End-column electrochemical detection for aromatic amines with high performance capillary electrophoresis

Electrochemical detection of five species of aromatic amines at a carbon fiber microdisk electrode after separation by capillary electrophoresis is described. Under the optimum conditions, the detection limit for 3,4-dihydroxybenzylamine, N,N-dimethylaniline, p-phenylenediamine, p-aminophenol and aniline sulfate was 0.9, 0.03, 0.075, 1.2 and 0.15 mu M (S/N = 3), respectively. The linear response range was 5-1000, 0.1-500, 0.5-500, 5-500 and 1-200 mu M, respectively The effect of the electrode position and buffer pH on the detection was also studied. This method is very simple, sensitive and stable for the detection of these compounds.

Separation of Th4+ and RE3+ with hollow fiber extraction

Separation of Th4+ and RE3+ was investigated by hollow fiber membrane extraction with N1923 in countercurrent recirculating operation. The effect of Hf concentration in aqueous phase and flow rates of aqueous and organic phases on mass transfer coefficient was tested. Then the extraction of Th4+ from RE sulfate obtained from Baotou ore was carried out. The results obtained show that the mass transfer coefficient of Th4+ changes with the flow rate of aqueous phase, but does not change with the flow rate of organic phase and H+ concentration in aqueous phase, which suggests that the mass transfer rate of Th4+ is controlled bg that in the water critical layer, The mass transfer rate of RE3+ does not change with the flow rate of water phase, changes a little with the flow rate of organic phase, and changes with H+ concentration, which suggests that the mass transfer rate is controlled by their reaction rate with N1923. Th4+ could be extracted completely in 8 h from RE sulfate solution of Baotou ore with relatively less extraction of RE3+. So the separation of radioactive element under the sealed condition could be done.

The application of matrix-assisted laser desorption jonization mass spectrometry in enzymes of Agkistrodon blomhoffii Ussurensis venom analysis

The matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS) spectra of four enzymes (PLA, AEase, Fibrolase, L-a.a. oxidase) in Agkistrodon blomhoffii Ussurensis venom, were given and interpreted. The experiment data showed that MALDI-TOF-MS can be used directly in enzyme analysis with high sensitivity and rapidity. In addition, the results were better than those from sodium dodecyl sulfate-polyacrylamide.

Purification and characterization of L-amino acid oxidase from Agkistrodon blomhoffii ussurensis snake venom of Changbai Mountains

The L-a. a, oxidase of Agkistrodon blomhof fii ussurensis of Changbai Mountains in northeast of China has been separated by using ion-exchange and gel filtration techniques, This enzyme is composed of two subunits, the molecular weight of one subunit is about 36 000, the another is about 57 000, determined by sodium dodecyl sulfate-polyacryamide gel electrophoresis and matrix assisted laser desorption ion/time of flight mass spectrometry, The activity of L-a, a. oxidase determined using L-Leu as substrate. The optimal pH of the enzyme is 4. 5 similar to 5. 5 and 8 similar to 9. The UV-Visible absorption spectrum of L-a, a. oxidase shows the characteristics of flavor-proteins.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of phospholipase A(2) and fibrinolytic enzyme, two enzymes obtained from Chinese Agkistrodon blomhoffii Ussurensis venom

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used to analyze two enzymes, phospholipase AZ and fibrinolytic enzyme isolated from Chinese Agkistrodon blomhoffii Ussurensis venom. Using sinapinic acid as the matrix, positive ion mass spectra of the enzymes were obtained, In addition to the dominant protein [M+H](+) ions, multimeric and multiply charged ions were also observed in the mass spectra, The higher the concentration of the enzymes, the more multiply charged polymer and multimeric ions were detected, Our results indicate that MALDI-TOFMS can provide a rapid and accurate method for molecular weight determination of snake venom enzymes, Mass accuracies of 0.1 and 0.3 % were achieved by analysis of highly dialyzed phospholipase A2 and fibrinolytic enzyme, and these results are much better than those obtained using sodium dodecyl sulfate-palyacrylamide gel electrophoresis. MALDI-TOFMS thus provides a reliable method to determine the purity and molecular weight of these enzymes, which are of potential use as therapeutants, Copyright (C) 1999 John Wiley & Sons, Ltd.

