Electrospray mass spectrometry of phospholipids

Phospholipids play a central role in the biochemistry of all living cells. These molecules constitute the lipid bilayer defining the outer confines of a cell, but also serve as the structural entities which confine subcellular components. Mass spectrometry has emerged as a powerful tool useful for the qualitative and quantitative analysis of complex phospholipids, including glycerophospholipids and the sphingolipid, sphingomyelin. Collision induced decomposition of both positive and negative molecular ion species yield rich information as to the polar head group of the phospholipid and the fatty-acyl substituents esterified to the glycerophospholipid backbone. This review presents the current level of understanding of the mechanisms involved in the formation of various product ions following collisional activation of molecular ion species generated by electrospray ionization of the common glycerophospholipids, including phosphatidic acid, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phosphatidylglycerol, phosphatidylserine, cardiolipin, and sphingomyelin. Recent advances in the application of matrix assisted laser desorption ionization is also considered. Several applications of mass spectrometry applied to phospholipid analysis are presented as they apply to physiology as well as pathophysiology.     

Structural analysis of organometallic compounds with soft ionization mass spectrometry

The analysis of organometallic compounds with mass spectrometry has some special features in comparison with organic and bioorganic compounds. The first step is the choice of a suitable ionization technique, where the electrospray ionization is certainly the best possibility for most classes of organometallic compounds and metal complexes. Some ionization mechanisms of organometallic compounds are comparable to organic molecules, such as protonation/deprotonation, and adduct formation with sodium or potassium ions; however, in many cases, different mechanisms and their combinations complicate the spectra interpretation. Organometallics frequently undergo various types of adduct and polymerization reactions that result in significantly higher masses observed in the spectra in comparison to molecular weights of studied compounds. Metal elements typically have more natural isotopes than common organic elements, which cause characteristic wide distributions of isotopic peaks; for example, tin has ten natural isotopes. The isotopic pattern can be used for the identification of the type and number of metal elements in particular ions. The ionization and fragmentation behavior also depend on the type of metal atom; therefore, our discussion of mass spectra interpretation is divided according to the different type of organometallic compounds. Among various types of mass spectrometers available on the market, trap-based analyzers (linear or spherical ion-traps, Orbitrap) are suitable to study complex fragmentation pathways of organometallic ions and their adducts, whereas high-resolution and high-mass accuracy analyzers (time-of-flight-based analyzers, or Fourier transform-based analyzers-Orbitrap or ion cyclotron resonance mass spectrometers) provide accurate masses applicable for the determination of the elemental composition of individual ions.     

An ultrashort-duration, high-repetition-rate pulse source for laser ionization/mass spectrometry

This paper describes a sample inlet system with several advantages over other pulsed valves, as applied to resonance-enhanced multiphoton ionization/time-of-flight mass spectrometry. The nozzle is based on online concentration by analyte adsorption/laser desorption (online COLD), where a capillary column with a narrowly synthesized tip is employed for sample introduction. The analyte molecules adsorbed at the tip are desorbed by a pulsed laser and are injected into a mass spectrometer as a packet. The online COLD nozzle can produce very short gas pulses on the order of 1 μs. Moreover, this nozzle is capable of operating over a wide range of repetition rates from 1 Hz to 1 kHz. In addition, this nozzle intrinsically possesses several unique characteristics; for instance, it can be heated to very high temperatures and has nearly zero dead volume. Therefore, the present sample introduction technique offers an ideal and versatile nozzle for laser ionization/mass spectrometry.     

