Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of phospholipase A(2) and fibrinolytic enzyme, two enzymes obtained from Chinese Agkistrodon blomhoffii Ussurensis venom

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used to analyze two enzymes, phospholipase AZ and fibrinolytic enzyme isolated from Chinese Agkistrodon blomhoffii Ussurensis venom. Using sinapinic acid as the matrix, positive ion mass spectra of the enzymes were obtained, In addition to the dominant protein [M+H](+) ions, multimeric and multiply charged ions were also observed in the mass spectra, The higher the concentration of the enzymes, the more multiply charged polymer and multimeric ions were detected, Our results indicate that MALDI-TOFMS can provide a rapid and accurate method for molecular weight determination of snake venom enzymes, Mass accuracies of 0.1 and 0.3 % were achieved by analysis of highly dialyzed phospholipase A2 and fibrinolytic enzyme, and these results are much better than those obtained using sodium dodecyl sulfate-palyacrylamide gel electrophoresis. MALDI-TOFMS thus provides a reliable method to determine the purity and molecular weight of these enzymes, which are of potential use as therapeutants, Copyright (C) 1999 John Wiley & Sons, Ltd.

Intensity exaggeration of endohedral metallofullerenes in laser-desorption-ionization time-of-flight mass spectrometry

The laser-desorption-ionization time-of-flight mass spectrometry and desorption-electron-ionization mass spectrometry were employed for the characterization of metallofullerenes extract. it was found that the relative intensities of metallofullerenes in this positive-ion, negative-ion LD-TOF MS and DEI MS were much different. This phenomenon should have relationship with the peculiar ionization energies and electron affinities of metallofullerenes.

Detection of specific noncovalent protein-fullerenols complexes by matrix-assisted laser desorption ionization mass spectrometry

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry is difficult for the characterization of noncovalent complexes hitherto because of the limitations in acidic matrix, sample preparation, laser-induced polymerization and adduct formation with matrix. Under our experimental conditions, sinapinic acid is used as a matrix, the specific noncovalent interactions of protein with fullerenols were observed by MALDI mass spectrometry. Some mass spectrometric features, such as mass shifts, broad adduct peaks and stoichiometries, showed that the specific non-covalent complexes between protein and fullerenols have been formed at a ratio of 1 : 4 for hemoglobin-fullerenols or 1 : 1 for myoglobin-fullerenols. The results implied that fullereneols could be used to protect partly hemoglobin from decomposition in acidic media, and therefore, it is possible to realize the molecular weight determination of a quaternary protein by MALDI mass spectrometry via the addition of specific organic compound in the matrix.

Matrixassisted laser desorption/ionization mass spectral study of saccharides

Matrixassisted laser desorption/ionization (MALDI) mass spectra of saccharides with three common matrixes were studied here. It is shown that Na+ and K+ play important roles in the determination of oligasaccharide molecule weight and the molecular weights (MW) of glucans(dextran) with MW more than 10 000 is determined successfully with the help of column chromatography, By comparing the effects of three matrixes in saccharide analysis and comparing positive-ion and negative-ion matrixassisted laser desorption/ionization mass spectra of oligo- and polysaccharides, the most suitable matrix for saccharide analysis, 2,5-dihydroxylbezonic acid (DHB), is selected and the ion formation processes of saccharides under matrix-assisted laser adsorption/ionization condition are clarified.

Synthesis and spectroscopic studies of endohedral metallofullerenes Tb@C-2n

Endohedral metallofullerenes Tb@C-2n were synthesized and extracted with high-yield by K-H carbon-are evaporation and an effective pyridine extraction technique at high-temperature high-pressure. Laser-desorption-ionization time-of-flight (LD-TOF) mass spectrometry, X-ray photoelectron spectroscopy (XPS), solid-state fluorescent emission spectroscopy and gas phase derivation reaction with the self-chemical ionization mass spectrometric ion system of vinyl acetate were employed for studying the electronic structures, fluorescent properties and gas phase reactivities of metallofullerenes Tb@C-2n. The experimental results suggest that endohedral metallofullerenes Tb@C-2n would have the approximate structures of Tb3+@C-2n(3-) similar to other metallofullerenes, good fluorescent emission properties and active reactivities in gas phase ion-molecular reactions.

