The fabrication of enzyme electrodes using self-assembled monolayers (SAMs) has attracted considerable interest because of the spatial control over the enzyme immobilization. A model system of glucose oxidase covalently bound to a gold electrode modified with a SAM of 3-mercaptopropionic acid was investigated with regard to the effect of fabrication variables such as the surface topography of the underlying gold electrode, the conditions during covalent attachment of the enzyme and the buffer used. The resultant monolayer enzyme electrodes have excellent sensitivity and dynamic range which can easily be adjusted by controlling the amount of enzyme immobilized. The major drawback of such electrodes is the response which is limited by the kinetics of the enzyme rather than mass transport of substrates. Approaches to bringing such enzyme electrodes into the mass transport limiting regime by exploiting direct electron transfer between the enzyme and the electrode are outlined.

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Amperometric measurement of superoxide dismutase (SOD) was carried out at cytochrome c-immobilized monolayers and ascorbate oxidase (AOD)/xanthine oxidase (XOD)/cytochrome c- and (AOD, XOD)/cytochrome c-multilayers. Cytochrome c was covalently immobilized on mercaptopropionic acid-containing self-assembled monolayers on gold. A biopolymer membrane of poly-L-lysine confining XOD and AOD was cast on the monolayer of cytochrome c. While both the cytochrome c-immobilized monolayer and multilayer electrodes show anodic current responses to the generation of superoxide radical, the sensitivity of the multilayer system for the detection of superoxide radical was high relative to that of the monolayer system. In the case of the cytochrome c-multilayer electrodes, the generation of superoxide radical near the sensing element, cytochrome c, resulted in high sensitivity for the detection of superoxide. The use of a XOD and AOD-incorporated poly-L-lysine membrane enabled the detection of the generation of superoxide radical in the presence of L-ascorbic acid. Though L-ascorbic acid could scavenge superoxide radical, the biopolymer membrane confined with AOD will oxidize any L-ascorbic acid that permeated into the membrane. By using the multilayer electrodes, one could measure the activity of SOD in the presence of L-ascorbic acid.

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A novel electrochemical system has been developed for monitoring the cyclic AMP-dependent protein kinase (PKA) activity. In this method, PKA activity was monitored as the change in the redox current of a ferrocene-pendant PKA substrate peptide (Fe-LRRASLG) on a gold electrode, which had been modified with thioctic acid, using cyclic voltammetry. The phosphrylation of the ferrocene-pendant substrate with PKA changed the net charge from +1 to -1. This caused a decrease in the redox current of the ferrocene unit due to an electrostatic repulsion between the substrate and the anionic surface of the electrode. We expect that this method is potentially useful for monitoring the enzyme activity in medical or pharmacological fields.

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The voltammetric detection of soybean agglutinin (SBA) was investigated on the basis of an interaction between the lectin and a sugar. Because galactose and lactose combined with SBA, the sugars were labeled by a Schiff base with an electroactive daunomycin. After the labeled sugar and SBA were mixed, measurements were carried out by voltammetry. When SBA-sugar binding occurs, a part of daunomycin of the labeled sugar is taken to the binding sites. As a result, SBA is detected by a change in the peak current of daunomycin, and the SBA-sugar interaction is evaluated. The length of the alkyl chain between daunomycin and the sugar was also considered. The electrode response to the concentration of SBA was linear over the range of 0.04 - 0.8 µg ml^-1. The merits of this procedure are the convenient preparation of labeled sugar and a rapid measurement without separation. On the other hand, the detection of sugar at the 10^-9 mol dm^-3 level was achieved by a competitive reaction to limited binding sites of the lectin between the sugar and the labeled sugar.