Characterization of proteins by ambient mass spectrometry

Proteins play important roles in living systems and are topics of many fundamental and applied research projects. With the introduction of electrospray ionization and matrix‐assisted laser desorption/ionization for analysis of biomacromolecules in the late 1980s, mass spectrometry has become an important tool for characterization of proteins. Characterization of proteins in raw samples by these mass spectrometric techniques, however, usually requires extensive sample pretreatment. Ambient ionization techniques are new mass spectrometric techniques that allow direct analysis of samples with no or little sample preparation. Can these techniques facilitate or even eliminate sample preparation for mass spectrometric analysis of proteins? Apart from sample preparation, do these techniques offer any new features for characterization of proteins as compared with conventional ESI or MALDI? Recent advances in characterization of proteins by ambient mass spectrometry are summarized and commented in this article. © 2011 Wiley Periodicals, Inc. Mass Spec Rev 31:437–447, 2012     

Electrospray mass spectrometry (ESI-MS) in the study of metal-ligand solution equilibria

In the 20 years, since the introduction of electrospray mass spectrometry (ESI-MS), the use of this technique in various fields of inorganic, organometallic, and analytical chemistry has been steadily increasing. In this study, the application of ESI-MS to the study of metal-ligand solution equilibria is reviewed (till 2004 included). In a first section, advantages and drawbacks of ESI-MS in this type of application are described. Subsequently, a list of ca. 300 studies is reported, in which ESI-MS was used to give number and stoichiometry of the species at equilibrium, or also to estimate their stability constants. All studies are classified according to the metal ions under examination. Other related applications, such as host-guest interactions and metal ion-protein binding studies, are briefly reviewed as well.     

Peptide mapping of proteins in human body fluids using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Human body fluids have been rediscovered in the post-genomic era as great sources of biological markers and perhaps particularly as sources of potential protein biomarkers of disease. Analytical tools that allow rapid screening, low sample consumption, and accurate protein identification are of great importance in studies of complex biological samples and clinical diagnosis. Mass spectrometry is today one of the most important analytical tools with applications in a wide variety of fields. One of the fastest growing applications is in proteomics, or the study of protein expression in an organism. Mass spectrometry has been used to find post-translational modifications and to identify key functions of proteins in the human body. In this study, we review the use of human body fluids as sources for clinical markers and present new data that show the ability of Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) to identify and characterize proteins in four human body fluids: plasma, cerebrospinal fluid (CSF), saliva, and urine. The body fluids were tryptically digested without any prior separation, purification, or selection, and the digest was introduced into a 9.4 T FTICR mass spectrometer by direct-infusion electrospray ionization (ESI). Even though these samples represent complex biological mixtures, the described method provides information that is comparable with traditional 2D-PAGE data. The sample consumption is extremely low, a few microliters, and the analysis time is only a few minutes. It is, however, evident that the separation of proteins and/or peptides must be included in the methodology, in order to detect low-abundance proteins and other proteins of biological relevance.     

Investigation of intact protein complexes by mass spectrometry

Mass spectrometry has grown in recent years to a well-accepted and increasingly important complementary technique in structural biology. Especially electrospray ionization mass spectrometry is well suited for the detection of non-covalent protein complexes and their interactions with DNA, RNA, ligands, and cofactors. Over the last decade, significant advances have been made in the ionization and mass analysis techniques, which makes the investigation of even larger and more heterogeneous intact assemblies feasible. These technological developments have paved the way to study intact non-covalent protein-protein interactions, assembly and disassembly in real time, subunit exchange, cooperativity effects, and effects of cofactors, allowing us a better understanding of proteins in cellular processes. In this review, we describe some of the latest developments and several highlights.     

Electron ionization time-of-flight mass spectrometry: historical review and current applications

This review presents an overview of electron ionization time-of-flight mass spectroscopy (EITOFMS), beginning with its early development to the employment of modern high-resolution electron ionization sources. The EITOFMS is demonstrated to be ideally suited for analytical and basic chemical physics studies. Studies of the formation of positive ions by electron ionization time-of-flight mass spectroscopy have been responsible for many of the known ionization potentials of molecules and radicals, as well as accepted bond dissociation energies for ions and neutral molecules. The application of TOFMS has been particularly important in the area of negative ion physics and chemistry. A wide variety of negative ion properties have been discovered and studied by using these methods including: autodetachment lifetimes, metastable dissociation, Rydberg electron transfer reactions and field detachment, SF(6) Scavenger method for detecting temporary negative ion states, and many others.     

