The elution of particles from a coupled acoustic-gravity channel has been controlled by a phase-shift operation. Two ultrasound transducers pasted onto the top and bottom walls of a separation channel were driven at the same frequency, but with different phases. Changing the phases of the ultrasounds has allowed the formation of a node near the channel wall, and particles are retained in the channel. Two types of particles are separated under an appropriate condition.
An ultra-micro sample injector for gas chromatography (GC) was developed. An ink-jet microchip, originally used for industrial recorder, was modified at the edge near to an orifice, and fixed into the GC. In order to evaluate the characteristics of this injector, a sample injector and a thermal conductive detector (TCD) were connected directly, while water was used as the test sample. The volume of the droplet, the interval time and the back-pressure to the ink-jet microchip were investigated. Within the range of 1 - 5 nL volume injected sample, the TCD response according to the amount of the sample volume (the volume of one droplet from the ink-jet microchip was about 1 nL) was obtained. A good reproducibility of the peak area was obtained to be about 1.0% of the RSD value. In order to compare the injection method of the ink-jet chip with that using a micro-syringe, the method using the ink-jet chip could introduce 1/1000 of the amount of the sample and gave reproducible results.
The development of an NMR interface microchip and its applications to the real-time monitoring of chemical reactions are described. The microchip device was named “MICCS” (MIcro Channeled Cell for Synthesis monitoring), and the method using it was named “MICCS-NMR”. MICCS was inserted into a 5 mm Φ NMR sample tube. Thus standard solution NMR probes without any modifications can be used in MICCS-NMR measurements. A gap between MICCS and the sample tube was filled with a deuterated solvent for an NMR lock. The reaction temperature and reaction time in MICCS can be easily changed by adjusting the temperature of the NMR probe and changing the flow rates, respectively. The effectiveness of the MICCS-NMR was verified in the real-time monitoring of the Wittig reaction. Preliminary data on the direct detection of intermediates of the Grignard reaction is also reported. Besides real-time monitoring of chemical reactions, MICCS-NMR would be useful as a qualitative detection method for microchip-based synthesis.
A rapid DNA analysis has been developed based on a fluorescence intensity change of a molecular beacon in a PDMS microfluidic channel. Recently, we reported a new analytical method of DNA hybridization involving a PDMS microfluidic sensor using fluorescence energy transfer (FRET). However, there are some limitations in its application to real DNA samples because the target DNA must be labelled with a suitable fluorescent dye. To resolve this problem, we have developed a new DNA microfluidic sensor using a molecular beacon. By monitoring the change in the restored fluorescence intensity along the channel length, it is possible to rapidly detect any hybridization of the molecular beacon to the target DNA. In this case, the target DNA does not need to be labelled. Our experimental results demonstrate that this microfluidic sensor using a molecular beacon is a promising diagnostic tool for rapid DNA hybridization analysis.
Heteroaromatic compounds, such as 2,5-dimethylthiophene, 2,5-dimethylpyrrole, and 2,5-dimethylfuran, were found to act as reducing agents for the tris(2,2′-bipyridine)ruthenium(III), Ru(bpy)33+, chemiluminescent reaction. In order to characterize the chemiluminescent reaction of Ru(bpy)33+ with heteroaromatic compounds, we have investigated various mono-, di-, and tri-heteroaromatic compounds. The π-electron density and stability of aromatic rings influence the chemiluminescent efficiency of the reaction. Above all, 2,5-dimethylthiophene produced strong chemiluminescence under acidic conditions. In addition, we confirmed that the rate of the chemiluminescent reaction of Ru(bpy)33+ with 2,5-dimethylthiophene is very fast.
A novel reagent, acetoacetanilide (AAA), was introduced to the determination of formaldehyde based on Hantzsch reaction. A simple and highly sensitive fluorometric method was achieved by using AAA. The main advantages in the use of this reagent are: the reaction is carried out at room temperature without any heating system, the cyclization product based on Hantzsch reaction is soluble in water, and the product can be detected by spectrophotometry and fluorometry. The maximum absorption wavelength of the product occurs at 368 nm, and the maximum excitation and emission wavelengths are found at 370 and 470 nm, respectively. Several important experimental variables of the procedures were examined; particularly, the reaction temperature, reaction time, concentrations of reagents, and pH of the reagent solution were optimized for improving the detecting sensitivity. The calibration graph was linear in the range of 1 × 10-7 - 1 × 10-6 M or much higher concentrations. The limit of detection (LOD), based on three times of the standard deviation of the reagent blank, was 2.0 × 10-8 M. The proposed method was applied to the determination of formaldehyde in environmental water samples. Many foreign species commonly existing in water samples did not interfere with the determination of formaldehyde in the proposed method.
