Mass spectrometry mass spectrometry is a powerful analytical technique used to quantify known materials, to identify unknown compounds within a sample, and to elucidate the structure and chemical properties of different molecules. Specific labeling with stable isotopes 2 h, c, 15 n and 18 o is also of importance in mass spectrometry for. At higher than threshold laser fluences, proteins exhibit a series of fragment ions. Mass spectrometry has played a significant role in the identification of unknown phosphoproteins and sites of phosphorylation in biological samples. Ch 3 ch 2 oh ch 2 brcl ch 3 co 2 h ch 3 ch 2 nh 2 ch 3 2 nch 2 c. The leading scientist in this field was fred mclafferty who together with frantisek turecek has written a very. Size effect on fragmentation in tandem mass spectrometry article in analytical chemistry 826. Prasain 011607 lecture goals value of fragmentation in determining structure how peptides fragment interpreting the tandem mass spectrum automating identification of peptides from. For the analytical chemist, a mass spectrum is useful for two applications. Ionattachment mass spectrometry iams is a form of mass spectrometry that uses a soft form of ionization similar to chemical ionization in which a cation is attached to the analyte molecule in a reactive collision. Still the most widely used technique in mass spectrometry, electron ionization ei produces molecular ions from gasphase analytes. A few simple rules for fragmentation of ions are as follows.
The complete process involves the conversion of the sample into gaseous ions, with or without fragmentation, which are then characterized by their. Price, a fragmentation study of isoflavones in negative electrospray ionization by msn ion trap mass spectrometry and triple quadrupole mass spectrometry. The presence of a m et and m ch 3 peak indicates that this four carbon alcohol determined from its molecular mass is the secondary alcohol 2butanol. Nmr ms peaks are narrower than nmr peaks ms is much more 104 x more sensitive than nmr among most sensitive tools ms allows one to analyze much larger molecules 50 kd than nmr ms samples are more difficult to prepare. Fragmentation pattern and its rules and interpretation of mass spectroscopy guided by. The process typically involves either chemical or electron impact ionization of the analyte. Estimating the efficiency of phosphopeptide identification. Ms is an analytical tool to measure the mass tocharge ratios mz of ions in order to determine their molecular weight mw. The fragments of a molecule cause a unique pattern in the mass spectrum. In matrixassisted laser desorptionionization of proteins, there exists a certain amount of fast metastable decay immediately after laser irradiation. What are the general rules predicting prominent peaks in a mass spectrum. In mass spectrometry, fragmentation is the dissociation of energetically unstable molecular ions formed from passing the molecules in the ionization chamber of a mass spectrometer. Applications of insource fragmentation of protein ions. Separate the ions on the basis of their masstocharge ratio mz 3.
The mass spectra are unusually clearly correlated with the structures. What class of compounds is most likely to give a fragment ion at mz m18. The acceleration voltage was 20 kv and positive ion spectra were recorded in reflectron mode. Mass spectrometry of alditols as trimethylsilyl derivatives. Ion fragmentation of small molecules in mass spectrometry uab. Apr 20, 2014 basic components of mass spectrometer four basic components sample inlet ionization source mass analyzer ion detector 12. Mass spectrometry dont call it spectroscopy absorption of electromagnetic radiation electron impact mass spectrometry 70 ev 1614 kcalmol contrast with energy from ir 110 kcalmol or nmr 0. Quadrupole mass filters timeofflight tof mass analyzer ion trap mass analyzer fouriertransform mass spectrometry ftms electron impact ionization magnetic sector separation single focusing mass spectrometer evacuated system 106 torr bi f the interior of the mass spectrometer must be evacuated.
Joseluis jimenez 2 business items last real lecture is today material from today, art ms, protein ms lectures included in exam. Among these methods, mass spectrometry represents the most advanced technique because of its high sensitivity and speci. Corporation, usa and a mass spectrometry toolkit version3. Order of radicalcarbocation loss tertiary secondary primary methyl hydrogen loss of longest is preferred 4.
