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A paper-based device, based on the fluorescence switching of water-soluble thioglycolic acid (TGA)-capped CdTe QDs, was developed for the simple, rapid and sensitive detection of organophosphorus (OP) and carbamate (CM) insecticides. A new design platform for the developed device was created, comprising a foldable sheet with a detection zone and a buffer solution loading channel, to simplify the multi-step reaction of a bi-enzyme assay with a signal enhancement system. This device was very useful for fast screening and on-site detection, without requiring many reagents. The bi-enzyme includes acetylcholinesterase (AChE), choline oxidase (ChOx) and TGA-capped CdTe QDs, which were coated on the paper-based device at the detection zone in a separated sheet, starting with an acetylcholine (ACh) substrate coating. After incubation of the sample at the detection zone, the device was closed to initiate enzyme catalyzed hydrolysis of the pre-spotted ACh. Subsequently, the buffer solution was applied to the device, allowing for the pre-spotted chemicals to mix well and importantly, result in signal enhancement. The hydrolysis of ACh was catalyzed by AChE followed by ChOx to yield H2O2, which can quench the fluorescence of the TGA-capped CdTe QDs. In the presence of OP and CM insecticides, the AChE activity was inhibited, resulting in decreased quenching of the TGA-capped CdTe QDs. The change in the fluorescence intensity of the TGA-capped CdTe QDs can be observed using the naked eye under UV-black light. Under optimized conditions, the limit of detection for pirimicarb, dichlorvos and carbaryl were found to be 0.05, 0.01 and 0.01 ppm, respectively, using visual measurements. This developed device was successfully applied to the detection of OP and CM insecticide residues in agricultural products. The data correlated well with the data obtained using gas chromatography-tandem mass spectrometry (GC-MS/MS). This developed device shows good selectivity and sensitivity for the rapid screening of insecticides and could be useful for on-site applications.

A gold nanoparticles (AuNPs) enhancement strategy for lateral flow immunoassays (LFIAs) was previously reported to improve assay sensitivity. The multi-steps of detection resulted in a tedious procedure. In this report, to reduce this limitation, a sequential flow LFIA was introduced to perform the AuNPs enhancement in only single step operation using C-reactive protein (CRP) as a model analyte. The delay and non-delay channels created on nitrocellulose membrane (NCM) using wax-printing method were used to deliver immunoassay and AuNPs enhancement reagents to the detection zone sequentially. For the enhancing step, AuNPs were enlarged by the catalytic reaction of KAuCl4 and NH2OH⋅HCl. Under optimal conditions, a calibration curve was found in a range of 0.1−5 μg⋅mL−1 with a good linear correlation (R2 = 0.9953). The limits of detection (LOD) were found to be 0.1 and 0.001 μg⋅mL-1 for naked eye observation and calculation (3SD/slope), respectively. Furthermore, the device shows no cross-reactivity with common interferences for CRP. The platform was successfully applied to detect CRP in certified human serum samples within 15 min, demonstrating the simplicity and speed of the AuNPs enhancement strategy platform. These promising results suggest that the new device may be used as an alternative tool for CRP detection and other biomarkers for POCT analysis.

A novel paper-based analytical device (PAD) was fabricated and developed for the simple and rapid determination of the albumin to creatinine ratio (ACR) in urine samples. The detection was based on a colorimetric reaction using bromocresol green (BG) in a phosphate buffer (PB) at pH 4 for the determination of albumin (AL) and creatinine (CR). BG changes color from greenish-yellow to bluish-green in the presence of AL and/or CR. Picric acid (PA) in 0.25 M NaOH was used to detect CR, and PA changes color from yellow to orange. Under the optimal conditions, the working range was 10 to 350 mg dL-1 with a detection limit of 7.1 and 5.4 mg dL-1 for AL + CR and CR detection, respectively. The repeatability was evaluated, and the %RSD value was less than 8.23 (n = 10). The ACR was obtained by calculating the AL and CR colorimetric results. Finally, the proposed devices were applied to the determination of AL, CR, and ACR in urine samples. The results obtained by the developed PADs were in good agreement with the standard method and demonstrated the method could reliably measure AL, CR, and ACR. The proposed method provides a low-cost, simple, sensitive, and promising tool for diagnostic identification assay for chronic kidney disease (CKD).

A simple paper-based sensor with electrochemical detection was first developed for rapid, selective, and sensitive determination of acetylcholinesterase (AChE). The screen-printed graphene electrode was used as working electrode providing sensitivity for the sensor. The amperometric detection of AChE is based on the determination of thiocholine (TCh) produced from hydrolysis of acetylthiocholine chloride (ATCh) by AChE. To detect TCh, the ATCh immobilized sheet was stacked onto the detection sheet using double adhesive tape, then samples of AChE were dropped on the back side of an ATCh immobilized sheet with only 1min of incubation time. To avoid interference, glutathione (GSH), the potential of 0.5V vs. Ag/AgCl was applied onto a graphene electrode and the current, which depends on AChE concentration, was measured. Under optimized conditions, the limit of detection (LOD) from the experiment of AChE determination was 0.1U/mL with AChE concentration in range of 0.1-15U/mL. The data correlated well with the data obtained using spectrophotometric method. The developed sensor had been successfully applied to detect AChE in blood samples.

