间歇电喷雾的初步研究

高灵敏度是电喷雾质谱应用最重要的需要之一[1].虽然电喷雾与基体辅助激光解吸技术的发展已经极大地提高了生物质谱的分析性能[2],但是这样的灵敏度对于许多生物样品的测试还是远远不够的,而现在采用了间歇电喷雾的方法,将灵敏度提高到了500 amol.这种质谱方法显著地改善了信噪比,提高了溶解、输出和离子化的效率,降低了样品消耗和仪器污染.间歇电喷雾的方法很容易在各种质谱仪上实现,尤其适合分子较大的蛋白质的质谱测定.     

蛋白质非共价化合物的电喷雾质谱研究

介绍了近几年来利用电喷雾离子化质谱技术研究蛋白质的基本构象及蛋白质非共价化合物的基本化学信息的进展。许多研究表明,电喷雾质谱在研究蛋白质的复杂结构和功能方面有十分广阔的前景。文中列举了若干实例说明了电喷雾离子化质谱技术在分析蛋白质非共价化合物的应用和一般方法。     

基质辅助激光解吸离子化质谱及其在生物大分子分析中的应用

近年来质谱离子化技术方面有两项重要成果：基质辅助激光解吸离子化(matrix-assisted laser desorption ionization，MALDI)和电喷雾离子化(electrospray ionization，ESI)。MALDI和ESI的应用使质谱在生物大分子研究方面取得重大突破。本文仅就MALDI的原理、特点、样品准备方法、基质的选择、仪器条件及其在生物大分子应用方面的最新进展进行简要的综述。     

基于双极性离子阱内质子转移反应的多肽离子电荷调控方法研究

电喷雾是质谱分析多肽和蛋白质等生物分子时最常用的离子化技术。电喷雾产生的多肽和蛋白质离子通常带有多电荷,从而产生多个谱峰,导致谱图复杂。通过气相离子/离子间的质子转移反应可以有效调节经电喷雾离子化后蛋白和多肽等生物大分子的电荷价态,简化谱图,对于复杂蛋白和多肽生物样品的分析具有重要意义。本研究通过研制的一套双极性离子阱质谱装置,利用阱内正、负离子间的质子转移反应,实现了多肽离子的电荷调控。以氧化型谷胱甘肽、缩宫素、强啡肽分子作为典型样品对方法的检测效果进行了验证。结果表明,此方法可去除多肽正离子中多余的正电荷,当注入足够的负离子进行反应时,可以将带多个电荷的多肽阳离子的价态降至最低,从而有效简化谱图。     

生物质谱

质谱已成为生物和生物化学研究的一个重要的分析工具,特别是在蛋白质组学研究的作用更显突出.它的分析速度、准确性和灵敏度都是传统分析技术所不可比拟的.主要介绍了两种近年来发展最为迅速、应用最为广泛的软离子化质谱技术:即基体辅助激光解吸离子化质谱(MALDI-MS)和电喷雾离子化质谱(ESI-MS)的原理、技术的最新进展,并简单介绍了它们在蛋白质和多肽分析中的应用.     

质谱电喷雾离子源中电化学与电晕放电氧化还原反应的研究进展

质谱电喷雾离子化过程中包含两类氧化还原反应:电化学氧化还原和电晕放电氧化还原。一方面,这两类反应干扰谱图解析、降低分析物的检测灵敏度;另一方面,利用氧化还原的特性可发展新型离子源,提高电喷雾离子化过程中难离子化化合物的离子化效率,研究蛋白质相互作用等。该文系统地介绍了国内外对于电化学氧化还原反应和电晕放电氧化还原反应的最近研究进展,主要包括此两类反应的弊端、应用价值,以及控制两类反应的方法。最后总结了区分两种反应的方法,并对电喷雾离子源的发展进行了展望。     

基质辅助激光解吸离子化质谱及其在生物大分子分析中的应用

近年来质谱离子化技术方面有两项重要成果:基质辅助激光解吸离子化(matrix-assisted laser desorptionionization,MALDI)和电喷雾离子化(electrospray ionization,ESI)。MALDI和ESI的应用使质谱在生物大分子研究方面取得重大突破。本文仅就MALDI的原理、特点、样品准备方法、基质的选择、仪器条件及其在生物大分子应用方面的最新进展进行简要的综述。     

