDTA are often used as models for physiological low-molecular weight iron complexes in biochemical studies, or for common biochemical protocols. In the biochemical literature there are apparent conflicts as to whether EDTA and DTPA are pro-oxidant or antioxidant additives. This apparent conflict is puzzling since in chemical systems FeIIEDTA and FeIIDTPA are well known Fenton reaction reagents. In this investigation we examined the voltammetric characteristics of the iron complexes of EDTA, DTPA, and citrate and the effect of the ligand:metal ratio (L:M) on the electrocatalytic (EC') waves that result from reduction of H2O2 by this complex. At a ratio of 1:1, the cyclic voltammetric waves of the complexes indicate the presence of a reversible species corresponding to the Fe(II/III)L couple, along with a second irreversible reduction peak. The second irreversible voltammetric peak decreases at higher L:M ratios for EDTA and citrate. The 1:1 iron complexes of EDTA, DTPA, and citrate clearly induce the catalytic reduction of H2O2. In the presence of a greater than 100 fold excess of H2O2 relative to iron, higher L:M ratios greatly reduced the catalytic EC' wave compared to the 1:1 ratios. At H2O2:Fe ratios less than 50, the L:M ratio has very little effect of the EC' current. These observations may explain the apparent discrepancies in the biochemical literature. Addition of EDTA or DTPA may enhance oxidative processes if the L:M is low (less than unity), whereas rates of on-going oxidative processes may decrease if that ratio, along with the relative amount of H2O2, are both high (excess ligand). The impact of this study is of particular importance given the widespread use of these ligands in biochemical studies.

Supercritical fluid extraction and high-performance liquid chromatography-diode array-electrochemical detection of signature redox compounds from sand and soil samples.

A supercritical fluid extraction procedure and a chromatographic separation/detection method were developed for the detection of Earth-based microorganisms. After microbes in a sand or a soil sample were hydrolyzed in a diluted NH(4)OH/acetone solution, several redox compounds from bacteria could be effectively extracted with trimethylamine-modified supercritical CO(2) at 35 degrees C and 300 atm. These signature redox-active compounds were separated by a reversed-phase HPLC column in an ion-pair mode and then monitored with a diode array detector and an electrochemical detector. The analytical results demonstrated the feasibility of using the reported techniques to detect the chemical signature of life in barren desert sand samples.

Dechlorination of chlorinated phenols by catalyzed and uncatalyzed Fe(0) and Mg(0) particles.

Uncatalyzed, and palladium-catalyzed Fe(0) and Mg(0) systems were examined for their efficiencies of dechlorination of 2.86 mM 4-chlorophenol (4-CP), 2.52 mM 2,6-dichlorophenol (2,6-DCP), 3.03 mM 2,4,6-trichlorophenol (2,4,6-TCP), and 2.48 mM pentachlorophenol (PCP) in 50/50 (v/v) 2-propanol/water under room temperature and pressure conditions. Previous investigators have found that PCP is extremely recalcitrant under these conditions. In this investigation, complete dechlorination of 5.0 ml of 2.48 mM PCP was observed for 1.0 g of 2659 ppm Pd/Mg (20 mesh) after 48 h. The only detectable products were cyclohexanol and cyclohexanone at 25% yield. No other chlorinated or otherwise products were observed by mass spectral analysis. It is hypothesized that volatile low molecular weight species were formed from the Pd/Mg dechlorination of PCP. Under conditions of equal surface area (0.0786 m2), the approximate order of PCP dechlorination power of these systems followed as 2659 ppm Pd/Mg>319 ppm Pd/Mg>Mg approximately 4856 ppm Pd/Fe>Fe. Degradation of the other chlorinated phenols by all metallic systems was more facile than PCP.