An L-DNA, the mirror-image isomer of natural DNA, has extraordinary nuclease resistance, and thus the molecules should be promising reagents for many applications, such as antisense technology. However, little is known about the structural and thermodynamic properties of DNAs with this modified nucleotide. In this study, we prepared the L-nucleotide (L-dA) and introduced it into oligodeoxyribonucleotides to assess the ability of the L-nucleotide as a functional molecule for many applications based on the DNA hybridization. Two decamers with an L-dA at the center were synthesized and duplexes with the complementary DNA strand were applied to structural and thermodynamic analyses. The structural study by CD spectra showed that the structures of both modified “L/D-D” duplexes were the typical B-form. This result suggests that the global structure of DNA was not collapsed by the introduction of an L-DNA. Thermodynamic parameters (ΔH°, ΔS°, and ΔG°37) of the duplex formation, determined by UV melting experiments, indicated that the both duplexes were destabilized at about 2.5 to 3.0 kcal mol-1 by the introduced L-dA, mainly due to an unfavorable enthalpic effect. In conjunction with information by other researchers, these results suggest that the L-DNA affect on the duplex structure and the stability vary locally; thus, the thermodynamic stability of modified L/D-D duplexes should be predictable by the nearest-neighbor thermodynamic parameters.
In the dye-binding method based on protein error of a pH indicator, the color development has been reported to be markedly affected by the buffer concentration of the color reagent. In this study, the author analyzed this phenomenon by a theoretical calculation based on the chemical equilibrium of protein error. The calculation was performed on the assumption that both the dissociated dye anion and the anion contained in the buffer solution react with protein, forming a dye-protein complex and an anion-protein complex, respectively. The calculated results were compared with those obtained by the experiments using bromophenol blue, bromocresol green and bromocresol purple that are employed widely for determining the human serum albumin concentration clinically. The calculated results of this method are summarized as follows: (1) the color development decreases with the increase in the concentration of the anion contained in the buffer solution; (2) the calibration curve is more linear in the higher concentration of the anion than in the lower one. These calculated results agreed well with the experimental ones. From these results, it was concluded that the change in the color development by the buffer concentration of the color reagent is due to the change in the concentration of the buffer anion.
Ambient particulate matter and gas in Kyoto were investigated by gravimetric analysis, X-ray fluorescence spectrometry, and ion chromatography in order to clarify their behavior and origin. The size distribution and characteristics of the chemical components in ambient particulates collected on PTFE membrane filters using an Andersen air sampler were examined from August 2001 to April 2004. A four-stage filter pack method was used to sample the atmosphere for the determination of gas (SO2, HNO3, HCl, NH3) and particulate matter (SO42-, NO3-, Cl-, Na+, K+, Ca2+, Mg2+, NH4+) concentrations from October 2002 to April 2004. The concentration of SPM mass was in the range of 6.7 - 80.2 µg/m3. The size distributions of SPM mass were bimodal, peaking at around 0.65 - 1.1 and 3.3 - 4.7 µm, and 40 - 85% of SPM mass was fine particles (< 2.1 µm). Na, Mg, Al, Si, Ca, Cl, and Fe were mainly present in coarse particles (2.1 to 11.0 µm), while S was present in fine particles. The concentrations of Al, Si, Ca, Mg, and Fe in fine particles increased from March to April in 2002, and those in coarse particulates increased in November 2002 and from March to April in 2004. This may be the effect of the continental yellow sand “Kosa.” The differences in the size distributions of Al, Si, Ca, Mg, and Fe in particles may depend on differences in their place of occurrence and course of transport from China to Japan. The concentration of HCl gas was higher than that of particulate chloride ion in summer. Nitric acid gas existed at higher concentrations in summer, but fine particulate nitrate ion was observed in winter. The gaseous-to-fine aerosol nitrate fraction became higher at warmer temperatures. Coarse sulfate was below 10%, and SO2 gas and fine particulate sulfate were above 90%.
A new modified method was applied to the determination of chloride anion in brackish water by using a common copper(II) sulfate solution and a spectrophotometer. The result of the quantitative analysis demonstrated that chloride concentrations were 1.61% w/v and 1.74% w/v at ebb tide and flood tide, respectively, of the Muya River. On the other hand, chloride concentrations were 1.17% w/v at ebb tide and 1.65% w/v at flood tide of the Yoshino River. The new method proved that chloride concentration corresponded well with results from Mohr and conductive methods. Therefore, the newly developed method can be exploited as common means to measure chloride concentration in aqueous solutions. Moreover, it is also valuable for application to other types of environmental samples.
