Optimization of geometry and surface modification of microchip input reservoirs were performed to achieve uninterferenced pressure-induced sample injection of multiple samples into microreactors using a single syringe pump. Nine samples of 3.5 µL were pipetted onto input reservoirs and loading of PCR mixture into 260 nL microreactors was achieved followed by successful PCR amplification, confirming that no cross-contamination occurs during injection.

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Reduction currents for H₂O₂ at a heme peptide (HP)-modified electrodes are suppressed by inhibitors, such as imidazole derivatives. Although this inhibition effect allows determinations of the total inhibition ability of imidazole derivatives, it has no selectivity. In this study the selectivity control of HP-modified electrodes for imidazole derivatives was performed utilizing the thermoresponsive phase transition of poly(N-isopropylacrylamide), which was chemically immobilized on HP-modified electrodes. The inhibition ratios for imidazole derivatives appeared to be small at temperatures below the lower critical solution temperature (LCST), and to be large above the LCST. This change was ascribed to a steric hindrance caused by a phase transition of the polymer. On the other hand, the inhibition ratio for histamine, which has a larger molecular size relative to imidazole, was not significantly changed by the phase transition. Thus, the selectivity of the HP-modified electrode was found to be controllable using an immobilized phase-transition polymer.

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A composite self-excited millimeter-sized lead zirconate titanate (PZT) glass cantilever (2 mm × 1.8 mm; sensing area of 6 mm²) was fabricated for the detection of Escherichia coli (E. coli) O157:H7. The fundamental and second mode resonance in air was 10.95 ± 0.05 kHz and 43.45 ± 0.05 kHz, respectively. Affinity purified monoclonal antibody (anti-E. coli O157:H7) specific to the pathogen E. coli O157:H7 was immobilized at the cantilever glass tip, and then immersed in liquid containing the pathogen (70 to 7 × 10⁷ cells/mL). The resonant frequency showed a reduction and reached a steady state shift of 0 ± 5, 46 ± 5, 260 ± 5, and 1010 ± 5 Hz corresponding to 0, 700, 7000, and 7 × 10⁷ cells/mL. From the experiments conducted, the detection limit of the sensor was 700 cells/mL.

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This paper reports on an inorganic reaction that performs complete discrimination of D- and L-enantiomers from each other in an aqueous solution at room temperature. This is the first finding of an inorganic reaction that acts like an organic biosystem in the sense that such a kind of complete discrimination of enantiomers is a matter solely done through biosystems in nature. We also assume that they suggest significant concerns with not only analytical chemistry in regards to the discrimination of isomers, but also with cosmo/geo-chemistry.

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Lipoylamino- β- and γ-cyclodextrin (LP- γ-CD and LP- γ-CD, respectively) were adsorbed at the surface of gold electrodes by sulfur-gold bonding. The resultant electrodes exhibited quasi-reversible voltammograms for the redox reaction of Fe(CN)₆^3-/4- in aqueous solutions, with peak-to-peak separation (ΔE_p) being 85 mV at 20 mV s^-1 as a potential sweep rate. When bile acids are added to the solution, ΔE_p values increased to 200 - 300 mV with increasing the concentration of bile acids. A Langmuir-type adsorption analyses satisfactorily afforded the binding constants (K_surf) of the surface-confined LP- β-CD and LP- γ-CD with the bile acids. The obtained K_surf values of LP- γ-CD are 5.0 - 50 times larger than the corresponding binding constants of γ-CD in homogenious aqueous solutions. Cyclic voltammetric experiments with positively, negatively, and non-charged adamantane derivatives as well as pH titration experiments revealed that the retardation of the electrode reaction of negatively charged Fe(CN)₆^3-/4- caused by bile acids was attributable (1) to electric potential changes due to the accumulation of the negative charges at the electrode surface, and (2) to an increase in the hydrophobicity of the electrode surface due to the binding of hydrophobic bile acids to the LP- β-CD and LP- γ-CD membranes.

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An amperometric glucose biosensor is developed that is based on immobilization of glucose oxidase (GOD) in a composite film of poly(o-aminophenol) (POAP) and carbon nanotubes (CNT), which are electrochemically co-polymerized at a gold (Au) electrode. Because of the high surface per volume ratio and excellent electrical conductivity of CNT, the biosensor based on an Au/POAP/CNT/GOD electrode has lower detection limit (0.01 mM), larger maximum response current (0.24 mA cm^-2) and higher sensitivity (11.4 mA M^-1 cm^-2) than the values of the biosensor based on an Au/POAP/GOD electrode. Additionally, the biosensor shows fast response time, large response current, and good anti-interferent ability for ascorbic acid, uric acid and acetaminophen. Good reproducibility and stability of the biosensor are also observed.

