A simple assay of DNA was developed based on the measurements of enhanced signals of Resonance Light Scattering (RLS) of cetyltrimethylammonium bromide (CTMAB) by DNA. The enhanced RLS signals, measured by simultaneously scanning the excitation and emission monochromators of a common spectrofluorometer with λex = λem, was optimized for the DNA assay with CTMAB. On the conditions of pH 2.21 and ionic strength 0.002, the enhanced RLS intensity at 470.0 nm, ΔI, was found to be proportional to the concentration of DNA in the range 0 - 2.5 μg/ml if 1.5 × 10-5 M CTMAB was used. Limits of determination for calf thymus DNA and fish sperm DNA were 4.9 ng/ml and 9.2 ng/ml, respectively. Synthetic samples were determined with the recovery ratio ranging from 93.2% to 105.1%, and the RSD is lower than 2.7%.
A highly sensitive and selective method for simultaneous determination of some hydroxyl group-containing endocrine disruptors, including bisphenol A (BPA), bisphenol B (BPB), bisphenol E (BPE), bisphenol F (BPF) and 4-nonylphenol (4-NP), was developed. The method consists of precolumn derivatization of the analytes, solid-phase extraction (SPE) and subsequent chromatographic analysis by high-performance liquid chromatography (HPLC) with fluorescence detection. 4,4′-Cyclohexylidenebisphenol (BPZ) was used as an internal standard. Derivatization was carried out using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) as a label. Parameters of the derivatization reaction (temperature, time, concentration of reagent, stability, etc.) and of the solid-phase extraction (recovery, solvent, etc.) were studied in detail. Detection limits of compounds studied in standard solutions ranged from 0.08 - 1.3 ppb (ng/ml). The proposed method was successfully applied to plastic samples; BPA was found in both polycarbonate and polyvinyl chloride plastics, while 4-NP was found in plastics made of polyvinyl chloride and another polymer.
A micro-flow enzyme system with a microdialysis probe is proposed for the amperometric detection of trace amounts of neurotransmitter L-glutamate released from rat brain cells. The L-glutamate oxidase (EC 22.214.171.124)/glutamate dehydrogenase (EC 126.96.36.199) coimmobilized reactor was used to enhance the sensitivity of L-glutamate as an on-line amplifier based on substrate recycling. A poly(1,2-diaminobenzene) film-coated platinum electrode was also used to selectively detect only the hydrogen peroxide generated into a upstream enzyme reactor, without interference from oxidizable species, such as L-ascorbate, the adsorption of low molecular-weight proteins in a dialysate, and NADPH added to the carrier solution to initiate substrate recycling. By the present in vivo system, L-glutamate was selectively assayed with about a 600-fold increase in sensitivity compared with the unamplified responses. The detection limit was 0.08 μmol dm-3. This method was applied to an in vivo assay of L-glutamate in the extracellular space of rat brain; also, monitoring of the L-glutamate level changed after a continuous stimulation of KCl to demonstrate the reliability of the system.
A new analytical method employing liquid chromatography/tandem mass spectrometry (LC/MS/MS) with a column-switching system was developed for quantitative determination of leukotriene E4 (LTE4) in human urine. A column-switching system using a trapping column, which concentrates the analyte and removes salts and other water-soluble contaminants, allowed direct injection of human urine. Because simultaneously eluted endogenous contaminants suppressed the ionization efficiency of LTE4, good liquid chromatographic separation was very important for establishing this method, notwithstanding the high selectivity of MS/MS. The calibration curve was linear over the range from 10 to 3000 pg/mL, and the method showed good accuracy and precision. This method should therefore be very useful for determination of LTE4 amounts in human urine in studies on leukotriene metabolism and the efficacy of antileukotriene drugs.
A new method that belongs to the differential category for determining the end points from potentiometric titration curves is presented. It uses a preprocess to find first derivative values by fitting four data points in and around the region of inflection to a non-linear function, and then locate the end point, usually as a maximum or minimum, using an inverse parabolic interpolation procedure that has an analytical solution. The behavior and accuracy of the sigmoid and cumulative non-linear functions used are investigated against three factors. A statistical evaluation of the new method using linear least-squares method validation and multifactor data analysis are covered. The new method is generally applied to symmetrical and unsymmetrical potentiometric titration curves, and the end point is calculated using numerical procedures only. It outperforms the “parent” regular differential method in almost all factors levels and gives accurate results comparable to the true or estimated true end points. Calculated end points from selected experimental titration curves compatible with the equivalence point category of methods, such as Gran or Fortuin, are also compared with the new method.
