高效毛细管电泳前沿分析法研究酸性药物那格列奈与人血浆白蛋白的结合

用高效毛细管电泳前沿分析法研究了酸性药物那格列奈与人血浆白蛋白的结合常数、结合位点和结合率。使用未涂层的毛细管柱 (4 0cm× 5 0 μmi.d .;有效柱长 32cm) ,磷酸盐缓冲溶液 (pH 7.4 ,离子强度0 .17)为背景溶液 ,在紫外检测波长 2 14nm、运行电压 18kV和重力进样 10 0s的条件下 ,利用那格列奈谱峰的平台高度和游离药物浓度的良好线性关系 (r>0 .999,n =6 ) ,测定了那格列奈的游离药物浓度。固定药物浓度 (2 0 0 μmol L ,2 5 0 μmol L) ,考察不同的蛋白质浓度对结合的影响 ;固定蛋白质浓度 (10 0 μmol L) ,考察不同的药物浓度对结合的影响。实验数据采用非线性拟和程序进行处理 ,得到了那格列奈的蛋白质结合参数。高效毛细管电泳前沿分析法测定的数据重现性良好 (RSD <2 .5 % ,n =3) ,在药代动力学和药效学研究方面具有简便、准确的优点。     

高效毛细管电泳在药物分析中的应用

对高效毛细管电泳（ＨＰＣＥ）在药物制剂及原料药分析中的应用（测定药物中主成分及杂质含量;手性药物拆分;微量制备及成分鉴别等）作一综述,并用表１,２列出了部分应用实例。     

碱性药物与人血清白蛋白结合参数的毛细管区带电泳/前沿分析

将毛细管区带电泳／前沿分析技术用于测定碱性药物ｖｅｒａｐａｍｉｌ（ＶＥＲ）和ｐｒｏｐｒａｎｏｌｏｌ（ＰＲＯ）与人血清白蛋白（ＨＳＡ）平衡体系中结合参数的研究。通过压力进样将药物白蛋白混合液引入石英毛细管电泳柱（３２ｃｍ×５０μｍｉ．ｄ．,聚丙烯酰胺涂渍柱）,在ｐＨ值为７．４、离子强度为０．１７的磷酸缓冲液及运行电压为１０ｋＶ的条件下,未结合的碱性药物的浓度可以通过电泳平台峰的高度定量,并且具有良好的线性关系（相关系数ｒ＝０．９９９）。     

高效毛细管电泳在糖分析中的应用

本文评述了近年来毛细管电泳在糖分析中的发展状况和一些常见的分析方法，引用文献５１篇。     

色谱技术在药物-血浆蛋白相互作用研究中的应用进展

小分子药物进入人体血液循环系统后与人血清白蛋白(HSA)、α1-酸性糖蛋白(AGP)等血浆蛋白存在广泛的相互作用,这些相互作用深刻影响药物在体内的分布及其与靶标蛋白的结合,进而影响药物效应的发挥.深入探究药物与血浆蛋白间的相互作用对于候选药物的成药性优化、新药研发、联合用药的风险评控等意义重大.而发展高效、灵敏、准确的...     

高效液相色谱法在药物分析中的应用

对高效液相色谱法在天然药物、复方药物、手性药物及临床治疗药物监测中的应用进行了综述。并展望了高效液相色谱法的发展-联用技术在药物分析中的应用前景。     

《现代色谱法在药物分析中的应用》

本书是现代色谱技术的专著。全书共10章，可分为4部分：1．绪论与样品预处理；2．色谱基础理论；3．色谱基本方法包含气相色谱法、高效液相色谱法及薄层色谱法；4．色谱新技术包含液相色谱溶剂优化导论、毛细管电泳法、微流控分析系统及色谱-光谱联用技术等。     

高效毛细管电泳在核酸、蛋白质分析中的新进展

高效毛细管电泳以其分离效率高，分析速度快，样品和试剂用量少，易于实现自动化等优点，在核酸、蛋白质等生物样品的分析方面发挥着重要的作用并具有巨大的潜力。本文介绍了近两年来高效毛细管电泳技术的进展，特别是PCR／CE、CE／MS以及电泳芯片技术等方面的新发展，并综述了高效毛细管电泳在核酸、蛋白质分析方面的应用，同时对其前景进行了展望。     

酮洛芬与蛋白结合作用的高效液相色谱-迎头分析法研究及其与高效毛细管电泳-迎头分析法的比较

采用高效液相色谱-迎头分析法(HPLC-FA),以67 mmol/L (pH 7.4, I=0.17 mol/L) 的等渗磷酸盐缓冲液为流动相,Pinkerton GFF　Ⅱ-S5-80内表面反相柱(150 mm×4.6 mm i.d., 5 μm)为固定相,254 nm下检测,研究了酮洛芬与人血清白蛋白(HSA)的结合作用,通过非线性回归参数估算求得酮洛芬与HSA的结合参数。与高效毛细管电泳-迎头分析法(HPCE-FA)相比,HPLC-FA法具有高灵敏度的优势,但进样量较大,分析时间较长。HPLC-F     

Protein Binding Study of Isoflavones by High-Performance Frontal Analysis

High-performance frontal analysis was used for protein binding study of isoflavones (daidzein, genistin, and genistein) to human serum albumin. The analysis was performed on a Develosil 100-Diol-5 column (10 cm × 4.6 mm). Sodium phosphate solution (pH 7.4, ionic strength 0.17) was used as the mobile phase at a flow rate of 1 mL min^–1. UV wavelength was set at 260 nm. To ensure the drug to be eluted as a trapezoidal peak with a plateau, injection volumes of 700 L for daidzein and genistin, 900 L for genistein were chosen, respectively. Experimental data were fitted by Scatchard equation. The binding constants (K) and binding affinities (nK) of isoflavones to HSA were: K=1.581 × 10⁵ (L mol^–1), nK=0.77 × 10⁴ (L mol^–1) for daidzein, K=1.082 × 10⁵ (L mol^–1), nK=0.32 × 10⁴ (L mol^–1) for genistein, and K=3.533 × 10⁵ (L mol^–1), nK=0.70 × 10⁴ (L mol^–1) for genistin, respectively.     