In the expression of bioaccumulated elemental mercury (Hg 0) toxicity, the first Hg 0 oxidation step is crucial. Therefore, to clarify the mechanism underlying the interactions of sulfhydryl (SH) compounds and Hg 0 in the present study, we analyzed the oxidation of reduced glutathione (GSH) and L-cysteine (Cys) in the presence of Hg 0 in aqueous solution by cyclic voltammetry (CV). Production of Hg2+ in the reaction mixture was found to increase along with a decrease in free SH residues. CV showed that the oxidation of Cys increased after a 4-h incubation in the presence of Hg(0), but the oxidation of Cys after a 24-h incubation was suppressed. Conversely, GSH oxidation increased with incubation time in the absence of Hg(0). In the presence of Hg(0), the oxidation of GSH was suppressed. The different reactivities of Cys and GSH with Hg(0) may arise from differences in their oxidation/reduction potentials and pH. The important SH compound interactions with Hg(0) oxidation were as follows: (i) oxidation of Hg(0) to form either mercurous ion (Hg+) or mercuric ion (Hg2+) which both form stable complexes in aqueous solution as Hg I (RS) or Hg II(RS)2; (ii) catalyzed oxidation of SH compounds in the presence of Hg 0; and (iii) suppression of the oxidation of SH compounds due to the reduced concentration of free SH compounds through the binding of SH compounds with Hg+ or Hg2+ The present results demonstrate the chemical reaction processes by which Hg 0 dissolves in aqueous solution in the presence of SH compounds, and contribute to our understanding of SH compounds in non-enzymatic Hg 0 oxidation in vivo.

A novel, highly selective and sensitive paper-based colorimetric sensor for trace determination of copper (Cu(2+)) ions was developed. The measurement is based on the catalytic etching of silver nanoplates (AgNPls) by thiosulfate (S2O3(2-)). Upon the addition of Cu(2+) to the ammonium buffer at pH 11, the absorption peak intensity of AuNPls/S2O3(2-) at 522 nm decreased and the pinkish violet AuNPls became clear in color as visible to the naked eye. This assay provides highly sensitive and selective detection of Cu(2+) over other metal ions (K(+), Cr(3+), Cd(2+), Zn(2+), As(3+), Mn(2+), Co(2+), Pb(2+), Al(3+), Ni(2+), Fe(3+), Mg(2+), Hg(2+) and Bi(3+)). A paper-based colorimetric sensor was then developed for the simple and rapid determination of Cu(2+) using the catalytic etching of AgNPls. Under optimized conditions, the modified AgNPls coated at the test zone of the devices immediately changes in color in the presence of Cu(2+). The limit of detection (LOD) was found to be 1.0 ng mL(-1) by visual detection. For semi-quantitative measurement with image processing, the method detected Cu(2+) in the range of 0.5-200 ng mL(-1)(R(2)=0.9974) with an LOD of 0.3 ng mL(-1). The proposed method was successfully applied to detect Cu(2+) in the wide range of real samples including water, food, and blood. The results were in good agreement according to a paired t-test with results from inductively coupled plasma-optical emission spectrometry (ICP-OES).

To the best of our knowledge, this is the first report on paper-based devices for automating the sequential multistep procedures of a sandwich-type enzyme-linked immunosorbent assay (ELISA) that require only a single-step application of the sample solution. The device was based on a piece of nitrocellulose (NC) membrane with specially designed channels, where all the reagents are applied at different locations in order to control the fluid travel to the detection region. The inkjet printing method, a simple and low-cost process, was used to create the flow channel and device barrier patterns. The fabricated barrier was found to be an efficient boundary for the liquid along the printed design in the NC membrane, enabling direct control of the reagent flow time. ELISA results were obtained with a single-step sample application. The developed devices (so-called automated paper-based devices) provided a simple procedure for the sandwich ELISA, while reducing assay time and reagent consumption. Colorimetric results were measured using digital camera imaging with software processing. The capability of the method developed herein was successfully used to determine the levels of human chorionic gonadotropin (hCG) by ELISA.

A novel lab-on-paper device combining electrochemical and colorimetric detection for the rapid screening of Au(III) in the presence of a common interference, Fe(III), in industrial waste solutions is presented here. With dilute aqua regia (0.1 M HCl + 0.05 M HNO(3)) as the supporting electrolyte, square wave voltammetry on paper provided a well-defined reduction peak for Au(III) at approximately 287 +/- 12 mV vs Ag/AgCl. Under the optimized working conditions, the calibration curve showed good linearity in the concentration range of 1-200 ppm of Au(III) with a correlation coefficient of 0.997. The limit of detection (LOD) of the proposed method is 1 ppm. Interferences from various cations were also studied. Fe(III) is the only metal that affects the electrochemical determination of Au(III) when present above a 2.5-fold excess concentration of that of the Au(III). To overcome this limitation, a colorimetric method was used to simultaneously detect Fe(III) as a screening tool. The procedure was then successfully applied to determine Au(III) in gold-refining waste solutions. The results are in agreement with those obtained from inductively coupled plasma-atomic emission spectrometry (ICP-AES).

The electrochemical behaviors of native and thermally denatured fish DNA was investigated using boron-doped diamond (BDD) film electrode by cyclic voltammetry. The BDD electrode afforded us to measure weak current less than μA for the DNA solution in 100 μl. The mixture of acetic acid and sodium acetate solution (0.2 M) was used as a supporting electrolyte. Two oxidation peaks were observed at about + 1.1 V and + 1.3 V at pH 4.6 for thermally denatured fish DNA. This is due to the oxidation of guanine and adenine in the denatured fish DNA, respectively. In contrast, the native fish DNA showed ill-defined peaks at + 1.1 V. Furthermore, the electrochemical behaviors of thermally denatured fish DNA were studied in the presence of cytosine, cytidine, cytidine-5-monophosphate, tetrakis(1-methypyridinium-4-yl)porphyrin (H2(TMPyP)4+) and RuII(TMPyP)4+. The oxidation peak intensity at + 1.1 V gradually decreased with the increase of the concentrations of the above compounds. Based on the above studies, electrochemical behaviors of the thermally denatured fish DNA at BDD electrode is discussed.