A study of Aconitum alkaloids from aconite roots in Aconitum carmichaeli Debx using matrix-assisted laser desorption ionization mass spectrometry

In the present paper a study of C-19-diterpene type of aconitum alkaloids, extracted from aconite roots in Aconitum carmichaeli Debx has been made using matrix-assisted laser desorption/ionization time of Eight mass spectrometry (MALDI-TOFMS), The results demonstrated that the aconitum alkaloids from aconite roots can be determined simultaneously by this method, which was found to be superior to other analytical methods with regard to speed and sensitivity. Fourteen known aconitum alkaloids, including aconitines, benzoylaconitines and lipoaconitines, were assigned in the methanol extract and three compounds not reported before have been targeted separation. The evaluation of the efficiency of different extractions has been studied. These results suggested that the differences of the polarity and basicity of aconitine, and benzoylaconitines and lipoaconitines result from the C-8 constituent groups that are easily lost under MALDI, (C) 1998 John Wiley & Sons, Ltd.

Positive- and negative-ion matrix-assisted laser desorption/ionization mass spectrometry of saccharides

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) of oligosaccharides and polysaccharides has been investigated in detail, It is demonstrated that cationized species of oligosaccharides, [M+Na](+) and [M+K](+), are dominant products under the MALDI condition, and negative ions of oligosaccharides are not formed to any significant extent in this process, The molecular masses of polysaccharides are similarly determined by positive- and negative-ion MALDI-MS with the help of column chromatography. The distinction between positive- and negative-ion MALDI mass spectra of oligo-and polysaccharides indicates that the MALDI processes for saccharides vary with molecular mass. The matrix plays a more important role in the ionization process for oligosaccharides, while in the desorption process for polysaccharides. (C) 1998 John Wiley & Sons, Ltd.

Matrix-assisted laser desorption/ionization of ginsenosides

Molecular weight of 8 ginsenosides and the component of total saponions in American ginseng have been determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The average error of the molecular weight of each ginsenoside was found less than 0.05%. The results demonstrate that MALDI-MS is a very simple and useful method to measure the molecular weight of some high polar, thermal unstable small molecules with high sensitivity and reproducibility.

Behavior of silver cluster on laser desorption/ionization time-of-flight mass spectrometry

A successful analysis of silver was reported utilizing laser desorption/ionization time-of-flight mass spectrometry (LDI/TOF-MS) in this paper, The silver cluster ions Ag-n(+) and AgnO+ (n=2 similar to 5) were formed during laser desorption/ionization. In the presence of I-, K+ and Na+, the peaks corresponding, to the cluster ions [AgnIn-1](+) (n=2 similar to 6) and the adduct ions [AgI](+), [AgI]Na+ and [AgI]K+ were observed in the positive ion spectrum; the peaks corresponding to [AgnIn+1](-) (n=1 similar to 3) were found in the negative ion spectrum, all of which accompanied by sliver isotope distribution, The formation of silver cluster ions was accomplished through two-stage reaction: the first step was the generation of clusters, which was followed by the processes of photoionization and ion/molecule reaction.

3种羧酸类基质的基质辅助激光解吸/电离质谱行为

引言基质辅助激光解吸/电离质谱(MALDI-MS)这种新的“软电离”质谱技术，能够在短短10年间，得到广泛应用与迅速发展，很大程度上要归功于基质的辅助效应。基质在样品的解吸/电离过程中，起着关键作用。对基质本身MALDI-MS的研究，不仅可以、刻认识M

人参皂苷的基质辅助激光解吸质谱研究

利用MALDI-TOFMS测定了八种人参皂苷的分子量,并分析了西洋参总皂苷的组成.同时,进行了灵敏度实验,并探讨了基质及碱金属离子的影响,证明该方法灵敏度高,重复性好,结果准确.是测定极性小分子分子量的有效方法.