A New Approach for Titration Calorimetric Data Analysis on the Binding of Magnesium Ion with Myelin Basic Protein

The interaction of the myelin basic protein (MBP) from the bovine central nervous system with divalent magnesium ion was studied by isothermal titration calorimetry at 27 °C in aqueous solution. A simple rapid method for determination of the dissociation binding constants for Mg²⁺-MBP interaction was introduced using the isothermal titration calometric data. The binding isotherm for Mg²⁺-MBP interaction is easily obtained by carrying out a titration calorimetric experiment using only one set of concentrations of MBP. There are two identical independent intrinsic association constants equal to 0.021 μmol⋅L⁻¹ in the first- and second-binding sites, respectively.     

A Thermodynamic Study on the Binding of Calcium Ion with Myelin Basic Protein

The interaction of myelin basic protein (MBP) from the bovine central nervous system with divalent calcium ion was studied by isothermal titration calorimetry at 27 °C in aqueous solution. The extended solvation model was used to reproduce the enthalpies of Ca²⁺-MBP interaction over the whole range of Ca²⁺ concentrations. The solvation parameters recovered from the solvation model were attributed to the structural change of MBP due to the metal ion interaction. It was found that there is a set of two identical and non-interacting binding sites for Ca²⁺ ions. The association equilibrium constant is 0.021 μmol⋅dm⁻³. The molar enthalpy of binding is ΔH=−15.10 kJ⋅mol⁻¹.     

白色念珠菌N-肉豆蔻酰基转移酶中药物结合位点的MCSS分析

采用多拷贝同时搜寻方法(MCSS)分析得到了CaNMT活性位点的疏水区域、氢键结合位点和负电性区域. MCSS计算结果显示, CaNMT活性位点有两个疏水性比较强的区域: 一个由Tyr107, Tyr109, Val108, Phe117, Phe123, Ala127, Phe176和Leu337等残基组成; 另一个由Phe115, Phe240和Phe339组成. CaNMT活性位点发现有两个氢键作用区域, 其中Tyr119, His227, Asn392和Leu451是与已有抑制剂的氢键结合位点, Tyr107, Asn175, Thr211和Asp412是新发现的氢键结合位点, 而且在NMT家族中高度稳定, 它们对设计新结构类型的CaNMT抑制剂具有重要作用. Leu451是负电性兼氢键作用位点, 是抑制剂设计时所必需考虑的位点.     

Study on Binding Properties of Poorly Soluble Drug Trimethoprim in Anionic Micellar Solutions

Binding and distribution properties of trimethoprim (TMP) in the presence of various anionic surfactants; sodium octyl sulfate (C₈SO₄Na), sodium decyl sulfate (C₁₀SO₄Na), sodium lauryl sulfate (C₁₂SO₄Na), and sodium tetradecyl sulfate (C₁₄SO₄Na) has been studied by conductivity, spectrophotometry and surface tension measurements. The surface properties of anionic surfactants, that is, maximum surface excess concentration (Γ _max ) and minimum area per surfactant molecule (A _min ) at the air/water interface have been evaluated in the absence and presence of TMP using Gibbs adsorption isotherm. From conductivity data the ionization degree and counterion binding parameter have been obtained. Spectrophotometric experiments were used to determine binding constants of TMP to anionic micelles. With the increasing alkyl chain of surfactants, the interaction becomes stronger, which shows the importance of hydrophobic forces and incorporation of TMP molecules to the pure micelles of anionic surfactants increased. The results obtained from the surface tension and conductometric studies have been correlated with those obtained from the spectroscopic studies and binding tendency of TMP to anionic micelles followed the order as: C₁₄SO₄Na > C₁₂SO₄Na > C₁₀SO₄Na > C₈SO₄Na. From these results, the study of the interaction TMP in different anionic micellar solutions provided information about the characteristics of binding properties of poorly soluble drugs.     

纳米材料在蛋白磷酸化分析中的应用研究

蛋白质磷酸化修饰是一种重要的蛋白质翻译后修饰,在细胞代谢过程中发挥着重要作用。当蛋白质的正常磷酸化调节发生异常时,会导致癌症、糖尿病、心脏病等各种疾病的发生。因此,蛋白磷酸化分析对于疾病的早期快速诊断、药物筛选和治疗等方面具有重大的意义。由于蛋白质磷酸化过程是动态的,并且磷酸化肽段或蛋白在生物样品中的含量较低,因此高灵敏的蛋白磷酸化分析面临着巨大的挑战。该文依据在检测过程中,选择性识别或捕获磷酸化的肽段或蛋白的主要机理,综述了近几年纳米材料对磷酸化肽段的富集和信号放大作用在蛋白磷酸化分析中的研究进展,并对其未来研究方向进行了展望。