液相色谱-离子阱质谱对水华蓝藻提取物中微囊藻毒素的定性研究

本文研究了野外蓝藻水华样品甲醇提取物的液相色谱和电喷雾质谱特征图谱,利用电喷雾离子化-串联质谱(ESI-MSn)离子阱技术对特征图谱中主要微囊藻毒素的分子离子峰进行了二级质谱分析,获得相应的子离子质谱图,并对其进行了结构解析,确定了野外蓝藻水华样品中微囊藻毒素种类,为准确鉴定和分析水华蓝藻中微囊藻毒素提供了基础.     

毛细管电泳-电喷雾质谱联用技术及其在蛋白质分析领域中的应用

综述了毛细管电泳与电喷雾质谱联用的接口技术、分离模式及其在蛋白质分析领域中的应用，特别是毛细管等电聚焦与电喷雾质谱联用在蛋白质组学中研究进展。     

水凝胶涂覆纸喷雾离子化用于生物流体样品直接质谱分析

基于琼脂糖水凝胶涂覆的纸基质,实现了纸喷雾离子化,建立了一种新的纸喷雾离子化质谱技术,直接分析生物流体样品。利用纸表面涂覆的水凝胶材料限制液滴的扩散,使互不相溶的喷雾溶剂液滴与生物样品液滴之间形成液液萃取,提高了检测灵敏度。同时通过限制液滴扩散,减少了喷雾溶剂挥发面积,降低了挥发速度,获得了时间更长的稳定质谱信号,可对生物流体样品中的低丰度药物快速、准确定量。相较于常规纸喷雾离子化,该方法喷雾时间延长近10倍,分析物信号提高近100倍,定量分析全血和尿液中的药物,检出限可降低7~33倍。方法材料制备简单,方便快捷,灵敏度高,在现场临床诊断上具有很大的应用潜力。     

Ambient imaging mass spectrometry by electrospray ionization using solid needle as sampling probe

Although being an atmospheric pressure ion source, electrospray ionization (ESI) has rarely been used directly for ambient imaging mass spectrometry because the sample has to be introduced as liquid solution through the capillary. Instead of capillary, probe electrospray ionization (PESI), which has been developed recently, uses a solid needle as the sampling probe, as well as the electrospray emitter, and has been applied not only for liquid solutions but also for the direct sampling on wet samples. Biological tissues are composed of cells that contain 70–90% water, and when the surface is probed by the needle tip, the biological fluid adhering to the needle can be electrosprayed directly or assisted by additional solvent added onto the needle surface. Here, we demonstrate ambient imaging mass spectrometry of mouse brain section using PESI, incorporated with an auxiliary heated capillary sprayer. The solvent vapor generated from the sprayer condensed on the needle tip, re‐dissolving the adhered sample, and at the same time, providing an indirect means for needle cleaning. The histological sections were prepared by fixation using paraformaldehyde, and the spatial analysis was automated by maintaining an equal sampling depth into the sample in addition to raster scan. Phospholipids and galactosylceramides were readily detected from the mouse brain section in the positive ion mode, and were mapped with 60 µm lateral resolution to form mass spectrometric images. Copyright © 2009 John Wiley & Sons, Ltd.     

A mathematical model for optimisation of flow injection-hydride generation atomic fluorescence spectrometry

A mathematical model was developed to predict signal attentuation and signal broadening in flow injection-hydride generation atomic fluorescence spectrometry owing to gas–liquid separation in order to optimise the performance with regard to sensitivity and throughput. Experimental data for arsenic measurements matched well with the model so that theoretical optimisation of various mutually dependent instrumental parameters (sample loop volume, carrier solution flow rate, GLS headspace and purge gas flow rate) was possible. A minimal GLS headspace and a maximal carrier solution flow rate resulted in the highest sensitivity and throughput for the system set-up under study.     