Formation and special solubility of endohedral thulium metallofullerenes

Endohedral metallofullerenes Tm@C-2n were synthesized with the carbon-arc discharge method of activating the Tm2O3-containing graphite anode in situ and back-burning technique, Toluene soxhlet extraction, toluene and pyridine high-temperature and high-pressure extraction technique were employed for the effective extraction of Tm@C-2n. The formation of Tm@C-2n was confirmed by LD-TOF and DEIMS characterization. The mass spectrometric results indicate that Tm@C-2n have a better solubility in toluene and worse solubility in pyridine than other metallofullerenes.

一种新型阻性板室（RPC）探测器的研制

气体探测器在核物理及粒子物理领域有着广泛的应用，尤其是阻性板室探测器，以其低廉的造价、优异的性能在各个大型实验室中都发挥着重要的作用。日本国家高能物理实验室(KEK)的Belle探测器，斯坦福大学斯坦福直线加速器中心(SLAC)的BaBar大型探测器，欧洲核子中心(CERN)的大型强子对撞机(LHC)的CMS与ATLAS谱仪，北京高能物理研究所(IHEP)的BESⅢ谱仪都使用阻性板室作为触发系统或缪子探测器。 随着HIRFL-CSR项目的完工。RIBLL2开展高能放射性束物理实验成为可能，而为了鉴别高能重离子碰撞实验中出射的高能轻粒子，有必要测量粒子的飞行时间(TOF)。为此，我们研制了单隙定时RPC，并作了简单性能测试。在CSR实验终端上，它可以作为传统塑料闪烁晶体+PMT时间探测器的替代，成为反应出射产物测量的大型探测器系统(4π)中的重要成员。 本论文对RPC的发展历史以及被广泛应用的原因做了深入而详细的研究，并对RPC的结构、性能、工作模式、特点、气体系统等进行了深入探讨。研制了一套单气隙RPC探测器样机，测设了一些相关性能，并对所遇到问题的产生原因以及解决方案进行了分析。为了测试RPC的需要，搭建了一套多组分工作气体配气系统，论文中还对包括质量流量计的选定，相关配件的选择，以及后期整个配气系统的组装与调试进行了详细描述

Biochemical and electron microscopic characterization of the F1F0 ATP Synthase from the hyperthermophilic eubacterium Aquifex aeolicus

The F1F0 ATP synthase has been purified from the hyperthermophilic eubacterium Aquifex aeolicus and characterized. Its subunits have been identified by MALDI-mass spectrometry through peptide mass fingerprinting and MS/MS. It contains the canonical subunits alpha, beta, gamma, delta and epsilon of F-1 and subunits a and c of F-0. Two versions of the b subunit were found, which show a low sequence homology to each other. Most likely they form a heterodimer. An electron microscopic single particle analysis revealed clear structural details, including two stalks connecting F-1 and F-0. In several orientations the central stalk appears to be tilted and/or kinked. It is unclear whether there is a direct connection between the peripheral stalk and the 6 subunit. (c) 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

