Development of Pipetteless Paper-Based Analytical Devices with a Volume Gauge

In this work, we propose a new design for paper based analytical devices (PADs) that eliminate the need to use a micropipette for sample introduction. With this design, a PAD is equipped with a distance-based detection channel that is connected to a storage channel that indicates the volume of a sample introduced into the PAD. The analyte in the sample solution reacts with a colorimetric reagent deposited into the distance-based detection channel as the sample solution flows into the storage channel where the volume is measured. The ratio of the lengths of the detection channel and that of the storage channel (D/S ratio) are constant for a sample containing a certain concentration, which is independent of the introduced volume. Therefore, the PADs permit volume-independent quantification using a dropper instead of a micropipette because the length of the storage channel plays the role of a volume gauge to estimate the introduced sample volume. In this study, the D/S ratios obtained with a dropper were comparable to those obtained with a micropipette, which confirmed that precise volume control is unnecessary for this PAD system. The proposed PADs were applied to the determinations of iron and bovine serum albumin using bathophenanthroline and tetrabromophenol blue as colorimetric reagents, respectively. The calibration curves showed good linear relationships with coefficients of 0.989 for iron and 0.994 for bovine serum albumin, respectively

All-Step-in-One Test Kit for Paraquat Detection in Water and Vegetable Samples

This work presents the first development of an all-steps-in-one test kit for the determination of paraquat in natural water, and vegetable and agricultural samples. A handheld photometer incorporated with a magnetic stirrer was used to complete the steps of extraction, mixing, and detection. Paraquat produces a blue free radical ion via a reduction with sodium dithionite in alkaline conditions. Sodium dithionite powder was investigated for the enhancement of reagent stability duration, which was added directly into sample solution that showed insignificant difference in sensitivity as compared with that of the solution format of sodium dithionite. The developed test kit showed good performance with the linear calibration of 0.5 to 10 mg L−1 with a high coefficient of determination (r2 = 0.9947). The lower limit of quantitation (LLOQ = 3SD of intercept per slope) carried out from the method using the handheld photometer was 0.50 mg L−1. The limit of detection (LOD) by naked eye was 0.30 mg L−1. The recovery study was acceptable in the range of 101–115%. Intraday (n = 3) and interday (n = 3) precision was less than 1%. On the basis of the significance test at the 95% confidence interval, quantitative results of the developed test kit agreed well with those from high-performance liquid chromatography (HPLC). To the best of our knowledge, this is the first report demonstrating an online extraction for vegetables incorporated into a test kit, applicable for on-site analysis. Single-point calibration based on the Beer–Lambert law also demonstrated the measurement of paraquat. In testing with a nominal standard solution of 5.00 mg L−1 paraquat, the reading concentration was 5.09 ± 0.03 mg L−1 paraquat (n = 20) with a K value of 0.0967 (close to the slope of multipoint calibration). This research is a direct benefit to agricultural products and the health of a population for the analysis of pesticides and herbicides

Iodoform Reaction-Based Turbidimetry for Analysis of Alcohols in Hand Sanitizers

This work presents the first development of an analytical turbidimetric method for the determination of legal alcohols in alcohol-based hand sanitizer products. A typical iodoform reaction is exploited to form a yellow product in the form of precipitates. An iodoform test shows a positive result as yellow precipitates in the presence of ethanol and isopropanol; therefore, the test can only be used to distinguish between methanol and those legal alcohols. In the presence of molecular iodine (I2) and a strong alkaline solution, the legal alcohol is converted to the corresponding carbonyl compound (i.e., ethanol to acetaldehyde, isopropanol to acetone). The susceptibility of this intermediate towards the reaction with hydroxide ions (strong alkaline condition) results in formations of yellow precipitation of iodoform (CHI3) and a water-soluble carboxylate salt in the solution. Therefore, this change allows for the detection of legal alcohols through either naked-eye observation (as semi-quantitative analysis) or a common benchtop/portable photometer/spectrophotometer (as quantitative analysis) by means of turbidimetric analysis. In this work, turbidimetry is employed, which is a useful alternative detection method in analytical practice, especially with colored samples in hand sanitizing products. This is because they can employ wavelengths at which the colored solution does not absorb light. As a result of our developed method, the calibration plots are in the range of 30 to 100% (v/v) for both ethanol and isopropanol. The limit of detection (LOD) (3SD of y-intercept/slope) was found to be 7.4% (v/v) ethanol and 6.5% (v/v) isopropanol. Direct analysis of the non-pretreatment of the sample is achieved. The results indicate that our new proposed analytical method is fit for purpose and valid to detect the legal alcohols in alcohol-based hand sanitizing products for both international and Thai regulations (at least 70% (v/v)). Our quantitative results were also comparable to a standard analytical method, such as the use of a gas chromatography-flame ionization detector (GC-FID). Our developed method and analytical operation could potentially be developed into a practically portable analysis

Sequential Injection System for Analysis of Degree Brix, Orthophosphate and pH in Raw Sugarcane Juice Applicable to Sugar Industry

