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Miniaturized chromate-free chloride assay in fish sauce based on linear height calibration of AgCl precipitate in inverted microtubes
(2026-06-01)
Chantiwas, Rattikan
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Wilairat, Prapin
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Putthasa, Papawarin
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Cheotchinda, Benjamapohn
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Teerasong, Saowapak
A miniaturized, chromate-free methodology for chloride determination in high-salt food matrices is presented in accordance with the principles of Green Analytical Chemistry. Classical argentometric precipitation is adapted to a microvolume, instrument-light assay by confining silver chloride formation within inverted microcentrifuge tubes and directly measuring the height of the compact precipitate layer using a simple ruler. Because the cylindrical tube region has a constant cross-sectional area, precipitate height is directly proportional to chloride concentration, enabling straightforward linear calibration without optical detection, signal transduction, or complex data processing. Controlled tube inversion and low-speed centrifugation ensure that precipitation occurs exclusively within this cylindrical zone, producing a sharp, reproducible solid–liquid interface. Under optimized conditions (150 µL sample volume and 180 µL of AgNO₃ in nitric acid; centrifugation at 600 rpm for 3 min followed by 5 min settling), the method provides a linear working range of 50–500 mM chloride (r² = 0.9988), a detection limit of 14 mM, and relative standard deviations of 3–4 % at 200–300 mM. Validation of 12 commercial Thai fish sauce samples showed no statistically significant differences compared to AOAC potentiometric titration at the 95% confidence level, with recoveries of 100–103 % and %RSD values of 1–4.2 %. The primary advantage of this method is its strictly linear height–concentration relationship, enabled by geometry-controlled confinement of the precipitate. Unlike other miniaturized or paper-based assays, it eliminates the need for image analysis or device fabrication. From a green chemistry perspective, the assay eliminates the use of potassium chromate indicators and disposable paper-based devices, operates at the microliter scale using reusable plastic microtubes, and requires only low-speed centrifugation. Analytical Greenness (AGREE), Analytical Eco-scale, Green Analytical Procedure Index (GAPI), and White Analytical Chemistry (WAC) evaluations (AGREE score: 0.76; Eco-scale: 83; WAC score: 88.9) classify the method as an excellent example of green analysis. The proposed approach offers a robust and sustainable screening tool for chloride determination in high-salt foods.
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A portable and stable dry starch-KI sensor for headspace iodine vapor detection: A green approach for sensitive ascorbic acid quantification in colored juices
(2025-03-01)
Paojinda, Ananya
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Sribunpeng, Tanaporn
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Kongsatjaviwat, Aumpika
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Kusumaningtyas, Nadia Mira
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Nuihuaykaew, Pichayaporn
This study introduces a stable and portable dry starch-KI paper sensor for detecting iodine vapor in the headspace of a microwell plate, enabling sensitive quantification of ascorbic acid (AA) in colored fruit and vegetable juices. The method leverages the reaction between AA and potassium iodate (KIO<inf>3</inf>) to generate iodine vapor, producing a strong colorimetric response upon interaction with the starch-KI coating. The sensor maintains consistent color intensity for up to 90 min, ensuring reliable on-site analysis. Uniformity testing of 96-spot absorbance for 0.5 mM AA across three batches (n = 3) yielded a mean absorbance of 0.264 ± 0.016 with a %RSD of 6.1 %. Stability assessments at three AA concentrations produced %RSD values of 9.4 %, 5.0 %, and 5.1 %, respectively. The microwell plate format facilitates rapid analysis with minimal sample preparation, effectively minimizing interference from juice color and matrix effects. The sensor demonstrated a linear detection range of 0.1–0.75 mM (r2 = 0.9942) with an LOD of 0.05 mM. Application to fresh juice samples yielded a %recovery of 80–120 % (n = 13). This approach addresses critical challenges such as stability, portability, and matrix interference by effectively separating analytical signals from juice pigments. With a high AGREE score of 0.7, reflecting strong alignment with green analytical chemistry principles, this innovative method offers a practical, eco-friendly solution for on-site AA quantification in nutritional analysis.
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Semi-quantitative analysis of formaldehyde in food using calibration chart based on number of colored wells of microwell plate titration
(2024-10-30)
Tongdee, Mintra
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Wilairat, Prapin
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Praditweangkum, Wiboon
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Chantiwas, Rattikan
Microplate titration quantifies sodium hydroxide generated from formaldehyde reacting with excess sulfite in a 96-microwell plate. Phenolphthalein indicators change from red to colorless when all hydroxide ions react. Methodology optimized reagent concentrations, and reaction time and created a Calibration Chart for semi-quantitative determination. The chart shows formaldehyde concentration ranges corresponding to red well counts from 0 to 200 mM in 20 mM increments. Inter-operator repeatability demonstrates precision (3 replicates), correlating red wells with standard formaldehyde concentrations. This instrument-free technique uses readily available commercial plates, eliminating the need for specialized equipment and calibration. The methodology offers simplicity with its reliance on readily available commercial plates and minimal specialized equipment, hence it is cost-effective and easily transportable 96-microwell plates enhancing the methodology's portability, and efficient semi-quantitative analysis of formaldehyde. The analysis of twelve solutions from food samples agrees with the quantitative values using titration.