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The scanning electrochemical microscope (SECM) is used to image the activity of enzymes immobilized on the surfaces of disk-shaped carbon-fiber electrodes. SECM was used to map the concentration of enzymatically produced hydroquinone or hydrogen peroxide at the surface of a 33-µm diameter disk-shaped carbon-fiber electrode modified by an immobilized glucose-oxidase layer. Sub-monolayer coverage of the enzyme at the electrode surface could be detected with micrometer resolution. The SECM was also employed as a surface modification tool to produce microscopic regions of enzyme activity by using a variety of methods. One method is a gold-masking process in which microscopic gold patterns act as mask for producing patterns of chemical modification. The gold masks allow operation in both a positive or negative process for patterning enzyme activity. A second method uses the direct mode of the SECM to produce covalently attached amine groups on the carbon surface. The amine groups are anchors for attachment of glucose oxidase by use of a biotin/avidin process. The effect of non-uniform enzyme activity was investigated by using the SECM tip to temporarily damage an immobilized enzyme surface. SECM imaging can observe the spatial extent and time-course of the enzyme recovery process.

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A glass capillary ultramicroelectrode (tip diameter ≈ 1.2 µm) having an electrokinetic sampling ability is described. It is composed of a pulled glass capillary filled with an inner solution and three internal electrodes (Pt working and counter electrodes and an Ag/AgCl reference electrode). The voltammetric response of the capillary electrode is based on electrokinetic transport of analyte ions from the sample solution into the inner solution across the conical tip. It was found that the electrophoretic migration of analytes at the conical tip is faster than electroosmotic flow, enabling electrokinetic transport of analyte ions into the inner solution of the electrode. By using [Fe(CN)₆]^4- and (ferrocenylmethyl)trimethylammonium (FcTMA⁺) ions as model analytes, differential pulse voltammetric responses of the capillary electrode were investigated in terms of tip diameter of the capillary, sampling voltage, sampling time, detection limit and selectivity. The magnitude of the response depends on the size and charge of analyte ions. With a capillary electrode having a ≈ 1.2-µm tip diameter, which minimizes non-selective diffusional entry of analytes, the response after 1 h sampling at +1.7 V is linearly related to [Fe(CN)₆]^4- concentration in the range of 0.50 - 5.0 mM with the detection limit of 30 µM. Application of a potential of the same sign as that of the analyte ion forces the analyte to move out from the electrode to the solution, enabling reuse of the same capillary electrode. The charge-selective detection of analytes with the capillary electrode is demonstrated for [Fe(CN)₆]^4- in the presence of FcTMA⁺.

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The half wave potential of reduction of Sb⁵⁺ in 16Na₂O·10CaO·74SiO₂ glass melts was examined by differential pulse voltammetry up to 1873 K. The half wave potential shifted to the positive direction with an increase in temperature. The results indicate that the equilibrium of Sb⁵⁺/Sb³⁺ shifted to negative direction with an increase in temperature. The half wave potential shifted to positive direction (48 mV at 1473 K) when the atmosphere over the melts changed from pure oxygen gas to air, in agreement with the theoretical prediction. The reversibility of Pt:O₂ reference electrode is confirmed.

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The electrode reaction of Pu³⁺/Pu couple at the interface between LiCl-KCl eutectic melt containing PuCl₃ and liquid Bi phase was investigated by cyclic voltammetry at 723, 773 and 823 K. For comparison, the system was also studied using a solid Mo electrode in place of the liquid Bi electrode. It was found that the electrode reaction of Pu³⁺/Pu couple at the Bi electrode was almost reversible. The redox potentials of Pu³⁺/Pu couple at the liquid Bi electrode in the LiCl-KCl eutectic melt containing 0.87 wt% PuCl₃ (0.0014 mole fraction) at 723, 773 and 823 K were observed to be more positive by 0.575, 0.572 and 0.566 V, respectively, than those at the Mo electrode. These differences in potential were thermodynamically analyzed by assuming a lowering of the activity of Pu in Bi phase according to the alloy formation of PuBi₂.

