Data processing in on-line laser mass spectrometry of inorganic, organic, or biological airborne particles

A general solution for data processing of large numbers of micrometer- or submicrometer-particle mass spectra in aerosol analysis is described. The method is based on immediate evaluation of bipolar laser desorption ionization mass spectra acquired in an on-line (impact-free) time-of-flight instrument. The goal of the procedure is a characterization of the particle population under investigation in terms of chemical composition of particle classes, particle distributions, size distributions, and time courses, rather than an investigation of each individual particle. After automatic peak analysis of each newly acquired bipolar mass spectrum, the mass spectral information is statistically evaluated by a fuzzy clustering algorithm, providing for an immediate attribution of the particle to predefined particle classes. The particle distributions over these classes can be monitored as a function of time and particle size range. Definition of the particle classes as used for on-line evaluation is performed in an earlier step, either by manual approach, or by selection from a particle class database, or, as in most cases, by fuzzy clustering of a set of particle mass spectra from the population (the aerosol) under investigation. Definition of the particle classes is depending only on the distinguishability of the spectra patterns of different particles. It is not necessary for the clustering approach to fully “understand” the mass spectra. The range of possible applications of the method is therefore very broad. Particles dominated by inorganic components, as typically observed in aerosol chemistry for example, can be investigated the same way as organic particles (e.g., from smoke or automobile exhaust) or even biological particles such as bacteria, yeast, or pollen. The data processing method has been successfully applied in several fields of stationary applications and will be employed in mobile instruments for large scale field studies in atmospheric chemistry, engine combustion research, and the characterization of house dust.     

Characterizing uranium oxide reference particles for isotopic abundances and uranium mass by single particle isotope dilution mass spectrometry

Uranium and plutonium particulate test materials are becoming increasingly important as the reliability of measurement results has to be demonstrated to regulatory bodies responsible for maintaining effective nuclear safeguards. In order to address this issue, the Institute for Reference Materials and Measurements (IRMM) in collaboration with the Institute for Transuranium Elements (ITU) has initiated a study to investigate the feasibility of preparing and characterizing a uranium particle reference material for nuclear safeguards, which is finally certified for isotopic abundances and for the uranium mass per particle. Such control particles are specifically required to evaluate responses of instruments based on mass spectrometric detection (e.g. SIMS, TIMS, LA-ICPMS) and to help ensuring the reliability and comparability of measurement results worldwide. In this paper, a methodology is described which allows quantifying the uranium mass in single micron particles by isotope dilution thermal ionization mass spectrometry (ID-TIMS). This methodology is characterized by substantial improvements recently achieved at IRMM in terms of sensitivity and measurement accuracy in the field of uranium particle analysis by TIMS. The use of monodisperse uranium oxide particles prepared using an aerosol generation technique developed at ITU, which is capable of producing particles of well-characterized size and isotopic composition was exploited. The evidence of a straightforward correlation between the particle volume and the mass of uranium was demonstrated in this study. Experimental results have shown that the uranium mass per particle can be measured via the ID-TIMS method to a relative expanded uncertainty of about 10% (coverage factor k = 2). The availability of reliable and validated methods for the characterization of uranium particles is considered to be essential for the establishment of SI-traceable measurement results. It is therefore expected that the method developed in this study is valuable for the certification of particulate materials in which the isotopic composition and the content of uranium must be accurately known.     

On-line direct determination of the second virial coefficient of a natural polysaccharide using size-exclusion chromatography and multi-angle laser light scattering

By combining a size-exclusion chromatographic (SEC) separation and an on-line multi-angle light scattering (MALLS) analysis, we have elaborated an original methodology permitting on-line direct determination of the second virial coefficient of molar mass fractions of polydisperse polysaccharides. By assimilating the SEC-MALLS data to a batch mode acquisition, we have obtained on-line the complete Zimm plot of the eluted fractions, leading to knowledge of their weight-average molar mass Mw, radius of gyration r(g) and second virial coefficient A2. Our methodology was successfully applied to a iota carrageenan sample in LiCl 100 mM, EDTA 1 g/l.     