Calix-bis-2,3-naphtho-crown-6 can be used to discriminate between methylammonium and other organic ammonium ions. An electrode based on this ionophore, potassium tetrakis(p-chlorophenyl)borate (20 mol% relative to the ionophore) as an ionic additive and bis(2-ethylhexyl) sebacate as a solvent mediator in a poly(vinyl chloride) membrane matrix, displayed higher selectivity for methylammonium than for various other organic ammonium ions. However, there was considerable interference by inorganic cations, especially Cs+. Similar calixarene-crown-6 conjugates, such as calix-bis-1,2-benzo-crown-6, calix-bis-crown-6, 1,3-dioctyloxy-calixarene-crown-6, 1,3-diisopropoxy-calixarene-crown-6 and 1,3-dimethoxy-calixarene-crown-6 were less effective in discriminating methylammonium.
A hydrogen ion-selective solid-contact electrode based on N,N,N′,N′-tetrabenzylethanediamine has shown the best Nernstian slope and selectivity and the widest response range in a Tris buffered pH sample solution. Its linear dynamic range was pH 3.50 - 11.94, and the Nernstian slope showed 52.1 mV/pH (at 20 ± 0.2°C). When it was directly applied to human whole blood (in pH range 6.0 - 8.5) we could obtain the same satisfying results. This electrode continuously contacted a Tris 7.47 buffered solution, human whole blood and a hydrofluoric acid solution for one month without any loss of performance. Also, hydrofluoric acid did not influence the surface of this electrode, and thus it was maintained without showing any changes in potentials after being used in a hydrofluoric acid solution. The standard deviation in the determined e.m.f. differences was 1.5 mV (N = 5) for Tris buffer solutions of pH 6.5 and 1.1 mV at a Tris buffer solutions of pH 8.5. The 90% response time of the electrodes obtained by injecting of hydrochloric acid into the Tris buffer sample solution was less than 10 s. Especially, in the this paper, with these potential response characteristics of hydrogen ion selective poly(aniline) solid contact electrode, we have also presented the pH response mechanism of this electrode and the role of poly(aniline) and a doped anion in a poly(aniline) layer.
A theory is presented that describes the matched potential method (MPM) for the determination of the potentiometric selectivity coefficients (KpotA,B) of ion-selective electrodes for two ions with any charge. This MPM theory is based on electrical diffuse layers on both the membrane and the aqueous side of the interface, and is therefore independent of the Nicolsky.Eisenman equation. Instead, the Poisson equation is used and a Boltzmann distribution is assumed with respect to all charged species, including primary, interfering and background electrolyte ions located at the diffuse double layers. In this model, the MPM-selectivity coefficients of ions with equal charge (zA = zB) are expressed as the ratio of the concentrations of the primary and interfering ions in aqueous solutions at which the same amounts of the primary and interfering ions permselectively extracted into the membrane surface. For ions with unequal charge (zA ≠ zB), the selectivity coefficients are expressed as a function not only of the amounts of the primary and interfering ions permeated into the membrane surface, but also of the primary ion concentration in the initial reference solution and the ΔEMF value. Using the measured complexation stability constants and single ion distribution coefficients for the relevant systems, the corresponding MPM selectivity coefficients can be calculated from the developed MPM theory. It was found that this MPM theory is capable of accurately and precisely predicting the MPM selectivity coefficients for a series of ion-selective electrodes (ISEs) with representative ionophore systems, which are generally in complete agreement with independently determined MPM selectivity values from the potentiometric measurements. These results also conclude that the assumption for the Boltzmann distribution was in fact valid in the theory. The recent critical papers on MPM have pointed out that because the MPM selectivity coefficients are highly concentration dependent, the determined selectivity should be used not as “coefficient”, but as “factor”. Contrary to such a criticism, it was shown theoretically and experimentally that the values of the MPM selectivity coefficient for ions with equal charge (zA = zB) never vary with the primary and interfering ion concentrations in the sample solutions even when non-Nernstian responses are observed. This paper is the first comprehensive demonstration of an electrostatics-based theory for the MPM and should be of great value theoretically and experimentally for the audience of the fundamental and applied ISE researchers.