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The electrolyte effects on the electrochemical behaviors of osmium complex polymer modified electrodes were investigated by a comparison between two osmium complexes, [Os(bpy)₂(PVI)₁₀Cl]Cl (Os-PVI₁₀) and [Os(bpy)₂(PVP)₁₀Cl]Cl (Os-PVP₁₀). The electrode process at Os-PVI₁₀ modified electrodes is reaction-controlled, while a diffusion-controlled electrode process exists at Os-PVP₁₀ modified electrodes. Both the cation and anion in supporting electrolytes strongly affect their electrochemical behaviors, such as the redox potential, wave shape and peak current. These phenomena are attributed to a change in the film structure and polymer swelling in various supporting electrolytes. The influence of electrolyte anions on the electrochemical behaviors is related to their hydrophobicity. The electrode process of Os-PVP₁₀ depends on the pH value of solutions, exhibiting different electron transfer mechanisms.

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The ion-transfer reaction of local anesthetics at an organic solvent/water interface has been studied using cyclic voltammetry (CV) with a stationary nitrobenzene (NB)/water (W) interface. Procaine and seven other local anesthetics gave reversible or quasi-reversible voltammograms at the NB/W interface in the pH range between 0.9 and 9.6. These drugs are present in aqueous solution in either neutral or ionic form, or both forms. The half-wave potential, as determined by the midpoint potential in CV, vs. pH curves, were determined and analyzed to determine the partition coefficients of both neutral and ionic forms of the drugs between NB and W. The partition coefficients of the ionic forms were derived from their formal potential of transfer at an NB/W interface. The dissociation constants of ionic forms of the drugs in NB were also deduced. A high correlation between the pharmacological activity and the partition coefficient of the ionic form of amide-linked local anesthetics has been shown.

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Fog and rain water samples were collected at the same time in the Akita Hachimantai mountain range in northern Japan from June to September in 1998 and 1999. The various ion concentrations in these samples were analyzed, and the fog droplet sizes were measured for each fog event. As the fog droplet size increased, the ion concentration decreased. The slope of log-log plots of the concentration versus the droplet size differed with the kind of ion. In order to characterize the air pollutant, moreover, these data were quantitatively analyzed by an oblique rotational factor analysis. We found that three factors were extracted as the air pollutant source: (NH₄)₂SO₄, acids (HNO₃ + H₂SO₄) and sea-salt. Combining the factor analysis with the 72 h back-trajectory at 850 hPa level, we found that the contribution of each factor varied with the transport pattern of air masses.

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Dissolved organic carbon (DOC) in water samples from Lake Biwa was chemically characterized by two inorganic adsorbents with completely different surface characteristics. The two adsorbents were HIO (hydrous iron oxide) and SG (silica gel). Solutions of reference standard materials were analyzed concerning their adsorption behavior to HIO and SG for bovine serum albumin (BSA), fulvic acid extracted from the bottom sediments of Lake Biwa, phthalic acid, and starch. The adsorption of DOC to HIO was mainly controlled by ligand exchange and electrostatic interaction; that of SG was by electrostatic interaction. It was found that in a weak acid solution of around pH 5, BSA adsorbs to both HIO and SG, but that fulvic acid, phthalic acid and starch only show adsorption to HIO. Using these characteristics, DOC samples in natural water samples were characterized into pro-DOC, which adsorbs to both HIO and SG at pH 5, and car-DOC, which only adsorbs to HIO at pH 5. The DOC samples in Lake Biwa on October 7, 1997, at sampling sites Nb-2 and Nb-5 (south basin of Lake Biwa, the depths were about 2 and 4 m), and Ie-1 (north basin of Lake Biwa, the depth was about 75 m) were characterized. The pro-DOC has different values, depending on their sampling sites and depths, and had the maximum value of 0.42 mg C l^-1 at the surface water of Ie-1, and had the lowest values at middle to deeper water depths (0.18 - 0.27 mg C l^-1). The car-DOC showed a relatively stable value at Ie-1 regardless of the depth (0.63 - 0.83 mg C l^-1), and the maximum value was observed in Nb-2 and Nb-5 (1.2 and 1.3 mg C l^-1). The ratios between car-DOC and pro-DOC concentrations were 0.2 - 0.5, and had different values for different sampling sites and depths. The ratios were significantly different for surface water samples where the biological activities are high and for bottom water samples where decomposition predominates.