Real-time,single-particle measurements of ambient aerosols in Shanghai

As one of the major components of the earth’s atmosphere, airborne particulate matter (or aerosol) has strong effects on air quality, regional and global climate, and human health. In ambient atmosphere, the different sources and complex evolutionary history of aerosol particles make the study of their chemical and physical properties extremely challenging. The invention of an online single-particle aerosol mass spectrometer provides a powerful technique to determine the size and chemical composition of individual aerosol particles in real time. We deployed an aerosol time-of-flight mass spectrometer (ATOFMS) to carry out single particle measurement in the urban area of Shanghai in the past few years. In this review paper, we summarize our recent work on the identification of particle type, mixing state and aging process, and the application of the individual particle information to the source apportionment of primary aerosol, and the investigation of the formation mechanism of secondary aerosol in Shanghai. The special capabilities of single particle mass spectrometry are proven essential to these studies. Multi-functional technique combinations of ATOFMS with other state-of-art aerosol instruments are also discussed for future studies.     

单颗粒-电感耦合等离子体质谱(SP-ICP-MS)法数据处理算法的技术发展现状

单颗粒-电感耦合等离子体质谱法(Single particle-inductively coupled plasma-mass spectrometry, SP-ICP-MS)将高通量的粒子计数技术与ICP-MS时间分辨分析相结合,进行快速的单个颗粒逐一表征。此技术不仅可以获取单个颗粒的质量和粒径信息,还能获得溶解态和颗粒态的元素含量信息。经过二十余年的发展,SP-ICP-MS法正逐渐成为表征单颗粒的重要方法之一,其显著的技术优势获得了广泛关注,业内诸多学者对其仪器运行条件优化、数据采集方法、数据处理算法等关键技术方面的研究也取得了长足进展。本文综述了SP-ICP-MS表征纳米颗粒的技术路线、数据处理算法及研究进展,重点讨论了单颗粒定量分析的关键问题及其未来研究方向。但必须指出的是,当前业内在不同应用场景下所建立的颗粒事件鉴别算法、溶液信号的判定阈值算法、颗粒信号质量控制和精确的颗粒定量表征方法尚未明确统一,仍然缺乏足够普适的数据处理方式,这使得SP-ICP-MS技术在广泛的纳米颗粒应用中仍需面对准确定量的挑战。     

Advances in the analysis of proteins and peptides by capillary electrophoresis with matrix-assisted laser desorption/ionization and electrospray-mass spectrometry detection

High throughput, outstanding certainty in peptide/protein identification, exceptional resolution, and quantitative information are essential pillars in proteome research. Capillary electrophoresis (CE) coupled to mass spectrometry (MS) has proven to meet these requirements. Soft ionization techniques, such as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI), have paved the way for the story of success of CE-MS in the analysis of biomolecules and both approaches are subject of discussion in this article. Meanwhile, CE-MS is far away from representing a homogeneous field. Therefore the review will cover a vast area including the coupling of different modes of CE (capillary zone electrophoresis, capillary isoelectric foscusing, capillary electrochromatography, micellar electrokinetic chromatography, nonaqueous capillary electrophoresis) to MS as well as on-line preconcentration techniques (transient capillary isotachophoresis, solid-phase extraction, membrane preconcentration) applied to compensate for restricted detection sensitivity. Special attention is given to improvements in interfacing, namely addressing nanospray and coaxial sheath liquid design. Peptide mapping, collision-induced dissociation with subsequent tandem MS, and amendments in mass accuracy of instruments improve information validity gained from MS data. With 2-D on-line coupling of liquid chromatography (LC) and CE a further topic will be discussed. A special section is dedicated to recent attempts in establishing CE-ESI-MS in proteomics, in the clinical and diagnostic field, and in the food sector.     

Structural characterization of silicone polymers using compositional ultra-high performance liquid chromatography separation,electrospray ionization,and high resolution/accurate mass

In this article, the development of an on-line ultra-high performance liquid chromatography (UHPLC)-high resolution/accurate mass (HR/AM) methodology for timely characterization of a broad range of silicone polymers is described. Electrospray ionization was optimized for silicone polymers to enable the direct coupling of the separation with mass spectrometry. Reverse-phase UHPLC was optimized to provide a high resolution separation of oligomers by molar mass and by end-group composition. With the high resolution compositional separation, multiple pseudo-molecular ion species were indexed with retention time and assisted in rapid identification of oligomer species. Relative retention times of different oligomer end-group species separated based on polarity provided additional information to aid data interpretation. These additional pieces of information were combined with accurate mass to provide a high information content analysis in a relatively short time frame. This methodology was applied to polymer end-group characterization, as well as comparative analysis examples.     