A method for calculating the solution resistance of an integrated dual ultramicroelectrode was introduced, and then tested by experiments using dummy cells. Then, with the reduction of anthracene on a gold ultramicroelectrode in a 0.1 M tetraethylammonium tetrafluoroborate/acetonitrile solution as the test system, it could be found that the solution resistance of this integrated dual ultramicroelectrode was much lower than that of the two-electrode system conventionally used. Thus, the compensation level could be improved up to nearly 100% at a scan rate of 1.34 MV s-1, while the latter could not. This showed that the integrated dual ultramicroelectrode was more suitable for acting as the electrode system in ultrafast cyclic voltammetry, especially in a high-resistance solution.
Novel miniaturized polyurethane (PU) membrane sensors in an all-solid state graphite support were developed, electrochemically evaluated and used for the assay of thiopental drug. The thiopental (T) sensors are based on the formation of ion-association complexes of thiopental with copper(II) and cobalt(II)-bathophenanthroline (bphen) counter anions as electroactive materials dispersed in a polyurethane matrix. The sensors show a linear response for thiopental over the range of 1 × 10-1 - 5 × 10-5 M thiopental at 25°C over the pH range 6 - 11 with anionic slopes of -28.7 and -28.3 mV decade-1 with Cu- and Co-bphen thiopental membrane sensors, respectively. These sensors exhibit a fast response time (25 - 45 s), a low detection limit (5 × 10-6 M), a long lifetime (7 weeks) and good stability. The selectivity coefficients for thiopental sensors relative to the number of interfering anions, were investigated. These sensors were used for the direct potentiometry of thiopental in a pharmaceutical formulation and human serum. Results with mean accuracy of 99.8 ± 0.5% of nominal were obtained, which compare well with data obtained using spectrophotometric (UV-Vis) and British Pharmacopoeia (BP) methods.
A chemiluminescence (CL) method using flow injection (FI) was developed for the determination of uric acid based on the enhancement chemiluminescence intensity of luminol-hexacyanoferrate(III)-hexacyanoferrate(II) in the presence of cetyltrimethylammonium bromide and the uric acid species. The linear range was 7.0 × 10-10 - 9.0 × 10-7 M with a detection limit (3σ) of 2.58 × 10-10 M, which was about two orders of magnitude lower than those reported. The proposed method was used for the determination of uric acid in real samples.
An enantiomeric separation of dopamine-derived neurotoxins by capillary electrophoresis has been developed. Tetrahydroisoquinoline (TIQ), dopamine (DA), (R/S)-1-benzyl-TIQ (BTIQ), (R/S)-6,7-dihydroxy-1-methyl-TIQ (salsolinol, Sal), and (R/S)-6,7-dihydroxy-1, 2-dimethyl-TIQ (N-methyl-salsolinol, NMSal) were studied as model compounds. The CE running buffer (50 mM phosphate buffer at pH 3.0) contained 1.5 M urea and 12 mM β-CD as a chiral selector. During separation, the (R)-enantiomers formed more stable inclusion complexes with β-CD, and thus had a longer migration time than their optical antipodes. It was noticed that the recovery rates of these TIQ derivatives were very poor (< 15%) during protein precipitation, a procedure widely used for cleaning up biological samples. The recovery was significantly improved by pre-mixing the sample with a surfactant (e.g., sodium hexanesulfonate or Triton X-100) to reduce the co-precipitation. The present method in combination with electrospray ionization tandem mass spectrometry (ESI-MS/MS) was applied to study samples obtained from in vitro incubation of two catecholamines, dopamine and epinine, with aldehydes forming neurotoxins including (S)- and (R)-NMSal enantiomers. The later is known to induce Parkinsonism in rats.
A simple, sensitive and rapid ion-exclusion chromatographic method for the simultaneous separation of carbonic acid and short chain organic acids (formic acid, acetic acid, and propionic acid) has been developed. A weakly acidic cation exchange resin column TSKgel OApak-A was used for the separation; pure water, owing to its lower background conductance, was employed as eluent in order to obtain high detection sensitivity. A good separation of these weak acids was achieved in 16 min. The linear range of the peak area calibration curve for carbonic acid was from 3.0 mg/L to 500 mg/L. The conductivity detection limit calculated at S/N = 3 was 0.084 mg/L for carbonic acid. The method developed in this work was successfully applied to the determination of carbonic acid in several environmental water samples without any pretreatment.
In the present work, an amylose tris(3,5-dimethylphenylcarbamate) (ADMPC) chiral stationary phase (CSP) was prepared by coating ADMPC on small-particle silica gel. This ADMPC-CSP was for the first time successfully applied to separate a series of novel chiral metal tetrahedrane-type clusters. Furthermore, the influence of a mobile-phase modifier (various alcohols added in the mobile phase), including its concentration and structure, and the structures of the clusters on the chiral separation and retention was investigated. The results suggest that not only the structure and concentration of alcohol in the mobile phase, but also the subtle structural differences in racemate can have a pronounced effect on the enantiomeric separation and retention.
