Transparent Cross-Flow Platform as Chemiluminescence Detection Cell in Cross Injection Analysis.

This work presents the use of a transparent 'Cross Injection Analysis' (CIA) platform as a flow system for chemiluminescence (CL) measurements. The CL-CIA flow device incorporates introduction channels for samples and reagents, and the reaction and detection channels are in one acrylic unit. A photomultiplier tube placed above the reaction channel detects the emitted luminescence. The system was applied to the analysis of (i) Co(II) via the Co(II)-catalyzed H2O2-luminol reaction and (ii) paracetamol via its inhibitory effect on the catalytic activity of Fe(CN)63- on the H2O2-luminol reaction. A linear calibration was obtained for Co(II) in the range of 0.002 to 0.025 mg L-1 Co(II) (r2 = 0.9977) for the determination of Co(II) in water samples. The linear calibration obtained for the paracetamol was 10 to 200 mg L-1 (r2 = 0.9906) for the determination of pharmaceutical products. The sample throughput was 60 samples h-1. The precision was ≤4.2% RSD. The consumption of the samples and reagents was ca. 170 µL per analysis cycle.

Microfluidic paper-based analytical device for determination of sucrose in sugarcane juice using Benedict's reagent.

This work presents a microfluidic paper-based analytical device (µPAD) for the determination of sucrose using the Benedict's test. An asymmetric dumbbell-shaped hydrophobic barrier was produced by rubber stamping the barrier pattern onto a laboratory filter paper. Hydrochloric acid and solution containing sucrose were successively deposited onto the sample reservoir of the µPAD attached to a glass slide. The device was placed in a plastic bag and dipped into boiling water for accelerating the hydrolysis of sucrose into the reducing sugars. Then the Benedict's reagent was added at the narrow straight channel connecting the two circular zones of the µPAD, which was replaced in the plastic bag and heated again for reduction of Cu(II) by the reducing sugars. Precipitate of brick-red copper(I) oxide was formed. The image of the µPAD was recorded by a smartphone. The ratio of the red to blue intensities gave linear correlation with the concentration of sucrose in the range of 0.5-10% w/v. The relative standard deviation of the measurement was less than 5% for 2 and 4% w/v sucrose (n = 10), with limit of determination, calculated using standard deviation of regression divided by slope of calibration, of 0.26% w/v sucrose. The method was successfully validated using the dinitrosalicylic acid method for sucrose measurement. Percent recoveries of sucrose were evaluated using ten sugarcane samples. The recoveries were in the range of 89 to 101%, demonstrating that there were no significant sample matrix effects on the quantification.

Dual-Purpose Photometric-Conductivity Detector for Simultaneous and Sequential Measurements in Flow Analysis.

This work presents a new dual-purpose detector for photometric and conductivity measurements in flow-based analysis. The photometric detector is a paired emitter-detector diode (PEDD) device, whilst the conductivity detection employs a capacitively coupled contactless conductivity detector (C4D). The flow-through detection cell is a rectangular acrylic block (ca. 2 × 2 × 1.5 cm) with cylindrical channels in Z-configuration. For the PEDD detector, the LED light source and detector are installed inside the acrylic block. The two electrodes of the C4D are silver conducting ink painted on the PEEK inlet and outlet tubing of the Z-flow cell. The dual-purpose detector is coupled with a sequential injection analysis (SIA) system for simultaneous detection of the absorbance of the orange dye and conductivity of the dissolved oral rehydration salt powder. The detector was also used for sequential measurements of creatinine and the conductivity of human urine samples. The creatinine analysis is based on colorimetric detection of the Jaffé reaction using the PEDD detector, and the conductivity of the urine, as measured by the C4D detector, is expressed in millisiemens (mS cm-1).

Simultaneous determination of iodide and creatinine in human urine by flow analysis with an on-line sample treatment column.

This work presents the first flow system for direct analysis of iodide and creatinine suitable for screening of human urine samples. The system had a mini-column packed with strong anion exchange resin for on-line extraction of iodide. After injection of a sample on the column the unretained urine sample was analyzed for creatinine in one section of the flow system using the Jaffe's reaction with spectrometric detection at 520 nm. Iodide was eluted off with 1.42 mL 5 M NaNO3. A 150 µL fraction of the eluate was analyzed in another section of the same flow system for iodide using the kinetic-spectrometric Sandell-Kolthoff reaction. At the optimum condition, the sample throughput was 12 samples per h. The linear working range covered the normal levels of iodide and creatinine in human urine: 0-200 µg I L(-1) and 50-1200 mg creatinine L(-1), respectively. Recoveries tested in 10 samples were 87-104% for iodide and 89-104% for creatinine. Bland-Altman plots (n = 50) showed that the scatter of the differences between values obtained by this method and those of reference methods, for both iodide and creatinine, was within mean ± 2SD.