On-line solid phase extraction fast liquid chromatography–tandem mass spectrometry for the analysis of bisphenol A and its chlorinated derivatives in water samples

In this study an on-line column-switching fast LC–MS/MS method was developed to analyze bisphenol A (BPA) and its chlorinated derivatives in water. Fast liquid chromatographic separation was performed on a C18 reversed phase column based on fused-core particle technology (2.7 μm particle size) providing analysis times shorter than 3 min and high peak efficiencies. The main benefit of this LC system is that it can easily be hyphenated to a conventional on-line preconcentration device allowing the direct analysis of water samples without any pretreatment at concentrations levels down to 60 ng L⁻¹ and preventing contaminations frequently reported in the analysis of BPA. This on-line SPE fast LC system was coupled to a triple quadrupole mass spectrometer operating in enhanced mass resolution mode (Q1 FWHM = 0.7 Th, Q3 FWHM = 0.1 Th) in order to minimize interferences and chemical noise. This highly sensitive and selective method was successfully employed to analyze BPA and its chlorinated derivatives in water samples.     

Advances in mass spectrometry-based post-column bioaffinity profiling of mixtures

In the screening of complex mixtures, for example combinatorial libraries, natural extracts, and metabolic incubations, different approaches are used for integrated bioaffinity screening. Four major strategies can be used for screening of bioactive mixtures for protein targets—pre-column and post-column off-line, at-line, and on-line strategies. The focus of this review is on recent developments in post-column on-line screening, and the role of mass spectrometry (MS) in these systems. On-line screening systems integrate separation sciences, mass spectrometry, and biochemical methodology, enabling screening for active compounds in complex mixtures. There are three main variants of on-line MS based bioassays: the mass spectrometer is used for ligand identification only; the mass spectrometer is used for both ligand identification and bioassay readout; or MS detection is conducted in parallel with at-line microfractionation with off-line bioaffinity analysis. On the basis of the different fields of application of on-line screening, the principles are explained and their usefulness in the different fields of drug research is critically evaluated. Furthermore, off-line screening is discussed briefly with the on-line and at-line approaches.     

飞行时间质谱仪新技术的进展及应用

本文介绍了近几年来发展应用于飞行时间质谱(TOF-MS)仪中的软电离技术。质子转移电离实现了可挥发性有机物的高灵敏度分析;真空紫外灯电离源体积小、简单,利于便携式仪器;电喷雾解吸电离在线、无损和灵敏度高,在公共安全方面具有很大的发展潜力,而且还可以直接用于活的生物体表面分析;大气压下的在线直接分析电离技术利用载气分子的激发态使得被分析化合物电离得到分子离子。针对不同的电离方法简单评述了其性能及应用,同时介绍了飞行时间质谱在串联方面的新发展,以及TOF-MS在仪器微型化方面的进展及其应用,并对飞行时间质谱仪今后的发展作了展望。     

Analysis of anions in ambient aerosols by microchip capillary electrophoresis

We describe a microchip capillary electrophoresis method for the analysis of nitrate and sulfate in ambient aerosols. Investigating the chemical composition of ambient aerosol particles is essential for understanding their sources and effects. Significant progress has been made towards developing mass spectrometry-based instrumentation for rapid qualitative analysis of aerosols. Alternative methods for rapid quantification of selected high abundance compounds are needed to augment the capacity for widespread routine analysis. Such methods could provide much higher temporal and spatial resolution than can be achieved currently. Inorganic anions comprise a large percentage of particulate mass, with nitrate and sulfate among the most abundant species. While ion chromatography has proven very useful for analyzing extracts of time-integrated ambient aerosol samples collected on filters and for semi-continuous, on-line particle composition measurements, there is a growing need for development of new compact, inexpensive approaches to routine on-line aerosol ion analysis for deployment in spatially dense, atmospheric measurement networks. Microchip capillary electrophoresis provides the necessary speed and portability to address this need. In this report, on-column contact conductivity detection is used with hydrodynamic injection to create a simple microchip instrument for analysis of nitrate and sulfate. On-column contact conductivity detection was achieved using a Pd decoupler placed upstream from the working electrodes. Microchips containing two Au or Pd working electrodes showed a good linear range (5-500 microM) and low limits-of-detection for sulfate and nitrate, with Au providing the lowest detection limits (1 microM) for both ions. The completed microchip system was used to analyze ambient aerosol filter samples. Nitrate and sulfate concentrations measured by the microchip matched the concentrations measured by ion chromatography.     

What does the future hold for top down mass spectrometry?

