Immobilization of DNAzyme catalytic beacons on PMMA for Pb2+ detection

Due to the numerous toxicological effects of lead, its presence in the environment needs to be effectively monitored. Incorporating a biosensing element within a microfluidic platform enables rapid and reliable determinations of lead at trace levels. A microchip-based lead sensor is described here that employs a lead-specific DNAzyme (also called catalytic DNA or deoxyribozyme) as a recognition element that cleaves its complementary substrate DNA strand only in the presence of cationic lead (Pb(2+)). Fluorescent tags on the DNAzyme translate the cleavage events to measurable, optical signals proportional to Pb(2+) concentration. The DNAzyme responds sensitively and selectively to Pb(2+), and immobilizing DNAzyme in the sensor permits both sensor regeneration and localization of the detection zone. Here, the DNAzyme has been immobilized on a PMMA surface using the highly specific biotin-streptavidin interaction. The strategy includes using streptavidin physisorbed on a PMMA surface to immobilize DNAzyme both on planar PMMA and on the walls of a PMMA microfluidic device. The immobilized DNAzyme retains its Pb(2+) detection activity in the microfluidic device and can be regenerated and reused. The DNAzyme shows no response to other common metal cations and the presence of these contaminants does not interfere with the lead-induced fluorescence signal. While prior work has shown lead-specific catalytic DNA can be used in its solubilized form and while attached to gold substrates to quantitate Pb(2+) in solution, this is the first use of the DNAzyme immobilized within a microfluidic platform for real time Pb(2+) detection.     

Lead as own luminescent sensor for determination

In our experiments, it was observed that adding bromide to Pb2+ solution of N,N'-dimethylformamide (DMF), the highly emissive cluster Pb4Br11(3-) can be formed and the fluorescence intensity of the formed cluster is proportional to the concentration of Pb2+, based on which, a novel, simple approach that uses the emission from itself as the sensor for determination of Pb2+ is proposed. Under the optimum conditions, the linear range and detection limit is 1.0 x 10(-7) to 1.0 x 10(-5) mol l(-1) (correlation coefficient r = 0.9997) and 7.6 x 10(-9) mol l(-1), respectively. Foreign substrates effects were also investigated. The proposed method has been successfully applied to the determination of lead in the synthetic samples. The mechanism of the reaction is also studied.     

Bismuth Film Voltammetric Sensor on Pyrolyzed Photoresist/Alumina Support for Determination of Heavy Metals

Voltammetric sensors based on bismuth film electrodes are an attractive alternative to other sensors for application in electroanalysis of heavy metals. Bismuth film electrodes can be formed by a similar method on the same substrates as mercury. These systems were used most frequently for simultaneous determination of heavy metals such as Pb, Cd and Zn by anodic stripping voltammetry. Our voltammetric sensor was fabricated on an alumina substrate. A photoresist film prepared by pyrolysis of positive photoresist S‐1813 SP15 on the alumina substrate was used as an electrode support for bismuth film deposition. The influence of the Nafion membrane on the measurement sensitivity of the sensor and mechanical stability of the bismuth film were investigated. The sensor was successfully applied for determination of Pb, Cd and Zn in an aqueous solution in the concentration range of 0.2 to 10 µg L^?1 by square wave anodic stripping voltammetry on an in‐situ formed bismuth film electrode with Nafion‐coating. Parameters of the sensor such as sensitivity, linearity, detection limit, repeatability and life‐time were evaluated. In the best case, the detection limits were estimated as 0.07, 0.11 and 0.63 µg L^?1 for Pb, Cd and Zn, respectively. Finally, the applicability of the sensor was tested in analysis of Pb, Cd and Zn in real samples of tap and river water using the method of standard additions.     

Sensitive Electrochemical Detection of Pb(II) and H2O2 via a Dual-functional Sn-doped Defective Bi2S3 Microspheres

In this work, a dual-functional electrochemical sensor has been proposed based on Sn-doped defective Bi₂S₃ (TDDB) microspheres, which exhibited the excellent electrochemical performance on Pb(II) and H₂O₂ detection. The TDDB offered a satisfied detection limit of 8.0 nM towards Pb(II) with a sensitivity of 96.7 μA ⋅ μM⁻¹. As a H₂O₂ sensor, a high sensitivity of 3540 μA mM⁻¹ cm⁻² was obtained in a linear range from 0.45 mM to 10 mM with a detection limit of 10 nM. Moreover, the electrochemical detection of Pb(II) in Taihu Lake and H₂O₂ in human serum was achieved with high reliability and good recovery.     