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A novel method for determination of trace amounts of tosufloxacin (TFLX) based on electrochemiluminescence (ECL) has been developed. The calibration graphs for TFLX were linear in the range of 5.0 × 10^-11 - 3.5 × 10^-7 mol/L, with the detection limit of 1.3 × 10^-11 mol/L. Key factors affecting the determination of TFLX were investigated. TFLX amounts in capsule and serum samples were successfully detected by this method. A possible mechanism of energy transfer, and thus the explanation of ECL in the Tb³⁺-TFLX-Na₂S₂O₄ system are discussed.

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A glassy carbon electrode modified with platinum nanoparticle-decorated carbon nanotubes (Pt-CNT/GCE) was prepared. The electrochemical behaviors for the catalysis oxidations of hydrogen peroxide and cysteine were studied. The Pt-CNT/GCE showed catalytic activity for electro-oxidation of hydrogen peroxide at 0.6 V in PBS (pH = 7.0) and for that of cysteine at 0.55 V in sulfuric acid medium (pH ≤ 2). The results indicated that the peak currents were proportional to the concentrations of both hydrogen peroxide and cysteine; the currents at Pt-CNT/GCE were almost 3-times higher than that at Pt electrode. The electrocatalytic oxidation mechanisms for hydrogen peroxide and cysteine at Pt-CNT/GCE surface were briefly studied.

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A procedure for separation and preconcentration of trace amounts of cadmium has been proposed. A column of analcime zeolite modified with benzyldimethyltetradecylammonium chloride and loaded with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP) was used for retention of cadmium. The cadmium was quantitatively retained on the column at pH ∼9 and was recovered from column with 5 ml of 2 M nitric acid with a preconcentration factor of 140. Anodic stripping differential pulse voltammetry was used for determination of cadmium. A 0.05 ng/ml detection limit for the preconcentration of aqueous solution of cadmium was obtained. The relative standard deviation (RSD) for eight replicate determinations at the 1 µg/ml cadmium levels was 0.31% (calculated with the peak height obtained). The calibration graph using the preconcentration system was linear from 0.01 to 150 µg/ml in final solution with a correlation coefficient of 0.9997. For optimization of conditions, various parameters such as the effect of pH, flow rate, instrumental conditions and interference of number of ions, were studied in detail. This method was successfully applied for determination of cadmium in various complex samples.

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A simple and efficient method for the selective separation and preconcentration of Ag⁺ using homogeneous liquid-liquid extraction was developed. Tetraspirocyclohexylcalix[4]pyrrole (TSCC4P) was synthesized and investigated as a suitable selective complexing ligand for Ag⁺. Zonyl FSA (FSA) was applied as a phase-separator agent under mild pH conditions. Under the optimal conditions ([TSCC4P] = 3.4 × 10^-4 M, [THF] = 25.0% v/v, [FSA] = 1.25% w/v, and pH = 4.5), 5 µg of Ag⁺ in 6.0 ml aqueous phase could be extracted quantitatively into 20 µl of the sedimented phase. The maximum concentration factor was 300-fold. The limit of detection of the proposed method was 0.005 ng/ml. The reproducibility of the proposed method was at most 3.5%. The influence of the pH, type and volume of the water-miscible organic solvent, concentration of FSA, concentration of the complexing ligand and the effect of different diverse ions on the extraction and determination of Ag⁺ were investigated. The proposed method was applied to the extraction and determination of Ag⁺ in different water samples.

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Uniformly-sized molecularly imprinted polymers (MIPs) for (S)-nilvadipine have been prepared by a multi-step swelling and polymerization method using methacrylic acid or 4-vinylpyridine (4-VPY) as a functional monomer, ethylene glycol dimethacrylate (EDMA) as a cross-linker, and toluene, chloroform, cyclohexanol or phenylacetonitrile as a porogen. The chiral recognition abilities of the MIPs for nilvadipine were evaluated using aqueous and non-aqueous mobile phases. Among the MIPs, the (S)-nilvadipine-imprinted 4-VPY-co-EDMA polymers prepared using toluene as a porogen showed the highest recognition ability for nilvadipine in both aqueous and non-aqueous mobile phases. In addition to molecular shape recognition, hydrogen-bonding interactions of the NH proton of nilvadipine with a pyridyl group of the (S)-nilvadipine-imprinted 4-VPY-co-EDMA polymers could play an important role in the retention and chiral recognition of nilvadipine in aqueous and non-aqueous mobile phases. Furthermore, the MIP for (S)-nilvadipine gave the highest molecular recognition ability when a porogenic solvent during polymerization was used as the mobile phase modifier.

