Application of matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in determining arginine esterase from snake venom

Using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The homogeneities and molecular weights of three arginine esterases from snake venom, which possessing therapeutic use in myocardial infarction, were determined and compared, MALDI-TOF-MS is possessed of high accuracy, high sensitivity and rapidity. MALDI-TOF-MS and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) can provide complementary and confirmatory results information. MALDI-TOF-MS can be directly used as an important method for the purification of snake venom complexes successfully.

Self-gelatinizable graft copolymer of poly(vinyl alcohol) with 4-vinylpyridine as an immobilization matrix for the construction of a tyrosinase-based amperometric biosensor

A tyrosinase-based amperometric biosensor using a self-gelatinizable graft copolymer of poly(vinyl alcohol) with 4-vinylpyridine (PVA-g-PVP) as an immobilization matrix was constructed. The 4-vinylpyridine component of PVA-g-PVP enhances the adherence to a glassy carbon electrode surface. The content of 4-vinylpyridine in this immobilization matrix plays a key role in retaining the activity of tyrosinase. A simple, milder method was adopted by simply syringing the copolymer-tyrosinase aqueous solution on to the electrode surface and allowing water to evaporate at 4 degrees C in a refrigerator. Several parameters, including copolymer composition; pH, applied potential and enzyme membrane composition, ware optimized. The enzyme membrane composition can be varied to obtain higher sensitivity or a wider linear detection range. The biosensor was used for the determination of phenol, p-cresol and catechol. The biosensor exhibited excellent reproducibility, stability and sensitive response and can be used in flow injection analysis. The biosensor showed an extended linear range in hydrophilic organic solvents and it can be used in monitoring organic reaction processes. The analytical performance demonstrated this immobilization matrix is suitable for the immobilization of tyrosinase.

Study on the energy transfer between UO22+ and Eu3+ in sol-gel derived titania matrix by luminescence spectroscopy

The TiO2 gel doped with UO22+ and Eu3+ has been prepared by a sol-gel method. The quenching of the UO22+ emission by Eu3+ and the energy transfer from the excited state of UO22+ to the ground state oh Eu3+ have been investigated. The energy transfer has been studied by the measurement of luminescence lifetime tau, calculations of energy transfer efficiency eta(ET) and energy transfer rate W-ET The experimental results indicated that the quenching is combined static and dynamic mechanism, but the static mechanism is dominant.

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.

Preparation and luminescence properties of terbium-Sal complex in-situ synthesized in silica matrix by a two-step sol-gel process

In-situ synthesis of terbium complex with salicylic acid (Sal) in silica matrix was made by a two-step sol-gel process. The terbium complex with salicylic acid was formed in sol-gel derived silica gel, and confirmed by the luminescence excitation spectra and infrared(IR) spectra. As compared to the pure terbium complex powder, the silica gel containing terbium complex exhibits its characteristic emission and presents a longer fluorescence lifetime than that for the pure complex. The luminescence properties of the complex containing;silica gel were investigated and compared with that of both terbium doped the silica gel and thp pure complex powder. The reasons leading to the above results were also discussed.

Luminescence characteristics of europium and terbium complexes with 1,10-phenanthroline in-situ synthesized in a silica matrix by a two-step sol-gel process

In-situ synthesis of europium and terbium complexes with 1,10-phenanthroline (phen) in silica matrix by a two-step sol-gel process has been proposed. The formation of europium and terbium complexes with phen in sol-gel derived silica gel were confirmed by the luminescence excitation spectra. The silica gels that contain in-situ synthesized europium and terbium complex exhibit the characteristic emission bands of the rare earth ions. Furthermore. the rare earth ions present longer fluorescence lifetimes than the comparable pure complex powder and the complexes dissolved in ethanol solutions. The luminescence properties of the silica gels codoped with europium (or terbium) and phen were also investigated with respect to the gels doped with europium (or terbium). (C) 1999 Elsevier Science B.V. All rights reserved.

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.

In-situ synthesis of terbium complex with salicylic acid in silica matrix by a two-step sol-gel process

A process for in situ synthesis of terbium complex with salicylic acid by a two-step sol-gel method in silica matrix has been proposed. The luminescence properties of the silica gels codoped with terbium and salicylic acid have also been discussed with respect to that of the gel doped with terbium and that of pure terbium complex with salicylic acid.

Detection of specific noncovalent protein-fullerenols complexes by matrix-assisted laser desorption ionization mass spectrometry

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry is difficult for the characterization of noncovalent complexes hitherto because of the limitations in acidic matrix, sample preparation, laser-induced polymerization and adduct formation with matrix. Under our experimental conditions, sinapinic acid is used as a matrix, the specific noncovalent interactions of protein with fullerenols were observed by MALDI mass spectrometry. Some mass spectrometric features, such as mass shifts, broad adduct peaks and stoichiometries, showed that the specific non-covalent complexes between protein and fullerenols have been formed at a ratio of 1 : 4 for hemoglobin-fullerenols or 1 : 1 for myoglobin-fullerenols. The results implied that fullereneols could be used to protect partly hemoglobin from decomposition in acidic media, and therefore, it is possible to realize the molecular weight determination of a quaternary protein by MALDI mass spectrometry via the addition of specific organic compound in the matrix.