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The native oligosaccharides of lacto-N-neotetraose (Gal β1-4GlcNAc β1-3Gal β1-4Glc; LNnT) and lacto-N-tetraose (Gal β1-3GlcNAc β1-3Gal β1-4Glc; LNT) were analyzed by using curved-field reflectron matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). Since a curved-field reflectron TOFMS enables a simultaneous focusing of a wide mass range of metastable fragment ions, the relative ion intensities in the post-source decay (PSD) mass spectra can be discussed. The PSD mass spectra of LNnT and LNT were distinguishable in their relative ion intensities. In the case of LNT, β-elimination could occur in the N-acetyl glucosamine (GlcNAc) at the C-3 position, which was bonded by galactose (Gal); however, it did not occur in LNnT. The 3-O elimination caused a difference in the relative ion intensities in the PSD mass spectra of LNnT and LNT. The β1-3 glycosyl linkage cleaved more easily than the β1-4 glycosyl linkage in the MALDI-PSD fragmentation. An analysis of the relative ion intensities in the MALDI-PSD mass spectra of oligosaccharides was very useful for distinguishing the linkage isomers and for characterizing the types of glycosyl linkages.

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We utilized microchip technology and found that the multilayer flow of liquids can be formed in microchannels. Liquid/liquid interfaces were formed parallel to the side wall of the microchannels, because the surface tension and friction force are stronger than the force of gravity. A water/ethylacetate/water interface was formed in a 70-µm-wide and 30-µm-deep channel. The interface was observed to be quite stable and to be maintained for a distance of more than 18 cm. As an example of a multilayer flow application, we demonstrated the liquid/liquid extraction of Co-dimethylaminophenol complex in a microchannel. The solvent-extraction process of the complex into m-xylene in the multilayer flow was found to reach equilibrium in 4 s, while it took 60 s in a simple two-phase extraction.

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A pulsed laser-induced stationary wave capillary vibration detection method was applied to the sensitive detection of capillary gel electrophoresis, and the direct detection of non-labeled nucleic acids, such as DNA sequencing products, was demonstrated. An excimer laser operating at 248 nm was used as a CVL excitation source, and polynucleotides were sensitively detected without derivatization. From an investigation on the endurance of several matrixes to pulsed laser irradiation, a polyacrylamide without a cross-linker (0%C) was found to have adequate endurance, and it exhibited no serious damage during an analysis. A cytosine-terminated sequence reaction product was detected with a sensitivity close to that of laser-induced fluorometry (LIF). These results suggest the feasibility of the highly sensitive detection of ultramicro amounts of biological materials without a pre- or post-column derivatization, which has usually been required in sensitive detection procedures, such as LIF. Furthermore, the feasibility of a novel DNA sequencing method is also suggested.

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The electrophoretic detection of two kinds of p-quinone anion radicals arising from the electrolysis of benzoquinone and chloranil was achieved by employing an acetonitrile medium. Sufficient dehydration of a running solution was necessary for the detection of the benzoquinone anion radical. Oxygen in the running solution also caused a serious decrease in the amount of the benzoquinone anion radical during electrophoresis. The addition of methanol as a hydrogen-donor decreased the electrophoretic mobility of the benzoquinone anion radical significantly, while that of the chloranil anion radical was little changed. This result is interpreted in terms of hydrogen-bonding interaction between the p-quinone anion radicals and methanol, reflecting the magnitude of their proton-accepting ability (benzoquinone anion radical > chloranil anion radical).

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The simultaneous determination of betaines, the key compounds for osmotic regulation in plants, was established by capillary electrophoresis (CE). After four betaines, glycine betaine (GB), β-alanine betaine (AB), proline betaine (PB), and 2-hydroxyproline betaine (HPB), were esterified with p-bromophenacyl bromide, the esters were electrophoresed in 100 mM sodium phosphate at pH 3.0. A low-pH condition in CE and esterification gave an advantage of resolving each of the ester peaks as well as those of the unreacted reagent and other components. Furthermore, the addition of 4% polyethylene glycol (PEG) gave a better resolution of 4 peaks, resulting in the separation of the overlapped peaks of PB and AB. It was found from the standard addition method being applied to barley leaves that the GB content in plants could be evaluated by using a calibration curve of the GB standard solution. The extraction of GB from plant leaves was also improved by adopting water as the extraction solvent instead of a mixture of organic solvents. The present method was suitably applied to actual plant specimens collected from a saline area of China.

