The Evaluation of a Lateral Flow Strip Based on the Covalently Fixed "End-On" Orientation of an Antibody for Listeria monocytogenes Detection.

In this study, the covalently fixed "end-on" orientation of a monoclonal Listeria monocytogenes antibody (mAb-Lis) to amino terminated oligo (ethylene glycol)-capped gold nanoparticles (NH2-TEG-AuNPs) was used to fabricate an in-house lateral flow strip (LFS), namely, the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS. The aim was to evaluate the performance of the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS in detecting L. monocytogenes. The proposed LFS enabled the sensitive detection of L. monocytogenes in 15 min with a visual limit of detection of 102 CFU/mL. Quantitative analysis indicated an LOD at 10 CFU/mL. The fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS showed no cross-reactivity with other pathogenic bacteria and practical performance across different food matrices, including human blood, milk, and mushroom samples. Furthermore, the clinical performance of the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS for detecting L. monocytogenes was evaluated by using 12 clinical samples validated by the hemoculture method. It demonstrated excellent concordance with the reference methods, with no false-positive or false-negative results observed. Therefore, the fixed "end-on" Lis-mAb-NH-TEG-AuNPs LFS serves as a promising candidate for a point-of-care test (POCT), enabling the rapid, precise, and highly sensitive detection of L. monocytogenes in clinical samples and contaminated food.

Measurement of sucrose concentration using Imbibition length on paper: A device for equipment-free and environmentally-friendly detection.

The Lucas-Washburn equation is commonly used to predict the distance (L) that a liquid travels through paper. This equation establishes that L2 is linear with time and inversely proportional to the viscosity of the liquid. However, there is currently no theoretical equation connecting the viscosity of a solution to its concentration. In this study, the imbibition flow of a sucrose solution was measured along the length of a horizontal strip of filter paper, featuring a printed, thermometer-shaped hydrophobic boundary. A sample (38 µL) was dispensed onto the bulb area, and the solution's flow was visually tracked using a red dye added to the sample. The imbibition length (L) was measured by a vernier caliper at 10.0 min after the sample addition. An empirical equation, based on literature values of the viscosity (η) and concentration (C) of sucrose solutions, was proposed. By integrating this empirical equation with the Lucas-Washburn equation, the following equation was derived: L = a⋅exp{-(bC + cC2)}, where 'a', 'b' and 'c' are parameters. This equation was fitted to the dataset of L and C, covering C values from 0 to 60 % w/w standard sucrose solutions, resulting in a coefficient of determination of 0.9987. The plot of L against C was observed to closely follow a linear line, with a fitting providing a coefficient of determination of 0.9986. The sucrose contents in samples, such as soft drinks, syrups, and sugarcanes, determined using the imbibition length method and conventional refractometry, were in statistical agreement via the paired t-test at the 95 % confidence level. This method is simple, instrument-free, requiring only a small amount of safe red food dye, and can be conducted on-site.

A rapid cotton swab for on-site screening of coloring curcumin on durian skin: food safety aspects.

Exported durians from Thailand are sometimes immersed in curcumin to give the fruits a good appearance. Curcumin is regarded as non-toxic additive, however some importing countries prohibited use of any additive to fresh fruits and vegetables. This work aims to develop a rapid, low cost and convenient cotton swab device for curcumin detection. The detection principle involves a colorimetric acid-base characteristic of curcumin. Curcumin in an acidic/neutral solution presents a bright yellow color, while it displays an intense orange-red color in basic solution. A cotton swab acted for both sample collection and as a sensing platform. A pre-moistened swab was used to wipe a durian surface. Afterward, a NaOH solution was dropped onto the swab. A distinct orange-red color appearing on the swab indicates the presence of curcumin. The cotton swab was applied for qualitative analysis of curcumin contaminated on durian husks via visual detection. The developed device provided good reliability, 93.75% (36 samples). Furthermore, the device was demonstrated for quantitative determination using camera detection. Two linear calibrations were obtained in ranges of 10-75 and 75-250 mg L-1, with a detection limit of 3.2 mg L-1. The method was also successfully applied to quantification of curcumin in durians (three samples) and dietary supplements (two samples). The test can be done in a few minutes. The developed device was established as an useful tool for food safety and control of contamination by curcumin in an on-site application.

Insight into the Covalently Oriented Immobilization of Antibodies on Gold Nanoparticle Probes to Improve Sensitivity in the Colorimetric Detection of Listeria monocytogenes.