Electrochemistry of vanadium hexacyanoferrate film

Two stable redox couples, accompanying clear color switches between yellow green and blue, can be observed when the VHCF-coated film platinum electrodes are cyclic potential scanned in 3.6 M H2SO4 and 0.2 M K2SO4 electrolyte solution. Electrochemical results and in situ Fourier transfer infrared (FT-IR) spectroscopy demonstrate that the redox reaction of the electroactive iron sites is related to the first redox couple (E-1/2 = 0.81 V) while the second redox couple (E-1/2 = 1.01 V) is due to the redox reactions of the electroactive vanadyl ions. Under the proper conditions, such as in high acidic solutions or thin films (deposition time is less than 2 min) and so on, the third redox couple (E-1/2 = 0.89-0.94 V) can be observed on the cyclic voltammograms, which originates from the redox reactions of the interstitial vanadyl ions. This electrochemical reaction mechanism is investigated by in situ probe beam deflection technique, exchange of K+ ions accompanies with redox reaction of the iron sites, but for redox reaction of the vanadyl ions, both H+ ions, K+ ions and water molecules are involved.

Conversion from electronic conducting phase to redox conducting phase of polypyrrole

Square-wave voltommetry is used to study the oxidation of polypyrrole doped with dodecylsulfate. The net current curve in this experiment shows why the oxidation current does not display the capacitive-like shape common in cyclic voltammetry. In cyclic voltammetry, the redox behavior of polypyrrole is attributed to the size of dodecylsulfate, irreversible incorporation and the complete consumption of dodecylsulfate. After the polypyrrole film was scanned in aqueous NaCl solution, square wave voltammetric measurements show different results, indicating the change of the polymer nature with regard to the charge transport. This is explained by anion replacement, exclusion and the change of the charge transport mechanism.

APPLICATION OF POLYBUTADIENE-BASED POLYMERIC SURFACTANTS IN LIQUID MEMBRANE SEPARATION

New polybutadiene-based surfactants (LYF) were synthesized by sulfonation of liquid polybutadiene with acetal sulfate at an elevated temperature, and their properties in a liquid surfactant membrane (LSM) separation process were examined by comparison with the two polyisobutylene-based surfactants ECA4360 and EM301. It was found that LYF surfactants had satisfactory overall properties as regards stability, swelling, and demulsification Of the W/O emulsion in the cases of both acidic and caustic internal aqueous phases.

ORDERED ARRAYS OF MYOGLOBIN ADSORBED ON THE SURFACTANT MODIFIED SURFACE BY SCANNING-TUNNELING-MICROSCOPY

Myoglobin molecules were deposited on a surfactant sodium dodecyl sulfate modified HOPG surface and imaged in air with a high resolution scanning tunneling microscope (STM) for the first time. STM images exhibit not only ordered arrays of the surfactant m

FACTOR-ANALYSIS SPECTROPHOTOMETRY FOR THE SIMULTANEOUS DETERMINATION OF VALENCE STATES OF METALS

Two M(n+)-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol systems for the simultaneous determination of the valence states of Cr and Fe using factor analysis were studied. (1) At pH 4.0, Cr(III) and Cr(VI) react with the reagent to form stable complexes and a slight difference in the wavelengths of maximum absorption (lambda(max.)) between the two complexes is observed when the sodium lauryl sulfate, which also acts as a solubilizing and sensitizing agent, is added, viz., 590 nm for Cr(III) and 593 nm for Cr(VI) complexes. (2) In the presence of ethanol, both Fe(II) and Fe(III) form 1:2 complexes with the reagent at pH 2.5-3.5 and the lambda(max.) of the Fe(II) and Fe(III) complexes is at 557 and 592 nm, respectively. In the target transformation factor analysis, the K coefficients calculated from the standard mixtures by classical least-squares analysis and a non-zero intercept added to each wavelength are used as the target vector instead of the pure component standards; this can decrease the analysis errors introduced by the interaction between the two species and by deviations from Beer's law.

