Imitation of biomembranes on the basis of cholic acid and endogenic thermostable protein complex in biopartitioning micellar chromatography

The structure of biomembranes was imitated by introducing nonionic surfactant polyoxyethylene (23) dodecylether, cholic acid and endogenic thermostable protein complex (14–65 kDa) into the mobile phase. The influence of concentration of these additives on the retention of the model compounds was studied. The competing interaction of cholic acid and endogenic thermostable protein complex in the lipid bilayer model was revealed on the basis of chromatographic data. The values of efficiency of the chromatographic column regarding solutes were increased by addition of endogenic thermostable protein complex to the mobile phase containing Brij‐35 and cholic acid. Copyright © 2011 John Wiley & Sons, Ltd.     

N-linked oligosaccharide analysis of rat brain Thy-1 by liquid chromatography with graphitized carbon column/ion trap-Fourier transform ion cyclotron resonance mass spectrometry in positive and negative ion modes

We have previously described the site-specific glycosylation analysis of rat brain Thy-1 by LC/multistage tandem mass spectrometry (MS(n)) using proteinase-digested Thy-1. In the present study, detailed structures of oligosaccharides released from Thy-1 were elucidated by mass spectrometric oligosaccharide profiling using LC/MS with a graphitized carbon column (GCC-LC/MS). First, using model oligosaccharides, we improved the oligosaccharide profiling by ion trap mass spectrometry (IT-MS) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS). Sequential scanning of a full MS(1) scan with FT-ICR-MS followed by data-dependent MS(n) with IT-MS in positive ion mode, and a subsequent full MS(1) scan with FT-ICR-MS followed by data-dependent MS(n) with IT-MS in negative ion mode enabled the monosaccharide composition analysis as well as profiling and sequencing of both neutral and acidic oligosaccharides in a single analysis. The improved oligosaccharide profiling was applied to elucidation of N-linked oligosaccharides from Thy-1 isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It was demonstrated that Thy-1 possesses a significant variety of N-linked oligosaccharides, including Lewis a/x, Lewis b/y, and disialylated structure as a partial structure. Our method could be applicable to analysis of a small abundance of glycoproteins, and could become a powerful tool for glycoproteomics.     

A practicable strategy for enrichment and separation of four minor flavonoids including two isomers from barley seedlings by macroporous resin column chromatography,medium‐pressure LC,and high‐speed countercurrent chromatography

Separation of minor compounds especially with similar polarities and structures from complex samples is a challenging work. In the present study, an efficient method was successfully established by macroporous resin column chromatography, medium‐pressure liquid chromatography, and high‐speed countercurrent chromatography for separation of four minor flavonoids from barley seedlings. Macroporous resin column chromatography and medium‐pressure liquid chromatography were used for enrichment of these four flavonoids. High‐pressure liquid chromatography analysis showed the total content of these four flavonoids increased from 2.2% in the crude extract to 95.3% in the medium‐pressure liquid chromatography fraction. It was indicated that the combination of macroporous resin column chromatography and medium‐pressure liquid chromatography could be a practicable strategy for enrichment of minor compounds from complex sample. Then, high‐speed countercurrent chromatography was employed for separation of these four flavonoids using ethyl acetate/n‐butanol/water (0.1% glacial acetic acid) (4:1:5, v/v/v) as solvent system. As a result, four flavonoids including two isomers with purities higher than 98% were obtained. Interestingly, two flavonoids existing in one high‐pressure liquid chromatography peak were also successfully separated. All these indicated high‐speed countercurrent chromatography had great potential for separation of compounds with similar structures and polarities. This study provides a reference for efficient enrichment and separation of minor compounds from complex sample.     

Electrodeposited copolymer electrolyte into nanostructured titania electrodes for 3D Li-ion microbatteries

The electrochemical synthesis of a copolymer electrolyte (PEO-PMMA) into titania nanotubes is described and studied. Compared with the electrochemical systems based on solid electrolytes deposited by top-down techniques, the copolymer/titania nanotube material reveals high electrochemical performance, opening new perspectives for the fabrication of 3D all-solid-state microbatteries.     