Groups of dioxadicarboxylic diamides, which were developed as potential ionophores for inorganic cations, were found to act as ionophores for a stimulant, phentermine. Especially, N,N-dioctadecyl-N′,N′-dipropyl-3,6-dioxaoctanediamide, which was originally developed as a lead ionophore and is commercially available from Fluka as lead ionophore I, was suitable for making a phentermine-selective electrode. The electrode constructed using this ionophore and bis(2-ethylhexyl) sebacate as a solvent mediator in a poly(vinyl chloride) membrane matrix discriminated between phentermine and analogous compounds more effectively than an electrode based on dibenzo-18-crown-6, a representative ionophore for organic ammonium ions. Moreover, the present electrode showed remarkably little interference by inorganic cations, such as Na+ and K+, as well as lipophilic quaternary ammonium ions including (C2H5)4N+ and (C3H7)4N+. The electrode exhibited a near-Nernstian response to phentermine in the concentration range of 2 × 10-6 to 1 × 10-2 M with a slope of 54.8 mV per concentration decade in 0.1 M MgCl2. The lower limit of detection was 7 × 10-7 M. This electrode was applied to determine phentermine in a cationic-exchange resin complex of this stimulant, which is the general dosage form in medical use.
In this paper, the reflection-absorption infrared (IR) spectroscopic method combined with the principle of solid-phase micro-extraction (SPME) is proposed to detect chlorinated aromatic amines in aqueous solutions. This proposed method provides simplicity in both the optical system and equipment setup. Compared to the SPME/attenuated total reflection-IR method, this method reduces the cost for internal-reflection elements and optical systems. Meanwhile, it has no SPME/transmission IR method problems, which require high polymer film preparation techniques to obtain a standing film that has no physical/chemical property changes when immersed in an aqueous solution. The typical linear coefficients obtained using this method for chloroanilines in aqueous solutions are around 0.995 and the detection can be lower than 100 ppb. The thickness of the hydrophobic film is relatively important in the SPME/ATR-IR method, but the uncertainty caused by the film thickness can be partially eliminated in the proposed method. This is because the IR signals are proportional to the film thickness and can be corrected using hydrophobic film signals. The low detection limits have also indicated that this proposed method can compete with the currently existing IR methods, but allowing much simpler detection.
Time-of-flight secondary ion mass spectrometry (TOF-SIMS) provides a method of elemental analysis that can distinguish among automotive paint samples of the same or nearly the same color. TOF-SIMS survey spectra were employed to determine the relative abundances of elements in the surface layers of the paint chips. The depth profile of paint samples permitted the analysis of small paint chips, the reproducible results for specific elements, and the identification of each car paint. Seventy-three samples of blue, red, white, and silver automobile paints from the major manufacturers in Korea were investigated using high resolution TOF-SIMS technique. It was found that paints of the same color produced by different manufacturers could be distinguished by this technique. TOF-SIMS is a reliable, nondestructive, and small area analyzing method for characterization of the elemental composition of automotive paint chips.
An application of capillary electrophoresis (CE) using sulfated β-cyclodextrin (SCD) has been investigated for separating alkylphenols with different chain lengths, as well as bisphenol A and bisphenol S. In the absence of SCD in running buffer, all the phenols migrated at the same velocity as the electroosmotic flow (EOF), whereas the addition of SCD effectively led to the baseline separation of alkylphenols on the basis of the difference in the abilities to bind into the hydrophobic cavity of CD. The host-guest binding constants between analyte phenols and SCD were evaluated from Benesi-Hildebrand plots of the data obtained by two independent methods, CE and UV-visible measurements, demonstrating that the greater the hydrophobicity of the phenols, the larger the binding constants. The effects of organic solvents on the resolution for alkylphenols and bisphenols were also examined. This system using SCD was effective for the separation of 4-octylphenol and 4-nonylphenol isomers having longer alkyl chains.
A new, simple, sensitive and selective catalytic method is developed for the determination of vanadium in natural and sea waters. The method is based on the catalytic effect of VV and/or VIV on the bromate oxidative-coupling reaction of metol with 2,3,4-trihydroxybenzoic acid (THBA). The reaction is followed spectrophotometrically by tracing the oxidation product at 380 and/or 570 nm after 5 min of mixing the reagents. The optimum reaction conditions are 6.4 × 10-3 mol l-1 of metol, 2.0 × 10-3 mol l-1 of THBA and 0.16 mol l-1 of bromate at 35°C and in the presence of an activator-buffer solution of 1 × 10-2 mol l-1 of tartrate (pH = 3.10). Following the recommended procedure, VV and/or VIV can be determined with linear calibration graphs up to 0.75 ng ml-1 and detection limits, based on the 3Sb criterion, of 0.008 and 0.018 ng ml-1 at 380 and 570 nm, respectively. The developed method was successfully applied, without any separation or preconcentration processes, to the determination of vanadium in natural and seawaters following the direct calibration and standard addition techniques, respectively.