Development of a Potential-Modulated Electrochemiluminescence Measurement System for Selective and Sensitive Determination of the Controlled Drug Codeine.

Codeine is a common analgesic drug that is a pro-drug of morphine. It also has a high risk of abuse as a recreational drug because of its extensive distribution as an OTC drug. Therefore, sensitive and selective screening methods for codeine are crucial in forensic analytical chemistry. To date, a commercial analytical kit has not been developed for dedicated codeine determination, and there is a need for an analytical method to quantify codeine in the field. In the present work, potential modulation was combined with electrochemiluminescence (ECL) for sensitive determination of codeine. The potential modulated technique involved applying a signal to electrodes by superimposing an AC potential on the DC potential. When tris(2,2'-bipyridine)ruthenium(II) ([Ru(bpy)3]2+) was used as an ECL emitter, ECL activity was confirmed for codeine. A detailed investigation of the electrochemical reaction mechanism suggested a characteristic ECL reaction mechanism involving electrochemical oxidation of the opioid framework. Besides the usual ECL reaction derived from the amine framework, selective detection of codeine was possible under the measurement conditions, with clear luminescence observed in an acidic solution. The sensitivity of codeine detection by potential modulated-ECL was one order of magnitude higher than that obtained with the conventional potential sweep method. The proposed method was applied to codeine determination in actual prescription medications and OTC drug samples. Codeine was selectively determined from other compounds in medications and showed good linearity with a low detection limit (150 ng mL-1).

Sensitive screening of methamphetamine stimulant using potential-modulated electrochemiluminescence.

Methamphetamine (MA) is one of the most commonly abused recreational stimulants; thus, rapid and sensitive screening methods for MA are of great importance in forensic analytical chemistry. In the present work, potential modulation was combined with electrochemiluminescence (ECL) for the sensitive determination of MA. The potential modulated (PM) technique involved applying a signal to electrodes by superimposing an alternating current potential on the direct current potential. When tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)32+) was used as an ECL emitter, the sensitivity of MA detection by PM-ECL was over 100 times that obtained with in conventional potential sweep mode. The radical on the α-carbon of the amine moiety is thought to play an important role in the ECL reaction mechanism involving amine-containing species. However, in the case of MA-type stimulants, density functional theory calculations suggest that the generated α-carbon radicals induce further intramolecular proton transfer. On the basis of the proposed ECL reaction route, we clarified the conditions under which MA could be selectively detected in the presence of the similar substance methoxyphenamine. The proposed method was applied to MA determination in a spiked human urine sample and showed good linearity with a low detection limit (100 nM, ca. 15 ng mL-1).

Electrochemiluminescence of Tris(2,2'-bipyridine)ruthenium(II)/Tri-n-propylamine with an Electric Contactless Power Transfer System.

An electrochemiluminescence (ECL) analytical device was developed using an electric contactless power transfer system. A mutually induced electromotive voltage was generated by wrapping an enameled wire around a commercial contactless charger. There was no electrical contact between the power supply and the electrochemical cell. For the tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)32+)/tri-n-propylamine system, a weak ECL signal was observed. When an inexpensive rectifier diode was introduced between the coil and the working electrode, the ECL intensity detection sensitivity increased by more than 100 times. The relationship between the waveform of the applied voltage and the ECL response was clarified, and the optimum conditions were determined. The intensity of the induced electromotive voltage was easily controlled by changing the number of turns in the coil. The proposed method is a safe, simple, and inexpensive technique without electrical contact.

Potential-modulated electrochemiluminescence of a tris(2,2'-bipyridine)ruthenium(II) / lidocaine system under 430 kHz ultrasound irradiation.

