Real‐Time Monitoring of Mass‐Transport‐Related Enzymatic Reaction Kinetics in a Nanochannel‐Array Reactor

To understand the fundamentals of enzymatic reactions confined in micro‐/nanosystems, the construction of a small enzyme reactor coupled with an integrated real‐time detection system for monitoring the kinetic information is a significant challenge. Nano‐enzyme array reactors were fabricated by covalently linking enzymes to the inner channels of a porous anodic alumina (PAA) membrane. The mechanical stability of this nanodevice enables us to integrate an electrochemical detector for the real‐time monitoring of the formation of the enzyme reaction product by sputtering a thin Pt film on one side of the PAA membrane. Because the enzymatic reaction is confined in a limited nanospace, the mass transport of the substrate would influence the reaction kinetics considerably. Therefore, the oxidation of glucose by dissolved oxygen catalyzed by immobilized glucose oxidase was used as a model to investigate the mass‐transport‐related enzymatic reaction kinetics in confined nanospaces. The activity and stability of the enzyme immobilized in the nanochannels was enhanced. In this nano‐enzyme reactor, the enzymatic reaction was controlled by mass transport if the flux was low. With an increase in the flux (e.g., >50 μL min^?1), the enzymatic reaction kinetics became the rate‐determining step. This change resulted in the decrease in the conversion efficiency of the nano‐enzyme reactor and the apparent Michaelis–Menten constant with an increase in substrate flux. This nanodevice integrated with an electrochemical detector could help to understand the fundamentals of enzymatic reactions confined in nanospaces and provide a platform for the design of highly efficient enzyme reactors. In addition, we believe that such nanodevices will find widespread applications in biosensing, drug screening, and biochemical synthesis.     

A pH responsive electrochemical switch sensor based on Fe(notpH3) [notpH6 = 1,4,7-triazacyclononane-1,4,7-triyl-tris(methylene-phosphonic acid)

The electrochemistry of a macrocyclic metal complex Fe(notpH₃) [notpH₆ = 1,4,7-triazacyclononane-1,4,7-triyl-tris(methylene-phosphonic acid)] reveals that the protonation/deprotonation of the non-coordinated P-OH groups in Fe(notpH₃) affects its formal potential value (E⁰′) considerably. Plotting E⁰′ as function of solution pH gives a straight line with a slope of −585 mV pH⁻¹ in the pH range of 3.4-4.0, which is about ten times larger than the theoretical value of −58 mV pH⁻¹ for a reversible proton-coupled single-electron transfer at 20 °C. A sensitive pH responsive electrochemical switch sensor is thus developed based on Fe(notpH₃) which shows an “on/off” switching at pH ∼ 4.0.     

A two-photon excited luminescence of water-soluble rhodamine-platinum(II) complex: fluorescent probe specific for Hg2+ detection in live cell

The first example of cyclometalated platinum(II)-containing rhodamine probe (1) with two-photon induced luminescent properties was synthesized and investigated for mercury detection. A highly selective color change of 1, from light yellow to pink, is observed only in the presence of Hg²⁺ due to the formation of 1,3,4-oxadiazole ring in 2. This selectivity of Hg²⁺ with color changes can be observed easily by the naked-eye. Meanwhile, a remarkable turn-on and selective 20-fold fluorescent enhancement of 1 upon binding with Hg²⁺ over the other tested metal ions was observed. The water-soluble probe 1 was successfully applied in the visualizing of the site of Hg²⁺ accumulation as well as estimating of trace amounts of mercury ions in live HeLa cells by two-photon microscopy.     

