Mass Spectrometry,Review of the Basics: Electrospray,MALDI, and Commonly Used Mass Analyzers

Abstract

Mass spectrometry (MS) has progressed to become a powerful analytical tool for both quantitative and qualitative applications. The first mass spectrometer was constructed in 1912 and since then it has developed from only analyzing small inorganic molecules to biological macromolecules, practically with no mass limitations. Proteomics research, in particular, increasingly depends on MS technologies. The ability of mass spectrometry analyzing proteins and other biological extracts is due to the advances gained through the development of soft ionization techniques such as electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) that can transform biomolecules into ions. ESI can efficiently be interfaced with separation techniques enhancing its role in the life and health sciences. MALDI, however, has the advantage of producing singly charges ions of peptides and proteins, minimizing spectral complexity. Regardless of the ionization source, the sensitivity of a mass spectrometer is related to the mass analyzer where ion separation occurs. Both quadrupole and time of flight (ToF) mass analyzers are commonly used and they can be configured together as QToF tandem mass spectrometric instruments. Tandem mass spectrometry (MS/MS), as the name indicates, is the result of performing two or more sequential separations of ions usually coupling two or more mass analyzers. Coupling a quadrupole and time of flight resulted in the production of high-resolution mass spectrometers (i.e., Q-ToF). This article will historically introduce mass spectrometry and summarizes the advantages and disadvantages of ESI and MALDI along with quadrupole and ToF mass analyzers, including the technical marriage between the two analyzers. This article is educational in nature and intended for graduate students and senior biochemistry students as well as chemists and biochemists who are not familiar with mass spectrometry and would like to learn the basics; it is not intended for mass spectrometry experts.     

聚芳醚酮环状低聚物的基质辅助激光解吸电离质谱表征

采用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术分别对2种合成的聚芳醚酮环状低聚物进行了分析研究 ,并讨论了低聚物中不同聚合度离子组分的分布规律。实验结果表明MALDI-TOF-MS是分析环状低聚物直观、准确、快速的分析工具。     

基体辅助激光解吸电离飞行时间质谱和热裂解色谱/质谱研究聚酯型聚氨酯链结构

聚酯型聚氨酯样品用基体辅助激光裂解离子化质谱(MALDI-TOF/MS)和热裂解色谱质谱(PYGC/MS)方法进行了鉴定,PYGV/MS方法可检出PU的一些组成单元的化学结构,但PYGC/MS中大多数的峰,无法从常规的标准化合物的质谱数据库中检索到,而MALDI-MS方法,可明确测出PU试样各种单元的组合、聚合度及PU长链的序列分布,质量数范围可达2300.     

超高效液相色谱-四极杆-飞行时间质谱法快速筛查农药制剂中非法添加的化学杀菌剂

建立了超高效液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF MS)快速筛查农药制剂中35种非法添加化学杀菌剂的方法.农药制剂样品经甲醇超声溶解提取后,采用Zorbax C18色谱柱(100 mm×1.8 mm,2.1μm)进行分离,以0.1%甲酸-乙睛作为流动相进行梯度洗脱,采用正离子全扫描模式进行分析.在所建立的条件下,35种杀菌剂在0.2,0.4和2.0 mg/kg水平下的添加回收率为81.0%~101.3%,相对标准偏差为1.0%~4.4%.在100份样品有6份样品中筛查出添加了化学杀菌剂,并进行了含量分析.本方法分析效率高,结果准确可靠,适用于农药制剂中非法添加化学杀菌剂的定性筛查.     

Reliable screening of pesticide residues in maternal and umbilical cord sera by gas chromatography-quadrupole time of flight mass spectrometry

The widespread use of pesticides induces heavy adverse effects on human health,especially for the pregnant women and the newborns.In this study,a screening method has been developed for the determination of multi-pesticides in maternal and umbilical cord sera.All pesticides in sera were collected using solid phase extraction(SPE),and analyzed by gas chromatography-quadrupole time of flight mass spectrometry(GC-QTOF MS).To set up the quality criteria,a database of 50 pesticides was created and the accurate masses of 3 up to 5 representative ions with their intensity ratios were included for each pesticide.In addition,a novel"identification points"(IPs)system relying on the accurate MS1 and MS2 spectra was used to interpret the data for each suspected pesticide.The methodology was then applied to a pair of maternal and umbilical cord sera.A total of six pesticide residues were screened out successfully.In conclusion,GC-QTOF MS combined with an accurate mass database seemed to be one of the most efficient tools for systematic pesticide analysis.     

Analysis of phenolic choline esters from seeds of Arabidopsis thaliana and Brassica napus by capillary liquid chromatography/electrospray‐ tandem mass spectrometry

Total phenolic choline ester fractions prepared from seeds of Arabidopsis thaliana and Brassica napus were analyzed by capillary LC/ESI‐QTOF‐MS and direct infusion ESI‐FTICR‐MS. In addition to the dominating sinapoylcholine, 30 phenolic choline esters could be identified based on accurate mass measurements, interpretation of collision‐induced dissociation (CID) mass spectra, and synthesis of selected representatives. The compounds identified so far include substituted hydroxycinnamoyl‐ and hydroxybenzoylcholines, respective monohexosides as well as oxidative coupling products of phenolic choline esters and monolignols. Phenolic choline esters are well separable by reversed‐phase liquid chromatography and sensitively detectable using electrospray ionization mass spectrometry in positive ion mode. CID mass spectra obtained from molecular ions facilitate the characterization of both the type and substitution pattern of such compounds. Therefore, LC/ESI‐MS/MS represents a valuable tool for comprehensive qualitative and quantitative analysis of this compound class. Copyright © 2008 John Wiley & Sons, Ltd.     