Crystallization of micelles and matrix in dilute diblock copolymer/homopolymer solutions

The crystallization, morphology, and crystalline structure of dilute solid solutions of tetrahydrofuran-methyl methacrylate diblock copolymer (PTHF-b-PMMA) in poly(ethylene oxide) (PEO) and PTHF have been studied with differential scanning calorimetry (DSC), X-ray, and optical microscopy. This study provides a new insight into the crystallization behavior of block copolymers. For the dilute PTHF-b-PMMA/PEO system containing only 2 to 7 wt % of PTHF content, crystallization of the PTHF micellar core was detected both on cooling and on heating. Compared the crystallization of the PTHF in the dilute solutions with that in the pure copolymer, it was found that the crystallizability of the PTHF micellar core in the solution is much greater than that of the dispersed PTHF microdomain in the pure copolymer. The stronger crystallizability in the solution was presumably due to a softened PMMA corona formed in the solution of the copolymer with PEG. However, the "soft" micelles formed in the solution (meaning that the glass transition temperatures (T-g) of the micelle is lower than the T-m of the matrix phase) showed almost no effects on the spherulitic morphology of the PEO component, compared with that of the pure PEO sample. In contrast, significant effects of the micelles with a "hard" PMMA core (meaning that the T-g of the core is higher than the T-m of the PTHF homopolymer) on the nucleation, crystalline structure, and spherulitic morphology were observed for the dilute PTHF-b-PMMA/PTHF system. (C) 1998 John Wiley & Sons, Inc.

Luminescence properties of rare-earth (Eu3+ and Tb3+) complexes with paraaminobenzoic acid and 1,10-phenanthroline incorporated into a silica matrix by sol-gel method

Ternary complexes of europium and terbium with paraaminobenzoic acid and 1,10-phenanthroline (Eu(p-ABA)(3). phen . 2H(2)O and Tb(p-ABA)(3). phen . 2H(2)O, where p-HABA = paraaminobenzoic acid and phen = 1,10-phenanthroline) were introduced into a silica matrix by sol-gel method. The luminescence behavior of the complexes in silica gels was studied in comparison with the. corresponding solid-state complexes by means of emission, excitation spectra, and Lifetimes. Within the range of effective dopant concentrations, the luminescence intensities of rare-earth complexes in silica gel increase with the increasing of their dopant concentration. The lifetimes of rare-earth ions (Eu3+ and-Tb3+) in silica gel doped with europium and terbium complexes become longer than those in pure complexes. Very small amounts of rare-earth complexes doped in silica gel matrix can exhibit excellent luminescence properties, (C) 1998 Elsevier Science Ltd.

A study of Aconitum alkaloids from aconite roots in Aconitum carmichaeli Debx using matrix-assisted laser desorption ionization mass spectrometry

In the present paper a study of C-19-diterpene type of aconitum alkaloids, extracted from aconite roots in Aconitum carmichaeli Debx has been made using matrix-assisted laser desorption/ionization time of Eight mass spectrometry (MALDI-TOFMS), The results demonstrated that the aconitum alkaloids from aconite roots can be determined simultaneously by this method, which was found to be superior to other analytical methods with regard to speed and sensitivity. Fourteen known aconitum alkaloids, including aconitines, benzoylaconitines and lipoaconitines, were assigned in the methanol extract and three compounds not reported before have been targeted separation. The evaluation of the efficiency of different extractions has been studied. These results suggested that the differences of the polarity and basicity of aconitine, and benzoylaconitines and lipoaconitines result from the C-8 constituent groups that are easily lost under MALDI, (C) 1998 John Wiley & Sons, Ltd.

Luminescence properties of the rare earth (Eu3+ and Tb3+) complexes with 1,10-phenanthroline incorporated in silica matrix by a sol-gel method

Europium and terbium complexes with 1,10-phenanthroline were introduced into silica gel by the sol-gel method. The luminescence behavior of the complexes in silica gels was studied compared with the corresponding solid state complexes by means of emission, excitation spectra and lifetimes. (C) 1998 Published by Elsevier Science S.A. All rights reserved.

Characterization of cyclic (aryletherketone)s by matrix-assisted laser desorption ionization time of flight mass spectrometry

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was used to identify cyclic(aryl ether ketone) oligomers, using 2,5-dihydroxybenzoic acid as a matrix, and Na+ and K+ as cationization agents, The existence of different ring-sized cyclic oligomers up to 9 and their distribution were determined and compared with GPC results, The results indicated that MALDI-TOF MS was a powerful and rapid analytical tool for the cyclic oligomers.