Inorganic-organic Ag-rhodamine 6G hybrid nanorods: "turn on" fluorescent sensors for highly selective detection of Pb2+ ions in aqueous solution

Lead metal ions are of great concern and the monitoring of their concentration in the environment has become extremely important. In the present study, a new inorganic-organic hybrid assay of Ag nanorods (AgNR)-Rhodamine 6G (R6G) was developed for the sensitive and selective determination of Pb(2+) ions in aqueous solutions. To the best of our knowledge there is almost no literature on the use of silver nanorod sensors for determination of lead ions in aqueous solutions. The sensor is developed by the coating of R6G on the surface of AgNRs. The sensing is based on the photoluminescence of R6G. The sensor was rapid as the measurements were carried out within 3 min of addition of the test solution to the AgNR-R6G hybrid. Moreover, the system showed excellent stability at tested concentration levels of Pb(2+) ions. The naked eye detection of the colour was possible with 1 mg L(-1) of Pb(2+) ions. The present method has a detection limit of 50 μg L(-1) of Pb(2+) (for a signal/noise (S/N) ratio > 3). The selectivity toward Pb(2+) ions against other metal ions was improved using chelating agents. The proposed method was validated by analysis using different techniques.     

Lead(II) Potentiometric Sensor Based on 1,4,8,11‐Tetrathiacyclotetradecane Neutral Carrier and Lipophilic Additives

A potentiometric sensor for lead(II) ions based on the use of 1,4,8,11‐tetrathiacyclotetradecane (TTCTD) as a neutral ionophore and potassium tetrakis‐(p‐chlorophenyl)borate as a lipophilic additive in plasticized PVC membranes is developed. The sensor exhibits linear potentiometric response towards lead(II) ions over the concentration range of 1.0×10^?5–1.0×10^?2 mol L^?1 with a Nernstian slope of 29.9 mV decade^?1 and a lower limit of detection of 2.2×10^?6 mol L^?1 Pb(II) ions over the pH range of 3–6.5. Sensor membrane without a lipophilic additive displays poor response. The sensor shows high selectivity for Pb(II) over a wide variety of alkali, alkaline earth and transition metal ions. The sensor shows long life span, high reproducibility, fast response and long term stability. Validation of the method by measuring the lower limit of detection, lower limit of linear range, accuracy, precision and sensitivity reveals good performance characteristics of the proposed sensor. The developed sensor is successfully applied to direct determination of lead(II) in real samples. The sensor is also used as an indicator electrode for the potentiometric titration of Pb(II) with EDTA and potassium chromate. The results obtained agree fairly well with data obtained by AAS.     

Visual detection of lead(II) using a label-free DNA-based sensor and its immobilization within a monolithic hydrogel

Lead is highly toxic and its detection has attracted a lot of research interests. In recent years, DNA has been used for Pb(2+) recognition and many fluorescent sensors with low to sub-nM detection limits have been reported. These figures of merit were typically measured using a spectrophotometer that can detect nM DNA with a high signal-to-noise ratio. For visual detection, however, μM DNA or dye was required, making it difficult to detect low nM targets. We recently achieved a visual sensitivity of 10 nM Hg(2+) by immobilizing a DNA probe in a hydrogel. This was made possible because the gel was able to actively adsorb Hg(2+). In this work, we aim to test whether this method can be extended to the detection of Pb(2+). First, a new Pb(2+) sensor was designed based on a guanine-rich DNA and DNA binding dyes such as thiazole orange and SYBR Green I. The free DNA showed a detection limit of 8 nM Pb(2+) using 40 nM DNA. For visual detection in solution with 1 μM of the DNA probe, however, ～300 nM Pb(2+) was required. After immobilization in a monolithic polyacrylamide hydrogel, even 20 nM Pb(2+) could be visually detected with a sample volume of 50 mL. Therefore, sensitive detection without signal amplification was achieved. Finally, we demonstrated simultaneous detection of both Hg(2+) and Pb(2+) in the same water sample with shape encoded hydrogel sensors.     

Pb(2+)-introduced activation of horseradish peroxidase (HRP)-mimicking DNAzyme

A novel nucleic acid hairpin structure composed of Pb(2+)-dependent DNAzyme and HRP-mimicking DNAzyme was developed. This hairpin structure can be used as a sensor for the detection of Pb(2+) based on colorimetry.     