The interactions between 6-amino-4-aryl-3-methyl-1-phenyl-1H-pyrazolo[3,4-b]pyridine-5-carbonitrile and lysozyme (LYSO) were investigated by using tryptophane fluorescence quenching and 6-amino-4-(2-hydroxyphenyl)-3-methyl-1-phenyl-1H-pyrazolo[3,4-b]pyridine-5-carbonitrile (1) was studied in detail because of its high water solubility. At different temperatures, the quenching constants KSV, the binding constants K and the binding sites n of LYSO with 1 were determined and the thermodynamic parameters were calculated. The distance r between tryptophane residues and 1 was obtained according to the Forster mechanism of non-radiation energy transfer. Furthermore, synchronous fluorescence spectroscopy data indicated that the association between 1 and LYSO changed LYSO's conformation and that the hydrophobic interaction played a major role in 1-LYSO association. It was proved that the fluorescence quenching of LYSO by 1 was related to the formation of a 1-LYSO complex and to a non-radiation energy transfer.
A combined approach based on solid-phase optosensing and multicommutation principles has been applied to develop a method for the simultaneous analysis of two pharmaceuticals (naproxen and salicylic acid) in biological fluids. The multicommuted flow-through optosensor was based on direct native fluorescence measurements of both analgesics using a non-polar sorbent (C18 silica gel) as a solid sensing zone. The flow system was controlled by Java-written home-made software and designed using three-way solenoid valves for an independent automated manipulation of sample and carrier solutions. Using an optimized sampling time, the method was calibrated in the range of 1 - 25 and 5 - 200 ng mL-1. The obtained detection limits were 0.3 and 1.3 ng mL-1 for naproxen and salicylic acid, respectively, with RSD (%) values of better than 2% for both analytes. The proposed methodology was successfully applied to urine, serum and pharmaceutical preparations. Recovery percentages ranging from 96.1 to 104% were obtained for both analytes.
The interaction of tetrandrine with human serum albumin (HSA) was studied by measuring fluorescence quenching spectra, synchronous fluorescence spectra and ultra-violet spectra. The fluorescence quenching spectra of HSA in the presence of tetrandrine showed that tetrandrine quenched the fluorescence of HSA. The quenching constants of tetrandrine on HSA were determined using the Stern-Volmer equation. Static quenching and non-radiation energy transfer were the two main reasons leading to the fluorescence quenching of HSA by tetrandrine. According to the Förster theory of non-radiation energy transfer, the binding distances (r) and the binding constants (KA) were obtained. The thermodynamic parameters obtained in this study revealed that the interaction between tetrandrine and HSA was mainly driven by a hydrophobic force. The conformational changes of HSA were investigated by synchronous spectrum studies.
In this paper we describe our study on the characterization of cigarette samples according to their mineral content. Acid digestion assisted by microwaves was employed, and inductively coupled plasma optical emission spectrometry was the analytical technique used for the determination of Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Na, P and Sr in conventional, light, and flavorized cigarettes. Multivariate techniques, such as hierarchical clusters analysis (HCA) and principal-component analysis (PCA), were applied to discriminate among different types of cigarettes. Cluster analysis and principal-component analysis showed differences in cigarettes according to the type and mineral composition. The cigarette samples were divided within the 3 groups according to their mineral composition. Ca, Sr, Cu, K and Na were the most important elements for cigarette classification, and only these 5 variables were sufficient for the classification and discrimination of the evaluated types of cigarettes.
Kinetic assay of serum ethanol was investigated by predicting maximal product absorbance at 340 nm (Amk) through fitting to the yeast alcohol dehydrogenase reaction curve with the integrated rate equation, taking into account product inhibition in the presence of semicarbazide. Predicted Amk linearly responded to the preset constant of steady-state concentration of acetaldehyde (Cald). An exponential correlation function was established between desired Cald and putative Amk for authentic ethanol. For unknown samples, iterative fitting to reaction curve till preset constant Cald and resultant Amk satisfied this exponential correlation function yielded Amk with variation coefficient <4.3%. Variations in enzyme activity, data range and kinetic parameters showed negligible effects. The recovery was consistent to 100% with resistance to methanol and isopropanol. The upper limit of linear response for Amk was about 40 times of the lower limit. These results indicated that this kinetic method was reliable for serum ethanol assays with obvious advantages.