A new calixresorcinarene bearing eight hydroxamic acid groups (C4RAHA) has been synthesized and characterized by FT-IR, 1H-NMR and elemental analysis. Its analytical properties were investigated, and showed high affinity and selectivity toward thorium(IV) in the presence of large quantities of associated metal ions. Thorium(IV) was extracted from an ethyl acetate solution of C4RAHA at pH 4.5. The λmax and molar absorptivity (ε) for thorium(IV) were 341 nm and 7120 l mol-1 cm-1. The complexation of thorium(IV) with C4RAHA has a 4:1 metal:ligand stoichiometry, which is relatively rare. The extracts were directly aspirated in the plasma for ICP-AES measurements for thorium(IV) in the presence of a diversified matrix. The system obeyed Beer’s law over the range 0.1 - 6.5 µg ml-1 of thorium(IV) with a Sandell sensitivity of 0.0325 µg cm-2. The preconcentration factor and overall stability constant evaluated at 25°C for thorium(IV) were 133 and 15.86, respectively. The complexation was characterized by a favorable enthalpy change. A liquid-membrane transport study of thorium(IV) was carried out from the source to the receiving phase under controlled conditions, and a mechanism of transport proposed. To check the validity of the proposed method, thorium(IV) was determined in monazite sand, rare earth sand and water samples.
A method for the determination of total selenium in wheat and wheat flour using graphite furnace atomic absorption spectrometry (GFAAS) with palladium/ascorbic acid as a chemical modifier was studied. The effects of nickel nitrate, palladium/ascorbic acid, and palladium/magnesium nitrate as chemical modifiers on the sensitivity in the determination of selenite, selenate and selenomethionine by GFAAS were compared. The palladium/ascorbic acid modifier was used for the determination of total selenium in wheat and wheat flour, because the oxidation states of the selenium ion are not important in the determination. The detection limit was estimated to be 1 µg L-1 (calculated as 3 σ of the blank); the calibration curve was linear for the concentration range 5 - 50 µg L-1 and the recovery range was 96.66 - 101.80%. The optimal ashing and atomizing temperatures were 1300°C and 2250°C, respectively. The proposed method was successfully applied to the determination of total selenium in wheat and wheat flour.
The influence of analyte mass concentration on determination of detection limits in X-ray fluorescence spectrometry has been investigated experimentally. Both the total reflection X-ray fluorescence (TXRF) and the conventional energy-dispersive X-ray fluorescence techniques have been used to derive the dependence of analyte mass concentration on the values of detection limits. Results obtained indicate that values of detection limits are optimum, or in other words, they are closer to the true detection limit of the technique, when analyte concentrations are in the range of 10 times of the detection limit.
A simple spectrophotometric method is described for resolving binary mixtures of some food dyes: Amaranth, Brilliant Blue, Sunset Yellow and Tartrazine, using the first-derivative spectra with measurements at zero-crossing wavelengths. Analytical curves are linear up to 20 mg L-1. Standard deviations of 1.30, 2.22, 1.93 and 0.81% were obtained for synthetic binary mixtures of 2 mg L-1 of Amaranth, Brilliant Blue, Sunset Yellow and Tartrazine, respectively. Before the spectrophotometric measurements, the dyes were sorbed onto polyurethane foam and recovered in sodium dodecyl benzene sulfonate solution. Therefore, matrix complexity was eliminated and simple spectra were obtained. The method was very satisfactorily used for determining the colorants in synthetic mixtures, with recoveries in the 96 - 101% range. Detection limit values were dependent on the colorant combination investigated. Commercial products containing binary combinations of these dyes in different ratios (from 1:1 to 1:8) were analyzed. The results were compared with those obtained by HPLC; very similar values were found by the two methods.
A novel flow injection method has been developed for the indirect determination of uranium by the on-line reduction and subsequent fluorimetric detection of cerium(III). A sample solution containing uranium(VI), prepared as a sulfuric acid solution, was injected into a sulfuric acid carrier solution and passed through a column packed with metal bismuth to reduce uranium(VI) to uranium(IV). The sample solution was merged with a cerium(IV) solution to oxidize uranium(IV) to uranium(VI) and the cerium(III) generated was then monitored fluorimetricaly. The present method is free from interference from zirconium, lanthanides, and thorium, and has been successfully applied to the determination of uranium in monazite coupled with an anion-exchange separation in a sulfuric acid medium to eliminate iron(III). The sample throughput was 25 per hour and the lowest detectable concentration was 0.0042 mg l-1.