Evaluation of a new electrolytic cold vapor generation system for mercury determination by AFS

In this study we firstly report a new electrolytic cold vapor generation system for mercury determination on Pt/Ti cathode in the presence of organic acid catholyte. Comparing with the traditional inorganic acid, formic acid increased the signal intensity of Hg vapor from electrolytic generation on Pt cathode and reduced the impact of cathode erosion on the stability of signal intensity. Moreover, formic acid has better interference tolerance. The introduction location for carrier gas is probably the most important factor that influences the signal intensity of Hg from electrolytic vapor generation. The effects of the electrolytic conditions and interference ions on the ECVG have been studied. Under the optimized conditions, the detection limit (3σ) of Hg (II) in aqueous solutions is 1.4 ng L⁻¹; a relative standard deviation of 2.3% for 1 μg L⁻¹ Hg was obtained. The accuracy of this method was verified by the determination of mercury in the certified reference materials. This system has been applied satisfactorily to the determination of Hg in Traditional Chinese Medicines samples.     

Nonenzymatic Electrochemical Glucose Sensor Based on Pt Nanoparticles/Mesoporous Carbon Matrix

A nonenzymatic amperometric sensor for sensitive and selective detection of glucose has been constructed by using highly dispersed Pt nanoparticles supported onto mesoporous carbons (MCs). The Pt nanoparticles/mesoporous carbons (Pt/MCs) composites modified electrode displayed high electrocatalytic activity towards the oxidation of glucose. At an applied potential of 0.1 V, the Pt/MCs electrode has a linear dependence (R=0.996) in the glucose concentration up to 7.5 mM with a sensitivity of 8.52 mA M^?1 cm^?2. The Pt/MCs electrode has also shown highly resistant toward poisoning by chloride ions and without interference from the oxidation of common interfering species.     

New Guaipyridine Sesquiterpene Alkaloids from Artemisia rupestris L.

Three new guaipyridine sesquiterpene alkaloids, rupestine A, B, C ( 1 – 3 , resp.), and the new nor‐sesquiterpne alkaloid rupestine D ( 4 ) were obtained from the flowers of Artemisia rupestris L. Their structures were elucidated on the basis of sepectroscopic data and by comparison with those of the related compounds reported in the literature. In addition, the absolute configurations of 2 and 4 were determined by single‐crystal X‐ray diffraction analyses.     

Size dependent active effect of CdTe quantum dots on pyrogallol-H2O2 chemiluminescence system for chromium(III) detection

Water-soluble CdTe quantum-dots (QDs) of different sizes and capped with mercaptosuccinic acid were prepared by the microwave irradiation method. The QDs can significantly enhance the chemiluminescence (CL) of the pyrogallol-H₂O₂ system. Those with a diameter of 3.8 nm produce the most intense CL. UV-vis, photoluminescence, and CL spectra were acquired in order to explore the effect. The results showed that the chromium(III) ion in the concentration range from 20 pM to 30 µM enhances CL, and this is exploited for its trace determination.The limit of detection (3σ) is 6 pM, with a relative standard deviation (n?=?11) of 1.7%. A continuous flow injection CL method was developed and applied to the determination of chromium(III) in tap water and lake water samples with satisfactory results.     

A Proof for Substitution of Endogenous Iron (II) in Lipoxygenase by Exogenous Cu2+

Soybean lipoxygenase (LOX) contains endogenous iron (II) at the active site, which is important for the enzyme activity. The activity of LOX can be accelerated by some exogenous metal ions including Cu²⁺. However, the mechanism of the activity improvement caused by exogenous metal ions remains unclear, not only for LOX but for most other metalloenzymes. Meanwhile, the possibility that exogenous metal ions can displace endogenous iron (II) is still in discussion for a lack of a direct and quantitative proof. In this paper, a quantitative proof of replacing iron (II) inside LOX by exogenous Cu²⁺ was provided, simply using UV-Vis spectrometry with two indicators p-carboxylantipyrylazo and 9-(4-carboxyphenyl)-2,3,7-trihydroxyl-6-fluorine. A 0.56 μM free iron (II) was observed in the bulk solution after incubating 9.45 μM Cu²⁺ with 16.10 μM LOX at 20°C for 5 min, which is in coincidence with the decrement of Cu²⁺ in the bulk solution (0.53 μM), implying that iron (II) was replaced by Cu²⁺.