Study on the multiple sites binding of human serum albumin and porphyrin by affinity capillary electrophoresis

Equations to describe the two sites binding between proteins and ligands were deduced. According to these equations, not only the binding constants, but also the mole fraction of proteins in different forms could be obtained. Using the published data on the interaction between human serum albumin (HSA) and three kinds of porphyrin (coproporphyrin (CP), uroporphyrin I (UP) and protoporphyrin (PP)), a further study on their binding was carried out. It was concluded that there may exist two binding sites with the binding constants at the first site. proved to be the preferential one, being 6.50 x 10(5) 1.94 x 10(6) and 8.94 x 10(5). respectively. In addition. it was also demonstrated that the two binding sites of HSA with CP and UP might be of different kinds, though those of HSA and PP were of the same kind but at different positions. (C) 2002 Elsevier Science B.V. All rights reserved.

Abnormal phenomenon of the dependence of the retention factors for uncharged species on applied voltage in capillary electrochromatography

Polymethacrylate-based monolithic columns were prepared for capillary electrochromatography (CEC) by in situ copolymerization of butyl methacrylate (BMA), 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS), and ethylene dimethacrylate (EDMA) in the presence of a porogen in fused-silica capillaries of 100 mum I.D. The abnormal phenomenon that retention factors for neutral species decreases with applied voltage in CEC was observed. Capillary electrophoresis (CE) instruments usually require a period of time to increase voltage from 0 kV to desired value, which is called as ramp time. Such ramp time and any error in the determination of dead time should be taken into account during the accurate calculation of retention factors. After the correction of the retention factors, the plots of the corrected factors for alkylbenzene versus applied voltage were made, the absolute value of the plot slopes are less than 1.8 X 10(-4), Which indicates that the corrected retention times for neutral species do not show any dependence on applied voltage. Further, the plots of the corrected retention times for acidic and basic compounds versus the reciprocal of applied voltage were drawn, where the target compounds were eluted in neutral form. The very nice linearity of the plots was obtained. The linear correlation coefficients are over 0.999. Here, the slopes of the plots represent

Analysis of aromatic amines by high-performance liquid chromatography with pre-column derivatization by 2-(9-carbazole)-ethyl-chloroformate

2-(9-Carbazole)-ethyl-chloroformate (CEOC), a novel pre-column fluorescence derivatization reagent, has been developed for the analysis of aromatic amines. Taking five monocyclic aromatic amines (o-toluidine, aniline, 3,4-dimethylaniline, N-ethyl-p-toluidine, and p-phenylenediamine) as testing compounds, derivatization conditions such as pH of borate buffer, reaction time and fluorescent tagging reagent concentration have been investigated. By a one-step procedure, CEOC reacts readily with the aromatic amines to form stable derivatives with excitation and emission wavelengths, respectively, at 293 and 360 nm. This derivatization reaction could be finished within 20 min even at room temperature. The peak shapes of the derivatized aromatic amines can be improved greatly without any addition of competition amines into the mobile phase. Furthermore, this method can offer excellent quantitative precision with high tolerance of the matrix of samples. (C) 2003 Elsevier B.V. All rights reserved.

Migration of neutral solutes by double stepwise gradient elution in capillary electrochromatography

Characteristics of electroosmotic flow (EOF) and the migration of neutral solutes under double stepwise gradient elution in capillary electrochromatography were studied systematically. EOF velocity proved to be the function of operation time changing with the introduction of the second mobile phase. Accordingly, the retention of components also changed. The migration of neutral solutes was studied under the following three situations; A, components eluted when the column was filled only with the first kind of mobile phase; B, solutes eluted still in the first kind of mobile phase while at that time two kinds of mobile phase coexisted in the column and C, samples eluted in the second kind of mobile phase. Equations to describe the retention times of components under these three kinds of conditions were deduced and applied to predict the retention times of 12 aromatic compounds. Relative errors between experimental and calculated values were below 5.0%, which proved the reliability of the equations. In addition, parameters that might affect the retention time of solutes, such as the transferring time of mobile phase vials, the capacity factors of components and EOF velocities two steps were studied systematically (C) 2001 Elsevier Science B.V. All rights reserved.

慢生大豆根瘤菌(Bradyrhizobiumjaponicum)聚羟丁酸合成酶基因(phbC)的克隆和序列测定

以苜蓿根瘤菌Rm10 2 1的 phaC基因突变体菌株Rm1114 4 (phaC ::Tn5 - 2 33)为受体菌 ,通过功能互补 ,成功地从构建的Bradyrhizobium japonicum USDA110基因文库中 ,筛查到能与Rm1114 4互补 ,使之恢复在以乙酰乙酸为唯一碳源的M9培养基 (M9-AA)平板上 5d形成明显可见菌落 ,以及在MOPS平板上形成粘液型菌落的表型的重组粘粒 pDC2 ;经证实 ,该粘粒带有 phbC基因 .完成了该基因的全序列测定并已在GenBank登记 ,登记号为AY0 775 80 .B .japonicumphbC基因由 180 3碱基对组成 ,GC含量 6 1.8% ,AT含量 38.2 % ;编码 6 0 0个氨基酸 ,Mr=6 6 .95× 10 3 .图 3表 3参 13

苜蓿化感作用的初步研究

利用生物测定法和气相色谱 /质谱 (GC/MS)联用技术对中国北方广为种植的紫花苜蓿的化感作用进行了初步研究。结果表明 ,紫花苜蓿全草的水和 95 %乙醇浸提液对玉米和小麦种子萌发均有显著的抑制作用 ,进一步用柱层析对苜蓿浸提液进行了初步分离 ,分离组份对玉米和小麦种子萌发的影响有所变化。抑制玉米萌发的活性物质主要在弱极性的二氯甲烷组份中 ,而抑制小麦萌发的活性物质不仅存在于二氯甲烷组份中 ,也存在于水中。采用GC/MS技术 ,初步推测苜蓿的化感物质是酚和皂甙两类化合物