Near-infrared (NIR) spectrometry will present a more promising tool for quantitative measurement if the robustness and predictive ability of the partial least square (PLS) model are improved. In order to achieve the purpose, we present a new algorithm for simultaneous wavelength selection and outlier detection; at the same time, the problems of background and noise in multivariate calibration are also solved. The strategy is a combination of continuous wavelet transform (CWT) and modified iterative predictors and objects weighting PLS (mIPOW-PLS). CWT is performed as a pretreatment tool for eliminating background and noise synchronously; then, mIPOW-PLS is proposed to remove both the useless wavelengths and the multiple outliers in CWT domain. After pretreatment with CWT-mIPOW-PLS, a PLS model is built finally for prediction. The results indicate that the combination of CWT and mIPOW-PLS produces robust and parsimonious regression models with very few wavelengths.
This paper theoretically derives a general rule that while the slope of the semi-logarithmic plot (Y vs. log X) of a calibration curve varies depending on analyte concentration, X, the slope takes a specific value at the detection limit (LD). This rule holds good irrespective of the shape of the calibration curve (linear or non-linear) and in this paper, is applied to competitive ELISA (enzyme linked immunosorbent assay). The following relationship is deduced: slope of log-dose B/B0 at LD = [relative standard deviation (RSD) of blank responses] ÷ 0.13. The LD obtained from the above-mentioned slope corresponds to the dose at which the RSD of dose estimates is 0.3 (= 30%). A commercial kit for 17α-hydroxyprogesterone is taken as an example.
The proximal region of the angiotensin II receptor (AT1A) carboxyl-terminus (known as helix VIII) is important for receptor function. In this study, we used surface plasmon resonance (SPR) to examine the interaction of helix VIII-derived peptides with three model lipid membranes. The membrane-binding properties of these synthetic peptides, as well as a series of peptide analogues with modified amino acid sequences, could be explained by both amino acid sequence and kinetic binding data by SPR. The helix VIII peptides showed a higher affinity for lipid membranes that contained negatively charged phospholipid, rather than zwitterionic phospholipid. The findings of an SPR study may be useful for estimating the cooperative binding of intracellular receptor domains with G proteins and the components of the lipid bilayer.
The tetrabutyl ester derived from 9,16,25,32-tetrahydroxy[18.104.22.168]metacyclophane was an excellent ionophore for constructing a K+-selective membrane electrode. This ionophore exhibited a much higher selectivity toward K+ than the structurally similar potassium ionophore IV commercially available from Fluka. In particular, the interference from organic ammonium ions decreased remarkably. Potassium ionophore IV possessed oxygen atoms in the ring structure, while the present ionophore changed the oxygen atoms to carbon atoms. Thus, the removal of oxygen atoms in ring constituents of the metacyclophane acted to reduce the interaction with the NH3+ group of organic ammonium ions. The size of the cavity of the present ionophore was between those of calixarene and calixarene derivatives, which act as Na+ and Cs+ ionophores, respectively, demonstrating that the ability to recognize alkali metal cations was strongly cavity size-dependent. The present K+-selective electrode had less interference from Rb+ and Cs+ than an electrode constructed using valinomycin, but suffered greater interference from Na+.
A poly(vinyl chloride)-based membrane of 3,4:11,12-dibenzo-1,6,9,14-tetraazacyclohexadecane with sodium tetraphenyl borate (NaTPB) as an anion excluder and dibutylphthalate (DBP), dibutyl(butyl)phosphonate (DBBP), tris(2-ethylhexyl)phosphate (TEHP) and tributyl phosphate (TBP) as a plasticizing solvent mediator was prepared and investigated as a Cd2+-selective electrode. The best performance was observed with the membrane having a ligand-PVC-DBP-NaTPB composition of 2:25:60:1, which worked well over a wide concentration range (1.6 × 10-6 - 1.0 × 10-1 M) with a Nernstian slope of 29.5 mV per decade of activity between pH 2.0 - 6.0. This electrode showed a fast response time of 13 s and was used over a period of 4 months with good reproducibility (S = 0.4 mV). The selectivity coefficient for mono-, di- and trivalent cations indicated excellent selectivity for Cd2+ ions over a large number of cations. Anions such as NO3- and SO42-, did not interfere. The membrane sensor has been successfully used to determine Cd2+ in real samples.
The adsorption and desorption properties of trans-Resveratrol (Res) on the cellulose cotton were investigated under various conditions, such as the pH, alcohol percentage, temperature and equilibrium times. Moreover, the acidic-dissociation constants were determined to be pKa1 = 8.01, pKa2 = 9.86 and pKa3 = 10.5 by a curve-fitting method. Also, it was found that the adsorption depended on the temperature and salting effect. On the other hand, the desorptions from cellulose were examined using several kinds of water-miscible organic solvents (methanol, ethanol, acetone and THF).