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The influence of temperature on the surface structure of the octadecylsilica (ODS) bonded phase was investigated with a molecular dynamics (MD) simulation. The MD simulation was applied to a molecular model consisting of three parts: amorphous silica base, dimethyloctadecylsilyl ligands and n-hexane as a mobile phase solvent. More detailed information on the effect of temperature was obtained at the low temperature region than that reported in our previous study. The motion of ODS ligands could be estimated by the mean square displacement (MSD) of the terminal carbon atoms of ODS ligands. The gauche fraction in the ODS ligand conformation can also be estimated to obtain the ligand conformation for each simulation condition in detail. It can be seen that an elevated temperature induced the more bent ligand conformation. The trend has a good agreement to that of the results experimentally observed by using various spectroscopic techniques such as nuclear magnetic resonance (NMR), Fourier-transform infrared spectroscopy (FT-IR), and Raman spectroscopy.

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The photoinduced changes of metal-ion extractability of crown ether derivatives bearing three or four spirobenzopyran moieties and their analogues were studied using 1,2-dichloroethane as the organic solvent. Under dark conditions, these compounds extracted Cu²⁺, Ag⁺, and Pb²⁺ with their counteranions from the aqueous phase to the organic phase. The extraction equilibrium constants of the photochromic crown ether derivatives for Cu²⁺ and Ag⁺ were determined successfully. Under UV-light irradiation conditions, the extraction of Cu²⁺ by crowned tris(spirobenzopyran) was enhanced, while that of Ag⁺ was suppressed. During the competitive metal-ion extraction of crowned oligo(spirobenzopyran)s between Cu²⁺ and Ag⁺, the metal-ion selectivity was reversed explicitly by photoirradiation.

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By coupling the Fenton reaction with the glucose oxidase (GOD)-catalyzed reaction, we have developed a fluorometric method for the determination of glucose; the linear response range and the detection limit are 0.108 - 2.59 µg/ml and 0.0432 µg/ml, respectively. This approach is selective and has the advantages of using inexpensive reagents and avoiding interferences. Satisfactory results were obtained for the determination of glucose in human serum samples.

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A micro-flow chemiluminescence (CL) system in vivo for glucose determination by the on-line microdialysis sampling is described in this paper. The micro-flow CL system uses discrete sample droplets, which formed at the tip of the capillary with the sampling volume of 4.5 µl. The sol-gel method is introduced to co-immobilize horseradish peroxidase (HRP) and glucose oxidase (GOD) on the inside surface of the micro-flow cell which was fabricated in polymethyl methacrylate (PMMA). The CL detection involved enzymatic oxidation of glucose to D-gluconic acid and H₂O₂, then H₂O₂ oxidizing luminol to produce CL in presence of HRP. The microdialysis probe was utilized for sampling in the rabbit blood; the sample throughput was 20 h^-1. The glucose level in blood of the rabbit was on-line monitored with good results.

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N-(9-Anthrylmethyl)amines which combine a fluorescent subunit and a chelate forming fragment have revealed a signal switching property in an aqueous solution upon complexation (off) with Cu(II) and liberation (on) of the probe molecule by substitution with an other ligand. The ligand exchange reaction between N-(phosphonomethyl)glycine (glyphosate), a typical herbicide, with N-(9-anthrylmethyl)amines on Cu(II) ion leads the fluorescence signal intensity to revive, providing an indirect detection system of glyphosate available in water of neutral pH region. The present system has been applied to the post column detection in the ion chromatographic separation of glyphosate and its metabolite aminomethylphosphonic acid (AMPA).

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The determination of trace water in gas samples, such as isobutene, chloromethane (polymeric staple gases) and SF₆ by a conventional Karl Fischer coulometer is very difficult, because of the adsorption of trace water on the surface of sample pipe, the gasification of the liquefied samples, and the migration of moisture into the measuring cell from the surroundings. To solve these problems, we improved a device for coulometric determination of water by Karl Fischer method. The improved coulometer was used to determine the trace water in isobutene, chloromethane and SF₆; RSD was less than 5%, and recoveries ranged from 94.1 to 109.1%, which is adequate for the analysis of industry.