Rapid analysis of raw solution samples by C18 pipette-tip electrospray ionization mass spectrometry

A C18 pipette-tip electrospray ionization mass spectrometry technique was developed for rapid analysis of raw solution samples. In this technique, a C18 pipette tip was employed for rapid purification and enrichment of analytes in raw sample solutions. The adsorbed analytes were eluted by solvents supplied by a syringe and a syringe pump, and a high voltage was applied onto the syringe needle to induce electrospray ionization at the pipette tip end for mass spectrometric analysis. This technique is simple, easy to assemble, enables generation of stable and reproducible signals, and can be conveniently used for qualitative and quantitative analysis of raw solution samples. Analysis by the technique only involved simple sample preparation procedures followed by direct mass spectrometric detection, all of which could be completed within minutes, while the analytical performances of the technique, including the limit of detection, limit of quantitation, liner range, accuracy and precision, were comparable to those by conventional methods.     

A polymeric microchip with integrated tips and in situ polymerized monolith for electrospray mass spectrometry

We describe the integration of a cyclo-olefin polymer based microchip with a sheathless capillary tip for electrospray ionization-mass spectrometry (ESI-MS). The microchip was fabricated by hot embossing and thermal bonding. Its design includes a side channel for adjusting the composition of the electrospray solution so that analytes in 100% water can be analyzed. The fused silica capillaries, used for sample introduction, and the electrospray tips for MS coupling were directly inserted into the microchannel before thermal bonding of the device. A microfabricated on-chip gold microelectrode was used to apply the electrospray voltage. Annealing the device after thermal bonding increased the pressure resistance of the microchip. The cross section of the microchannel was imaged by scanning electron microscopy to estimate the effects of the annealing step. The relationship between the applied electrospray voltages and MS signal was measured at different flow rates by coupling the device to an ion trap mass spectrometer. The performance of the microchip was evaluated by MS analysis of imipramine in ammonium acetate buffer solution by direct infusion. An alkylacrylate based monolith polymer bed for on-chip sample pretreatment and separation was polymerized in the microchannel and tested for ESI-MS applications.     

A new mass spectrometry electrospray tip obtained via precise mechanical micromachining

A new electrospray tip incorporating a beveled edge has been designed for use in plastic electrophoresis chip/electrospray mass spectrometry. Theoretical hydrodynamic analysis has been conducted to model the analytical sensitivity of the spray tip shape. A larger sample wall, that provides increased hydraulic pressure, is recommended in order to preserve the fluid stability at the tip outlet. A polymer with better hydrophobic characteristics than glass was used for the spray tip in order to restrict moisture accumulation at the spray tip outlet. Experimental results demonstrate that the analytical sensitivity of the proposed scheme is better than that obtained using the flat-head tip. Although a tapered capillary tip is commonly used in electrophoresis chip/electrospray mass spectrometry, the proposed tip offers a similar sensitivity while being more rugged and durable than the conventional tapered capillary tip. The cost of our design is also much lower than conventional spray tips.     

Silicon micropillar array electrospray chip for drug and biomolecule analysis

We have developed a lidless micropillar array electrospray ionization chip (microPESI) combined with mass spectrometry (MS) for analysis of drugs and biomolecules. The microPESI chip, made of silicon, contains a sample introduction spot for a liquid sample, an array of micropillars (diameter, height, and distance between pillars in the range of 15-200, 20-40, and 2-80 microm, respectively), and a sharpened tip for direct electrospray formation. The microchips were fabricated using deep reactive ion etching (DRIE) which results in accurate dimensional control. The chip, providing a reliable open-channel filling structure based on capillary forces and a electrospray emitter tip for ionization, allows an easy operation and reliable, non-clogging liquid transfer. The microPESI chip can be used for a fast analysis using single sampling or for continuous infusion measurements using a syringe pump for sample introduction. The microPESI-MS shows high sensitivity, with limit of detection 30 pmol/L (60 amol or 28 fg) for verapamil measured with tandem mass spectrometry (MS/MS) and using a sample volume of 2.5 microL. The system shows also good quantitative linearity (r2 > 0.99) with linear dynamic range of at least six orders of magnitude and good ion current stability (standard deviation <5%) in 1-h continuous flow measurement. The microPESI-MS is shown to be a very potential method for direct analysis of drugs and biomolecules.     

