The contents of manganese (Mn), iron (Fe), cobalt (Co), zinc (Zn), and selenium (Se) in nuclear (NU), mitochondrial (MT), microsomal (MC), and cytosolic (CS) fractions of liver homogenates of normal and selenium-deficient (SeD) rats were determined by instrumental neutron activation analysis (INAA). The uptake rates of these elements in the liver cell fractions of both groups of rats were determined by multitracer analysis (MTA). The results indicated that Se-deficiency caused a significant increase in the content of Fe in the MC fractions. The MTA showed that the uptake rate of Fe was highest in the MC fraction, and that the uptake rate in the fraction was similar between the SeD and normal rats.
A highly sensitive time-resolved fluoroimmunoassay of human plasma cytokines is described. The cytokines such as interleukin-1α (IL-1α), tumor necrosis factor α (TNF α) are known to be acute inflammatory cytokines and it has been reported that these cytokines are secreted into blood by physical exercise. In this study, a sandwich-type immunoassay of cytokines was established using a europium chelate BHHCT-Eu3+ as a powerful labeling material. The minimum detection limits of cytokines, i.e. IL-1α, TNF α, and interferon γ (IFN γ) were about 1/10 smaller than those of enzymelinked immunosorbent assay currently used. By this immunoassay we investigated cytokine increase/decrease in plasma which was thought to derive from the myocytes damaged by bicycle exercise. Healthy young men performed two kinds of bicycle ergometer exercises, under conditions of an incremental and a constant loading. Blood samples were taken before, during, and after exercises, and the concentration levels of plasma IL-1α, TNF α, and IFN γ were determined. In the case of incremental exercise, IL-1α increased significantly at the first stage but decreased to the basal level from the second stage, in spite of heavier exercise. In the case of 30 min constant exercise, the level of plasma IFN γ increased in recovery period, 2 h after the light-exercise. TNF α level was significantly higher in a heavy-exercise. The concentration of IL-1α peaked at the early stage of the incremental exercise; this fact has not been reported in previous studies. This cytokine is unique in showing a sudden increase during the early stage, while others increase after the exercise. Our highly sensitive assay made it possible to detect a slight change in plasma cytokines.
A simple electrochemical method for estimating the antioxidant activity (AA) of flavonoids has been developed. The proposed method is based on a measurement of the half-wave potential (E1/2) of the first oxidation wave of flavonoids by using flow-through column electrolysis. At the same time, the lipid peroxidation (LPO) inhibiting effects of these flavonoids were determined. A quantitative structure-activity relationship was obtained to describe the AA of flavonoids: IC50(μM) = 30.36 + 151.50E1/2 (V) - 12.63log P (r = 0.852), where IC50 represents the concentration for 50% inhibition of LPO, and P represents the octanol/water partition coefficient. This method is expected to be useful for the quick screening of flavonoid antioxidants, and evaluating the AA of flavonoid-containing foods and medicinal plants.
The feasibility of fabricating lead-sensitive chemically modified electrodes (CMEs) for trace analysis in aqueous and 40%(v/v) ethanol-water media was investigated. Carbon paste electrodes modified with crown ethers were constructed by mixing the crown ethers into a graphite powder-paraffin oil matrix. The thus-formed electrodes were able to bind Pb(II) ions chemically, and gave better voltammetric responses than unmodified ones. The crown ethers studied and compared were 18-crown-6 and dibenzo-18-crown-6. With a 5% 18-crown-6 CME, Pb(II) could be quantified at subppm levels by differential pulse voltammetry with a detection limit of 0.02 ppm. It was possible to selectively pick up Pb(II) from a solution of several other ions at an open circuit through complexation. A simultaneous analysis of Cu(II) and Pb(II) was also attempted. By differential pulse anodic stripping voltammetry Pb(II) could be quantified over the range of 1 to 100 ppb. Interference from metal ions like Ni(II), Co(II), Mn(II), Zn(II), Cd(II), Ag(I), Fe(III), Ca(II) and Mg(II) was also studied. The method was successfully applied to artificial as well as commercial samples of alcoholic beverages.