Polymerized crystalline colloidal array chemical-sensing materials for detection of lead in body fluids

We have developed intelligent polymerized crystalline colloidal array (IPCCA) chemical-sensing materials for detection of Pb(2+) in high ionic-strength environments such as body fluids with a detection limit of <500 nmol L(-1) Pb(2+) (100 ppb). This IPCCA lead sensor consists of a mesoscopically periodic array of colloidal particles polymerized into an acrylamide hydrogel. The array Bragg-diffracts light in the visible spectral region because of the periodic spacing of the colloidal particles. This material also contains a crown ether chelating agent for Pb(2+). Chelation of Pb(2+) by the IPCCA in low-ionic-strength solutions results in a Donnan potential that swells the gel, which red-shifts the diffracted light in proportion to the Pb(2+) concentration. At high ionic strength the Donnan potential is, unfortunately, swamped and no static response occurs for these sensors. We demonstrate, however, that we can determine Pb(2+) at high ionic strength by incubating these IPCCA in a sample solution and then measuring their transient response on exposure to pure water. The non-complexed ions diffuse from the IPCCA faster than the bound Pb(2+). The resulting transient IPCCA diffraction red-shift is proportional to the concentration of Pb(2+) in the sample. These IPCCA sensors can thus be used as sensing materials in optrodes to determine Pb(2+) in high-ionic-strength solutions such as body fluids.     

Label-free DNAzyme-based fluorescing molecular switch for sensitive and selective detection of lead ions

We developed a label-free fluorescent Pb(2+) sensor utilizing a DNAzyme-based fluorescing molecular switch, which enables fluorescence detection of Pb(2+) in aqueous solution with high sensitivity and selectivity.     

2,1,3-Benzoxadiazole-based selective chromogenic chemosensor for rapid naked-eye detection of Hg2+ and Cu2+

We report a simple twisted intramolecular charge transfer (TICT) chromogenic chemosensor for rapid and selective detection of Hg(2+) and Cu(2+). The sensor was composed of an electron-acceptor 4-fluoro moiety and an electron-donor 7-mercapto-2,1,3-benzoxadiazole species where the S together with the 1-N provided the soft binding unit. Upon Hg(2+) and Cu(2+) complexation, remarkable but different absorbance spectra shifts were obtained in CH(3)CN-H(2)O mixed buffer solution at pH 7.6, which can be easily used for naked-eye detection. The sensor formed a stable 2:1 complex with Cu(2+), and both 2:1 and 3:1 complexes with Hg(2+). While alkali-, alkaline earth- and other heavy and transition metal ions such as Na(+), Mg(2+), Mn(2+), Co(2+), Ni(2+), Ag(+), Zn(2+), Pb(2+) and Cd(2+) did not cause any significant spectral changes of the sensor. This finding is not only a supplement to the detecting methods for Hg(2+) and Cu(2+), but also adds new merits to the chemistry of 4,7-substituted 2,1,3-benzoxadiazoles.     

A flow cell optosensor for lead based on immobilized gallocynin in chitosan membrane

Gallocynin immobilized in chitosan membrane has been studied as a sensor element of an optical sensor for lead using a flowing system. By using this set up, lead in solution has been determined in the concentration range from 1.0x10(-1) to 1.0x10(3) ppm with a detection limit of 0.075 ppm. The standard deviation of the method for the repeatability of lead detection at a concentration of 100 ppm was found to be 2.10%. The response of the sensor was reproducible and can be regenerated by using acidified saturated KNO(3) solution. Interference from foreign ions was also studied at 1:1 mole ratio of Pb(II):foreign ions.     

基于 G-四联体门控制效应的铅离子适配体传感器

将富鸟嘌呤(G)序列核酸适配体与门控制效应相结合,通过控制门的开关实现信号放大,构建了新型电化学生物传感器,用于铅离子(Pb2+)的高灵敏检测。首先将单-6-巯基-β-环糊精(6-SH-β-CD)自组装在金电极上,形成有序排列的分子自组装膜,且分子之间留有空隙,可作为探针通过的门结构。随后将发夹结构的富含 G 序列的核酸适配体修饰在环糊精次面端口,制得可特异性识别 Pb2+的电化学生物传感器。富 G 序列适配体结合 Pb2+后可折叠形成 G-四联体结构(G4-Pb2+),覆盖住电极表面的探针通道,产生关门效应,使探针氧化还原电流强度减小,进而形成门控制效应,利用该效应可进行 Pb2+的定量检测。门控制效应显著提高了信噪比和检测的灵敏度,在1×10-13~5×10-11 mol/ L 浓度范围内,Pb2+浓度的负对数与 DPV 响应电流呈良好的线性关系,检出限为3.6×10-14 mol/ L(DL=3δb / K)。传感器用于实际水样品中 Pb2+的测定,结果令人满意。     