An evaluation of the performance of porous polymer monolith assisted electrospray

A novel electrospray interface, which has distinct advantages over conventional pulled silica emitters, has been developed. This novel interface can be easily fabricated by forming a porous polymer monolith (PPM) at the end of a fused-silica capillary that facilitates a stable electrospray over a wide range of flow rates with only a modest increase in back-pressure. A comparison was made between the PPM-assisted electrospray and a commercial nanosprayer in terms of sensitivity, stability and robustness. A PPM-filled electrospray tip produced a day-to-day signal variation of 23% relative standard deviation (RSD) over a 3-day period when spraying a 1.0 microM test peptide solution. Furthermore, three different capillaries fabricated by the same process produced a signal variation of 17% RSD, indicating that the fabrication process shows good reproducibility. The multiple flow paths of the PPM function to split the flow and reduce clogging. Even following the accumulation of debris after prolonged use, a stable spray could still be generated with the PPM-filled capillary while the commercial nanosprayer ceased to function properly. In terms of sensitivity, PPM-assisted electrospray showed an enhancement in sensitivity at infusion flow rates between 100 to 1000 nL/min while commercial nanosprayers performed slightly better at flow rates below 100 nL/min. A sample purification step can be combined with the PPM-assisted sprayer, using the PPM as a stationary phase to desalt and preconcentrate samples prior to mass spectrometric detection.     

Factors influencing the determination of analyte ion surface partitioning coefficients in electrosprayed droplets

The observed response in mass spectrometry utilizing electrospray as a sample introduction technique can be affected by a number of factors. In this study a series of two-electrolyte systems was investigated and the mass spectrometric responses were modeled by the use of droplet surface partitioning coefficients and instrumental response factors according to a recently reported method (Sjöberg et al., Anal. Chem. 2001, 73, 23–28). The partitioning coefficient and the instrumental response factor were found to be affected by the chosen experimental conditions. Experimental parameters that were investigated include spray position relative to the orifice, spray potential, nebulizer and curtain gas flow rates, ionic strength, and organic content of the sprayed solution. The time history of the generated droplets turned out to be of importance to both the partitioning coefficients and the instrumental response factor. For example, a general increase in the surface partitioning coefficients for the tetrapentylammonium ion was initially observed when the spray was aiming closer to the sampling orifice. Furthermore, it was shown with a small amount of deuterium labeled electrolyte that the total ionic strength and not just the electrolyte concentration influence the instrumental response factor.     

Continuous flow infrared matrix‐assisted laser desorption electrospray ionization mass spectrometry

Continuous flow infrared matrix‐assisted laser desorption electrospray ionization (CF IR MALDESI) mass spectrometry was demonstrated for the on‐line analysis of liquid samples. Samples in aqueous solution were flowed through a 50 µm i.d. fused‐silica capillary at a flow rate of 1–6 µL/min. As analyte aqueous solution flowed through the capillary, a liquid sample bead formed at the capillary tip. A pulsed infrared optical parametric oscillator (OPO) laser with wavelength of 2.94 µm and a 20 Hz repetition rate was focused onto the capillary tip for sample desorption and ablation. The plume of ejected sample was entrained in an electrospray to form ions by MALDESI. The resulting ions were sampled into an ion trap mass spectrometer for analysis. Using CF IR MALDESI, several chemical and biochemical reactions were monitored on‐line: the chelation of 1,10‐phenanthroline with iron(II), insulin denaturation with 1,4‐dithiothreitol, and tryptic digestion of cytochrome c. Copyright © 2010 John Wiley & Sons, Ltd.     

Characteristics of probe electrospray generated from a solid needle

Probe electrospray ionization (PESI) has recently been developed, in which the electrospray was generated from a solid needle instead of by using a capillary. In this paper, the characteristics of probe electrospray ionization were studied based on the measurement of spray current, optical microscopy, and PESI mass spectrometry. In the experiment, the solid needle was moved up and down a vertical axis, and a small amount of sample was repeatedly loaded to the needle when the tip of the needle touched the surface of the liquid sample at the lowest position. After the application of high voltage, a liquid droplet was formed on the tip of the solid needle probe, with its size was determined by the size of the needle tip. The liquid flow rate to the tip, as indicated by the spray current, depends on the voltage applied to the needle as well as the loaded liquid amount. Stable electrospray can be maintained until the total consumption of liquid sample. The kilohertz current pulsation takes place in the case of overloading the sample to the needle. The influences of the applied voltage and the liquid flow rate on the PESI mass spectra were also examined.