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Precise ⁶⁶Zn/⁶⁴Zn and ⁶⁸Zn/⁶⁴Zn isotopic ratios of biochemical samples have been measured using multiple collector-ICP-mass spectrometry (MC-ICPMS). In order to eliminate the mass spectrometric interferences on Zn isotopes (e.g., ⁶⁴Ni⁺ and ¹³⁶Ba²⁺), we chemically purified the analyte using an ion chromatographic technique. The resulting precisions of the ⁶⁶Zn/⁶⁴Zn and ⁶⁸Zn/⁶⁴Zn ratio measurements were 0.05‰ and 0.10‰ (2SD), respectively, which were enough to detect the isotopic variation of Zn in nature. Red blood cell (RBC) samples were collected from five volunteers (four males and one female), including a series of 12 RBC samples from one person through monthly-based sampling over a year. These were analyzed to test possible seasonal changes and variations in ⁶⁶Zn/⁶⁴Zn and ⁶⁸Zn/⁶⁴Zn ratios among the individuals. The ⁶⁶Zn/⁶⁴Zn and ⁶⁸Zn/⁶⁴Zn ratios for a series of 12 RBC samples collected over a year were 0.43‰ and 0.83‰ higher than the values of highly purified Zn metal (JMC Zn), and no seasonal change could be found. The ⁶⁶Zn/⁶⁴Zn and ⁶⁸Zn/⁶⁴Zn ratios for RBC samples collected from five volunteers did not vary significantly. In order to investigate Zn isotopic heterogeneity in a human body, Zn isotopic ratios of a hair sample collected from one of the volunteers was also analyzed. The ⁶⁶Zn/⁶⁴Zn and ⁶⁸Zn/⁶⁴Zn ratios for the hair sample were 0.59‰ and 1.14‰ lower than the mean value of RBC samples. This result demonstrates that detectable isotopic fractionation occurs in the human body. The data obtained here suggest that the isotopic ratios of trace metals could provide new information about transportation of metal elements in vivo.

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The electrooxidation of benzylic alcohol derivative in acidic aqueous solution shows an oxidation pre-peak in the cyclic voltammogram, which means that the reaction is proceeding via an ECE mechanism where the second electron transfer occurs at a less positive potential. From the result of the rotating ring-disk electrode voltammetry, the initial oxidation response of the electrode reaction can be extracted.

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The adsorption of arsenic(V) was investigated using macroporous resin beads containing magnetite crystals. Arsenic(V) was favorably adsorbed at pH 2 - 9, where the distribution coefficients were larger than 10³. The maximum capacity was 0.050 mmol/g. Metal cations including Ca(II), Mn(II), Co(II), Ni(II), Cu(II), Zn(II) and La(III) did not give serious interference at 10^-4 M level. Diluted arsenic(V) was collected with a packed column, and the retained arsenic(V) was quantitatively eluted out with 1 M NaOH.

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We demonstrate the extended application of a simple bead injection-flow injection system with a modified simple colorimetric detection unit for the determination of low amount of Cu²⁺ in samples of different matrices (water and supplement tablet samples) with minimal sample pretreatment by employing ammonium pyrrolidinedithiocarbamate (APDC) reagent.

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The cyclic voltammetric behavior of acetaldehyde and the derivatized product with 2,4-dinitrophenylhydrazine (DNPHi) has been studied at a glassy carbon electrode. This study was used to optimize the best experimental conditions for its determination by high-performance liquid chromatographic (HPLC) separation coupled with electrochemical detection. The acetaldehyde-2,4-dinitrophenylhydrazone (ADNPH) was eluted and separated by a reversed-phase column, C₁₈, under isocratic conditions with the mobile phase containing a binary mixture of methanol/LiCl_(aq) at a concentration of 1.0 × 10^-3 M (80:20 v/v) and a flow rate of 1.0 mL min^-1. The optimum condition for the electrochemical detection of ADNPH was +1.0 V vs. Ag/AgCl as a reference electrode. The proposed method was simple, rapid (analysis time 7 min) and sensitive (detection limit 3.80 µg L^-1) at a signal-to-noise ratio of 3:1. It was also highly selective and reproducible [standard deviation 8.2% ± 0.36 (n = 5)]. The analytical curve of ADNPH was linear over the range of 3 - 300 mg L^-1 per injection (20 µL), and the analytical recovery was > 99%.