激光解吸电离飞行时间质谱技术及应用

本文简述了激光解吸电离飞行时间质谱(NALDI-TOF-MS)技术及其应用,同时介绍了MALDI-TOF-MS的构造、原理及特点。     

Monitoring 2-D gel-induced modifications of proteins by MALDI-TOF mass spectrometry.

In addition to more than 200 endogenously produced post-translational modifications, a detailed analysis of 2-D gel-separated proteins must also consider other modifications that a protein can experience during various steps of its separation. This review describes the use of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to investigate some of these modifications, which can originate during sample preparation and/or during the separation phase. The analyses described were mostly conducted at pH 9-9.5, and yielded reliable information on stable adduct formation that involved protein-bound amino acids and a number of gel components, including acrylamide derivatives, gel cross-linkers, and Immobiline chemicals. The -SH group of Cys was found to be the prime target of such adducts; however, longer reaction times revealed the involvement of the epsilon-NH2 of Lys. The same analysis revealed that the failure to achieve full reduction/alkylation prior to any electrophoretic step could result in protein-protein interaction, which could lead to a number of spurious spots in the final 2-D map. The implications of these modifications on the MS analysis in particular and on proteome research in general are discussed.     

基质辅助激光解吸电离飞行时间质谱法分析核糖核酸酶

应用基质辅助激光解吸电离飞行时间质谱法(MALD I-TOF-MS)成功地对核糖核酸酶相对分子量进行测定。探讨三种不同基质对其分析结果的影响,实验中发现α-氰基-4-羟基肉桂酸是适宜的基质,该方法是传统生物方法无法比拟的。     

Quantitative high-throughput analysis of drugs in biological matrices by mass spectrometry

To support pharmacokinetic and drug metabolism studies, LC-MS/MS plays more and more an essential role for the quantitation of drugs and their metabolites in biological matrices. With the new challenges encountered in drug discovery and drug development, new strategies are put in place to achieve high-throughput analysis, using serial and parallel approaches. To speed-up method development and validation, generic approaches with the direct injection of biological fluids is highly desirable. Column-switching, using various packing materials for the extraction columns, is widely applied. Improvement of mass spectrometers performance, and in particular triple quadrupoles, also strongly influences sample preparation strategies, which remain a key element in the bioanalytical process.     

Fragmentation of toxicologically relevant drugs in positive-ion liquid chromatography-tandem mass spectrometry

The identification of drugs and related compounds by LC-MS-MS is an important analytical challenge in several application areas, including clinical and forensic toxicology, doping control analysis, and environmental analysis. Although target-compound based analytical strategies are most frequently applied, at some point the information content of the MS-MS spectra becomes relevant. In this article, the positive-ion MS-MS spectra of a wide variety of drugs and related substances are discussed. Starting point was an MS-MS mass spectral library of toxicologically relevant compounds, available on the internet. The positive-ion MS-MS spectra of ～570 compounds were interpreted by chemical and therapeutic class, thus involving a wide variety of drug compound classes, such benzodiazepines, beta-blockers, angiotensin-converting enzyme inhibitors, phenothiazines, dihydropyridine calcium channel blockers, diuretics, local anesthetics, vasodilators, as well as various subclasses of anti-diabetic, antidepressant, analgesic, and antihistaminic drugs. In addition, the scientific literature was searched for available MS-MS data of these compound classes and the interpretation thereof. The results of this elaborate study are presented in this article. For each individual compound class, the emphasis is on class-specific fragmentation, as discussing fragmentation of all individual compounds would take far too much space. The recognition of class-specific fragmentation may be quite informative in determining the compound class of a specific unknown, which may further help in the identification. In addition, knowledge on (class-specific) fragmentation may further help in the optimization of the selectivity in targeted analytical approaches of compounds of one particular class.     