In this work, the covalently oriented conjugation of monoclonal Listeria monocytogenes antibody (mAb-Lis) to amino-terminated oligo(ethylene glycol)-capped gold nanoparticles (NH2-TEG-AuNPs) was studied. After NH2-TEG-AuNPs were synthesized, the amino-terminated ligands on the particles were then linked to the carboxyl groups in the mAb-Lis through EDC/NHS chemistry. By maintaining the pH of the solution at ∼5, the Fc region of the antibody could preferably attach to the particle surface, providing a specific Fab region that was available for binding with the target pathogen. The resulting mAb-NH-TEG-AuNPs could act as a colorimetric probe for L. monocytogenes based on a particular antigen-antibody interaction, which resulted in a drastic aggregation of particles. This caused the color of the colloidal solution to transition from red-pink to purple or even gray depending on the pathogen concentration. To perform quantitative analysis, the absorbance ratio of A650/A534 was monitored as a function of L. monocytogenes concentration using a spectrophotometer. The detection limit was very low at 11 CFU/mL. Furthermore, a lateral flow strip (LFS) was fabricated as a portable device for onsite utilization. LFS detection could be completed by the naked eye and by a smartphone. The detection limit of LFS was estimated to be 103 CFU/mL. Our methods exhibited a substantial improvement in sensitivity compared to that of previous studies on immuno-based nanoparticles. The assay could be completed in 15 min, and no cross reactivity by any pathogen was found. Hence, the designed AuNPs exhibit great promise for use in monitoring L. monocytogenes for food safety and in other applications.

Direct determination of ethanol in alcoholic beverages based on its anti-aggregation of melamine-silver nanoparticle assembly.

We report a new method for determination of ethanol based on anti-aggregation of silver nanoparticles (AgNPs) in the presence of melamine. In the system, ethanol and melamine act as protecting and aggregating agents, respectively. Melamine can induce citrate-stabilized AgNPs to aggregate, leading to a color change from yellow to green. However, if the AgNPs are pre-incubated in ethanol, ethanol readily surrounds the particles by forming hydrogen bonds with the citrate stabilizer. An external nanoshell of ethanol hinders particle aggregation caused by melamine. Minor aggregation of AgNPs was observed, the solution color maintained its yellow-orange color. Higher ethanol concentrations result in a lower degree of particle aggregation. The colorimetric response of AgNPs was monitored using a UV-vis spectrophotometer at 390 nm. The current method could determine ethanol concentrations over a wide dynamic range of 5-80% (v/v), with a detection limit of 3.1% (v/v) (3SD of blank/slope). This method was applied for direct quantification of ethanol in alcoholic drinks without sample pretreatment and the results are well correlated with those of gas chromatography. Our method is convenient and cost effective, making it auspicious for ethanol monitoring in alcoholic drink manufacture and control.

A new mechanism for resonance Rayleigh scattering detection of minoxidil based on catalytic oxidation of silver nanoparticles.

This work presents a new method for minoxidil detection based on silver nanoparticle (AgNP) oxidation. Minoxidil, which is a pyrimidine N-oxide, can be reduced to its corresponding pyrimidine via a redox reaction. In this system, acetate buffer serves as a proton source. AgNPs act as electron donors that contribute electrons to the reaction, producing Ag+. Consequently, the sizes and numbers of AgNPs in the system decrease, which results in a decline in their resonance Rayleigh scattering (RRS). By monitoring the RRS intensity at 409 nm, a change in intensity was linearly related to the minoxidil concentration over a concentration range of 0.5 - 5.0 mM. The detection limit was 0.35 mM. This approach is simple and rapid. It is done by directly mixing the drug and AgNPs in an acidic buffer. The reaction was completed within 2 min. This proposed method was successfully utilized for quantification of minoxidil in topical hair-growth formulations.

Simple and green method for direct quantification of hypochlorite in household bleach with membraneless gas-separation microfluidic paper-based analytical device.

This work presents development of a microfluidic paper-based analytical device (µPAD) for direct determination of hypochlorite in household bleach. The recent design of a membraneless gas-separation microfluidic paper-based analytical device (MBL-GS µPAD) was employed to fabricate the hypochlorite-µPAD. Chlorine gas is generated in the µPAD via acidification of an aliquot of sample loaded on to the donor reservoir located at the bottom layer of the µPAD. The liberated chlorine gas diffuses through the air space to oxidize iodide ion previously impregnated in the acceptor reservoir at the top layer of the µPAD, leading to formation of the brown color of the tri-iodide ions. Digital image of the brown zone was captured at exactly 5 min after loading the acid. Image J program is used for analysis of the image for quantification of the hypochlorite in unit of g Cl2 L-1. It was found that employing a relatively large volume of the air space (ca. 270 µL) direct analysis of the high concentration of hypochlorite in the bleach was achieved without prior dilution. The method thus provides a linear working range of 25-100 g Cl2 L-1, which is suitable for most commercial household products. The calibration line has a coefficient of determination of 0.999. The precision of measurements is 0.96% RSD and 0.30% RSD at 30 g Cl2 L-1 and 80 g Cl2 L-1 (n = 10), respectively. Using the paired t-test (P = 0.05, n = 8), the method agreed well with the iodometric titration method. Our µPAD for hypochlorite is portable and cost-effective. The method is also "green" since there is a significant reduction in use of reagents compared to other conventional methods.

Poly(vinyl alcohol) capped silver nanoparticles for antioxidant assay based on seed-mediated nanoparticle growth.