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A highly sensitive and selective fluorometric determination method for ornithine and lysine has been developed. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase liquid chromatography (LC). The analytes, containing two amino moieties in a molecule, were converted to the corresponding dipyrene-labeled derivatives by reaction with PSE. The derivatives afforded intramolecular excimer fluorescence (450 - 550 nm) which can clearly be discriminated from the normal fluorescence (370 - 420 nm) emitted from PSE and monopyrene-labeled derivatives of monoamines. The structures of the derivatives and the emission of excimer fluorescence were confirmed by LC with mass spectrometry and with three-dimensional fluorescence detection system, respectively. The PSE derivatives of ornithine and lysine could be separated by reversed-phase LC on ODS column with isocratic elution. The detection limits (signal-to-noise ratio = 3) for ornithine and lysine were 3.5 and 3.7 fmol, respectively, for a 20-µl injection. Furthermore, this method had enough selectivity and sensitivity for the determination of ornithine and lysine in normal human urine.

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A molecular-dynamics simulation method has been applied to investigate the influence of the mobile-phase composition on the retention of solutes in HPLC. The distribution profiles of the distance between two atoms in ODS ligands were constructed to characterize the conformation of ODS ligand molecules. The distinct difference of ODS conformation is observed by comparing molecular models consisting of solvent molecules at each solvent composition. The distribution profiles of the distance between the mobile-phase solvent molecules and ODS ligand molecules were also constructed to characterize the distribution of the solvent molecules at each composition. In all distribution profiles, the difference in the distribution due to a change in the solvent compositions was very clearly found, and the facts seem to be very reasonable. The distribution profiles of the distance between the solute, n-propylbenzene, and the terminal carbon atom in the ODS ligand, and between the solute and the silicon atom in the ODS ligand have been also constructed to see the distribution of the solutes in the separation system. The calculated solute distribution in the ODS-methanol/water system is very consistent with the actual chromatographic retention behaviors.

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An azobenzene-modified γ-cyclodextrin stationary phase (Azγ-CDSP) was prepared and its photo- and temperature-responses for the retention of perylene and pentacene were investigated using a mixture of methanol and water as the mobile phase in micro-HPLC. The retention of perylene slightly increased, whereas that of pentacene significantly decreased by UV light irradiation to Azγ-CDSP. These retentions recovered upon irradiation with visible light. Both retentions decreased upon an increase in the column temperature. It was presumed that the trans-azobenzene moiety acts as a preventive cap for perylene and a spacing for pentacene in filling the CD cavity. An azobenzene-modified stationary phase changed its retention behavior with the column temperature and the light irradiation. An improvement in the micro-HPLC system and the optimization of the molecular structure of the photo-responsive stationary phase would provide selective retention control by the irradiation of light in micro-separation systems.

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In situ solid-state NMR (SSNMR) methodologies have been used to investigate the surface properties and photooxidative reactivities of a number of metal oxide photocatalysts. Adsorption of ethanol on single monolayers of TiO₂, SnO₂, V₂O₅, and WO₃ supported on porous Vycor glass results in the formation of hydrogen-bonded ethanol species and metal-bound ethoxide species. The chemical shift of the metal-bound ethoxide species varies with the metal oxide catalyst while the chemical shift of the hydrogen-bonded species is independent of the metal oxide. X-ray powder diffraction, UV-VIS spectroscopy, and SSNMR investigations of ethanol adsorption show that increasing the number of monolayers of TiO₂ on the Vycor surface changes the morphology of the catalyst from amorphous at a single monolayer coverage to anatase at a four monolayer coverage. The rate of photocatalytic oxidation of ethanol, acetone, and 2-propanol also increases with increasing TiO₂ monolayer coverage.