Factors that effect antioxidant activity of C-phycocyanins from Spirulina platensis

Currently, antioxidants are added in the human diet to prevent free radical-induced cell damage, and there has been an explosive interest in the use of antioxidant nutritional supplements. The effects of different factors on the antioxidant activity of phycocyanins (PCs) were studied. The results showed that PCs generated hydroxyl radicals in the light, while scavenging them in the dark. When PCs were denatured by sodium dodecyl sulfate, urea and in alkaline condition, their ability to generate hydroxyl radicals disappeared and that of scavenging them greatly increased. This showed that the phycobilin moiety is the main part of PC involved in scavenging hydroxyl radicals. Trypsin hydrolysis of PCs showed that the apoprotein portion of the molecule also made a significant contribution to the antioxidant activity.

Purification and partial characterization of Mn superoxide dismutase from muscle tissue of the shrimp Macrobrachium nipponense

Superoxide dismutase (SOD; EC 1.15.1.1) is an enzyme that protects against oxidative stress from superoxide radicals in living cells. This enzyme had been isolated, purified and partially characterized from muscle tissue of the shrimp Macrobrachium nipponense. The purification was achieved by heat treatment, ammonium sulfate fractionated precipitation and column chromatograph on DEAE-cellulose 32. Some physiological and biochemical characterization of it was tested. The molecular weight of it was about 21.7 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The purified enzyme had an absorption peak of 278 nm in ultraviolet region, and the enzyme remained stable at 25-45 degreesC within 90 min. However, it was rapidly inactivated at higher temperature. Treatment of the enzyme with 1 mM ZnCl2, SDS and 1 mM or 10 mM mercaptoethanol showed some increasing activity. However, the enzyme activity was obviously inhibited by 10 mM CaCl2, CuSO4, ZnCl2 and 1 mM CaCl2 and 10 mM K2Cr2O7. SOD activity did not show significantly variation after incubated with 1 mM CaCl2, EDTA and 10 MM SDS. The enzyme was insensitive to cyanide and contained 1.03 +/- 0.14 atoms of manganese per subunit shown in atomic absorption spectroscopy, which revealed that purified SOD was Mn superoxide dismutase. (C) 2004 Elsevier B.V. All rights reserved.

Expression of TNF-alpha gene in Anabaena sp PCC 7120 and purification of its product by affinity chromatography

The effects of N (NaNO3) and C (NaAc) source in medium on the expression of tumor necrosis factor-alpha (TNF-alpha) gene in transgenic Anabaena sp. PCC 7120 were compared. The data showed that N source stabilized the expression of foreign protein and C source altered the synthesis of cell walls. Comparing several methods for breaking the cells, supersonic was able to extract TNF-alpha better than others. For purification of TNF-alpha, transgenic Anabaena cells were broken, the extracts were precipitated with ammonia sulfate, and the impure TNF-alpha was eluted from DEAE ion exchange chromatography. Electrophoresis (PAGE-SDS) showed a single band at 17 kD position.

Algal biotechnology industries and research activities in China

In old China there were very few people engaged in the study of the algae, but in new China, freshwater and marine algae are studied by over one hundred old and new phycologists. There is now an algal biotechnology industry consisting of an aquaculture industry, producing large amounts of the seaweeds Laminaria, Porphyra, Undaria, Gracilaria, eucheumoids, and the microalgae Dunaliella and Spirulina. There is also a phycocolloid industry, producing algin, agar and carrageenan; an industry producing chemicals and drugs, such as iodine, mannitol, phycocyanin, beta -carotene, PSS (propylene glycol alginate sulfate) and FPS (fucose-containing sulfated polysaccharides) and an industry producing food, feed and fertilizer. The Laminaria cultivation industry produces about 900,000 t dry Laminaria, probably the largest producer in the world and 13,000 t algin, undoubtedly one of the largest algin producer in the world.