Direct detection of Pb in urine and Cd, Pb, Cu, and Ag in natural waters using electrochemical sensors immobilized with DMSA functionalized magnetic nanoparticles

Urine is universally recognized as one of the best non-invasive matrices for biomonitoring exposure to a broad range of xenobiotics, including toxic metals. Detection of metal ions in urine has been problematic due to the protein competition and electrode fouling. For direct, simple, and field-deployable monitoring of urinary Pb, electrochemical sensors employing superparamagnetic iron oxide (Fe3O4) nanoparticles with a surface functionalization of dimercaptosuccinic acid (DMSA) has been developed. The metal detection involves rapid collection of dispersed metal-bound nanoparticles from a sample solution at a magnetic or electromagnetic electrode, followed by the stripping voltammetry of the metal in acidic medium. The sensors were evaluated as a function of solution pH, the binding affinity of Pb to DMSA-Fe3O4, the ratio of nanoparticles per sample volume, preconcentration time, and Pb concentrations. The effect of binding competitions between the DMSA-Fe3O4 and urine constituents for Pb on the sensor responses was studied. After 90 s of preconcentration in samples containing 25 vol.% of rat urine and 0.1 g L(-1) of DMSA-Fe3O4, the sensor could detect background level of Pb (0.5 ppb) and yielded linear responses from 0 to 50 ppb of Pb, excellent reproducibility (%RSD of 5.3 for seven measurements of 30 ppb Pb), and Pb concentrations comparable to those measured by ICP-MS. The sensor could also simultaneously detect background levels (<1 ppb) of Cd, Pb, Cu, and Ag in river and seawater.     

Simultaneous and Sensitive Detection of Cd(II) and Pb(II) Using a Novel Bismuth Film/Ordered Mesoporous Carbon‐molecular Wire Modified Graphite Carbon Paste Electrode

An electrochemical sensor for the simultaneous and sensitive detection of Cd(II) and Pb(II) is proposed on the basis of square‐wave anodic stripping voltammetry (SWASV) experiments using a novel bismuth film/ordered mesoporous carbon‐molecular wire modified graphite carbon paste electrode (Bi/OMC‐MW/GCPE). Ordered mesoporous carbon (OMC) and molecular wire (MW) (diphenylacetylene) were used as the modifier and binder, respectively. The Bi/OMC‐MW/GCPE was prepared with the addition of graphite powder, OMC and DPA at the ratio of 2 : 1 : 1. The electrochemical properties and morphology of the electrode were characterized by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), SWASV and scanning electron microscopy (SEM). The parameters affecting the stripping current response were investigated and optimized. The experimental results show that the prepared electrode exhibited excellent electrochemical performance, good electrical conductivity and a high stripping voltammetric response. Under optimized conditions, a linear range was achieved over a concentration range from 1.0 to 70.0 μg/L for both Cd(II) and Pb(II) metal ions, with detection limits of 0.07 μg/L for Cd(II) and 0.08 μg/L for Pb(II) (S/N=3) with the deposition time 150 s. Moreover, the sensor exhibited improved sensitivity and reproducibility compared to traditional CPEs. The fabricated electrode was then successfully used to satisfactorily detect Cd(II) and Pb(II) in real soil samples.     

Electroanalytical Determination of Cd(II) and Pb(II) in Bivalve Mollusks using Electrochemically Reduced Graphene Oxide‐based Electrode

An electrochemical sensor for the simultaneous determination of Cd(II) and Pb(II) by square wave anodic stripping voltammetry (SWASV) in bivalve mollusks using a glassy carbon electrode modified with electrochemically reduced graphene oxide has been developed. The modified surface was characterized by cyclic voltammetry, high resolution scanning electron microscopy (HR‐SEM), and Raman spectroscopy. The optimum conditions were optimized and a linear range was observed from 15–105 μg L^?1 with a limits of detection of 15 μg L^?1 for Cd(II) and Pb(II). The methodology was validated and applied in different samples of commercial bivalve mollusks with satisfactory results. The high conductivity and greater surface area of the modifying agent improves the preconcentration capacity of the electrochemical sensor, allowing to develop a simple, rapid and sensitive analysis in the detection of lead and cadmium in marine resources.     