Determination of 3-hydroxyanthranilic acid in the sweat of healthy older adults.

3-Hydroxyanthranillic acid (3HAA) is one of the key metabolites from the tryptophan (TRP) metabolism pathway and is associated with aging, age-associated diseases, and healthy lifespan extension. This study aims to detect 3HAA in the sweat of healthy older adults using simple, high-performance liquid chromatography (HPLC) method. Chromatographic separation using 20 mmol/L sodium acetate, 3 mmol/L zinc acetate, and 7% (v/v) acetonitrile as mobile phase is possible to simultaneous detect 3HAA, KYN, and TRP with UV and fluorescence detection, respectively, under 6 min. This method demonstrated excellent linearity with coefficient of determination (r2) greater than 0.998 for all analytes. The linear range were 0.05-6 µg/mL for TRP, 0.1-6 µg/mL for KYN and 0.2-6 µg/mL for 3HAA. Percentage recoveries from spiked in human sweat ranged from 90 ± 7-101 ± 3 for TRP, 86 ± 1-92 ± 3 for KYN, and 96 ± 1-103 ± 4 for 3HAA. The precision (%RSD) of repeatability and reproducibility is less than 3% and 6%, respectively. This method was used in a cross-sectional study with 81 participants aged 50-79 years, selected randomly from a local primary healthcare hospital's sampling frame. A detectable amount of 3HAA was observed in all sweat samples, marking the first report of 3HAA presence in human sweat. Additionally, the results revealed that the 3HAA sweat levels increased with age analyzed in three different age groups ranging from 50-59, 60-69, and 70-79. These findings enhance our understanding of sweat profiles and their correlation with aging, potentially further improving early diagnosis, disease monitoring, and development of customized treatment programs for older adults.

Carbon dots derived from citric acid and urea as fluorometric probe for determining melamine contamination in infant formula sample.

Melamine has been intentionally added into food products to increase the protein count at less cost, especially in dairy products for infant resulting in serious adverse effects on health of consumers. Therefore, this study aimed to develop a method to quantify melamine in dairy products based on the change of fluorescent properties of carbon dots (CDs) as sensing probe. CDs with green-fluorescent emission were synthesized from citric acid and urea under microwave irradiation. The synthesized CDs emitted fluorescence at the maximum wavelength of 538 nm with excitation wavelength of 410 nm. Thus, they provided high sensitivity and selectivity on melamine detection by which fluorescent emission of the CDs was increasingly quenched upon increasing melamine concentrations. Optimal conditions for melamine determination using the CDs was under pH 6, volume ratio between CDs and sample of 2:8 and reaction time of 15 min. The developed method provided high precision of melamine determination with less than 5% of %RSD (n = 5), wide detection range from 1.0 to 200.0 ppm, and high sensitivity with limit of detection (LOD) of 0.47 ppm and limit of quantification (LOQ) of 1.56 ppm, which is within the regulated level by the Food and Drug Administration of the United States for melamine in dairy products. Several analytical characterization techniques were conducted to elucidate the reaction mechanism between CDs and melamine, and the hydrogen bonding interaction was proposed.

Enantiomeric separation of some common controlled stimulants by capillary electrophoresis with contactless conductivity detection.

CE methods with capacitively coupled contactless conductivity detection (C(4)D) were developed for the enantiomeric separation of the following stimulants: amphetamine (AP), methamphetamine (MA), ephedrine (EP), pseudoephedrine (PE), norephedrine (NE) and norpseudoephedrine (NPE). Acetic acid (pH 2.5 and 2.8) was found to be the optimal background electrolyte for the CE-C(4)D system. The chiral selectors, carboxymethyl-ß-cyclodextrin (CMBCD), heptakis(2,6-di-O-methyl)-ß-cyclodextrin (DMBCD) and chiral crown ether (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid (18C6H(4)), were investigated for their enantioseparation properties in the BGE. The use of either a single or a combination of two chiral selectors was chosen to obtain optimal condition of enantiomeric selectivity. Enantiomeric separation of AP and MA was achieved using the single chiral selector CMBCD and (hydroxypropyl)methyl cellulose (HPMC) as the modifier. A combination of the two chiral selectors, CMBCD and DMBCD and HPMC as the modifier, was required for enantiomeric separation of EP and PE. In addition, a combination of DMBCD and 18C6H(4) was successfully applied for the enantiomeric separation of NE and NPE. The detection limits of the enantiomers were found to be in the range of 2.3-5.7 µmol/L. Good precisions of migration time and peak area were obtained. The developed CE-C(4)D method was successfully applied to urine samples of athletes for the identification of enantiomers of the detected stimulants.

