Capillary surface modification using millimolar levels of aminosilane reagent for highly efficient separation of phenolic acids and flavonols by capillary electrophoresis with UV detection.

INTRODUCTION: Phytochemical analysis of phenolic acids and flavonols poses a challenge, necessitating the development of an efficient separation method. This facilitates the quantification of these compounds, yielding valuable insights into their benefits. OBJECTIVE: To develop a highly effective separation of phenolic acids and flavonols by capillary electrophoresis and ultraviolet (UV) detection through the modification of the capillary surface using 3-aminopropyltriethoxysilane (APTES) at millimolar concentrations. METHODS: The capillary surface is modified with 0.36 mM-APTES solution. The electrolyte is 20.0 mM borate buffer (pH 9.0). Separation performance (plate number N, resolution Rs ), stability, and reproducibility of the coating procedure are evaluated using the analysis of phenolic acids, rutin and quercetin. RESULTS: The modified capillary provided efficient separation with plate numbers N ≥ 1.0 × 104 m-1 and resolution Rs ≥ 0.8 for all pairs of adjacent peaks of the separation of five selected phenolic acids, rutin, quercetin, caffeine and methylparaben (as internal standard). The precisions of the relative migration times for 17 consecutive analyses of samples over 3 h were 1% relative standard deviation (RSD) for rutin and 7% RSD for quercetin. The analysis of rutin and quercetin in 12 dietary supplement product samples only required a simple dilution step for sample preparation. CONCLUSION: A straightforward modification technique utilising millimolar concentrations of APTES resulted in highly efficient separation of phenolic acids, rutin and quercetin, accompanied by high precision and surface stability. The modified capillary proved successful in analysing rutin and quercetin content in dietary supplements.

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.

Disposable Microchamber with a Microfluidic Paper-Based Lid for Generation and Membrane Separation of SO2 Gas Employing an In Situ Electrochemical Gas Sensor for Quantifying Sulfite in Wine.

This work presents a fully disposable microchamber for gas generation of a sample solution. The microchamber consists of a cylindrical well-reactor and a paper-based microfluidic lid (µFluidic lid), which also serves as the reagent loading and dispensing unit. The base of the reactor consists of a hydrophobic membrane covering an in-house graphene electrochemical gas sensor. Fabrication of the gas sensor and the three-layer µFluidic lid is described. The µFluidic lid is designed to provide a steady addition of the acid reagent into the sample solution instead of liquid drops from a disposable syringe. There are three steps in the procedure: (i) acidification of the sample in the reactor to generate SO2 gas by the slow dispensing of the acid reagent from the µFluidic lid, (ii) diffusion of the liberated SO2 gas through the hydrophobic membrane at the base of the reactor, and (iii) in situ detection of SO2 by cathodic reduction at the graphene electrode. The device was demonstrated for quantitation of the sulfite preservative in wine without heating or stirring. The selectivity of the analysis is ensured by the combination of the gas-diffusion membrane and the selectivity of the electrochemical sensor. The linear working range is 2-60 mg L-1 SO2, with a limit of detection (3SD of intercept/slope) of 1.5 mg L-1 SO2. This in situ method has the shortest analysis time (8 min per sample) among all voltammetric methods that detect SO2(g) via membrane gas diffusion.

Rapid measurement of indole levels in Brassica vegetables using one millilitre binary organic extraction solvent and capillary electrophoresis-UV analysis.

INTRODUCTION: Brassica vegetables contain high levels of indole compounds which have been found to provide health benefits, especially as cancer-preventive agents. An efficient and rapid method using solvent extraction with capillary electrophoresis (CE) and ultraviolet (UV) detection was developed for the determination of four major indoles from four types of Brassica vegetables. MATERIALS AND METHODS: Freeze-dried samples of four Brassica vegetables, i.e. broccoli, cauliflower, Chinese cabbage and cabbage, were selected. Hence, 1 mL of the binary solvent dimethylformamide (DMF)-methanol, 4:1 (v/v), was used for sample extraction. The extracts were diluted with the running buffer and directly analysed using CE with UV detection of four indole compounds. RESULTS: The binary solvent DMF-methanol, 4:1 (v/v) was selected from studies of the extraction efficiency of standard indoles spiked in ivy gourd (as the negative control sample) and using diphenylamine as the internal standard. Recovery was 80(±10)-120(±3)% for the four indoles: indole-3-carbinol (I3C), indole-3-acetonitrile (I3A), indole-3-acetic acid (IAA), and 3,3'-diindolylmethane (DIM). For direct analysis suitable dilution of the extract with the running buffer was required. The linear range of the quantitation is 0.75-25.0 µg/mL, limit of detection (LOD) of 0.14-0.52 µg/mL and r2 > 0.998. The amount of indole in the Brassica vegetables are in the order I3C > > IAA, I3A > DIM. CONCLUSION: A rapid method for extraction and quantitation of four indoles in four Brassica vegetables using CE with UV detection was developed. It has the potential as an efficient technique for generating data for use in agricultural and nutritional studies.

