Fumiki Takahashi

Enzymatic flow injection method for rapid determination of choline in urine with electrochemiluminescence detection

In order to determine trace choline in human urine, a flow injection analysis (FIA) system has been developed by coupling an enzyme reactor with an electrochemiluminescence (ECL) detector. The enzyme reactor is prepared by covalently immobilizing choline oxidase (ChOx) onto the aminopropyl-controlled pore glass beads, which are then carefully packed into a micro column. The enzyme reactor catalyzes the production of hydrogen peroxide that is in direct proportion to the concentration of choline. In this study, the enzymatically produced hydrogen peroxide was detected by an ECL detector positioned at the down stream of an enzyme reactor based on the luminol/H(2)O(2) ECL system. Under the optimized condition, the enzymatic FIA/ECL provided high sensitivity for the determination of choline with the detection limit as low as 0.05 microM (absolute detection limit was at sub pmol level). The method was successfully applied in the determination of choline in the samples of human urine, and the analytical results were in good agreement with those obtained by using the microbore HPLC with an immobilized enzyme reactor-electrochemical detection system.

Electrochemical Determination of Chromium(VI) in River Water with Gold Nanoparticles–Graphene Nanocomposites Modified Electrodes

In this study, nanocomposites of ligand-free gold nanoparticles that are anchored onto the graphene surface (Graphene/AuNPs) were synthesized by a sonochemical method in a single reaction step. A highly sensitive amperometric sensor using Graphene/AuNPs is proposed for the determination of trace hexavalent chromium Cr(VI) in environmental water samples. Compared with a gold electrode, a glassy carbon electrode and a AuNPs modified glassy carbon electrode, the Graphene/AuNPs modified glassy carbon electrode exhibits the highest electrocatalytic activity and stability towards the reduction of Cr(VI), based on the results by cyclic voltammetry and electrochemical AC impedance studies. This study shows that the Graphene/AuNPs-based sensor can detect Cr(VI) with a low detection limit of 10 nM (∼0.5 μg/L), a wide dynamic range of 0 to 20 μM (R = 0.999) and very good selectivity and reproducibility. The electrode is applied to the determination of Cr(VI) in river samples with satisfactory recovery values.

Sonochemical preparation of gold nanoparticles for sensitive colorimetric determination of nereistoxin insecticides in environmental samples

A simple colorimetric method using gold nanoparticles (AuNPs) was developed as an efficient strategy for specific and sensitive detection of insecticides that are analogs of nereistoxin (NRT). The AuNPs were synthesized by a surfactant-free sonochemical reaction with ultrasonication at 430 kHz. A color change occurred in the presence of NRT because the AuNPs aggregated if they were coated with a small amount of thioctic acid (TA). At a pH of around 5, the TA adsorbed on the AuNPs was deprotonated, whereas NRT was protonated (NRT-H+). Adsorption of NRT-H+ onto the TA-coated AuNPs surface would decrease the surface charge of the AuNPs, and this resulted in aggregation. Because the aggregation of the TA-coated AuNPs could not be induced by amine compounds without thiol groups, this provided a surface-limited aggregation mechanism for specific sensing of NRT. The absorbance at 700 nm was dependent on the concentration of NRT, and the calibration curve was linear over the concentration range 85 nM (12 ng/mL) to 1000 nM (140 ng/mL). The applicability of the proposed method to detection of trace levels of NRT in environmental water samples was successfully demonstrated using a simple liquid-liquid reverse extraction technique.

Sonochemiluminescence of lucigenin: Evidence of superoxide radical anion formation by ultrasonic irradiation

The sonochemiluminescence (SCL) behavior of lucigenin (Luc2+) has been studied in aqueous solutions irradiated with 500 kHz ultrasound. Compared with the SCL of a luminol system, a tremendously increased SCL intensity is observed from 50 µM Luc2+ aqueous solution (pH =11) when small amounts of coreactants such as 2-propanol coexist. It is shown that SCL intensity strongly depends on the presence of dissolved gases such as air, O2, N2, and Ar. The highest SCL intensity is obtained in an O2-saturated solution, indicating that molecular oxygen is required to generate SCL. Since SCL intensity is quenched completely in the presence of superoxide dismutase (SOD), an enzyme that can catalyze the disproportionation of O2 •−, the generation of O2 •− in the ultrasonic reaction field is important in the SCL of Luc2+. In this work, the evidence of O2 •− production is examined by a spectrofluorometric method using 2-(2-pyridyl)benzothiazoline as the fluorescent probe. The results indicate that the yield of O2 •− is markedly increased in the O2-saturated solutions when a small amount of 2-propanol coexists, which is consistent with the results of SCL measurements. 2-Propanol in the interfacial region of a cavitation bubble reacts with a hydroxyl radical (•OH) to form a 2-propanol radical, CH3C•(OH)CH3, which can subsequently react with dissolved oxygen to generate O2 •−. The most likely pathways for SCL as well as the spatial distribution of SCL in a microreactor are discussed in this study.