高分辨率电喷雾电离飞行时间质谱仪测定大豆磷脂中的磷脂酰胆碱

首次实现了国内用自制高分辨率电喷雾电离飞行时间质谱仪(ESI-TOF-MS)检测大豆磷脂中的磷脂酰胆碱(PC),共检测出了52种磷脂酰胆碱.实验证明,高分辨率ESI-TOF-MS具有定性速度快、灵敏度高、精确度好的优点,是目前磷脂检测的最为有效的工具.     

Determination of polychlorinated dibenzo-p-dioxins,dibenzofurans, and biphenyls by gas chromatography/mass spectrometry in the negative chemical ionization mode with different reagent gases

Reagent gases that are used in mass spectrometry in the NCI mode for the determination of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs) are discussed. Ion-molecule reactions and respective characteristic ions that form while using reagent gases (CH(4), O(2), i-C(4)H(10), NH(3), H(2), He, Ar, Xe, SF(6)) or gas mixtures (CH(4)/O(2), Ar/CH(4), CH(4)/H(2)O, Ar/O(2), i-C(4)H(10)/CH(2)Cl(2)/O(2)) are reviewed. It is shown that only CH(4), O(2), CH(4)/O(2), and CH(4)/N(2)O are widely used and well studied, even though-in the case of these reagent gases-there are contradictions between the publications of various authors. Such reagent gases as NH(3) and He are not well studied, but further investigations of their use for the determination of organochlorine pollutants could be of interest. The possibilities of more sensitive and selective determination of PCDDs, PCDFs, and PCBs are discussed.     

电喷雾质谱新方法及在生命分析化学中的应用

电喷雾质谱是目前应用最广泛的生物质谱技术之一。复杂生物基质对质谱分析的选择性和准确性提出挑战 ,迫切需要在质谱方法学上有所发展。本文介绍具有广泛应用前景的几种电喷雾质谱新方法的原理和特点及若干应用实例 ,对生物质谱的发展趋势和在生命分析化学中的应用前景进行展望     

Laser ablation with resonance-enhanced multiphoton ionization time-of-flight mass spectrometry for determining aromatic lignin volatilization products from biomass

We have designed and developed a laser ablation∕pulsed sample introduction∕mass spectrometry platform that integrates pyrolysis (py) and∕or laser ablation (LA) with resonance-enhanced multiphoton ionization (REMPI) reflectron time-of-flight mass spectrometry (TOFMS). Using this apparatus, we measured lignin volatilization products of untreated biomass materials. Biomass vapors are produced by either a custom-built hot stage pyrolysis reactor or laser ablation using the third harmonic of an Nd:YAG laser (355 nm). The resulting vapors are entrained in a free jet expansion of He, then skimmed and introduced into an ionization region. One color resonance-enhanced multiphoton ionization (1+1 REMPI) is used, resulting in highly selective detection of lignin subunits from complex vapors of biomass materials. The spectra obtained by py-REMPI-TOFMS and LA-REMPI-TOFMS display high selectivity and decreased fragmentation compared to spectra recorded by an electron impact ionization molecular beam mass spectrometer (EI-MBMS). The laser ablation method demonstrates the ability to selectively isolate and volatilize specific tissues within the same plant material and then detect lignin-based products from the vapors with enhanced sensitivity. The identification of select products observed in the LA-REMPI-TOFMS experiment is confirmed by comparing their REMPI wavelength scans with that of known standards.     

电喷雾电离质谱及其在蛋白质化学研究中的应用

本文介绍了电喷雾电离质谱并综述其在蛋白质化学研究中的应用.由于电喷雾电离质谱可产生多电荷峰,因此扩大了检测的分子质量范围,同时灵敏度较高,另外它可与HPLC及高效毛细管电泳分离技术联用,扩大了质谱在蛋白质化学研究中的应用.