超高效液相色谱-串联质谱法鉴定猪肉的特异性肽生物标志物

以亲缘关系较近的猪、牛和羊3个物种的肌肉组织为研究对象,采用超高效液相色谱-串联质谱(UPLC-MS),筛选并确认了猪物种肉特异性肽生物标志物.3种纯肉样品经蛋白质提取、胰蛋白酶消化和UPLC-TripleTOF-MS分离鉴定,得到的总离子流图谱(TIC)与Uniprot蛋白质数据库对比分析,筛选出3个物种肉的3种高丰度同源蛋白和8种潜在的肽生物标志物;潜在的肽生物标志物经QTRAP-MS质谱的多反应模式(MRM)分析,最终确认了猪物种肉的5种肽生物标志物,其中3种肽生物标志物未见文献报道.     

基于MP-AES的在线检测技术在地表水中重金属监测中的应用

通过集成在线富集和在线热消解技术,建立了基于微波等离子体原子发射光谱法(MP-AES)的地表水中重金属的在线检测技术,对珠江干流之一的西江水样中重金属元素(Cd,Cu,Cr,Ni,Pb,Fe,Mn和Zn)进行现场同时在线监测。结果表明,该在线检测技术对这些重金属元素的定量检测能力满足地表水环境质量标准(GB 3838-2002)的限量要求;据环境标准样品中重金属元素分析结果,测定值与配制标准值一致;自来水加标样品的回收率为81.5%~102%。该检测技术对重金属的检出限为1.14~5.34μg/L,检测结果的相对标准偏差(RSD)为0.79%~9.4%,方法可满足地表水中重金属的现场、快速、连续、准确监测需求。     

Nanomaterial Based Biosensors for Detection of Biomarkers of Exposure to OP Pesticides and Nerve Agents: A Review

Organophosphorus (OP) pesticides are primarily used as insecticides and chemical warfare agents worldwide. Due to their impact on the environment and health, it is important to develop prompt and accurate pesticide analysis method. This review addresses recent advances and new trends in nanotechnology‐based biosensors for biological monitoring of exposures to OP pesticides and nerve agents. In order to determine them, we have to find the corresponding biomarkers. In 1989, the national academy of sciences (NAS)divided biomarkers into the following three categories: biomarker of exposure, biomarker of effect and biomarker of susceptibility (Figure 1A). The unique chemical and physical properties of nanomaterial have paved the way to new and improved sensing devices, in general, and electrochemical/optical biosensors, in particular. In this paper, background information and a general overview of electrochemical/immunoassay detection techniques are provided. Various nanomaterial labels are discussed. Usually nanomaterials can be roughly divided into nanometer powder, nanometer fiber, nanometer film, nanometer block and so on four classes, such as colloidal gold, semiconductor nanoparticles and carbon nanomaterial (Figure 1B). In addition, we discuss some future considerations and opportunities for advancing the use of biosensors for environmental and health studies.     

A sensitive quantitative assay for the determination of propafenone and two metabolites, 5‐hydroxypropafenone and N‐depropylpropafenone,in human K2EDTA plasma using LC–MS/MS with ESI operated in positive mode

Propafenone is a potent antiarrhythmic agent; clinically propafenone has been used for a number of cardiac arrhythmias because it possesses multiple modes of action, via beta adrenergic receptor blockade and calcium antagonistic activity. Propafenone (PPF) exhibits extensive saturable presystemic biotransformation (first‐pass effect) resulting in two active metabolites: 5‐hydroxypropafenone (5‐OH PPF) formed by CYP2D6 and N‐ depropylpropafenone (NDP) formed by both CYP3A4 and CYP1A2 enzymes. A specific and sensitive LC–MS/MS method was developed and validated for quantitation of PPF, 5‐OH PPF and NDP using turboion spray in a positive ion mode. A solid‐phase extraction was employed for the extraction from human plasma. Chromatographic separation of analytes was achieved using an ACE‐5 C₈ (50 × 4.6 mm) column with a gradient mobile phase comprising ammonium acetate containing 0.01% TFA in purified water and acetonitrile. The retention times achieved were 1.36, 1.23, 1.24 min and 1.34 min for PPF, 5‐OH PPF, NDP and IS (carbamazepine), respectively. Quantitation was performed by monitoring multiple reaction monitoring transition pairs of m /z 342.30 to m /z 116.20, m /z 358.30 to m /z 116.20, m /z 300.30 to m /z 74.20 and m /z 237.20 to m /z 194.10, respectively. The developed method was validated for various parameters. The calibration curves of PPF and 5‐OH PPF showed linearity from 1 to 500 ng/mL, with a lower limit of quantitation of 1.0 ng/mL and for NDP linearity from 0.1 to 25 ng/mL with a lower limit of quantitation of 0.1 ng/mL. The bias and precision for intra‐ and‐inter batch assays were <10 and 5%, respectively. The developed assay was used to evaluate pharmacokinetic properties of propafenone and its major metabolites in healthy human subjects.     