Automated portable analyzer for lead(II) based on sequential flow injection and nanostructured electrochemical sensors

A fully automated portable analyzer for toxic metal ion detection based on a combination of a nanostructured electrochemical sensor and a sequential flow injection system has been developed in this work. The sensor was fabricated from a carbon paste electrode modified with acetamide phosphonic acid self-assembled monolayer on mesoporous silica (Ac-Phos SAMMS) which was embedded in a very small wall-jet (flow-onto) electrochemical cell. The electrode is solid-state and mercury-free. Samples and reagents were injected into the system and flowed through the electrochemical cell by a user programmable sequential flow technique which required minimal volume of samples and reagents and allowed the automation of the analyzer operation. The portable analyzer was evaluated for lead (Pb) detection due to the excellent binding affinity between Pb and the functional groups of Ac-Phos SAMMS as well as the great concern for Pb toxicity. Linear calibration curve was obtained in a low concentration range (1-25 ppb of Pb(II)). The reproducibility was excellent; the percent relative standard deviation was 2.5 for seven consecutive measurements of 10 ppb of Pb(II) solution. Excess concentrations of Ca, Ni, Co, Zn, and Mn ions in the solutions did not interfere with detection of Pb, due to the specificity and the large number of the functional groups on the electrode surface. The electrode was reliable for at least 90 measurements over 5 days. This work is an important milestone in the development of the next-generation metal ion analyzers that are portable, fully automated, and remotely controllable.     

Highly Sensitive and Selective Detection of Pb(II) by NH2−SiO2/Ru(bpy)32+−UiO66 based Solid‐state ECL Sensor

In this study, a novel electrochemiluminescence (ECL) sensor for highly sensitive and selective detection of Pb(II) was developed based on Ru(bpy)₃²⁺ encapsulated UiO66 metal‐organic‐framework (Ru(bpy)₃²⁺?UiO66 MOF) and ?NH₂ group functionalized silica (NH₂?SiO₂). The NH₂?SiO₂ with large surface area provided an excellent platform for the ECL sensor. As numerous exposed carboxyl groups were present on UiO66 backbone, the Ru(bpy)₃²⁺?UiO66 could be steadily immobilized to NH₂?SiO₂ by forming amide bonds. Meanwhile, the introduced UiO66 MOF which used for the encapsulation of Ru(bpy)₃²⁺, significantly enhanced the ECL efficiency of the proposed sensor, as it possessed a large specific surface area and porosity for the loading of Ru(bpy)₃²⁺. Moreover, a high quenching effect on ECL intensity was obtained in the presence of Pb(II) in the electrolyte. Under the optimal conditions, the quenched ECL intensity showed a good linear relationship within Pb(II) concentration in the range from 1.0×10^?6 to 1.0×10² μM, with a detection limit of 1.0×10^?7 μM (S/N=3). The proposed sensor for Pb(II) detection was simple in operation, rapid in testing, stable in signal, and showed a good anti‐interference ability to some other metal ions. Besides, its application for detecting Pb(II) in a real sample was also investigated here. This work provides a potential platform for metal ions detection in environmental monitoring field.     

A novel 5-mercapto triazole Schiff base as a selective chromogenic chemosensor for Cu2+

A novel colorimetric cation sensor bearing phenol, thiol and HCN groups was designed and synthesized. In a DMSO/H2O (9:1, v/v) solution, the sensor exhibited highly selective recognition of Cu2+ among a range of metal ions tested. In the presence of Cu2+, solutions of the sensor underwent a dramatic color change from colorless to yellow, while the presence of other metal cations such as Zn2+, Pb2+, Cd2+, Ni2+, Co2+, Fe3+, Hg2+, Ag+ and Ca2+ had no effect on the color. The detection limit of the sensor toward Cu2+ is 8.0×10(-7) M and an association constant Ka of 4.3×10(5) M(-1) was measured. The sensing of Cu2+ by this sensor was found to be reversible, with the Cu2+-induced color being lost upon addition of EDTA.     

Bis(2-hydroxyacetophenone)ethylenediimine as a neutral carrier in a coated-wire membrane electrode for lead(II).

A coated-wire ion-selective electrode (CWISE), based on a Schiff base as a neutral carrier, was successfully developed for the detection of Pb(II) in aqueous solution. CWISE exhibited a linear response with a Nernstian slope of 29.4 +/- 0.5 mV/decade within the concentration range of 1.0 x 10(-5) - 1.0 x 10(-1) M lead ion. CWISE has shown detection limits of 5.0 x 10(-6) M. The electrode exhibited good selectivity over a number of alkali, alkaline earth, transition and heavy metal ions. This sensor yielded a steady potential within 10 to 20 s at a linear dynamic range. The electrode was suitable for use in aqueous solutions in a pH range of 2.0 to 5.0. Applications of this electrode for the determination of lead in real samples and as indicator electrode for potentiometric titration of Pb2+ ion using K2CrO4 are reported.