An application of miniaturized electrochemical sensing for determination of arsenic in herbal medicines.

This study aimed to create a miniaturized electrochemical platform for detecting As(III) contamination in herbal medicines. To reduce the operational steps of modification and determination, only a single drop of mixed standard Au(III) and sample solution is proposed to perform the electrochemical measurements using a screen-printed graphene electrode (SPGE). Square wave anodic stripping voltammetry was employed to integrate the simultaneous modification and determination processes. To perform the measurement, As(III) and Au(III) migrate to the SPGE surface while the reduction potential is held at -0.5 V, forming an Au-As intermetallic alloy. Then, As is stripped off for the electrochemical determination of As(III). The total assay time is less than 3 min. Under suitable conditions, the electrochemical sensing system can detect As(III) at concentrations ranging from 0.1 to 3.0 ppm, with a limit of quantification and limit of detection of 0.1 and 0.03 ppm, respectively. The applicability and accuracy of the proposed sensor were verified by determining As(III) in herbal medicinal samples, and they were found to be in line with the standard method (ICP-OES). The benefits of simple operation, rapid detection, portability, and low cost (<1 USD) make this a more powerful tool for routine monitoring and on-site analysis applications.

Contactless conductivity detector from printed circuit board for paper-based analytical systems.

This work presents a capacitively coupled contactless conductivity detector (C4D) etched out from a printed circuit board (PCB) as potential sensor for paper-based analytical systems. Two lines of any desirable pattern forming 35-µm thick planar copper electrodes were produced on a PCB plate (40 mm × 60 mm) by photolithography. The final PCB plate was covered with polypropylene film to serve as the insulating layer for the C4D detector. The film also protected the copper electrodes from corrosion. Electrodes made in this planar geometry make the PCB-C4D suitable as sensor for flat devices such as paper-based analytical devices. For this work, plain paper strips were employed as sample reservoir and as fluidic channel without hydrophobic pattern. A dried paper strip was first placed over the sensor, followed by dispensing a fixed volume of the liquid sample onto the paper. Entrapment of the liquid sample in the paper strip leads to reproducible size and position of the detection zone of the sample liquid for the capacitive coupling effect. High precision was obtained with %RSD ≤1% (n = 18) for standard solutions of KCl. Soil suspensions could be analyzed without prior filtration by placing a drop onto the paper strip extending away from the detector zone. The paper strip filtered out soil particles at the surface of the paper. Therefore, only soil filtrate moved towards the detection zone by lateral flow. The C4D detection using paper strip showed high tolerance to soil suspension with turbidity up to 6657 NTU, offering direct analysis of soil salinity. Cleaning with moist tissue paper between samples is adequate even for dirty samples such as soil suspension. We also monitored conductivity of acid-base reaction in the microfluidic paper channels, which was later applied to the quantification of bicarbonate in water and in antacid tablet ("Soda Mint Tablet").

Simultaneous and direct determination of urea and creatinine in human urine using a cost-effective flow injection system equipped with in-house contactless conductivity detector and LED colorimeter.

This work presents a cost-effective and simple flow injection analysis (FIA) system for simultaneous and direct determination of urea and creatinine in human urine. The FIA system comprises two in-house detectors, a contactless conductivity detector and a light emitting diode (LED) detector. The contactless detector was built as a flow-through detection cell with axial electrodes, commonly known as capacitively coupled contactless conductivity detector (C4D) and the diode detector was fabricated based on the concept of paired emitter detector diodes (PEDD). With appropriate dilution of urine, the sample is directly injected into a stream of glycine-NaOH buffer pH 8.8 (the gas donor stream) and is carried by the carrier through a urease minicolumn for on-line enzymatic hydrolysis. The generated NH3 diffuses from the carrier stream through a porous polytetrafluoroethylene (PTFE) membrane into a stream of deionized water (the acceptor stream) leading to an increase in the signal at the C4D due to the NH3 dissolution in the water. In this system, creatinine is determined based on the Jaffé reaction by merging a stream of alkaline picrate with the gas donor stream. The change in color is detected using the PEDD equipped with two green LEDs. Under the optimum condition, the linear range of urea and creatinine were 30-240 mg L-1 and 10-500 mg L-1, with limits of detection of 9.0 mg L-1 and 0.9 mg L-1, respectively. The proposed system provides satisfactorily good precision (RSD < 3%), with sample throughput of 31 sample h-1 for the two analytes. The FIA system tolerates potential interference commonly found in human urine. The system was successfully applied and validated with selected reference methods.

