We report the development of a fast and accurate fluorescence-based assay for amidine linked to cellulose membranes and Sepharose gel. The assay is founded on the glyoxal reaction, which involves reaction of an amidine group with glyoxal and an aromatic aldehyde, leading to the formation of a fluorophore that can be analyzed and quantified by fluorescence spectroscopy and imaging. While the assay has been reported previously for aromatic amidine estimation in solution phase, here we describe its adaptation and application to amidine linked to diverse forms of solid matrices, particularly benzamidine Sepharose and benzamidine-linked cellulose membranes. These functionalized porous matrices find important application in purification of serine proteases. The efficacy of a protein separation device is determined by, among other factors, the ligand (amidine) density. Hence, a sensitive and reproducible method for amidine quantitation in solid phase is needed. The glyoxal reaction was carried out on microbead-sized Sepharose gel and cellulose membranes. Calibration curves were developed for each phase, which established linearity in the range of 0–0.45 μmol per mL amidine for free amidine in solution, 0–0.45 μmol amidine per mL Sepharose gel, and 0–0.48 μmol per mL cellulose membrane. The assay showed high accuracy (~ 3.4% error), precision (RSD < 2%), and reproducibility. Finally, we show how this fluorescent labeling (glyoxal) method can provide a tool for imaging membranes and ligand distribution through confocal laser scanning microscopy.
Feasibility of several sorbents, Amoco PX-21, Carbosphere, Carbopack B and C and 2-(1-pyrenyl) ethyldimethylsilylated silica gel (PYE), for the fractionation of polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and furans was investigated. Selection was based on their suitability for quantitative isolation of the target compounds with special attention for the most toxic mono- and non-ortho-CBs to prevent coelution with other congeners during the final determination by gas chromatography. Cost-effectiveness in terms of solvent and time consumption as well as feasibility for routine analysis and automation were considered additional merits of the methods compared. Final evaluation of the procedures providing the best results was done by comparison of the results obtained from the analysis of real-life samples. The results showed that, among the sorbents tested, Carbopack B and PYE were the most suitable for routine analysis. In particular, these sorbents allowed a more reliable determination of the toxic congeners and, consequently, of the toxic equivalents of 2,3,7,8-tetrachlorodibenzo-p-dioxin content in environmental samples. 相似文献
The stability constant of the EuCl2 + complex was determined in perchloric acid media at 0.2, 0.5, and 1.0 M ionic strength by means of a solvent extraction method with dinonyl naphthalene sulfonic acid at 30°C. The values for 1,Cl were: 2.1 ± 0.1, 1.45 ± 0.01, and 0.84 ± 0.07, respectively, for each of the media. The Brønsted–Guggenheim–Scatchard specific ion interaction theory (SIT) was applied to these data and to some literature data to calculate a 1,Cl of 23.9 at infinite dilution. The calculated specific ion interaction coefficient for the pair EuCl2 +, ClO4– was 0.52. Species-distribution diagrams have been analyzed. 相似文献
The first reported use of a regenerable chemiluminescent polymer for chemical analysis is described. Ruthenium tris(4-methyl-4′-vinyl-2,2′-bipyridine) was electropolymerised onto the surface of a Pt electrode. This system was optimised for detection of oxalate by investigating the effect of both scan rate and the pH of the analyte solution. A chemically modified electrode successfully completed over 200 regeneration cycles over a 6-month period, demonstrating the stability of the system. A range of other species has also been tested for activity with the chemiluminescent polymer. The system functions in a similar way to traditional homogeneous chemiluminescent methods, but the active compound is retained and re-used rather than discarded. This results in both environmental and cost savings. 相似文献
Herein, we describe how the conformational differences exhibited by aminoglycosides in the binding pockets of the ribosome and those enzymes involved in bacterial resistance can be exploited in the design of new antibiotic derivatives with improved activity in resistant strains. The simple modification shown in the figure, leading to the conformationally restricted 5, provides an effective protection against aminoglycoside inactivation by Staphylococcus aureus ANT4, both in vivo and in vitro. 相似文献
The use of modulated temperature differential scanning calorimetry (TMDSC) and differential scanning calorimetry (DSC) in the measurement of the glass transition temperature (Tg) in polymer-water systems presents several important problems. These include the presence of water evaporation endotherms, partial water evaporation during scanning, changes in pan integrity due to vapour pressure developing in the pan headspace during analysis, and possible interaction between water and polymer at high temperatures. As a result, in most of the cases, only apparent Tg values can be obtained. In this study, TMDSC and DSC were used to determinate the thermal behaviour of methylmethacrylate copolymer-water systems. The samples were previously equilibrated at different relative humidities (RH) from 0 to 97% RH. Three different pan arrangements were used. In addition, thermogravimetric analysis (TG) was carried out to determine the initial amount of water in the sample. None of the pan arrangements was entirely suitable for the study of these systems. When sealed pans were used, the plastification effect of water was observed. Some evidence of degradation was also observed in which water and methylmethacrylate appeared to play roles.This revised version was published online in November 2005 with corrections to the Cover Date. 相似文献
The equilibrium solubilities of naproxen (NAP), ketoprofen (KTP), and ibuprofen (IBP) in methanol + water binary mixtures at 298.15 K were determined and the preferential solvation parameters were derived by means of the inverse Kirkwood–Buff integrals (IKBI) method. These drugs are very sensitive to specific solvation effects. The preferential solvation parameters by methanol δx1,3 are negative in water-rich mixtures but positive in compositions from 0.32 in mole fraction of methanol to pure methanol. It is conjecturable that in the former case the hydrophobic hydration around aromatic rings and/or methyl groups plays a relevant role in the solvation. The higher solvation by methanol in mixtures of similar co-solvent compositions and in methanol-rich mixtures could be explained in terms of the higher basic behaviour of this co-solvent interacting with the hydroxyl group of the drugs. Moreover, drug solubilities were correlated by using the modified nearly ideal binary solvent/Redlich–Kister model obtaining average percentage deviations (APDs) lower than 9.0%. 相似文献
<正>Recent years have seen a significant surge in energy storage research and development across the globe.This is due to a number of factors including the increasingly grave environmental concerns,increased energy consumption,rising fuel prices,limited fossil fuel resources and the growing interest in the expansion of renewable 相似文献