Copper(II) was selectively accumulated on a hanging mercury-drop electrode by using 2-mercaptobenzimidazole. Ensuing measurements were carried out by differential pulse adsorption stripping voltammetry. Factors affecting the accumulation, reduction, and stripping steps were investigated and a procedure was developed. The optimum conditions for the analysis of copper were pH 5.0, 5.33 × 10-5 M 2-mercaptobenzimidazole and an accumulation potential of -0.10 V (vs. Ag/AgCl). A linear range was obtained in the concentration range 0.2 - 100 ng/ml with a 90 s accumulation time and a scan rate of 16 mV/s. For ten successive determinations of 1.0, 20.0 and 70.0 ng/ml copper(II), relative standard deviations of 4.2, 3.5 and 2.8%, respectively, were obtained. The developed method was applied to the determination of copper in commercial salts and aluminium alloys.
The equivalent-circuit parameters of the 9-MHz piezoelectric quartz crystal (PQC) resonance were measured in situ during the galvanostatic polymerization of aniline on 4-aminothiophenol(4-ATP)-modified and bare Au electrodes for ca. 2000 s, respectively. Two polymerization media, 0.100 mol L-1 aniline in 1.0 mol L-1 H2SO4 and in 2.0 mol L-1 HClO4 aqueous solutions, and two values of the current density, 12 and 36 μA cm-2, were used. At identical levels of the resonant frequency shifts in the solutions, obviously greater increases in the motional resistance (R1) were found after aniline polymerization on bare Au electrodes, though the absolute values of Δf0/ΔR1 were all large; also, the resonant frequency shifts in air (Δf0g) were considerably smaller for PANI films grown on bare Au electrodes. It is thus concluded that, under identical polymerization conditions, (1) the PANI film grown on a bare gold electrode is rougher, less compact, and can entrap solution more notably; (2) the deposition efficiency of PANI is higher on a 4-ATP-modified Au electrode, owing to a significantly greater observed “dry” frequency shift, and thus a greater “net” mass value of the polyaniline backbone. SEM observations have confirmed that PANI films on 4-ATP-modified Au electrodes were smoother and more compact than those grown on bare Au ones under identical polymerization conditions. In addition, a technique of simultaneous measurements of the electroacoustic admittance of the PQC resonance and the electrochemical impedance was used to monitor the adsorption of 4-ATP onto a PQC gold electrode.
The effect of hydrophobicity of acidic chelating agents as sensing materials on the potentiometric responses of polymeric liquid membranes was investigated. The chelating agents tested were 8-quinolinol (HOx), dithizone (HDz), 1-(2- pyridylazo)-2-naphthol (PAN) and their alkylated analogues, 5-octyloxymethyl-8-quinolinol (HO8Q), di(p-hexylphenyl) thiocarbazone (C6HDz), 7-pentadecyloxy-1-(2-pyridylazo)-2-naphthol (C15PAN) and a series of N-alkylcarbonyl- N-phenylhydroxylamines (CnPHA, n = 3, 6, 9, 12). The distribution coefficients between membrane solvent and water were determined to evaluate the hydrophobicity of the agents. The potential-pH profiles of the membranes containing hydrophobic chelating agents demonstrated the generation of potentiometric responses, while less hydrophobic agents gave no response. A possible model for the generation of membrane potential is proposed. The charge separation is attained by the permselective uptake of metal cations by the chelating agent anion at membrane/solution interface, where the high hydrophobicity of the agent enables the anionic or deprotonated form of the agents to remain at the membrane/solution interface.
A spectrophotometric enzymatic flow injection (FI) system for the determination of diethyl-p-nitrophenylphosphate (paraoxon) is proposed. The method was based on the determination of the acetic acid formed by the enzymatic reaction of the acetylcholinesterase, immobilized on glass beads, with the substrate acetylcholine. The acetic acid formed permeates through a PTFE membrane and is received by a solution (pH 7.0) containing the acid-base indicator Bromocresol Purple (B. C. P.), leading to a pH change and therefore to a color change. The variation of the absorbance of the solution is detected spectrophotometrically at 400 nm. The determination of paraoxon is related to its inhibitory action on the enzyme. Therefore the analytical signal is the difference between the signal that corresponds to the free and the one that corresponds to the inhibited enzyme, considering a fixed acetylcholine concentration. The correlation between the peak height and paraoxon concentration at a given acetylcholine concentration is linear in the range from 5.0 × 10-7 mol L-1 to 5.0 × 10-5 mol L-1 (r = 0.998) of paraoxon, with a relative estimated standard deviation (R.S.D.) of ±1.7% (n = 10) considering a solution containing 5.0 × 10-6 mol L-1 of paraoxon and a solution containing 5.0 × 10-3 mol L-1 of acetylcholine. Therefore, the quantitative limit detection is about 2.5 × 10-7 mol L-1 of paraoxon (3 σ). A 1,1′- trimethylene-bis(4-formylpyridinium bromide)dioxime (TMB-4) solution was used to reactivate the enzyme.