The electrochemiluminescence (ECL) of tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)32+) in the presence of lidocaine was investigated under ultrasound (US) irradiation. The sonoelectrochemical experiments are conducted by indirect irradiation of ultrasound with a piezoelectric transducer operating at 430 kHz. In a supporting electrolyte at pH 11, the Ru(bpy)32+/lidocaine system gave weak ECL peaks around +1.2 V and +1.45 V, respectively. The ECL signal at +1.2 V was attributed to redox reactions of the oxidative intermediates of Ru(bpy)32+ and lidocaine, while the signal at +1.45 V was assumed to be caused by an advanced oxidation process due to the generation of hydroxyl radicals (OH) at the electrode surface. In this study, the potential modulation approach is employed in the study of ECL process upon US irradiations because it can suppress the noise components from sonoluminescence effectly and improve the resolution of ECL-potential profiles. It is found ECL signals were greatly enhanced upon US irradiation at the output power of 30 W, however, the relative intensity of ECL signal at +1.2 V was larger than that obtained with a rotating disk electrode even though the mass transport effect is equilvalent. The experiment results suggest that the chemical effect (i.e., generation of OH) by 430 kHz US becomes remarkable in the electrochemical process. Detailed ECL reaction routes under US are proposed in this study.

High-Frequency Heating Extraction Method for Sensitive Drug Analysis in Human Nails.

Background: A simple, sensitive, and rapid extraction method based on high-frequency (H-F) heating was developed for drug analysis in human nails. Methods: A human nail was placed in a glass tube with an extraction solvent (methanol and 0.1% formic acid; 7:3, v/v), and a ferromagnetic alloy (pyrofoil) was wrapped in a spiral around the glass tube. Then, the glass tube was placed in a Curie point pyrolyzer, and a H-F alternating voltage (600 kHz) was applied. The sample and extraction solvent were heated at the Curie temperature for 3 min. Different Curie temperatures were applied by changing the pyrofoil (160 °C, 170 °C, 220 °C, and 255 °C). Results: The caffeine in the nail was effectively and rapidly extracted into the extraction solvent with the pyrofoil at 220 °C. The peak area obtained for the caffeine using liquid chromatography mass spectrometry (LC-MS/MS) was five times that of what was obtained after conventional ultrasonic irradiation extraction. Because the extraction uses high-pressure and high-temperature conditions in a test tube, the drugs that were strongly incorporated in nails could be extracted into the solvent. The amount of caffeine extracted was independent of the size of the pieces in the sample. Conclusions: Therefore, the sensitive determination of target drugs in nails is possible with rapid (20 min, including H-F extraction for 3 min) and simple sample preparation. The developed method was applied to a nail from a patient with hypertension.

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.

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.

Development and application of ultrasound-assisted microextraction to analysis of fenitrothion in environmental samples.

A microextraction technique based on ultrasonic emulsification and demulsification was developed for detecting pesticides at trace levels in environmental water samples. In this ultrasound-assisted microextraction (UAME), chloroform was emulsified with an aqueous sample solution containing trace fenitrothion (MEP) by ultrasonic irradiation (48 kHz) for 5 min. The emulsion was then demulsified by ultrasonic irradiation (2.4 MHz) for 10 min. This resulted in phase separation of the water and chloroform without centrifugation. The demulsified chloroform was collected by a microsyringe and submitted to gas chromatography-mass spectrometry. In conventional extraction with mechanical stirring, the extraction recovery (ER) of MEP was strongly dependent on the sample/chloroform volume ratio. However, in UAME, the ER was independent of the volume ratio and the ER was >80 % when the enrichment factor was 40. In UAME, MEP was rapidly extracted into the chloroform because of the large specific surface areas of the small chloroform droplets in the oil-in-water (O/W) emulsion. This gave a high extraction efficiency for MEP. UAME is a simple method requiring only a change in the ultrasound frequency and with no pretreatment steps that could contaminate the sample. The suitability of UAME was demonstrated by application to the detection of trace levels of pesticides in a spiked water sample from a fish tank. Graphical abstract Schematic diagram of the ultrasound-assisted microextraction (UAME) method.