光致清洁电极可见光在线更新与细胞实时监测

电化学传感器在用于细胞实时监测过程中,电极界面污染严重影响其检测性能.通过将纳米光催化剂与电化学传感材料复合,构建光致清洁电化学传感器,为电极界面的高效及无损更新提供了新思路.然而光催化产生的活性氧自由基导致细胞损伤,限制了细胞培养及检测过程中电极界面的实时更新.为此,我们在PEDOT@CdS/TiO₂/ITO可见光致更新电极表面旋涂明胶薄层,在保持电极良好的光致清洁和电化学传感性能同时,利用明胶薄层阻碍光催化产生的活性氧自由基扩散至细胞表面,显著降低了细胞损伤.此外,明胶优良的生物相容性有利于细胞的黏附及增殖.利用该电极,我们实现了人脐静脉内皮细胞（HUVECs）培养过程中,电极的在线更新以及细胞释放一氧化氮的实时监测.     

One‐Step Labeling of Collagen Hydrogels with Polydopamine and Manganese Porphyrin for Non‐Invasive Scaffold Tracking on Magnetic Resonance Imaging

Biomaterial scaffolds are the cornerstone to supporting 3D tissue growth. Optimized scaffold design is critical to successful regeneration, and this optimization requires accurate knowledge of the scaffold's interaction with living tissue in the dynamic in vivo milieu. Unfortunately, non‐invasive methods that can probe scaffolds in the intact living subject are largely underexplored, with imaging‐based assessment relying on either imaging cells seeded on the scaffold or imaging scaffolds that have been chemically altered. In this work, the authors develop a broadly applicable magnetic resonance imaging (MRI) method to image scaffolds directly. A positive‐contrast “bright” manganese porphyrin (MnP) agent for labeling scaffolds is used to achieve high sensitivity and specificity, and polydopamine, a biologically derived universal adhesive, is employed for adhering the MnP. The technique was optimized in vitro on a prototypic collagen gel, and in vivo assessment was performed in rats. The results demonstrate superior in vivo scaffold visualization and the potential for quantitative tracking of degradation over time. Designed with ease of synthesis in mind and general applicability for the continuing expansion of available biomaterials, the proposed method will allow tissue engineers to assess and fine‐tune the in vivo behavior of their scaffolds for optimal regeneration.     

质谱法定量测定高密度脂蛋白结合蛋白的糖基化水平

采用质谱法对4种高密度脂蛋白(HDL)的结合蛋白重组人载脂蛋白血清淀粉样蛋白A(SAA)、 α1-抗胰蛋白酶(A1AT)、 α2-人体血清糖蛋白(A2HSG)和A载脂蛋白C3(Apo C3)从蛋白质含量(蛋白的绝对定量)、 位点特异性糖基化(糖肽的相对定量)及聚糖位点占有率等方面进行了研究. 利用四极杆-飞行时间质谱仪(Q-TOF)测量糖蛋白标样酶解产物的二级质谱碎片离子, 用Byonic软件发现了新的糖基化位点信息, 即增加了原位点处聚糖糖型的种类. 对于A2HSG, 新增了N-糖基化156位点上的4种糖型, N-糖基化176位点上的6种糖型, O-糖基化319位点的4种O-聚糖和O-糖基化346位点上的1种糖型. 对于Apo C3, 只有O-糖基化94一个位点, 在此位点上新增了9种糖型. 同时, 调整了用于定量蛋白的多肽, 使得定量更加准确. 采用三重四极杆串级质谱仪(UPLC-ESI-QQQ)研究了4种结合蛋白中多肽和糖肽的多反应监测(MRM)行为, 并重新计算了每种聚糖的位点占有率, 优化了现有的定量方法.     

高效液相色谱/串联质谱法同时测定虾中氯霉素、甲砜霉素和氟甲砜霉素残留量

用高效液相色谱／串联质谱(LC／MS,／MS)同时测定虾中的氯霉素(CAP)、甲砜霉素(TAP)和氟甲砜霉素(FF)。均质后的虾样品,采用碱化乙酸乙酯提取。浓缩提取物经液．液分配(LLP)去除脂肪,C18固相萃取(SPE)柱净化后,采用LC／MS／MS电喷雾电离(ESI),负离子,多反应监测(MRM)模式检测,外标法定量。检出限为：氯霉素和氟甲砜霉素0．01ng/g;甲砜霉素为0．05ng／g。在添加浓度0．1～2．0ng/g范围内,氯霉素回收率为73．9％～96．0％;甲砜霉素回收率为78．6％～99．5％;氟甲砜霉素回收率为74．9％～103．7％;相对标准偏差(RSD)均小于6．4％。     

Kinetics of cationic photopolymerizations of divinyl ethers characterized using in situ Raman spectroscopy

In situ Raman spectroscopy experiments were used to determine effective kinetic propagation constants for a series of unsteady-state divinyl ether polymerizations at different isothermal temperatures and light intensities. Raman spectroscopy was found to be ideally suited for monitoring cationic photopolymerizations because the technique allows isothermal experiments to be performed with excellent time resolution and allows several spectral features to be observed simultaneously. In addition, the Raman experiments provided direct information about the vinyl bond concentration in situ as the reaction takes place. For these cationic photopolymerizations, the reaction rate and limiting conversion were both found to increase as the reaction temperature was increased. At all temperatures, the profile for the propagation rate constant, k_p, exhibited a dramatic increase at the start of the reaction, plateaued at a value between 10 and 40 l/mol s (depending upon temperature), and then decreased as the reaction reached a limiting conversion due to trapping of the active centers. Finally, the overall activation energy for polymerization was found to be 25.1 ± 6.1 kJ/mol. © 1996 John Wiley & Sons, Inc.