N，N＇－硫代磷酰基芳酰胺基硫脲的合成及生物活性

Ｎ，Ｎ＇－硫代磷酰基芳酰胺基硫脲的合成及生物活性金桂玉，解敏雨，赵国锋（南开大学元素有机化学研究所天津３０００７１）关键词芳酰肼，硫代磷酰基异硫氰酸酯，硫代磷酰基芳酰胺基硫脲，合成，生物活性含（硫代）磷酰基的硫脲衍生物具有广泛生物活性，如杀虫［１，２...     

Subminute and sensitive determination of the neurotransmitter serotonin in urine by capillary electrophoresis with laser-induced fluorescence detection

In this work, a sub-minute and sensitive capillary electrophoresis with laser-induced fluorescence (CE-LIF) method was developed for the analysis and quantitation of the neurotransmitter 5-hydroxytryptamine (5-HT) or serotonin in urine. The method involves precolumn derivatization with fluorescein isothiocyanate isomer I (FITC) using an excitation light from an argon ion laser of 488 nm and a 520 nm band pass emission filter. Different variables that affect derivatization (pH, FITC concentration, reaction time and temperature) and separation (buffer concentration, pH, applied voltage and injection time) were studied. The linear dynamic range obtained was between 0 and 188 nM with a detection limit of 16 nm with a RSD between 2 and 9%. The applicability of the proposed method was demonstrated by analysis of 5-HT in human urine, establishing a concentration of 57 nM in control urine. The method was validated by standard-addition methodology.     

Sulfonate-based fluorescent probes for imaging hydrogen peroxide in living cells

Based on the mechanism of H₂O₂-mediated hydrolysis of sulfonates, two fluorescein disulfonates compounds (FS-1 and FS-2) were designed and synthesized as the highly selective and sensitive fluorescent probes for imaging H₂O₂ in living cells. The probes were detected with elemental analysis, IR, ¹H NMR and ¹³C NMR. Upon reaction with H₂O₂, the probes exhibit strong fluorescence responses and high selectivity for H₂O₂ over other reactive oxygen species and some biological compounds. Furthermore, the sulfonate-based probes, as novel fluorescent reagents, are cell-permeable and can detect micromolar changes in H₂O₂ concentrations in living cells by using confocal microscopy. Supported by the National Basic Research Program of China (Grant No. 2007CB936000), the National Natural Science Funds for Distinguished Young Scholar (Grant No. 20725518), Major Program of the National Natural Science Foundation of China (Grant No. 90713019), the National Natural Science Foundation of China (Grant No. 20875057), the Natural Science Foundation of Shandong Province, China (Grant No. Y2007B02), and the Science and Technology Development Programs of Shandong Province, China (Grant No. 2008GG30003012)     

基于磺酸酯的荧光探针用于活细胞内过氧化氢的成像检测

基于过氧化氢（H2O2）特异性催化水解磺酸酯,设计合成了新型绿色荧光探针：荧光素二磺酸酯（FS—1）和二氯荧光素二磺酸酯（FS-2）两种螺环内酯型化合物,用于活细胞内过氧化氢的检测．探针结构由元素分析、IR、^1H NMR及^13C NMR表征．实验表明：探针FS-1和FS-2在模拟生物体系中检测过氧化氢具有良好的选择性和灵敏度,且线性范围较宽．细胞成像显示：探针FS-1和FS-2用于PMA刺激或外加不同浓度H2O2孵育的小鼠腹膜巨噬细胞均呈现明亮的绿色荧光,且能对细胞内H2O2微摩尔级浓度变化产生响应,证明两探针均具有良好的膜渗透性、高的选择性及良好的灵敏度．该方法的建立对研究生物体内H2O2的产生,H2O2导致的各种疾病机制及H2O2介导的信号转导途径具有重要的理论及实际意义．     

异硫氰基孔雀石绿受激拉曼光谱研究

本文报道了具有时间分辨能力的全频宽带受激拉曼（BBSRS）系统和关于异硫氰基孔雀石绿（MGITC）受激拉曼光谱（sRs）的研究．BBSRS系统的探测光为450-800nm宽带连续白光,泵浦光为280～900nm范围内连续可调谐的ps窄带可见光（带宽≈7．5cm-1,脉宽≈2．5ps）．在合适的泵浦波长下,该系统可同时获取拉曼损失和拉曼增益光谱．MGITC的SRS研究结果表明,当拉曼损失谱峰出现在最大吸收波长（≈627nm）时,共振SRS谱峰强度最大;当泵浦或增益谱峰在最大吸收波长附近时,未观察到明显的共振拉曼信号;共振峰强度随浓度增大而增大,随泵浦功率增大而迅速增大,后趋于饱和;共振和非共振峰强在延时零点附近达到最大值,并随延时绝对值的增大而减小．     