Mass spectrometry (MS) research has revolutionized modern biological and biomedical fields. At the heart of the majority of mass spectrometry experiments is the use of Bottom Up mass spectrometry methods where proteins are first proteolyzed into smaller fragments before MS interrogation. The advent of electron capture dissociation and, more recently, electron-transfer dissociation, however, has allowed Top Down (analysis of intact proteins) or middle down (analysis of large polypeptides) mass spectrometry to both experience large increases in development, growth, and overall usage. Nevertheless, for high-throughput large-scale proteomic studies, Bottom Up mass spectrometry has easily dominated the field. As Top Down mass spectrometry methodology and technology continue to develop, will it genuinely be able to compete with Bottom Up mass spectrometry for whole proteome analysis? Discussed here are the current approaches, applications, issues, and future view of high-throughput Top Down mass spectrometry.     

A new multi-axial particle shape factor--application to particle sampling

Because for a given sample size the sampling uncertainty increases with increasing particle mass, the mass of a representative sample depends on the particle mass during chemical, physical and biological analysis. Sampling theory can be used to formulate the quantitative relationship between the particle mass and the corresponding mass or weight of a representative sample. But in practice, especially for small particles, it is often easier to evaluate the particle size in dimension of length (e.g. μm) rather than in dimension of mass (e.g. μg). In order to be able to apply sampling theory to predict the mass or weight of a representative sample, a well-defined methodology that relates the mass of a particle to its size is required. We here propose a new multi-axial shape factor which requires information of multiple sizes of the particle of interest, whereas a uniaxial shape factor only needs one. In view of the information loss that is implicit in the use of a one-dimensional shape factor like the Brunton shape factor, the here-proposed new multi-axial shape is expected to perform better. Experimental data confirm the better performance of the new shape factor. A multi-segment generalisation of the new multi-axial shape factor is proposed.     

Improvements in ion signal reproducibility obtained using a homogeneous laser beam for on-line laser desorption/ionization of single particles

A major factor limiting on-line single particle mass spectrometry techniques from becoming more quantitative is the large shot-to-shot variability in ion intensities observed in the laser desorption/ionization (LDI) mass spectra.1,2 In previous work, lab-generated particles showed fluctuations of up to 152% in the absolute ion intensities in averaged spectra of 200-300 'identical' particles.2 Most of these fluctuations were attributed to inhomogeneities in the laser beam profile, leading to significant differences in the power each particle encountered depending on the position in the LDI laser beam where it underwent analysis. The goal of the work presented herein is to determine whether a fiber optic actually reduces the observed variability in single particle LDI mass spectral data. Initial results are presented for individual single component organic particles composed of 2,4-dihydroxybenzoic acid (2,4-DHB) analyzed using a low-power flat-top laser beam profile created by sending an ultraviolet (266 nm) DI laser through a fiber optic. Relative standard deviations of the total ion intensities for peaks in individual spectra are reduced to 31%. Single particle spectra, compared with and without laser homogenization at the same nominal laser fluence, show a marked enhancement. Specifically, the ion signal patterns of the 2,4-DHB particle spectra obtained using a homogenous LDI beam look identical to one another (i.e. only one particle type was produced with a commonly used neural network grouping algorithm), whereas without beam homogenization 25 different particle types (based on ion intensity patterns) were obtained. Future publications will explore more particle types and matrices but the initial results described herein are quite encouraging.     

Present state and future development of instrumental techniques for solution thermochemical (thermometric) analysis

A review of instruments for thermochemical (thermometric) solution analysis is given, including new types of isoperibol reaction of bomb calorimeters, the design of which can influence the future development of thermochemical measuring techniques. The new type of Dithermanal (Vaskut-EMG, Hungary, and Herrmann- Morris, France) controls the individual steps of the thermochemical analysis, adding memory and computing capabilities. Similar programming units with microprocessors for bomb and/or reaction calorimetry have been developed by Parr Comp. and Leco Comp. (USA). The latest modification of the Technicon flow analyzer permits on-line analysis of solid samples. For thermochemical titrations, the Sanda titrator (USA) and Vaskut automatic titrator (Hungary) are available. In the USA, the Tronac isothermal or isoperibol calorimeters are widely used. At the Technical University of Brno, ?SSR, smaller table instruments with a water-bath are applied for different modes of solution thermochemical analysis, and also for the reactions of solid samples in liquids. For the measurement of very small temperature differences, PTC thermistors are used. The possibilities of the future development of measuring techniques are outlined.     