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The capabilities of using a confocal fluorescence microscope (CFM) for the interface-selective observation of dye molecules at an oil-water interface were evaluated. A simple expression for a criterion of interface selectivity was derived. The adsorptive behavior of dioctadecyl-rhodamine B at a cyclohexane-water interface was investigated experimentally to demonstrate the interface-selective observation with a CFM. The minimum number of detectable molecules in the probe area at the interface was estimated to be 82 for the present system. Preliminary experimental results of co-adsorption systems composed of adsorbed dye molecules and charged surfactants are also presented.

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Sum frequency generation (SFG) spectroscopy is a valuable tool for studying interfaces such that the boundary between two adjoining phases can be probed with minimal interference from the adjacent bulk material. More recently, broad-bandwidth sum frequency generation (BBSFG) techniques are being explored. This technique using IR broad-bandwidth fs pulses overlapped with narrow-bandwidth ps pulses to obtain BBSFG spectra is described. In the BBSFG system design presented here, the fs pulse and the ps pulse that are generated in separate regenerative amplifiers are overlapped temporally. This temporal overlap process is discussed. In addition, images of the sum frequency response demonstrate its viability. The new approach in experimental design described here for this emerging technology, BBSFG, has application for studying time-dependent processes at interfaces that inherently produce low SFG signal levels such as air-aqueous interfaces.

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A dual-beam transient thermal-lens technique was employed for the determination of absolute fluorescence quantum-yield measurements of Rhodamine B laser dye in different solvents. We investigated the effect of excitation on the absolute fluorescence quantum yield of Rhodamine B. 514 nm radiation from an argon ion laser was used as a cw excitation source and 532 nm pulses from a Q-switched Nd:YAG laser were used as a pulsed excitation source. The fluorescence quantum-yield values were found to be strongly influenced by environmental effects as well as the transient nature of the excitation beam. Our results also indicate that parameters, like the concentration of the dye solution, aggregate formation and excited state absorption, affect the absolute values of the fluorescence yield significantly.

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Electron-probe x-ray micro analysis (EPMA) and particle-induced x-ray emission analysis (PIXE) were performed under grazing-exit conditions. To control the exit angle (take-off angle), a new sample holder having a stepping motor was developed for grazing-exit EPMA. A carefully polished surface of a stainless-steel sample was measured. The surface of stainless steel is normally covered with a thin native oxide layer. The intensity ratio of Cr K_α to Fe K_α increases significantly at the grazing angle, becoming about 5-times larger at 0.2° than at 40°. This result indicates that grazing-exit EPMA is useful for surface analysis. In addition, a new PIXE equipment was developed for grazing-exit x-ray measurements. The sample is fixed and the x-ray detector is moved by applying a linear stage. Preliminary experimental results of grazing-exit PIXE are also shown.

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Ni L- and Ti L-edge as well as Ti K-edge X-ray absorption experiments for TiO₂ thin films and Ni-doped TiO₂ thin films coated on glass plates were performed using synchrotron radiation to investigate the structures around Ni and Ti ions in the films. The obtained spectra were compared with the results of theoretical calculations. It has consequently been found that the spectral features were affected by a change in the oxidizing form of Ni ions due to hydrogen reduction, by the charge variation and/or slight orbital splitting of Ti ions, and by the magnitude of the interaction between the center Ti ion and neighboring Ti ions.

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Fluorescently labeled oligonucleotide probes have been widely used in biotechnology, and fluorescence quenching by the interaction between the dyes and a nucleobase has been pointed out. This quenching causes big problem in analytical methods, but is useful in some other cases. Therefore, it is necessary to estimate the fluorescence quenching intensity under various conditions. We focused on the redox properties of some commercially available fluorescent dyes, and investigated dye-nucleotide interactions between a free dye and a nucleotide in aqueous solution by electrochemical and spectroscopic techniques. Our results suggested that the quenching was accompanied by photoinduced electron transfer between a thermodynamically quenchable excited dye and a specific base. Several kinds of fluorescent dyes labeled to the 5′-end of oligonucleotide C₁₀T₆ were prepared, and their quenching ratios compared upon hybridization with the complementary oligonucleotide A₆G₁₀. The quenching was completely reversible and their efficiencies depended on the attached fluorophore types. The fluorescence of 5-FAM, BODIPY FL or TAMRA-modified probe was strongly quenched by hybridization.