A simple and rapid method for measurement of total antioxidant capacity (TAC) was developed. In this work, gallic acid was used as the antioxidant standard. Poly(vinyl alcohol) embedded silver nanoparticles (PVA-AgNPs) were employed as a colorimetric sensor. The detection principle was based on the seed-mediated nanoparticle growth technique. The PVA-AgNPs act as a catalyst in the reduction of Ag+ by gallic acid by providing nucleation seeds. Ag+ was reduced to Ag° and accumulated on the PVA-AgNP surface, leading to an increase in the size of particles. The absorbance of the colloidal solution was drastically enhanced with a small red shift. Under optimal conditions, a linear response was established between the change in absorbance and the TAC value expressed in terms of gallic acid equivalents. The linear range was from 25 to 200µM with a detection limit of 22.1µM. Satisfactory precision was obtained with % relative standard deviation (RSD) of 2.17. The developed sensor was successfully applied for TAC assessment of commercial ginger products. The PVA-AgNP sensor offers rapid analysis (within 5min) compared to other nanoparticle-based antioxidant assays. Synthesis of the particles and assay involved less-toxic chemicals, and is therefore a "greener" method.

A Simple and Rapid Method Based on Anti-aggregation of Silver Nanoparticles for Detection of Poly(diallyldimethylammonium chloride) in Tap Water.

A simple and rapid method was developed for the detection of poly(diallyldimethylammonium chloride) (PDADMAC) using citrate-capped silver nanoparticles (AgNPs). Detection was based on anti-aggregation of AgNPs in phosphate buffer caused by PDADMAC. Due to its positive charges, PDADMAC was adsorbed onto AgNPs via electrostatic interaction with citrate, which resulted in the charges at the particle surfaces to become positive and caused repulsion among particles. Furthermore, long-chain PDADMAC provided steric hindrance. These two effects promoted the dispersion of AgNPs in the phosphate buffer. A change in the state of dispersion influenced the surface plasmon resonance (SPR) of AgNPs. Therefore, in this work, the concentration of PDADMAC was determined by monitoring changes in absorbance (at 396 nm) caused by SPR of AgNPs. Under optimal conditions, the calibration was linear over the range of 1 to 100 mg L(-1) with a detection limit of 0.7 mg L(-1). Satisfactory precision was obtained (RSD = 2.8%). This method was successfully applied to the determination of PDADMAC in tap water samples. The recoveries ranged from 86.0 - 107.5%.

Zone fluidics for measurement of octanol-water partition coefficient of drugs.

A novel zone fluidics (ZF) system for the determination of the octanol-water partition coefficient (Pow) of drugs was developed. The ZF system consisted of a syringe pump with a selection valve, a holding column, a silica capillary flow-cell and an in-line spectrophotometer. Exact microliter volumes of solvents (octanol and phosphate buffer saline) and a solution of the drug, sandwiched between air segments, were sequentially loaded into the vertically aligned holding column. Distribution of the drug between the aqueous and octanol phases occurred by the oscillation movement of the syringe pump piston. Phase separation occurred due to the difference in densities. The liquid zones were then pushed into the detection flow cell. In this method, absorbance measurements in only one of the phase (octanol or aqueous) were employed, which together with the volumes of the solvents and pure drug sample, allowed the calculation of the Pow. The developed system was applied to the determination of the Pow of some common drugs. The log (Pow) values agreed well with a batch method (R(2)=0.999) and literature (R(2)=0.997). Standard deviations for intra- and inter-day analyses were both less than 0.1log unit. This ZF system provides a robust and automated method for screening of Pow values in the drug discovery process.

A multiple processing hybrid flow system for analysis of formaldehyde contamination in food.

This work proposes a flow system suitable for the rapid screening of formaldehyde contaminated in food. The system is based on the concept of a flow analyzer with a Hantzsch reaction. An operating procedure was developed for multiple tasking and high sample throughput. This resulted in a significant sample throughput of 51 samples h(-1). Under the optimized conditions, linear calibration from 10 to 100 microM was obtained. The system gave a limit of detection and a limit of quantitation of 0.06 and 0.10 mg kg(-1), respectively. The system was successfully applied to re-hydrated dry squids, vegetables and mushrooms.

A low-cost method for determination of calcium carbonate in cement by membraneless vaporization with capacitively coupled contactless conductivity detection.

This work presents a flow analysis method for direct quantitation of calcium carbonate in cement without pretreatment of the sample. The method is based on online vaporization of CO(2) gas following acidification of the sample inside a small chamber that has a flow of acceptor solution passing around it. Solubilization of the CO(2) gas into the acceptor stream changes the conductivity of the acceptor solution causing an increase of signal at the capacitively coupled contactless conductivity detection (C(4)D) placed at the outlet of the vaporization chamber. This chamber is an adaption from previous work reported on 'membraneless vaporization' (MBL-VP). The method can be used in the quality control of production of mixed cement. These cement materials usually have calcium carbonate contents at high concentration range (e.g., 33-99% (w/w) CaCO(3)). Analysis of samples by this method is direct and convenient as it requires no sample pretreatment. The method is low-cost with satisfactory accuracy and acceptable precision.