A large computer system for on-line data acquistion and analysis of data from analytical instruments

Summary Experience with a unique large dual processor system designed to acquire and analyze data from about 140 instruments in a research installation will be described. The system has been in operation for nearly 4 years serving a variety of analytical instruments including gas chromatographs, liquid chromatographs, tensile testing machines, differential thermal analyzers, and other miscellaneous instruments. The system also serves a large community of scientific time-sharing users, a substantial portion of which reduce data and carry out scientific calculations on data acquired off-line on paper or magnetic tape. Unique features of the system are long distance transmission of analog data, retention of raw data for reanalysis, post processing of data into material balance, kinetic or statistical equations, interactive graphical display of the data acquired on-line and the ability to monitor instrument performance. Another unique features is the ability to develop new analysis programs in time-sharing without fear of jeopardizing the integrity of the data collection process.     

Copolymer solutions as separation media for DNA capillary electrophoresis

Capillary electrophoresis techniques offer high plate numbers and are highly suited for the efficient separations of a wide variety of chemical components in diverse matrices. Because of the small capillary and detection cell dimensions, together with the minute volumes of samples to be injected, sensitive detection schemes based on different physicochemical principles are being developed. One logical approach to increased sensitivity in capillary electrophoresis detection has been the development of chemiluminescence-based detectors. The development of on-line ultrasensitive chemiluminescence detection (referred to the concentration detection limit of nM order of magnitude or mass detection limit of amol order of magnitude) in capillary electrophoresis system is reviewed. The applications and limitations of the current detection methodology are briefly considered and future prospects for the development are discussed.     

甲醛监测方法及仪器

介绍了甲醛检测方法的新进展和常用的仪器,指出发展灵敏度高、选择性好、能够实现现场分析或长期稳定地实时在线监测的新方法和新仪器仍然是空气中甲醛监测研究的一个重要方向。     

Rapid protein identification using a microscale electrospray LC/MS system on an ion trap mass spectrometer

A methodology has been developed for the rapid identification of gel separated proteins. Following in gel protein digestion with trypsin, the resulting peptide mixture is analyzed by on-line liquid chromatography, electrospray mass spectrometry (LC/MS). The mass spectral data containing either accurate mass values or sequence specific fragment ion information is then matched to a database of known protein sequences. Key features of the LC/MS system are the use of a novel integrated, microscale LC column-electrospray interface and variable flow solvent delivery to optimize the efficiency of sample loading and gradient elution. With these enhancements, only 10 min is required to analyze each sample. The method is routine for sample amounts ranging from 50 to 500 fmol. The analysis parameters for the ion trap mass spectrometer have to be carefully adjusted in order to keep pace with the rapidly eluting LC peaks. Although designed for rapid LC separations, the integrated column-electrospray interface is also able to provide extended analyses of selected components using a technique known as “peak parking. ”     

Inverse 15N-metabolic labeling/mass spectrometry for comparative proteomics and rapid identification of protein markers/targets

The inverse labeling/mass spectrometry strategy has been applied to protein metabolic (15)N labeling for gel-free proteomics to achieve the rapid identification of protein markers/targets. Inverse labeling involves culturing both the perturbed (by disease or by a drug treatment) and control samples each in two separate pools of normal and (15)N-enriched culture media such that four pools are produced as opposed to two in a conventional labeling approach. The inverse labeling is then achieved by combining the normal (14)N-control with the (15)N-perturbed sample, and the (15)N-control with the (14)N-perturbed sample. Both mixtures are then proteolyzed and analyzed by mass spectrometry (coupled with on-line or off-line separation). Inverse labeling overcomes difficulties associated with protein metabolic labeling with regard to isotopic peak correlation and data interpretation in the single-experiment approach (due to the non-predictable/variable mass difference). When two data sets from inverse labeling are compared, proteins of differential expression are readily recognized by a characteristic inverse labeling pattern or apparent qualitative mass shifts between the two inverse labeling analyses. MS/MS fragmentation data provide further confirmation and are subsequently used to search protein databases for protein identification. The methodology has been applied successfully to two model systems in this study. Utilizing the inverse labeling strategy, one can use any mass spectrometer of standard unit resolution, and acquire only the minimum, essential data to achieve the rapid and unambiguous identification of differentially expressed protein markers/targets. The strategy permits quick focus on the signals of differentially expressed proteins. It eliminates the detection ambiguities caused by the dynamic range of detection. Finally, inverse labeling enables the detection of covalent changes of proteins responding to a perturbation that one might fail to distinguish with a conventional labeling experiment.