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The reaction of Ce(IV) with pyrogallol caused chemiluminescence, which was enhanced by dissolved oxygen. Dissolved oxygen in water was able to help in the determination by enhancing the chemiluminescence intensity. The limit of detection calculated from 3σ was 43 µmol/dm³, and the relative standard deviation was 1.2% at 613 µmol/dm³ (n = 5). The results obtained for natural and tap water samples were compared with those provided by conventional methods; the agreement between them corroborated the usefulness of the proposed method. The chemiluminescence mechanism was studied by examining the effect of interference with Cl^- and measuring the chemiluminescence spectrum. The chemiluminescence emitter, however, could not be identified.

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Urinary 1-hydroxypyrene is a biomarker in the measurement of human exposure to polycyclic aromatic hydrocarbons. A rapid and simple derivative isopotential synchronous fluorescence method was developed for the direct determination of 1-hydroxypyrene in urine. A length of iso-intensity route was scanned on the three-dimensional fluorescence spectrum of urine and this result was combined with that from derivative technique. Thus the strong background signals of urine were removed and the 1-hydroxypyrene can be determined directly in urine without tedious pre-separation. The derivative isopotential synchronous fluorescence spectrum was directly obtained from a single scan on a spectrofluorometer, which further simplified isopotential synchronous fluorescence technique. The recoveries of 93% to 115% were obtained for 1-hydroxypyrene added to urine.

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A combined method of electrothermal vaporization and low-pressure helium ICP-MS was developed for the determination of traces of arsenic in chloride solutions, because serious spectral interference occurred in conventional argon ICP-MS. On a tungsten filament was placed 5 µl of a sample and heated electrothermally to remove the solvent. The resulting residue on the filament was covered with a vaporization chamber, and after reducing the pressure to ca. 5 Torr, it was rapidly heated by discharging a high-capacity condenser (0.22 F). The vapor of arsenic was transferred to a helium plasma with a stream of carrier gas for the determination. The background was not observed at m/z of 75, which allowed the determination of arsenic at ng/ml levels in chloride solutions. Sodium chloride significantly enhanced the intensity of the arsenic signal, whereas other chlorides, including KCl, MgCl₂, CaCl₂ and NH₄Cl, suppressed the analytical signals. This enhancement due to sodium ions seems to be a peculiar phenomenon to the helium ICP. The proposed method can be applied to direct determinations of as low as 0.9 ng/ml of arsenic in seawater.

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In this work, tungsten coil (W-Coil) devices are used as atomizers for electrothermal atomization atomic absorption spectrometry (ETAAS), electrothermal atomization laser excited atomic fluorescence spectrometry (ETA-LEAFS), and electrothermal vaporization inductively coupled plasma atomic emission spectrometry (ETV-ICP-AES). For most cases in ETAAS and ETA-LEAFS, limits of detection (LODs) using the W-Coil are within a factor of ten of those observed with commercial graphite furnace systems. LOD for Cd by W-Coil AAS is 10 pg, while LODs for As, Se, Cr, Sb and Pb by W-Coil LEAFS are 950, 320, 1400, 330, and 160 fg, respectively. The compact W-Coil device makes it an ideal atomizer for portable atomic spectrometry instrumentation, especially when coupled with a miniature charge coupled device spectrometer. Alternatively, the atomizer can be used as an inexpensive, modular add-on to an existing commercial ICP-AES system; and the thermal separation of Pb with interference elements Al, Mn, and Fe is demonstrated.

