Activation of large ions in FT-ICR mass spectrometry

The advent of soft ionization techniques, notably electrospray and laser desorption ionization methods, has enabled the extension of mass spectrometric methods to large molecules and molecular complexes. This both greatly extends the applications of mass spectrometry and makes the activation and dissociation of complex ions an integral part of these applications. This review emphasizes the most promising methods for activation and dissociation of complex ions and presents this discussion in the context of general knowledge of reaction kinetics and dynamics largely established for small ions. We then introduce the characteristic differences associated with the higher number of internal degrees of freedom and high density of states associated with molecular complexity. This is reflected primarily in the kinetics of unimolecular dissociation of complex ions, particularly their slow decay and the higher energy content required to induce decomposition--the kinetic shift (KS). The longer trapping time of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) significantly reduces the KS, which presents several advantages over other methods for the investigation of dissociation of complex molecules. After discussing general principles of reaction dynamics related to collisional activation of ions, we describe conventional ways to achieve single- and multiple-collision activation in FT-ICR MS. Sustained off-resonance irradiation (SORI)--the simplest and most robust means of introducing the multiple collision activation process--is discussed in greatest detail. Details of implementation of this technique, required control of experimental parameters, limitations, and examples of very successful application of SORI-CID are described. The advantages of high mass resolving power and the ability to carry out several stages of mass selection and activation intrinsic to FT-ICR MS are demonstrated in several examples. Photodissociation of ions from small molecules can be effected using IR or UV/vis lasers and generally requires tuning lasers to specific wavelengths and/or utilizing high flux, multiphoton excitation to match energy levels in the ion. Photodissociation of complex ions is much easier to accomplish from the basic physics perspective. The quasi-continuum of vibrational states at room temperature makes it very easy to pump relatively large amounts of energy into complex ions and infrared multiphoton dissociation (IRMPD) is a powerful technique for characterizing large ions, particularly biologically relevant molecules. Since both SORI-CID and IRMPD are slow activation methods they have many common characteristics. They are also distinctly different because SORI-CID is intrinsically selective (only ions that have a cyclotron frequency close to the frequency of the excitation field are excited), whereas IRMPD is not (all ions that reside on the optical path of the laser are excited). There are advantages and disadvantages to each technique and in many applications they complement each other. In contrast with these slow activation methods, the less widely appreciated activation method of surface induced dissociation (SID) appears to offer unique advantages because excitation in SID occurs on a sub-picosecond time scale, instantaneously relative to the observation time of any mass spectrometer. Internal energy deposition is quite efficient and readily adjusted by altering the kinetic energy of the impacting ion. The shattering transition--instantaneous decomposition of the ion on the surface--observed at high collision energies enables access to dissociation channels that are not accessible using SORI-CID or IRMPD. Finally, we discuss some approaches for tailoring the surface to achieve particular aims in SID.     

Collisional activation of peptide ions in FT-ICR mass spectrometry

In the last decade, the characterization of complex molecules, particularly biomolecules, became a focus of fundamental and applied research in mass spectrometry. Most of these studies utilize tandem mass spectrometry (MS/MS) to obtain structural information for complex molecules. Tandem mass spectrometry (MS/MS) typically involves the mass selection of a primary ion, its activation by collision or photon excitation, unimolecular decay into fragment ions characteristic of the ion structure and its internal excitation, and mass analysis of the fragment ions. Although the fundamental principles of tandem mass spectrometry of relatively small molecules are fairly well-understood, our understanding of the activation and fragmentation of large molecules is much more primitive. For small ions, a single energetic collision is sufficient to dissociate the ion; however, this is not the case for complex molecules. For large ions, two fundamental limits severely constrain fragmentation in tandem mass spectrometry. First, the center-of-mass collision energy-the absolute upper limit of energy transfer in a collision process-decreases with increasing mass of the projectile ion for fixed ion kinetic energy and neutral mass. Secondly, the dramatic increase in density of states with increasing internal degrees of freedom of the ion decreases the rate of dissociation by many orders of magnitude at a given internal energy. Consequently, most practical MS/MS experiments with complex ions involve multiple-collision activation (MCA-CID), multi-photon activation, or surface-induced dissociation (SID). This review is focused on what has been learned in recent research studies concerned with fundamental aspects of MCA-CID and SID of model peptides, with an emphasis on experiments carried out with Fourier transform ion cyclotron resonance mass spectrometers (FT-ICR MS). These studies provide the first quantitative comparison of gas-phase multiple-collision activation and SID of peptide ions. Combining collisional energy-resolved data with RRKM-based modeling revealed the effect of peptide size and identity on energy transfer in collisions-very important characteristics of ion activation from fundamental and the analytical perspectives. Finally, the combination of FT-ICR with SID was utilized to carry out the first time-resolved experiments that examine the kinetics of peptide fragmentation. This has lead to the discovery that the time-dependence of ion dissociation varies smoothly up to a certain collision energy, and then shifts dramatically to a time-independent, extensive dissociation. This near-instantaneous "shattering" of the ion generates a large number of relatively small fragment ions. Shattering of ions on surfaces opens up a variety of dissociation pathways that are not accessible with multiple-collision and multiphoton excitation.     