沈阳地区典型绿化树种生物源挥发性有机物的排放速率

采用二次热解析仪与GC-FID联用技术对沈阳地区8种典型绿化树种生物源挥发性有机物(BVOCs)的排放速率进行了观测和研究。结果表明:银中杨和垂柳的BVOCs排放量显著高于其它6种树木,排放速率分别为97.63和18.24μg.(g.h)-1。阔叶树中银中杨、垂柳、丁香、水腊主要排放异戊二烯,排放速率分别为97.33、17.71、3.78、0.13μg·(g·h)-1,榆树、皂角、银杏主要排放柠檬烯,排放速率分别为0.60、1.64、2.00μg·(g·h)-1;而针叶树油松以排放α-蒎烯为主,排放速率为2.25μg·(g·h)-1。

植物挥发性单萜经土壤载体的化感作用——以三裂叶豚草(AmbrosiatrifidaL.)为例

植物释放的挥发性单萜在生态系统中起着重要的作用,这些单萜不仅能以空气为载体对其他植物显示直接的化感作用,而且也能以土壤为载体进行间接的化感作用。通过对三裂叶豚草挥发物对植物种子萌发和土壤微生物种群影响及其化学成分的鉴定研究,验证了以单萜类物质为主的三裂叶豚草挥发物可以经土壤载体对其他植物及土壤微生物显示化感效应。还对植物挥发物化学成分鉴定的GC(气相色谱)和GC-MS(气相色谱-质谱联用)方法存在的一些误区进行了澄清,这将有助于对生态系统中植物挥发物化感作用的研究和认识。

电化学方法检测DNA碳纳米管修饰电极

在碳纳米管(CNTs)修饰玻碳(GC)后得到CNTs/GC电极的基础上,分别用吸附法和共价键合法制备了脱氧核糖核酸(DNA)修饰的CNTs/GC电极。以硫堇(Th)为电化学指示剂,用循环伏安、示差脉冲伏安、交流阻抗法等测试技术对制备的修饰电极进行了电化学表征。结果表明,用共价键合法制备的DNA修饰的CNTs电极上DNA的固定量多、稳定性好。制备的DNA修饰电极可用于电化学基因传感器等领域。

Properties of poly(sodium 4-styrenesulfonate)-ionic liquid composite film and its application in the determination of trace metals combined with bismuth film electrode

A new kind of bismuth film modified electrode to sensitively detect trace metal ions based on incorporating highly conductive ionic liquids 1-butyl-3-methyl-imidazolium hexafluorophosphate (BMIMPF6) in solid matrices at glassy carbon (GC) was investigated. Poly(sodium 4-styrenesulfonate) (PSS), silica, and Nafion were selected as the solid matrices. The electrochemical properties of the mixed films modified GC were evaluated. The electron transfer rate of Fe(CN)(6)(4-)/Fe(CN)(6)(3-) can be effectively improved at the PSS-BMIMPF6 modified GC.

Rhythmic Growth-Induced Ring-Banded Spherulites with Radial Periodic Variation of Thicknesses Grown from Poly(epsilon-caprolactone) Solution with Constant Concentration

Birefringent ring-banded spherulites with radial periodic variation of thicknesses were grown from poly(epsilon-caprolactone) (PCL) solutions under conditions for which the Solution concentration was held constant during the whole development of the morphology. The as-grown ring-banded spherulites were investigated by optical (OM) and atomic force (AFM) microscopies, by transmission electron microscopy (TEM) of samples sectioned parallel to the plane of film, and also by electron diffraction (ED) and grazing incidence X-ray diffraction (GIXD) techniques.

Direct electron transfer of horseradish peroxidase and its electrocatalysis based on carbon nanotube/thionine/gold composites

Horseradish peroxidase (HRP) was incorporated into multiwalled carbon nanotube/thionine/Au (MTAu) composite film by electrostatic interactions between positively charged HRP and negatively charged MTAu composite. The results of electrochemical impedance spectroscopy (EIS) confirmed adsorption of HRP on the surface of MTAu modified GC electrode.

The direct electrochemistry behavior of Cyt c on the modified glassy carbon electrode by SBA-15 with a high-redox potential

It is discovered that SBA-15 (santa barbara amorphous) can provide the favorable microenvironments and optimal direct electron-transfer tunnels (DETT) of immobilizing cytochrome c (Cyt c) by the preferred orientation on it. A high-redox potential (254 mV vs. Ag/AgCl) was obtained on glassy carbon (GC) electrode modified by immobilizing Cyt c on rod-like SBA-15. With ultraviolet-visible (UV-vis), circular dichroism (CD), FTIR and cyclic voltammetry, it was demonstrated that immobilization made Cyt c exhibits stable and ideal electrochemical characteristics while the biological activity of immobilized Cyt c is retained as usual.

A bifunctional structure for the direct electron transfer of Cytochrome c

It was found that silicon dioxide (SiO2) nanoparticles modified onto glassy carbon (GC) electrode exhibited a dramatic promotion on the direct electron transfer of Cytochrome c (Cyt c). The corresponding mechanism was discussed based on the electrochemical characteristics and a spatial geometrical model of the bifunctional structure. The model could offer insight to the study of biosensors and bioreactors without chemical mediator and serve as a basis for their fabrication. (c) 2008 Elsevier Ltd. All rights reserved.