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An optical-fiber chemical sensor (optode) is described for the simultaneous determination of Zn²⁺ and Cd²⁺ mixtures using 2-(5-bromo-2-pyridylazo)-5-(diethylamino)phenol immobilized onto XAD-4 (Br-PADAP/XAD-4). A simultaneous analysis was achieved by using a single-wavelength kinetic approach coupled with a regression model. Here, the regression model was derived for the slope response of Br-PADAP/XAD-4 to binary mixtures of the metal ions, Zn²⁺ and Cd²⁺, at pH 8. The model was used then to predict the concentration of one metal ion while the other was kept constant. This was found to be reasonably successful for Zn²⁺ and Cd²⁺, with deviations of 3% and 10%, respectively. The sensor was also found to provide excellent sensitivity for both metal ions with excellent LODs (< 0.1 ppm) and very short analysis times (< 30 s).

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The ligand substitution reaction of [Co(an)₆]²⁺ (an = acetonitrile) with 1,1,3,3-tetramethylurea (TMU) in the noncoordinating solvent, nitromethane, was spectrophotometrically investigated by titration. The observed spectral changes were analyzed using a model with the four steps of ligand substitution. The component complexes involved in the substitution were found to be 6-coordinate [Co(an)₆]²⁺ and [Co(an)₅(tmu)]²⁺, 5-coordinate [Co(an)₃(tmu)₂]²⁺ and [Co(an)₂(tmu)₃]²⁺, and 4-coordinate [Co(tmu)₄]²⁺. The logarithmic values of the stepwise equilibrium constant are 2.17 ± 0.26, 1.06 ± 0.15, 1.19 ± 0.06, and -0.4 ± 0.4 at 25°C. The decrease in the coordination number of the Co(II) ion from 6 to 5 during the formation of [Co(an)₃(tmu)₂]²⁺ and from 5 to 4 during the formation of [Co(tmu)₄]²⁺ is ascribed to the steric repulsion between the coordinating bulky TMU molecules.

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A novel mutual selectivity of lanthanoids in the N,N′-bis(5-nitrosalicylidene)ethylenediamine (H₂Nsalen)-KCl and N,N′-bis(5-nitrosalicylidene)-o-phenylenediamine (H₂Nsaloph)-KCl extraction systems was evaluated by the X-ray analysis of similar model complexes for extracted species. Cerium(IV) complexes with N,N′-bis(5-chlorosalicylidene)-ethylenediamine (H₂Clsalen), N,N′-bis(salicylidene)-o-phenylenediamine (H₂saloph) and N,N′-bis(3,5-dibromosalicylidene)-o-phenylenediamine (H₂Br₂saloph) were selected as the models. The result of the X-ray analysis suggested that [Ln(III)(Nsalen)₂]^- and [Ln(III)(Nsaloph)₂]^- are meridional type (two ligands are oriented perpendicular to each other) and sandwich type (two ligands are oriented parallel to each other), respectively. It was suggested that the selectivity of the meridional structure is superior to that of the sandwich structure in these extraction systems.

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Diazacrown ethers having two carbodithioate groups were synthesized, and their spectrophotometric properties were studied. Diammonium 1,4,10,13-tetraoxa-7,16-diazacycrooctadecane-N,N′-bis(carbodithioate) (DA18CC) and ammonium 1,4,10-trioxa-7,13-diazacycropentadecane-N,N′-bis(carbodithioate) (DA15CC) reacted with most heavy-metal ions through their two carbodithioate groups, and also reacted with alkali-metal ions or alkaline earth-metal ions through their diazacrown rings. Although the UV-VIS absorption spectra of DA18CC and DA15CC were hardly influenced by the addition of alkali-metal ions or alkaline earth-metal ions, the spectra of some heavy-metal chelates of DA18CC and DA15CC were influenced. The composition ratio (metal:ligand) of Cu(II) and Pb(II) chelates of DA18CC and DA15CC was altered from 1:1 to 1:2 by the addition of alkali-metal ions or alkaline earth-metal ions. It is suggested that the change in the absorption spectra and the composition ratio of the chelates can be attributed to an alteration of the equilibrium among species of the heavy-metal chelates of DA18CC and DA15CC due to the coordination of alkali-metal ions or alkaline earth-metal ions into the diazacrown ring.

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