New separation tools for comprehensive studies of protein expression by mass spectrometry

Mass spectrometry has emerged as a core technique for protein identification and characterization because of its high sensitivity, accuracy, and speed of analysis. The most widespread strategy for studying global protein expression in biological systems employs analytical two-dimensional polyacrylamide gel electrophoresis (2D PAGE) followed by enzymatic degradation of isolated protein spots, peptide mapping, and bioinformatics searches. Using this method, thousands of proteins can be resolved in a gel and their expression quantified. However, certain types of proteins possessing important cellular functions are not easily analyzed using this strategy. These proteins include membrane, low copy number, highly basic, and very large (> 150 kDa) and small (< 10 kDa) proteins. To meet the growing need to simultaneously monitor all types of proteins in a biological system, new separation strategies have emerged that are amenable to hyphenation to mass spectrometric techniques. This article will review these new techniques and examine their usefulness in studies of protein expression.     

Hydrogen exchange mass spectrometry for the analysis of protein dynamics

Hydrogen exchange coupled to mass spectrometry (MS) has become a valuable analytical tool for the study of protein dynamics. By combining information about protein dynamics with more classical functional data, a more thorough understanding of protein function can be obtained. In many cases, protein dynamics are directly related to specific protein functions such as conformational changes during enzyme activation or protein movements during binding. The method is made possible because labile backbone hydrogens in a protein will exchange with deuterium atoms when the protein is placed in a D2O solution. The subsequent increase in protein mass over time is measured with high-resolution MS. The location of the deuterium incorporation is determined by monitoring deuterium incorporation in peptic fragments that are produced after the labeling reaction. In this review, we will summarize the general principles of the method, discuss the latest variations on the experimental protocol that probe different types of protein movements, and review other recent work and improvements in the field.     

Charge derivatization of peptides for analysis by mass spectrometry

The analysis of peptide derivatives by fast atom bombardment, liquid secondary-ionization mass spectrometry, plasma desorption, electrospray ionization, and matrix-assisted laser desorption/ionization is reviewed. The fragmentation patterns of peptides and of charge-derivatized peptides are compared, and the proposed fragment ion structures are summarized. A variety of derivatization approaches and the distinguishing features of mass spectra produced from these derivatives are described. The most promising derivatization approaches are evaluated, and the strengths and limitations of these approaches are discussed.     

Reliability analysis for thermal cutting method based non-explosive separation device

In order to increase the reliability of a separation device for a small satellite, a new non-explosive separation device is invented. This device is activated using a thermal cutting method with a Ni-Cr wire. A reliability analysis is carried out for the proposed non-explosive separation device by applying the Fault tree analysis (FTA) method. In the FTA results for the separation device, only ten single-point failure modes are found. The reliability modeling and analysis for the device are performed considering failure of the power supply, the Ni-Cr wire burns failure and unwinds, the holder separation failure, the balls separation failure, and the pin release failure. Ultimately, the reliability of the proposed device is calculated as 0.999989 with five Ni-Cr wire coils.     