In the present work, five different spectrophotometric techniques for simultaneous determination of formulations containing atorvastatin calcium (ATOR) and fenofibrate (FENO) in various combinations are described. In ratio spectra derivative spectrophotometry, analytical signals were measured at wavelengths corresponding to either maximums or minimums for both drugs in first derivative spectra of ratio spectra obtained by using either spectrum as divisor. For the remaining four methods using chemometric techniques, namely, classical least squares (CLS), inverse least squares (ILS), principal component regression (PCR) and partial least squares (PLS), the calibrations were constructed by using the absorption data matrix corresponding to the concentration data matrix, with measurements in the range of 231 - 310 nm (Δλ = 1 nm) in their zero-order spectra. The linearity range was found to be 4 - 22 and 2 - 20 µg/ml for ATOR and FENO, respectively. The validity of the proposed methods was successfully assessed for analyses of both drugs in laboratory-prepared mixtures and in commercial tablet formulations.
An alternatively minimizing covariant matrix error (AMCME) algorithm, newly proposed by the present authors, was applied to the simultaneous fluorometric determination of pyridoxal, pyridoxamine and 4-pyridoxic acid without loss of sensitivity. The experimental results illustrate that the profiles of spectra and concentration can be accurately resolved using the AMCME algorithm with a high sensitivity and stable repeatability. That is to say, the closely overlapping problem of the spectra could be resolved owing to the characteristic features of the AMCME algorithm.
Water vapor diffusion coefficient (WVDC) and thermal diffusivity (α) were determined in gelatin-starch films through photothermal techniques. The effect of different variables in the elaboration of these films, such as starch and glycerol concentrations and pH, were evaluated through the response surface methodology. The results indicated that an increase in the glycerol concentration and pH favored the WVDC of the films. On the other hand, α was influenced principally by the starch content and pH of the film-forming solution. The minimum α value was 4.5 × 10-4 cm2/s, which is compared with α values reported for commercial synthetic polymers.
The determination of body fluid acidity using a wireless magnetoelastic pH-sensitive sensor is described. The sensor was fabricated by casting a layer of pH-sensitive polymer on a magnetoelastic ribbon. In response to an externally applied time-varying magnetic field, the magnetoelastic sensor mechanically vibrates at a characteristic frequency that is inversely dependent upon the mass of the pH polymer film, which varies as the film swells and shrinks in response to pH. As the magnetoelastic sensor is magnetostrictive, the mechanical vibrations of the sensor launch magnetic flux that can be detected remotely using a pickup coil. The sensor can be used for direct measurements of body fluid acidity without a pretreatment of the sample by using a filtration membrane. A reversible and linear response was obtained between pH 5.0 and 8.0 with a measurement resolution of pH 0.1 and a slope of 0.2 kHz pH-1. Since there are no physical connections between the sensor and the instrument, the sensor can be applied to in vivo and in situ monitoring of the physiological pH and its fluctuations.
A poly(vinyl chloride)-based membrane composed of dithio-tetraaza macrocyclic compound as a neutral carrier with sodium tetraphenylborate (NaTPB) as an anion excluder and nitrobenzene (NB) as plasticizer was prepared and investigated as a Th(IV)-selective electrode. The electrode exhibits a Nernstian slope of 14.2 ± 0.3 mV per decade over a wide concentration range (1.0 × 10-6 to 1.0 × 10-1 M) with a detection limit of 8.0 × 10-7 M between pH 3.5 and 9.5. The response time of the sensor is about 10 s and it can be used over a period of 5 months without any divergence in potential. The proposed membrane sensor revealed a good selectivity for Th(IV) over a wide variety of other metal ions and proved to be a better electrode in many respects than those reported in the literature. It was successfully applied as an electrode indicator as well as in the direct determination of thorium ions in standard and real samples.
A glassy carbon electrode modified with a coating of polypyrrole (Ppy) exhibited an attractive performance for the detection and determination of a non-steroidal and non-narcotic analgesic compound, ketorolac tromethamine (KT). Cyclic voltammetry, differential pulse and square wave voltammetry were used in a combined way to identify the electrochemical characteristics and to optimize the conditions for detection. For calibrating and estimating KT, square-wave voltammetry was mainly used. The drug shows a well-defined peak at -1.40 V vs. Ag/AgCl in the acetate buffer (pH 5.5). The existence of Ppy on the surface of the electrode gives higher electrochemical active sites at the electrode for the detection of KT and preconcentrate KT by adsorption. The square-wave stripping voltammetric response depends on the excitation signal and the accumulation time. The calibration curve is linear in the range 1 × 10-11 to 1 × 10-7 M with a detection limit of 1.0 × 10-12 M. Applicability to serum samples was also demonstrated. A detection limit of 1.0 ng ml for serum was observed. Square-wave voltammetry shows superior performance over UV spectroscopy and other techniques.