Genotype-Phenotype Correlation of ß-Thalassemia in Malaysian Population: Toward Effective Genetic Counseling.

Effective prevention of ß-thalassemia (ß-thal) requires strategies to detect at-risk couples. This is the first study attempting to assess the prevalence of silent ß-thal carriers in the Malaysian population. Hematological and clinical parameters were evaluated in healthy blood donors and patients with ß-thal trait, Hb E (HBB: c.79G>A)/ß-thal and ß-thal major (ß-TM). ß-Globin gene sequencing was carried out for 52 healthy blood donors, 48 patients with Hb E/ß-thal, 34 patients with ß-TM and 38 patients with ß-thal trait. The prevalence of silent ß-thal carrier phenotypes found in 25.0% of healthy Malaysian blood donors indicates the need for clinician's awareness of this type in evaluating ß-thal in Malaysia. Patients with ß-TM present at a significantly younger age at initial diagnosis and require more blood transfusions compared to those with Hb E/ß-thal. The time at which genomic DNA was extracted after blood collection, particularly from patients with ß-TM and Hb E/ß-thal, was found to be an important determinant of the quality of the results of the ß-globin sequencing. Public education and communication campaigns are recommended as apparently healthy individuals have few or no symptoms and normal or borderline hematological parameters. ß-Globin gene mutation characterization and screening for silent ß-thal carriers in regions prevalent with ß-thal are recommended to develop more effective genetic counseling and management of ß-thal.

Development of a Simple Reversible-Flow Method for Preparation of Micron-Size Chitosan-Cu(II) Catalyst Particles and Their Testing of Activity.

A simple flow system employing a reversible-flow syringe pump was employed to synthesize uniform micron-size particles of chitosan-Cu(II) (CS-Cu(II)) catalyst. A solution of chitosan and Cu(II) salt was drawn into a holding coil via a 3-way switching valve and then slowly pumped to drip into an alkaline solution to form of hydrogel droplets. The droplets were washed and dried to obtain the catalyst particles. Manual addition into the alkaline solution or employment of flow system with a vibrating rod, through which the end of the flow line is inserted, was investigated for comparison. A sampling method was selected to obtain representative samples of the population of the synthesized particles for size measurement using optical microscopy. The mean sizes of the particles were 880 ± 70 µm, 780 ± 20 µm, and 180 ± 30 µm for the manual and flow methods, without and with the vibrating rod, respectively. Performance of the flow methods, in terms of rate of droplet production and particle size distribution, are discussed. Samples of 180 µm size CS-Cu(II) particles were tested for catalytic reduction of 0.5 mM p-nitrophenol to p-aminophenol by 100-fold excess borohydride. The conversion was 98% after 20 min, whereas without the catalyst there was only 14% conversion.

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).

Solidification of floating organic droplet microextraction for determination of seven insecticides in fruit juice, vegetables and agricultural runoff using gas chromatography with flame ionization and mass spectrometry detection.