A new detector for mass spectrometry: direct detection of low energy ions using a multi-pixel photon counter

A new type of ion detector for mass spectrometry and general detection of low energy ions is presented. The detector consists of a scintillator optically coupled to a single-photon avalanche photodiode (SPAD) array. A prototype sensor has been constructed from a LYSO (Lu(1.8)Y(0.2)SiO(5)(Ce)) scintillator crystal coupled to a commercial SPAD array detector. As proof of concept, the detector is used to record the time-of-flight mass spectra of butanone and carbon disulphide, and the dependence of detection sensitivity on the ion kinetic energy is characterised.     

偏二甲肼自氧化产物的气质联用分析

本文应用GC/MS系统对黄色偏二甲肼进行了分析,实验合成了亚硝基二甲胺、四甲基四氮烯,从而确定了偏二甲肼发黄变质的主要产物,并推测偏二甲肼的自氧化机理。     

基于质谱的核酸指纹识别技术

基因转录是继基因组计划完成后基因功能研究的基本内容。基因转录研究迫切需要有新方法的支持。人们从蛋白的指纹分析技术中得到启发,开发基于质谱分析的核酸指纹识别技术。通过对核酸进行酶解处理,使其核酸片段分子量位于质谱检测的范围内,通过高分辨质谱技术获得所检测酶解核酸片段的分子量列表;利用基因组研究所获得的大量数据,通过计算机技术,模拟产生核酸酶解产物数据库,通过独特的算法以及基于这些算法开发的工具软件,把酶解的核酸质谱数据和虚拟的核酸酶解产物数据库进行比对,以发现和鉴别核酸。随着这一技术的逐渐应用和发展,将来还可能直接建立基于质谱数据的核酸数据库。相信这一技术在转录组研究中将发挥重要作用。     

Preparation of cells cultured on silicon wafers for mass spectrometry analysis

The distribution of specific atoms and molecules within living cells is of high interest in bio-medical research. Laser secondary neutral mass spectrometry (laser-SNMS) and time-of-flight secondary ion mass spectrometry (TOF-SIMS) detect atoms with high sensitivity and spatial resolution. The application of these methods to cultured cells requires special preparation techniques preserving morphological and chemical integrity of the living cells. The cells should, therefore, be grown on a conducting material preventing charging of the sample during ion bombardment. Silicon is currently used as the preferred support material for non-biological samples in mass spectrometry. This study investigates (1) the influence of silicon surfaces on cell growth and (2) the suitability of a sandwiched, rapid freezing method to analyse transmembrane ion gradients. Human melanoma cells were grown on silicon with polished or etched surfaces. Growth kinetics were studied using the Sulforhodamine-B assay. Number, shape, and morphology of the cells were assessed by epifluorescence microscopy of calcein AM- and DAPI-stained cells. Cells were subjected to rapid freezing, freeze-fracturing, and freeze-drying prior to analysis by TOF-SIMS and laser-SNMS. While cell numbers and morphology on the rough silicon wafers were impaired, morphology and growth kinetics of cells on polished silicon were identical to control cells on cell culture tested polystyrene. TOF-SIMS and laser-SNMS resulted in high-resolution elemental images and mass spectra. Measurement of the intracellular Na+ and K+ concentrations revealed a ratio as observed in living cells. In conclusion, culturing cells on polished silicon wafers followed by sandwiched, rapid freezing is an adequate preparation method to study intracellular ion distribution with mass spectrometry.     

生物质谱在基因组学研究中的应用

综述生物质谱用于分析核糖核酸的应用进展。概述电喷雾电离质谱(ESI-MS)和基质辅助激光解吸电离质谱(MALDI-MS)的原理及它们在核酸分析中的应用。总结质谱用于单核苷酸多态性分型分析(SNPgenotyping);对短的串联重复序列(STR)的分析;对寡核苷酸片断的序列分析等三个方面的研究成果。提出质谱用于基因组学研究存在的问题,并展望生物质谱未来的发展方向。     

基于ADAMS的运载火箭分离装置动力学研究

分离装置是运载火箭中的一个重要部件,直接影响运载器飞行任务成败,以系统多体动力学理论为基础,基于动力学仿真软件ADAMS建立了分离动力学仿真模型,对分离过程中导向机构的参数进行了研究,分析结构设计参数对分离过程的影响规律,分剐就分离导向机构支柱刚度、高度、滚轮与载荷部之间预压力对分离结果的影响进行了研究.结果表明,当刚度和高度的取值越大时,分离期间的最小分离间隙也就越大,对火箭与载荷部的成功分离越有利;预压力对分离机构的影响较小.     