Theoretical investigation on redox-switchable second-order nonlinear optical responses of push-pull Cp*CoEt2C2B4H3-expanded (metallo)porphyrins

The second-order nonlinear optical (NLO) properties of the Cp*Co(C(2)H(5))(2)C(2)B(4)H(3)-expanded (metallo)porphyrins (Cp* = C(5)Me(5)) have been investigated by using ab inito RHF and density functional theory (DFT) methods. The investigation shows that the compound with expand porphyrin possesses remarkable large molecular hyperpolarizability β(tot) value, ~414.1 × 10(-30) esu (at LC-ωPBE level), and might be an excellent second-order NLO material. From the character of charge transfer (CT) transition, it indicates that the -Cp*Co(C(2)H(5))(2)C(2)B(4)H(3) acts as an electron donor in this kind of systems. As a result of the redox behavior on expanded (metallo)porphyrin, the redox switching character of the NLO responses for the systems 2a-4a has also been studied. The results show that the β(tot) values of reduced forms are larger than that of neutral ones. Furthermore, the time-dependent DFT calculation illustrates that reduced forms have a significant difference on the CT patterns versus neutral ones. The present investigation provides insight into the comparison with DFT results on estimating first hyperpolarizability and the NLO properties of the series of push-pull compounds.     

Profiling analysis of oligosaccharides in antibody pharmaceuticals by capillary electrophoresis

Carbohydrate chains in glycoprotein pharmaceuticals have important roles for the expression of their biological activities. Therefore, development of an assessment method for the carbohydrate chains is an important parameter for quality control of glycoprotein pharmaceuticals such as newly developed therapeutic antibodies. In this report, we applied capillary electrophoresis with laser-induced fluorescence detection to the analysis of carbohydrate chains after releasing with glycoamidase followed by derivatization with 3-aminobenzoic acid. We found that four major oligosaccharides present in antibody pharmaceuticals were successfully separated with good resolution. The present method showed good precision in both migration times and relative peak areas, and gave comparable accuracy with that using a derivatization method with 8-aminopyrene-1,3,6-trisulfonate.     

13顶点金属碳硼烷结构和非线性光学性质的DFT研究

采用量子化学密度泛函理论(DFT)B3LYP/6-31G(d)方法,对4,1,6-MC2B10H1213顶点金属碳硼烷几何构型进行优化,结合有限场(FF)方法计算了它们的极化率和二阶非线性光学(NLO)系数.结果表明,十个13顶点金属碳硼烷分子中1a～6a的二阶NLO系数与其构型纵向扩张呈现相同的规律.分子的前线分子轨道能级差越小,其二阶NLO系数越大.对于不同自旋态的同种金属碳硼烷分子,其偶极矩值为高自旋态大于相应的低自旋态,极化率和二阶NLO系数与自旋多重度没有一致的对应规律,自旋多重度对NLO性质影响不大.     

柴胡舒肝丸的毛细管电泳指纹图谱及其黄芩苷含量的测定

建立了柴胡舒肝丸(Chaihu Shugan Pill, CHSGP)的毛细管区带电泳指纹图谱(capillary electrophoresis fingerprint, CEFP),并采用内标法测定了黄芩苷的含量。以50 mmol/L硼砂-150 mmol/L磷酸二氢钠-50 mmol/L磷酸氢二钠(1:1:1, v/v/v)(含5 mmol/L庚烷磺酸钠)为背景电解质(BGE)溶液,采用未涂层石英毛细管(总长度75 cm,有效分离长度63 cm,内径75 μm),以色谱指纹图谱分离量指数(RF)为目标函数优化实验条件,在紫外检测波长265 nm、运行电压11 kV条件下,以黄芩苷峰为参照物峰,确定了22个共有指纹峰,建立了CHSGP的CEFP,通过对20批样品聚类分析确定用其中13批生成对照CEFP(RCEFP),以此RCEFP为标准用系统指纹定量法鉴别20批柴胡舒肝丸质量。结果其中的4批化学成分数量和分布比例不合格,4批含量明显偏低,其他12批完全合格。采用内标法测定黄芩苷的含量,在5～200 mg/L范围内线性良好(r=0.9999),平均回收率(n=9)为98.2%。该法具有较好的精密度和重现性,为柴胡舒肝丸的质量控制提供了一种新的参考。     

Complete NMR analysis of oxytocin in phosphate buffer

Complete NMR analysis of oxytocin (OXT) in phosphate buffer was elucidated by one‐dimensional (1D)‐ and two‐dimensional (2D)‐NMR techniques, which involve the assignment of peptide amide NH protons and carbamoyl NH₂ protons. The ¹H? ¹⁵N correlation of seven amide NH protons and three carbamoyl NH₂ protons were also shown by HSQC NMR of OXT without ¹⁵N enrichment. Copyright © 2009 John Wiley & Sons, Ltd.