稠油中饱和烃复杂混合物成分解析及其意义

利用两根极性不同的毛细柱,在全二维气相色谱上分析辽河油田遭受严重生物降解形成的稠油饱和烃组分,可以将传统色谱分析时形成的“基线鼓包”即不可分辨的复杂混合物（Unresolved Complex Mixtures）分开．根据饱和烃全二维气相色谱谱图的族分离特点和瓦片效应,结合飞行时间质谱提供的质谱信息初步解析不可分辨的复杂混合物主要成分．发现常规色谱分析时形成的所谓“基线鼓包”是由成千上万、含量相对较低的不同取代基的环状化合物组成,这些化合物在一维色谱上以分子量递增的顺序排列,在二维色谱上以极性的差异或者环的多少排列．C24之前的第一组不可分辨的复杂混合物主要由环己烷为基本单元的单环、双环和三环烷烃类化合物组成,信噪比在100以上的化合物数量约为饱和烃总数量的75％,质量分数是饱和烃总量的80％以上,是饱和烃的主要组成部分．C24之后出现的第二组不可分辨的复杂混合物主要由四个环或者五个环为基本单元的化合物组成,信噪比在100以上的化合物数量约为饱和烃总数量的17％,质量分数是饱和烃总量的0．5％．对稠油中这些不可分辨的复杂混合物的解析有助于对其成因机理的认识和高效开采方案的制定．     

Capillary electrophoresis-mass spectrometry of intact basic proteins using Polybrene-dextran sulfate-Polybrene-coated capillaries: System optimization and performance

A capillary electrophoresis-mass spectrometry (CE-MS) method using sheath liquid electrospray ionization interfacing was studied and optimized for the analysis of intact basic proteins. To prevent protein adsorption, capillaries with a noncovalent positively charged coating were utilized. Capillaries were coated by subsequent rinsing with solutions of Polybrene, dextran sulfate and Polybrene. The coating proved to be fully compatible with MS detection, causing no background signals and ionization suppression. The composition of the sheath liquid and BGE was optimized using the model proteins α-chymotrypsinogen A, ribonuclease A, lysozyme and cytochrome c. A sheath liquid of isopropanol-water-acetic acid (75:25:0.1, v/v/v) at 2 μL min⁻¹ resulted in optimal signal intensities for most proteins, but caused dissociation of the heme group of cytochrome c. Optimum protein responses were obtained with a BGE of 50 mM acetic acid (pH 3.0), which allowed a baseline separation of the test protein mixture. Several minor impurities present in the mixture could be detected and provisionally identified using accurate mass and a protein modification database. The selectivity of the CE-MS system was investigated by the analysis of acetylated lysozyme. Eight highly related species, identified as non-acetylated lysozyme and lysozyme acetylated in various degrees, could be distinguished. The CE-MS system showed good reproducibility yielding interday (three weeks period) RSDs for migration time and peak area within 2% and 10%, respectively. With the CE-MS system, determination coefficients (R²) for protein concentration and peak area were higher than 0.996, whereas detection limits were between 11 and 19 nM.     

In-source fragmentation and accurate mass analysis of multiclass flavonoid conjugates by electrospray ionization time-of-flight mass spectrometry

In-source collision-induced dissociation (CID) fragmentation features of multiclass flavonoid glycoconjugates were examined using liquid chromatography electrospray time-of-flight mass spectrometry. Systematic experiments were performed to search for optimal conditions for in-source fragmentation in both positive and negative ion modes. The objective of the study was to attain uniformly appropriate conditions for a wide range of analytes independently of the aglycone, the attached sugar part and the type of bond between the aglycone and the glycan moieties (O- or C-glycosides). Studied substances included representatives of flavonols, flavones, flavanones and anthocyanins and, regarding their glycan parts, mono-, di- and triglycosides with varying distribution of carbohydrate moieties (di-O-glycosides, O-diglycosides, O,C-diglycosides). The breakdown properties of the analytes along with the abundances of the characteristic diagnostic ions required for structural elucidation of complex flavonoid derivatives were evaluated. An optimized value was found for the instrument parameter (fragmentor voltage) affecting the in-source CID fragmentation of the analytes [230 V (ESI+) and 330 V (ESI-)]. Thus, appropriate performance in terms of both highly sensitive full-scan acquisition and fragmentation information was obtained for all the investigated flavonoids. In addition, singularities in the abundance of selected diagnostic ions (e.g. Y(0), Y(1) and Y*) due to variations in the interglycosidic linkage (rutinoside-neohesperidoside) in the glycan part were found and are also evaluated and discussed in detail. The combination of in-source CID fragmentation with high mass accuracy MS detection establishes a working basis for the development of versatile and useful LC-MS methods for wide-scope screening, non-targeted detection and tentative identification of flavonoid derivatives.