A two-channel device for time-of-flight-based event selection in fast neutron spectrometry

A device that uses a time interval meter to provide concurrent recording of spectra from two detectors containing information on the radiation type and their correlation form is described. The block and schematic diagrams of the device are given, and versions of its application are discussed.     

Time-of-flight mass spectrometry for time-resolved measurements

A time-resolved time-of-flight mass spectrometer (TOF-MS) that can simultaneously monitor multiple species on the millisecond time scale has been constructed. A pulsed photolysis laser is used to initiate reaction, and then via a pinhole the reaction mixture is sampled by the TOF-MS. The ions are created by photoionization via either a discharge lamp or a pulsed laser. Comparison between the two ionization sources showed that the laser is at least an order of magnitude more efficient, based on the time to accumulate the data. Also, unlike the continuous lamp the pulsed laser is not mass limited. Frequency tripling the 355 nm output of a Nd:YAG laser provided a convenient laser ionization source. However, using a dye laser provided an equally intense laser ionization source with the ability to tune the vacuum ultraviolet (vuv) light. To show the versatility of the system the kinetics of the reaction of SO and ClSO radicals with NO(2) were simultaneously measured, and using the dye laser the vuv light was tuned to 114 nm in order to observe H(2)CO being formed from the reaction between CH(3)CO and O(2).     

Intact-protein based sample preparation strategies for proteome analysis in combination with mass spectrometry

The complexity of tissue and cell proteomes and the vast dynamic range of protein abundance present a formidable challenge for analysis that no one analytical technique can overcome. As a result, there is a need to integrate technologies to achieve the high-resolution and high-sensitivity analysis of complex biological samples. The combined technologies of separation science and biological mass spectrometry (Bio-MS) are the current workhorse in proteomics, and are continuing to evolve to meet the needs for high sensitivity and high throughput. They are relied upon for protein quantification, identification, and analysis of post-translational modifications (PTMs). The standard technique of two dimensional poly-acrylamide gel electrophoresis (2D PAGE) offers relatively limited resolution and sensitivity for the simultaneous analysis of all cellular proteins, with only the most highly abundant proteins detectable in whole cell or tissue-derived samples. Hence, many alternative strategies are being explored. Numerous sample preparation procedures are currently available to reduce sample complexity and to increase the detectability of low-abundance proteins. Maintaining proteins intact during sample preparation has important advantages compared with strategies that digest proteins at an early step. These strategies include the ability to quantitate and recover proteins, and the assessment of PTMs. A review of current intact protein-based strategies for protein sample preparation prior to mass spectrometry (MS) is presented in the context of biomedically driven applications.     

磁分离水处理装置的研制

为了实现富营养化水体的根治目的,在分析水处理工艺的基础上,结合磁分离技术设计了一种高效率的水处理装置,并且已经获得了国家专利1。在实际试验中验证了该装置的处理效果,经过处理的水体达到国家三类水质标准。     

Structural investigation of bacterial lipopolysaccharides by mass spectrometry and tandem mass spectrometry

Mass spectrometric studies are now playing a leading role in the elucidation of lipopolysaccharide (LPS) structures through the characterization of antigenic polysaccharides, core oligosaccharides and lipid A components including LPS genetic modifications. The conventional MS and MS/MS analyses together with CID fragmentation provide additional structural information complementary to the previous analytical experiments, and thus contribute to an integrated strategy for the simultaneous characterization and correct sequencing of the carbohydrate moiety. © 2010 Wiley Periodicals, Inc., Mass Spec Rev 29:606–650, 2010     

基于虚拟仪器的谐波测试分析系统的设计

随着电力电子技术的飞速发展,非线性装置被广泛应用于电力牵引系统,这就不可避免地引发了谐波危害问题.本文提出了一种有效的基于虚拟仪器的电力牵引系统谐波的测试分析系统的设计开发方案,可以对电网输出、变压器输出.逆变器输出三个主要环节的电力参数进行谐波测试分析,可以得出可靠的结论和制定有效的防治措施.