海洋环境中有机锡的形态分析及其代谢降解研究

有机锡化合物被广泛用作塑料制品中的稳定剂、船舶油漆的防污剂、工业催化剂、农林业杀虫杀菌剂以及用于木材的防腐保存等，已经引起严重的环境污染。世界上许多国家纷纷制定相应的法规对其使用加以禁止或限制。我国目前还没有明确的限制有机锡使用的法律法规，缺少有机锡污染的第一手资料，更没有长期的控制、监测与研究计划。由于有机锡的种类繁多，理化性质存在差别，所以在提取、分离和测定中均存在较大的困难。从我国这方面己有的工作来看，缺乏各种高选择性的分离方法和高灵敏度的检测方法是制约这项研究广泛开展的原因之一。有机锡的痕量与超痕量分析技术是当今环境和食品安全分析领域的前沿技术。 本论文利用高效液相色谱和电感耦合等离子体质谱联用技术建立了海洋环境中多种有机化合物的同时快速检测方法；发展了多种海洋环境样品中有机锡的前处理技术；研究了有机锡在海洋生物中的分布、代谢及降解过程中化学形态的变化；同时发展了海洋环境中多种痕量元素的快速检测方法。所建立的高效液相色谱和电感耦合等离子体质谱联用技术可同时、快速分析5种有机锡的形态(三甲基锡TMT、二苯基锡DPhT、二丁基锡DBT、三丁基锡TBT和三苯基锡TPhT)，其检出限均低于0.3μg/L。 用所建立方法对南海海洋生物样品中的有机锡污染进行了研究，利用SPSS软件对检测结果进行了探讨，发现在所研究海洋生物样品的97.2%中可检出丁基锡和苯基锡化合物，其浓度分布处于该化合物检出限～1487.8ng/g范围内。其中，贝类样品中总有机锡的平均浓度为416.9ng/g，远远高于鱼类样品中总有机锡的平均浓度（211.9ng/g）。海洋生物中存在高浓度的有机锡说明本海域有机锡污染严重，已经对生态环境造成了严重影响，危害到人类生活。其主要的污染源是防污涂料的应用，目前紧迫的问题是采取必要的措施来控制有机锡的使用。 本工作建立了海水样品和沉积物样品中五种有机锡的简单快速萃取方法。采用加入2%的环庚三烯酚（tropolone）的二氯甲烷CH2Cl2对海水中的有机锡进行萃取，大大提高了有机锡的萃取率，减少了萃取的时间，二苯基锡（DPhT）、二丁基锡（DBT）、三丁基锡（TBT）和三苯基锡（TPhT）的萃取率均在80%以上，仅三甲基锡（TMT）的萃取率较低（在50%左右），究其原因，可能是因为在萃取的过程中三甲基锡（TMT）产生了降解。采用流动相和0.2%环庚三烯酚酮（tropolone）对沉积物国际标准物质PACS-2进行超声萃取及高速离心后，用所建方法进行了分析。结果表明，测定值与标准值吻合。研究表明，所建立的方法可用于实际环境沉积物中有机锡的形态分析。 本文建立了流动注射与电感耦合等离子体质谱联用技术直接同时测定海水中多种痕量元素的方法。该方法采用痕量进样技术，能够有效地减少海水中Na，Mg, Ca和Cl等大量基体元素对待测痕量元素测定的干扰，减少这些元素在电感耦合等离子体采样锥上的盐沉积，可以同时测量海水中的V、Cr、Mn、Fe、Co、Ni、Cu、Zn、As、Mo、Cd、Pb，Hg和U等痕量级元素。用所建的方法测定南海海域海水中的重金属元素，发现Cd，Cr，As等有毒有害元素的污染很轻，均符合Ⅰ级海水的限量。 在海洋沉积物样品处理研究中，本工作改进了不需要赶走HF酸就可以对沉积物消解完全的密闭容器消解法，由于减少了赶走HF酸的步骤，使消解的时间由原来的二十个小时降低为十个小时，大大降低了消解的时间。采用该样品消解方法，并用ICP-MS测定了南黄海海域沉积物中锡及其他重金属元素的含量。建立了微波消解-ICP-MS测定海洋生物中锡、砷、镉、汞及铅等有害重金属元素的分析方法，并用于南黄海7个及南海海域29个海产品中的测定。测定结果表明海洋生物中上述有毒有害元素有不同程度的超标问题；不同种类，不同产地的海洋生物中重金属元素的含量有一定的差别，这些研究结果为海产品安全质量控制提供了有价值的科学信息。 在上述各章工作的基础上，本文研究了有机锡在海洋生物中的分布、代谢及降解过程，并初步建立了高效液相-电喷雾-飞行时间质谱（LC-APCI-TOF-MS）测定有机锡的方法，可对未知的有机锡化合物进行结构表征。有机锡在贝类中不同的组织显示，其内脏中有机锡的含量高于肌肉中有机锡含量。常规的煮、炸、蒸及微波的烹饪方式并不能降解海产品中的有机锡化合物。