分光光度法测定钻井液中示踪剂荧光素钠

研究了钻井液示踪剂荧光素钠的分光光度法测定方法。在pH9.0的乙酸钠缓冲介质中,用5cm比色皿于波长485nm条件下,分光光度法测定钻井水示踪剂荧光素钠的测量下限为0.005μg/mL。对荧光素钠含量为0.012μg/mL的水样,测量相对标准偏差(RSD)为8.7%,可以满足钻井水示踪技术要求。     

Active bead-linked immunoassay on protein microarrays

Protein microarrays are becoming a powerful tool in proteome, biochemical, and clinical studies. In addition to the quality of arrayed immobilized probe molecules, sensitivity of the microarray-based assay is highly dependent on the detection technique. Here we suggest four simple techniques for rapid detection of analytes bound to protein microarrays. The techniques employ functionalized magnetic and non-magnetic beads moved to, from, or along the array surface by external forces. In contrast to other labeling techniques actively controlled physical labels: (i) make detection extremely fast to allow microarray reading in seconds; (ii) provide a low background due to active removal of weakly bound beads; and (iii) provide a highly sensitive detection, since one antigen-antibody bond is capable of holding bead immobilized on the array surface. In combination with the electrophoretically assisted active immunoassay we described recently such active reading allows to reduce total indirect immunoassay time to 7-10 min while having sensitivity in the femtomolar concentration range. High speed, sensitivity, and specificity make active bead-linked detection an ideal choice in rapid high-throughput screening and in emergency diagnostics.     

Analysis of kanamycin A in human plasma and in oral dosage form by derivatization with 1-naphthyl isothiocyanate and high-performance liquid chromatography

A simple and sensitive HPLC method has been developed for trace determination of kanamycin A by derivatization. Plasma proteins are precipitated by acetonitrile and chemical derivatization is performed on the supernatant containing kanamycin A with 1-naphthyl isothiocyanate in pyridine at 70 degrees C. After the derivatization reaction, a methylamine/acetonitrile solution was added to the reaction mixture to eliminate the excess of derivatizing agent and shorten the analysis time. The resulting derivative was separated using a Lichrocart Purospher STAR RP-18e column and water/methanol (33:67, v/v) as a mobile phase (detection at 230 nm). Optimization conditions for the derivatization of kanamycin A were investigated by HPLC. The linear range for the quantitation of kanamycin A in spiked plasma was over 1.2-40 microg/mL; the detection limit (signal to noise ratio = 3; injection volume, 10 microL) was about 0.3 microg/mL. The relative standard deviation was less than 2.9% for intra-day assay (n = 6) and inter-day assay (n = 6) and relative recoveries were found to be greater than 98%. Preliminary application of the method for monitoring kanamycin A in humans upon intramuscular injection of the injection product demonstrated the usefulness of the assay for clinical studies. The proposed method can also be used to analyze the compound in pharmaceutical formulations.     

纳米银对表面吸附荧光素的荧光增强和荧光猝灭效应及KCl的荧光猝灭释放效应

制备了两种不同粒径的纳米银溶胶，研究了在水溶液条件下其对表面吸附荧光素(FL)的荧光性能的影响及KCl电解质对该体系荧光性能的影响。FL溶液中加入纳米银，FL分子吸收峰位发生红移。随着纳米银浓度的增加，FL分子荧光先出现增强，而后又逐渐猝灭。粒径较大的纳米银产生最大荧光增强比率所需浓度较低。在纳米银猝灭FL分子荧光的溶液中加入KCl电解质，随着KCl浓度的增加，荧光逐渐增强，出现了荧光猝灭释放效应。研究结果表明，纳米银对表面吸附FL的荧光作用与FL分子附近局域电磁场增强和分子到金属表面无辐射跃迁能量转移过程决定并与纳米银的浓度、尺寸及电解质等密切相关。     

基于聚苯乙烯微球的拉曼增强效应及其应用于金单晶表面单层分子的检测

利用在样品表面上组装聚苯乙烯微球, 可以使得表面拉曼信号得到增强. 系统考察了增强效应与微球粒子尺寸的依赖关系, 发现当微球直径为3.00 μm 时, 拉曼信号的增强效应最强, 可以达到约5倍的增强. 进一步利用聚苯乙烯微球的增强效应, 获得了单层吸附在Au(111)表面上具有共振增强效应的异氰基孔雀石绿分子的拉曼信号, 得到约20倍的信号净增强, 相当于约3个数量级的拉曼增强效应, 表明利用这种方法可以显著提高单晶表面吸附分子的检测灵敏度. 这种增强效应主要是由于激光在透明微球的作用下, 在微球底部产生纳米光束流, 从而形成高度局域化的电磁场, 使拉曼散射过程得到极大的增强. 初步探讨了两种类型样品表面获得不同的增强效应的原因.