This work presents, for the first time, a new sequential injection analysis (SIA) method to simultaneously analyze degree Brix, orthophosphate and pH in raw cane juice. These key parameters relate to price of harvested sugarcane and quality of cane juice for sugar production. The SIA system employed two detectors: the first detector is a diode-array spectrophotometer, equipped with a regular flow cell, for measurements of degree Brix and orthophosphate. Quantitative of degree Brix (°Bx; ca. % (w/w) sucrose) was based on manipulation of the schlieren effect at the interface between plugs of sample and water. Orthophosphate analysis was carried out based on the molybdenum blue method with significant reduction in consumption of the reagents. Compensation of the schlieren effect from sucrose for determination of orthophosphate was achieved by using a dual-wavelength spectrometric detection. Second detector is a pH-sensing device, called ion-selective field-effect transistors (ISFET). The ISFET is based on the current through the ISFET arising according to the H+ concentration in solution. Our developed SIA system provides linear calibration graphs fitting for purpose in analysis of sugarcane juice (pH: 0–14, °Bx: 1.0–7.0 and P2O5: 20–200 mg L−1). Simultaneous analysis of sugarcane juice for pH, °Bx and P2O5 is carried out within 5 min (12 sample per h). Precision of SIA system is acceptable (RSD < 3%). Our SIA system gave quantitative results insignificantly different, as compared with conventional methods for analysis of pH, °Bx and P2O5 in sugarcane juice

Simple Flow-Based System with an In-Line Membrane Gas–Liquid Separation Unit and a Contactless Conductivity Detector for the Direct Determination of Sulfite in Clear and Turbid Food Samples

This study presents a simple flow-based system for the determination of the preservative agent sulfite in food and beverages. The standard method of conversion of sulfite ions into SO2 gas by acidification is employed to separate the sulfite from sample matrices. The sample is aspirated into a donor stream of sulfuric acid. A membrane gas–liquid separation unit, also called a ‘gas-diffusion (GD)’ unit, incorporating a polytetrafluoroethylene (PTFE) hydrophobic membrane allows the generated gas to diffuse into a stream of deionized water in the acceptor line. The dissolution of the SO2 gas leads to a change in the conductivity of water which is monitored by an in-line capacitively coupled contactless conductivity detector (C4D). The conductivity change is proportional to the concentration of sulfite in the sample. In this work, both clear (wine) and turbid (fruit juice and extracts of dried fruit) were selected to demonstrate the versatility of the developed method. The method can tolerate turbidity up to 60 Nephelometric Turbidity Units (NTUs). The linear range is 5–25 mg L−1 SO32− with precision <2% RSD. The flow system employs a peristaltic pump for propelling all liquid lines. Quantitative results of sulfite were statistically comparable to those obtained from iodimetric titration for the wine samples

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

Interferon-inducible protein (IFI) 16 regulates Chikungunya and Zika virus infection in human skin fibroblasts.

International audienceChikungunya virus (CHIKV), a re-emerging infectious arbovirus, causes Chikungunya fever that is characterized by fever, skin rash, joint pain, arthralgia and occasionally death. Despite it has been described for 66 years already, neither potential vaccine nor a specific drug is available yet. During CHIKV infection, interferon type I signaling pathway is stimulated and releases hundreds of interferon stimulated genes (ISGs). Our previous study reported that IFI16, a member of ISGs, is up-regulated during CHIKV virus infection and the suppression of the gene resulted in increased virus replication. Furthermore, our group also found that inflammasome activation can inhibit CHIKV infection in human foreskin cells (HFF1). Concomitantly, it has been reported that IFI16 activates the inflammasome to suppress virus infection. Therefore, we have hypothesized that IFI16 could be involved in CHIKV infection. In this study, we confirmed the expression level of IFI16 by Western blotting analysis and found that IFI16 was up-regulated following CHIKV infection in both HFF1 and human embryonic kidney cells. We next investigated its antiviral activity and found that forced expression of IFI16 completely restricted CHIKV infection while endogenous silencing of the gene markedly increased virus replication. Furthermore, we have discovered that IFI16 inhibited CHIKV replication, at least, in cell-to-cell transmission as well as the diffusion step. Interestingly, IFI16 also exerted its antiviral activity against Zika virus (ZIKV) infection, the global threat re-emerging virus can cause microcephaly in humans. Taken together, this study provides the first evidence of an antivirus activity of IFI16 during in vitro arbovirus infection, thus expanding its antiviral spectrum that paves the way to further development of antiviral drugs and vaccines

Size-Based Characterization of Polysaccharides by Taylor Dispersion Analysis with Photochemical Oxidation or Backscattering Interferometry Detections

International audienceTaylor dispersion analysis (TDA) is a powerful sizing technique very well suited for (macro)molecules between angstroms and submicrons (typically up to 200 nm). However, new detection modes are required for non-UV-absorbing (macro)molecules such as most of the polysaccharides, including starches. In this work, two different detection modes were compared, backscattering interferometry (BSI) and UV-photooxidation detection (UV-POD). TDA-BSI measures the relative change of the refractive index between eluent and sample (water as eluent in this work), whereas TDA-UV-POD detects the UV-absorbing photo-oxidized products of polysaccharides/starches in strong alkaline media (130 mM NaOH or 1 M KOH). TDA-UV-POD detection was evaluated for linearity and sensitivity at two wavelengths, 214 and 266 nm. The mass-average hydrodynamic radius (Rh) obtained by TDA-BSI and TDA-UV-POD was found to be in excellent agreement, whereas higher average Rh values were obtained by batch dynamic light scattering (DLS) under the same conditions, because of the higher sensitivity of DLS to large-size solutes and aggregates. The hydrodynamic radius distributions obtained by TDA and DLS are intrinsically different but both techniques were found to be complementary, providing useful information on sample dispersity. Owing to the absence of the stationary phase, low sample consumption with straightforward sample preparation (no filtration), and no calibration, TDA is anticipated to become a method of choice for the size-based characterization of polysaccharides, including starches