The adsorptive and electrochemical behaviors of clozapine (CLZ) were investigated on a glassy carbon electrode that was electrochemically treated by anodic oxidation at +1.8 V, following potential cycling in the potential range from -0.8 to 1.0 V vs. Ag/AgCl reference electrode. Based on the obtained electrochemical results, an electrochemical-chemical (EC) mechanism was proposed to explain the electrochemical oxidation of CLZ. The resulting electrochemically pretreated glassy carbon electrode (EPGCE) showed good activity to improve the electrochemical response of the drug. CLZ was accumulated in a phosphate buffer (pH 6) at a certain time, and then determined by differential pulse voltammetry. The anodic and cathodic peak currents showed a linear function in the concentration ranges of 0.1 - 1, 1 - 10 and 10 - 100 µM with various accumulation times. The proposed method was successfully used for the determination of CLZ in pharmaceutical preparations. The preconcentration medium-exchange approach was utilized for the selective determination of the drug in spiked urine samples with satisfactory results. The recovery levels of the method reached 96% (RSD, 1.8%) and 90% (RSD, 2.8%) for urine and plasma samples, respectively.
A liquid chromatographic (LC) determination of catecholamines and indoleamines is described. This is based on intramolecular excimer-forming fluorescence derivatization with 4-(1-pyrene)butanoyl chloride, followed by reversed-phase LC. The analytes, containing an amino moiety and phenolic hydroxyl moieties in a molecule, were converted to the corresponding polypyrene-labeled derivatives by one-step derivatization. They afforded intramolecular excimer fluorescence, which can clearly be discriminated from the normal fluorescence emitted from reagent blanks. The detection limits (S/N = 3) for catecholamines and indoleamines were femto-mole levels per 20-µL injection. Furthermore, this method was applied to a urine assay.
Affinity capillary electrophoresis has become an established approach for performing interaction studies. In affinity electrokinetic chromatography the retention factor, as in liquid chromatography, is useful for describing the migration behavior of the analytes, and is instrumental for assessing the affinity of an analyte for the pseudo-stationary phase. Erroneous use of the retention factor concept in affinity capillary electrophoretic studies has appeared in a number of recent papers. The errors and their origin are pointed out, and the correct use of retention factors in affinity electrokinetic chromatography and capillary electrophoresis is summarized.
A capillary electrophoresis (CE) method is described for the simultaneous determination of copper and iron after complexation with a readily biodegradable chelating agent, [S,S′]-ethylenediaminedisuccinic acid (EDDS), in wood pulp. CE separation was performed in a fused-silica capillary (50 µm i.d.; total length, 65 cm) with an electrolyte containing 25 mM borate buffer and 0.5 mM CTAB at pH 7.0 and an applied voltage of -25 kV. The samples were introduced by applying a 50 mbar pressure for 2 s, and detection of the complexes was monitored at 245 nm. The methodology performance of the methods was evaluated in terms of the linearity, limit of detection (LOD), limit of quantitation (LOQ) and reproducibility. The applicability of the method was demonstrated for the analysis of copper and iron in wood pulp.
A passive air sampler, using 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole, was evaluated for the determination of formaldehyde in indoor environments. Chromatography paper cleaned using a 3% hydrogen peroxide solution was experimentally determined as being the optimum absorption filter for the collection of formaldehyde (0.05 µg cm-2 formaldehyde). From a linear-regression analysis between the mass of formaldehyde time-collected on a passive air sampler and the formaldehyde concentration measured by an active sampler, the sampling rate of the passive air sampler was 1.52 L h-1. The sampling rate, determined for the passive air sampler in relation to the temperature (19 - 28°C) and the relative humidity (30 - 90%), were 1.56 ± 0.04 and 1.58 ± 0.07 L h-1, respectively. The relationship between the sampling rate and the air velocity was a linear-regression within the observed range. In the case of exposed samplers, the stability of the collected formaldehyde decreased with increasing storage time (decrease of ca. 25% after 22 days); but with the unexposed samplers the stability of the blank remained relatively unchanged for 7 days (decrease of ca. 37% after 22 days). The detection limits for the passive air sampler with an exposure time of 1 day and 7 days were 10.4 and 1.48 µg m-3, respectively.