Electrochemiluminescence of Tris(2,2'-bipyridyl)ruthenium(II) with Ascorbic Acid and Dehydroascorbic Acid in Aqueous and Non-aqueous Solutions.

The electrochemiluminescence (ECL) of tris(2,2'-bipyridyl)ruthenium(II) (Ru(bpy)3(2+)) is studied in non-aqueous media using dehydroascorbic acid (DHA) as coreactant to validate the evidence for the mechanism of the ascorbic acid (H2A)/Ru(bpy)3(2+) ECL system in an aqueous media. DHA is electrochemically reduced around -1.2 V vs. Ag/Ag(+) in pure acetonitrile to generate the ascorbyl radical anion (A(•-)), which is confirmed by in-situ UV-visible absorption measurements using a thin-layer spectroelectrochemical cell. The ECL of the DHA/Ru(bpy)3(2+) system in non-aqueous media is not observed in the potential range from 0 to +1.4 V in anodic potential sweep mode; however, distinct ECL is detected using double potential step electrolysis from -1.2 to +1.4 V vs. Ag/Ag(+). The ECL may be generated by a homogeneous charge-transfer process between A(•-) produced during the first pulse potential step (-1.2 V) and Ru(bpy)3(3+) generated during the second pulse potential step (+1.4 V). The calculated standard enthalpy (-ΔH°) for the charge-transfer reaction between A(•-) and Ru(bpy)3(3+) is 2.29 eV, which is larger than the lowest excited singlet state energy of Ru(bpy)3(2+) (*Ru(bpy)3(2+); 2.03 eV, 610 nm). It is determined that the generated intermediate A(•-) is crucial in the Ru(bpy)3(2+) ECL reaction.

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.

Rapid determination of ascorbic acid, dehydroascorbic acid, and total vitamin C by electrochemiluminescence with a thin-layer electrochemical cell.

This paper reports on a rapid and sensitive method for the simultaneous determination of ascorbic acid (H(2)A), dehydroascorbic acid (DHA), and total vitamin C by electrochemiluminescence (ECL) using a thin-layer electrochemical cell. Significant ECL signals can be generated by the anodic oxidation of Ru(bpy)(3) (2+) in the presence of H(2)A or DHA in pH 8.8 phosphate buffer solution. Because of the extremely small dead volume of the thin-layer cell (approximately 1.5 microL), almost all amount of H(2)A is assumed to be completely oxidized to DHA with a short pre-electrolysis step. As a result, it is possible to determine the reductive vitamin C (H(2)A) by square wave voltammetry before the pre-electrolysis step, while total vitamin C (sum of H(2)A and DHA) is able to be determined at a subsequent ECL step. The method was employed for the determination of vitamin C in commercial beverages with the analytical results in good agreement with the certified values.

Self-quenching in the electrochemiluminescence of Ru(bpy)3 2+ using ascorbic acid as co-reactant.

In this study, electrochemiluminescence (ECL) of Ru(bpy)(3)(2+) (bpy = 2,2'-bipyridyl) using ascorbic acid (H(2)A) as co-reactant was investigated in an aqueous solution. When H(2)A was co-existent in a Ru(bpy)(3)(2+)-containing buffer solution, ECL peaks were observed at a potential corresponding to the oxidation of Ru(bpy)(3)(2+), and the intensity was proportional to H(2)A concentration at lower concentration levels. The formation of the excited state *Ru(bpy)(3)(2+) was confirmed to result from the co-reaction between Ru(bpy)(3)(3+)and the intermediate of ascorbate anion radical (A(-)*), which showed the maximum ECL at pH = 8.8. It is our first finding that the ECL intensity would be quenched significantly when the concentration of H(2)A was relatively higher, or upon ultrasonic irradiation. In most instances, quenching is observed with four-fold excess of H(2)A over Ru(bpy)(3)(2+). The diffusional self-quenching scheme as well as the possible reaction pathways involved in the Ru(bpy)(3)(2+)-H(2)A ECL system are discussed in this study.