Temperature-dependent schlieren effect in liquid flow for chemical analysis.

In flow analysis, such as flow injection analysis, liquid lens is formed at the boundary between two adjacent liquid media which have different refractive indices. Light refraction at the liquid interface gives the so-called 'schlieren signal'. Schlieren effect is both concentration-dependent and temperature-dependent. In this work, the schlieren signal from temperature difference was quantitatively investigated for application in enthalpimetric measurement. The schlieren phenomena was then exploited for chemical analysis. A thermal insulated single flow line manifold was constructed using deionized water at 23 °C as the carrier. Deionized water at various temperatures in the range of 5-85 °C was injected into the carrier flow. A correlation between the schlieren signal and sample temperature was observed. A heat exchanger unit (HEU), consisting of a small volume glass-reaction chamber with a surrounding water jacket, was constructed. The unit was thermally insulated in a double layer cylindrical PVC unit. For demonstrating the applicability of temperature-dependent schlieren effect in chemical analysis, the exothermic oxidation reaction between acid dichromate and ethanol or ascorbic acid was employed with heat transferring to the surrounding water layer. When an aliquot of water from the HEU is injected into the constant temperature flow line the observed schlieren signal was dependent on the analyte concentration. Linear calibration (r2 > 0.99) were obtained covering the concentration range of ethanol and ascorbic acid as found in samples. The developed flow system provides good precision (RSD < 5%) with sample throughput of 4 sample h-1. The system were applied to the determination of ethanol in Thai white spirit and ascorbic acid in vitamin C tablets, respectively. The quantitative results obtained from the schlieren method were in agreement with the comparative methods.

Development of flow systems incorporating membraneless vaporization units and flow-through contactless conductivity detector for determination of dissolved ammonium and sulfide in canal water.

Use of membraneless vaporization (MBL-VP) unit with two cone-shaped reservoirs is presented for on-line separation and detection of non-volatile species. A flow system comprising two sets of MBL-VP units with a single in-house capacitively coupled contactless conductivity detector (C4D) was developed for dual determination of ammonium and sulfide ions. Using the continuous-flow section, two zones (280µL) of a sample, either mixed with sodium hydroxide (for ammonium) or hydrochloric acid (for sulfide), are separately delivered into the donor reservoir of the MBL-VP units. The acceptor reservoir contains either 150µL of 15µM HCl solution (for ammonia) or pure water (for hydrogen sulfide), respectively. Vaporization and trapping of the ammonia or hydrogen sulfide gas from the donor reservoir into the liquid acceptor cone occur concurrently in the two separate MBL-VP units. After trapping the gas for 3min, the two 150-µL liquid acceptors are sequentially aspirated through the C4D flow cell for recording the changes in the conductivity. Linear calibrations were obtained for ammonium from 5 to 80µM (Volt = (0.0134 ± 0.0003) [NH4+] - (0.01 ± 0.01), r2 = 0.998) and for sulfide from 5 to 200µM (Volt = (0.0335 ± 0.0009) [S2-] - (0.13 ± 0.09), r2 = 0.996). Analysis time for both analytes is only 320s. Our method was applied to analyze canal water samples. The results agree well with membrane gas-diffusion flow injection techniques, using bromothymol blue for ammonium and methylene blue for sulfide. Recoveries ranged from 95% to 104%.

Cross injection analysis: concept and operation for simultaneous injection of sample and reagents in flow analysis.

This work presents a new flow injection strategy, called 'cross injection analysis' or CIA, an alternative cost-effective approach in flow analysis. The flow platform is made from a rectangular acrylic block, approximately 5×3×1.5 cm (x×y×z), with crossing cylindrical channels drilled out along the x- and y-axis of the block. The outlet from the single x-axis channel is connected to a detector flow cell. This channel is filled with the carrier solution. The flow in the x-axis channel is driven by a computer controlled single-channel peristaltic pump. The multiple y-axis channels, running perpendicular to the x-channel, are connected to a multi-channel peristaltic pump. These channels contain the sample and reagent solutions that flow across the intersection zones of the channels. To mix the sample and reagent with subsequent detection of the reaction zone, flow is applied along the x-axis channel, while flow in the y-axis channels is stopped. We successfully demonstrated the validity of the CIA technique by the spectrometric determination of Fe(II) using 1,10-phenanthroline and the speciation of Fe(II) and Fe(III). To place the CIA technique within the context of flow analysis, a brief overview of the evolution of flow injection analysis and its later innovative development is included.