Liquid microextraction employing solidification of the floating organic droplet, with vortexing and heating to optimize extraction efficiency, was developed for the determination of seven insecticides in fruit juice, vegetables, and agricultural runoff water. The extracts were analyzed by gas chromatography with both flame ionization and mass spectrometry detection for the determination of chlorpyrifos, prothiofos, profenofos, ethion, λ-cyhalothrin, permethrin, and cypermethrin, respectively. Using 20 µL of 1-undecanol in 10 mL of aqueous solution containing 1% w/v sodium chloride provided preconcentration factor of 500. The enrichment factor of the analytes was in the range of 355 to 509 with extraction recovery >71%. The linearity ranges were 4-200 µg/kg for gas chromatography with flame ionization detection and 1-100 µg/kg for gas chromatography with mass spectrometry, with limits of detection ranging from 0.04 to 1.2 µg/kg, which are lower than the international maximum residue limits for vegetables and fruit juice. Intra-day and inter-day precisions are less than 5.4 and 7.0% relative standard deviation, respectively. The method was successfully applied to the determination of the seven insecticides in samples of vegetables, fruit juice and agricultural runoff, with recoveries ranging from 61.7 to 120.8%. The extraction method is simple, efficient and environmentally friendly.

Quadrupole-Time-of-Flight Mass Spectrometric Identification of Hemoglobin Subunits α, ß, γ and δ in Unknown Peaks of High Performance Liquid Chromatography of Hemoglobin in ß-Thalassemias.

This is the first report of quadrupole time-of-flight (Q-TOF) mass spectrometric identification of the hemoglobin (Hb) subunits, α, ß, δ and γ peptides, derived from enzymatic-digestion of proteins in the early unknown peaks of the cation exchange chromatography of Hb. The objectives were to identify the unknown high performance liquid chromatography (HPLC) peaks in healthy subjects and in patients with ß-thalassemia (ß-thal). The results demonstrate the existence of pools of free globin chains in red blood cells (RBCs). The α-, ß-, δ- and γ-globin peptides were identified in the unknown HPLC peaks. The quantification and role of the free globin pool in patients with ß-thal requires further investigation. Identification of all types of Hb subunits in the retention time (RT) before 1 min. suggests that altered Hbs is the nature of these fast-eluting peaks. Relevancy of thalassemias to the protein-aggregation disorders will require review of the role of free globin in the pathology of the disease.

Simple and fast analysis of iohexol in human serums using micro-hydrophilic interaction liquid chromatography with monolithic column.

A simple and rapid method based on micro-liquid chromatography using a synthetic monolithic capillary column was developed for determination of iohexol in human serums, a marker to evaluate the glomerular filtration rate. A hydrophilic methacrylic acid-ethylene dimethacrylate monolith provided excellent selectivity and efficiency for iohexol with separation time of 3 min using a mobile phase of 40:60 v/v 50 mM phosphate buffer pH 5/methanol. Four serum protein removal, methods using perchloric acid, 50% acetonitrile, 0.1 M zinc sulfate, and centrifuge membrane filter were examined. The method of zinc sulfate was chosen due to its simplicity, compatibility with the mobile phase system, nontoxicity, and low cost. Interday calibration curves were conducted over iohexol concentrations range of 2-500 mg/L (R(2) = 0.9997 ± 0.0001) with detection limit of 0.44 mg/L. Intra- and interday precisions for peak area and retention time were less than 2.8 and 1.4%, respectively. The method was successfully applied to serum samples with percent recoveries from 102 to 104. The method was applied to monitor released iohexol from healthy subject. Compared with the commercially available reversed-phase high-performance liquid chromatography method, the presented method provided simpler chromatogram, faster separation with higher separation efficiency and much lower sample and solvent consumption.

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.

The effect of acetone on the dynamics of temporal oscillations and waves in the ruthenium-catalyzed Belousov-Zhabotinsky reaction.

The effect of acetone on temporal oscillations and spatio-temporal patterns occurring in the ruthenium-catalyzed Belousov-Zhabotinsky (BZ) reaction was investigated in a closed batch system. The periods of temporal oscillations and waves significantly decrease with increasing acetone concentration. At low concentrations of acetone (0.01-0.05 M), regular wave patterns are observed with prolonged lifetimes of both temporal oscillations and waves. However, for higher concentrations (0.10-1.00 M acetone), the lifetime is shortened and irregular patterns are formed. The photosensitivity of waves of the Ru(bpy)3(2+)-catalyzed BZ reaction remains the same for all acetone concentrations. The results are discussed in terms of the proposed reaction mechanism.

Simple in-house flow-injection capillary electrophoresis with capacitively coupled contactless conductivity method for the determination of colistin.