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Multi-well plate as headspaces for paper-based colorimetric detection of sulfur dioxide gas: An alternative method of sulfite titration for determination of formaldehyde
(2023-01-25)
Yodpach, Nattapon
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Chantiwas, Rattikan
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Wilairat, Prapin
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Choengchan, Nathawut
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Praditweangkum, Wiboon
This work describes the analysis of formaldehyde using a 96-well microplate as multiple headspaces for the separation of sulfur dioxide gas generated from the sulfite remaining after its reaction with the formaldehyde in the sample. The quantitation of the gas is by colorimetric detection of an indicator paper placed over the microplate. The samples are aqueous extracts of various foods that are possibly adulterated with formaldehyde. A known excess amount of sulfite is added to the extract solution aliquoted in the well. The remaining sulfite is acidified with hydrochloric acid to generate sulfur dioxide gas which diffuses through the headspace above the solution to be absorbed at the moist strip of the indicator paper placed over the mouth of the wells. Anthocyanins extracted from the butterfly pea flower is used as the pH indicator giving a color change from the increase of hydrogen ions by hydrolysis of the absorbed sulfur dioxide gas. The exposed paper strip is scanned, and the digital images of the colored region analyzed using ImageJ software. The optimized method has a linear range of 200–1000 mg L−1 formaldehyde with limit of detection ((2.57*SD of intercept)/(slope of calibration line)) of the aqueous extract of 40 mg L−1 and coefficient of determination (r2) > 0.9979. Samples of fresh produce, such as seafood, meat, and vegetables, and various processed food were analyzed for their possible formaldehyde content. The results obtained from the headspace paper-based colorimetric detection are not statistically different from the values obtained from the titration method by paired t-tests.
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A ninety-six well plate as headspaces with moist starch indicator paper as a cover for the determination of ascorbic acid by iodate oxidation and formation of volatile iodine
(2022-02-21)
Nawalohakul, Thichaphat
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Charoenjiraroj, Pannarat
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Chantiwas, Rattikan
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Wilairat, Prapin
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Praditweangkum, Wiboon
This work presents the use of a 96-well plate as headspaces for the determination of ascorbic acid in samples loaded in the 96-well plate. Ascorbic acid in the sample is oxidized to iodide by the addition of excess acidic iodate solution into the well. The iodide is further oxidized by the remaining iodate to molecular iodine. A single sheet of moist starch indicator paper is immediately placed over the 96-well plate after the addition of the iodate with the moisture forming a gas seal. The iodine gas in each well diffuses through the headspace to react with the starch paper producing circular areas of a colored starch-iodine complex. After 15 min the indicator paper is scanned, and the digital images of the complex are analyzed by using ImageJ software to obtain blue intensity values. The precision of the intensity values from 12 wells containing 20 μL of 2.84 mM standard ascorbic acid is <2% relative standard deviation. Optimal conditions for detection were investigated, including the starch concentration, the acidic iodate reagent, and the measurement time. The linear calibration range of ascorbic acid is 0.284-2.84 mM, based on the plot of concentration vs. -log(reflectance). The coefficient of determination (r2) is >0.998. Samples of fruit juice and dietary supplements were analyzed for their ascorbic acid contents. The results obtained from the headspace reflectance method are not statistically different from values obtained from the titration method using paired t-tests (α = 0.05). This journal is
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Microfluidic analysis with front-face fluorometric detection for the determination of total inorganic iodine in drinking water
(2018-01-01)
Inpota, Prawpan
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Strzelak, Kamil
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Koncki, Robert
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Sripumkhai, Wisaroot
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Jeamsaksiri, Wutthinan
A microfluidic method with front-face fluorometric detection was developed for the determination of total inorganic iodine in drinking water. A polydimethylsiloxane (PDMS) microfluidic device was employed in conjunction with the Sandell-Kolthoff reaction, in which iodide catalyzed the redox reaction between Ce(IV) and As(III). Direct alignment of an optical fiber attached to a spectrofluorometer was used as a convenient detector for remote front-face fluorometric detection. Trace inorganic iodine (IO <inf>3</inf> - and I - ) present naturally in drinking water was measured by on-line conversion of iodate to iodide for determination of total inorganic iodine. On-line conversion efficiency of iodate to iodide using the microfluidic device was investigated. Excellent conversion efficiency of 93 - 103% (%RSD = 1.6 - 11%) was obtained. Inorganic iodine concentrations in drinking water samples were measured, and the results obtained were in good agreement with those obtained by an ICP-MS method. Spiked sample recoveries were in the range of 86%(±5) - 128%(±8) (n = 12). Interference of various anions and cations were investigated with tolerance limit concentrations ranging from 10-6 to 2.5 M depending on the type of ions. The developed method is simple and convenient, and it is a green method for iodine analysis, as it greatly reduces the amount of toxic reagent consumed with reagent volumes in the microfluidic scale.

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