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A novel fluorimetric method for the determination of peroxynitrite (ONOO^-) using hemoglobin (Hb) as a catalyst is described. The method employs the reaction of ONOO^- with thiamine (TM), a colorless, non-fluorescent reagent in a glycine-NaCl-NaOH buffer solution (pH 12.7), to generate a highly fluorescent product, thiochrome (TC). The fluorescent product was monitored by fluorimetry. A linear calibration graph was obtained over an ONOO^- concentration range from 4.95 × 10^-7 mol L^-1 to 2.97 × 10^-5 mol L^-1, with a detection limit of 9.78 × 10^-9 mol L^-1 ONOO^-. The relative standard deviation at an ONOO^- concentration of 2.11 × 10^-6 mol L^-1 was 4.15% (n = 9).

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The characterization of water molecules bound to ribonuclease T₁ (RNase T₁) was carried out using cold-spray ionization mass spectrometry (CSI-MS). CSI-MS is a variant of electrospray ionization mass spectrometry (ESI-MS) operating at low temperature, and is particularly suitable for investigating the weaker molecular associations, since the temperature at the spray interface is much lower than that in the conventional ESI-MS. In this approach, ion peaks due to the addition of nine water molecules were identified at a spray temperature of 48°C. This result showed good agreement with that inferred by the combinational analysis of NMR and X-ray crystallography, indicating that CSI-MS is capable of rapidly providing reliable information to characterize the number of water molecules bound to a macromolecule.

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A column packed with red blood cells (RBCs) was prepared for electrochromatography as a separation and reaction column. RBCs were kept inside a piece of fused silica capillary tubing with 2% agarose gel. In the column, RBCs were uniformly distributed in the agarose gel matrix and their electrophoretic movements due to an applied voltage were suppressed well. The durability of the biological function of the column under applied voltage was about 1 h, although it could remain for 2 - 3 days without applied voltage. The column could not be used when hemolysis of the RBCs was observed in the column. When the developed “RBC-gel column” was used, both pyridoxamine and serotonin were converted to other compounds through their direct contact with RBCs.

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A new chemiluminescence method using flow injection is described for the determination of four penicillins, namely: phenoxymethylpenicillin potassium, amoxicillin, ampicillin, and ampicillin sodium. The method is based on sensitizing effect of these drugs on the chemiluminescence reaction of potassium permanganate in sulfuric acid with glyoxal. The different experimental parameters affecting the chemiluminescence intensity were carefully studied and incorporated into the procedure. The method allows the determination of 0.1 - 1.0 µg/ml phenoxymethylpenicillin potassium, 0.1 - 1.0 µg/ml amoxicillin, 0.1 - 1.0 µg/ml ampicillin, and 0.1 - 1.0 µg/ml ampicillin sodium. The method was successfully applied to the determination of four penicillin antibiotics in pharmaceutical preparations.

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A new flow methodology exploiting the multi-pumping approach was developed for the spectrophotometric determination of ambroxol hydrochloride in pharmaceutical preparations. The flow manifold was implemented by using, exclusively, multiple solenoid-actuated micro-pumps, which acted simultaneously as sample insertion, solutions propelling and reagents commutation units. Linear calibration plots were obtained over an ambroxol concentration ranging from 10 to 200 mg l^-1 (r.s.d. < 0.5%, n = 15) and a sampling rate of about 60 samples per hour (flow rate = 1.92 ml min^-1, sample volume = 80 µl).

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An improved method of sample injection was demonstrated for introducing ultra-low volume liquid on a microfabricated device. In our previous study, a pressure-driven injection method has been introduced and was applied to on-chip electrophoresis. In this study, the need for control of the air vent, which was indispensable for sample injection in the previous study, was completely eliminated, facilitating sample injection with great simplicity and high accuracy. This was realized by altering the topology of the air vent channel, which is connected to a hydrophobic and narrow channel (called a passive valve). Several types of air vent channels were designed and their injection performances were tested. In addition, by modifying the shape and the position of air vent channel and passive valve, the residual liquid volume inside the passive valve after sample injection was decreased to approximately 0.5% of the injected volume, a value which showed high reproducibility.

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In scanning microscopy by total internal reflection with thermal lens spectroscopy, its spatial resolution depends on the distance between the sample and a converging lens, which corresponds to the objective lens in an ordinary optical microscope. It was found that the resolution was best when the signal induced by the thermal lens effect was maximum. The distance was precisely adjusted by monitoring the signal intensity, and the resolution became twice better than that previously reported. Using a shorter focal-length lens, a resolution of 1.9 µm was attained.

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