An in-house flow-injection capillary electrophoresis with capacitively coupled contactless conductivity detection method was developed for the direct measurement of colistin in pharmaceutical samples. The flow injection and capillary electrophoresis systems are connected by an acrylic interface. Capillary electrophoresis separation is achieved within 2 min using a background electrolyte solution of 5 mM 2-morpholinoethanesulfonic acid and 5 mM histidine (pH 6). The flow-injection section allows for convenient filling of the capillary and sample introduction without the use of a pressure/vacuum manifold. Capacitively coupled contactless conductivity detection is employed since colistin has no chromophore but is cationic at pH 6. Calibration curve is linear from 20 to 150 mg/L, with a correlation coefficient (r(2) ) of 0.997. The limit of quantitation is 20 mg/L. The developed method provides precision, simplicity, and short analysis time.

Data independent acquisition for gas chromatographic MS/MS analysis of volatile compounds.

This study presents a novel tandem mass spectrometry (MS/MS) approach utilizing a data independent acquisition (DIA) concept specifically designed with gas chromatography-electron ionization-triple quadrupole mass spectrometry (GC-EI-QqQMS). This allows compound identification based on comparison between all the experimental MS/MS product ion spectra and the simulated library data of >1,000 MS/MS transitions of 71 compounds. The simulation data were generated by using the Competitive Fragmentation Modeling (CFM-ID) 3.0 program. The approach for calculation of the DIA MS/MS library match scores was then established and applied for identification of a range of terpenoids and oxygenated compounds in perfume. The identity of each peak was confirmed using 4-241 MS/MS transitions. The established data collection and analysis methods are expected to be useful for increased confidence in volatile compound analysis.

Determination of promethazine in forensic samples using multi-walled carbon nanotube-gold nanoparticle electrochemical sensor.

An electrochemical sensor was developed based on a glassy carbon electrode (GCE) modified with multi-walled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) for the determination of promethazine (PMZ) in 'purple drank', pharmaceutical formulations, and synthetic saliva. The oxidation of PMZ at the modified electrode occurred at a higher cathodic potential and produced a higher sensitivity compared to the unmodified GCE. The morphology of the modified electrode was characterized using field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDS), and transmission electron microscopy (TEM). The presence of MWCNTs and AuNPs was confirmed. The optimized parameters included the concentration and pH of the supporting electrolyte, amount of modifiers used to fabricate the electrode, deposition potential, and time. Using these optimized conditions, the method has a linear range from 0.5 to 100 µmol L-1, with a R2 value of 0.9991. The limit of detection (3SDblank/slope) was 0.13 µmol L-1. The proposed electrochemical sensor was successfully applied for the determination of PMZ in 'purple drank', pharmaceutical formulations, and spiked synthetic saliva samples. The results obtained from this sensor were in statistical agreement with the values obtained using the reference gas chromatography-flame ionization method.

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.

Multi-well plate as headspaces for paper-based colorimetric detection of sulfur dioxide gas: An alternative method of sulfite titration for determination of formaldehyde.

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.

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.

A simple cost-effective paper-based electrochemical device for detection of adulterated sibutramine in slimming products.

This work presents the first paper-based electrochemical device, or ePAD, for direct detection of adulterated sibutramine in slimming products. The ePAD was fabricated using a screen-printing technique for defining the hydrophilic area for sample loading and for the working, reference and counter electrodes. The ePAD gave reproducible responses comparable to both conventional rod electrodes and commercial screen-printed electrodes (SPEs). Use of paper to fabricate the ePAD device provides advantages over the conventional SPE platforms (e.g. glass, ceramics and polymers) in terms of biocompatibility, strong capillary action and environmental friendliness. To detect sibutramine, square wave voltammetry was employed after sample loading on the circular hydrophilic area. The linear range is 2.51 to 83.7 mg L-1 sibutramine, with a precision of 6 %RSD (n = 3) and an instrumental limit of detection (3SD of intercept/slope) of 2.46 mg L-1 sibutramine. Recovery of spiked samples ranged from 83 to 116%. The samples were capsules, slimming coffee powders and nutraceutical beverages. The samples were appropriately diluted to give concentrations within the linear calibration range. Filtration of undissolved solids found with the capsules and coffee powder samples was not required, demonstrating that the method is not susceptible to solid particles. The ePAD is cost-effective (

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.

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).