A new matching algorithm for library searches of mass spectra is presented in this paper. The algorithm is based on the substructure similarity of substances. It emphasizes m/z positions rather than abundance values. 32 spectra, whose corresponding molecular weights are less than 200, were randomly selected from a mass library of 61993 spectra and taken as targets of library search to illustrate the availability of this algorithm. The results show that the algorithm is better than the one built in a commercial instrument when there is no spectrum of the unknown in the library but there are similar ones.
A simple and sensitive method for the determination of liquid methanol and ethanol at trace levels by an alkyl nitrite formation reaction has been established. Alcohol was allowed to react with nitrous acid, which was yielded from sulfuric acid and sodium nitrite in the solution, to form the corresponding alkyl nitrite in the hexane organic phase. Alkyl nitrites in hexane were analyzed by a gas chromatograph with an electron capture detector (GC-ECD). The detection limits, which were determined at a signal-to-noise ratio of 3, were 1.1 and 0.7 μg/L for methanol and ethanol, respectively, by 1 μL injection. The relative standard deviations for n = 8 were 4.0 and 3.3% for methanol and ethanol, respectively. The method was applied to determine the alcohol concentration in a rice paddy, pond water, tap water, and well water. Those aqueous samples were also spiked with standard alcohols; the average recoveries of spiked methanol and ethanol were 98 and 91% with relative standard deviations of 6.1 and 4.0%, respectively.
A novel method for the determination of residual solvents in pharmaceuticals by thermal desorption (TD)-GC/MS has been established. A programmed temperature pyrolyzer (double shot pyrolyzer) is applied for the TD. This method does not require any sample pretreatment and allows very small amounts of the sample. Directly desorbed solvents from intact pharmaceuticals (ca. 1 mg) in the desorption cup (5 mm × 3.8 mm i.d.) were cryofocused at the head of a capillary column prior to a GC/MS analysis. The desorption temperature was set at a point about 20°C higher than the melting point of each sample individually, and held for 3 min. The analytical results using 7 different pharmaceuticals were in agreement with those obtained by direct injection (DI) of the solution, followed by USP XXIII. This proposed TDGC/ MS method was demonstrated to be very useful for the identification and quantification of residual solvents. Furthermore, this method was simple, allowed rapid analysis and gave good repeatability.
A capillary electrophoretic method for a high-sensitivity analysis of cyanide has been developed. Cyanide was derivatized with 2,3-naphthalenedialdehyde and taurine to give a fluorescent product of 1-cyanobenz[f]isoindole. This compound was detected with high sensitivity by fluorescence detection. The detection limit was 0.1 ng/mL, and the calibration curve was linear over the range 0.1 - 200 ng/mL. The precision of the migration time of within-run assays (n = 6) of 1 ng/mL cyanide standard solution was 0.14%. The precision of the peak area for the same runs was 1.0%. This method was applicable to blood analysis. Detection of the cyanide derivative by UV was also examined.
The collection efficiency of metallic contaminants on four different types of silicon wafers was investigated. P, p+, n and n+-type polished silicon wafers were used for the substrate, and 14 metallic elements (Na, Mg, Al, K, Ca, Cr, Fe, Mn, Co, Ni, Cu, Zn, Mo and Ti) were contaminated on silicon wafer surface. Vapor-phase decomposition-droplet collection (VPD-DC) was employed as the sample preparation procedure. For the collecting solution, HNO3, HF and a mixture of HF and H2O2 were used, respectively. A liquid droplet collecting metallic contaminants during VPD-DC was analyzed by inductively coupled plasma-mass spectrometry (ICP-MS). As a result, it was found that HNO3 and HF were not suitable for collecting Cu. Copper was not collected completely in HNO3 and HF. A mixture of HF and H2O2 is the most effective to collect all of the tested metallic elements, regardless of the dopant concentration and type of substrate.