Evaluation of the mass spectrometric fragmentation of. Fragment screening by native state mass spectrometry sallyann poulsen eskitis institute for drug discovery, griffith university, brisbane, queensland 4111, australia. The complete process involves the conversion of the sample into gaseous ions, with or without fragmentation, which. Using the mass spectrum of 2,2,4trimethylpentane shown below, answer the questions that follow about its fragmentation. These molecular ions then fragment in a reproducible way, which results in a fingerprint of the analyte. Fragment ion stability takes precedence over radical 5. This section will ignore the information you can get from the molecular ion or ions. Analyses of protein phosphorylation, particularly large scale phosphoproteomic experiments, have recently been enhanced by efficient enrichment, fast and accurate instrumentation, and better software, but challenges remain because of. Pdf general fragmentation rules for isomerization and fragmentation processes based on firm thermochemical grounds are presented for. The fragment ions thus formed can be resolved and their mz values measured accurately by employing delayed extraction linear timeofflight mass spectrometry.
A fragmentation study of isoflavones in negative electrospray. The mass spectrometer was operated in the positive ion mode with the turboionspray heater set at 450 c api 4000 lcmsms system, applied biosystems, foster city, ca, usa. Explain rules of fragmentation in mass spectroscopy with one example each. Because of the uniqueness of these chemical fingerprints, commercially available libraries. Convert neutral atoms or molecules into a beam of positive or negative ions 2. We use a wileymclaren lineartype tof mass spectrometer which consists of a series of parallel plates, onto which voltages are pulsed at the repetition rate of the experiment 10 hz. Electrospray mass spectrometry and fragmentation of n. Ms ms mclafferty, 1978, tandem in space or time precursor ionparent ion. Basic principles of mass spectrometry ms ms is defined as the minimum scale in the world due to the dimensions of what it weighs chughtai et al. At least one question from each exam on tue 6nov during class period. General fragmentation rules fororganic and organometallic compounds under electron impact article pdf available in european mass spectrometry 86. Organic lecture series 3 mass spectrometry ms organic lecture series 4 a mass spectrometer a mass spectrometer is designed to do three things convert neutral atoms or molecules into a beam of positive or negative ions separate the ions on the basis of their masstocharge mz ratio measure the relative abundance of each ion. Basic components of mass spectrometer four basic components sample inlet ionization source mass analyzer ion detector 12. Obviously, this is not done with a conventional balance or scale.
Justesen examined the fragmentation rule of methoxylated flavones and flavonols in negative ion mode, and found that the loss of a. Mass spectrometry, infrared spectroscopy, and ultravioletvisible spectroscopy learning objectives. Mass spectroscopy mass spectrometry is the study of systems generating the formation of gaseous ions, with or without fragmentation. Which of the following compounds will give a molecular ion having mz an odd number.
The other lines in the mass spectrum are more difficult to explain. Assistant professor department of pharmaceuticcal chemistry dharmaj degree pharmacy college presented by. When working with mass spectrometry, particularly msms spectra, a number of tables are essential in order to understand these. Try writing the mechanisms with fish hook arrows using figures 8 and 9 as a guide. Try writing the mechanisms with fish hook arrows using. Remember that you are not expected to interpret all signals in a mass spectrum. An organic chemists guide to electrospray mass spectrometric. Some helpful reminders for mass spectrometry and compound. Table of common fragment ions wake forest university.
The mass spectrometer gives the mass to charge ratio mz, therefore the sample analyte must be an ion. Mass spectrometry fragmentation patterns last updated. Following are examples of compounds listed by functional group, which demonstrate patterns which can be seen in mass spectra of compounds ionized by electron impact ionization. Be able to describe what happens to a compound in a mass spectrometer 3. There are many ways in which the molecules might break apart, but a few of these pathways are especially useful to know. Size effect on fragmentation in tandem mass spectrometry. Lecture notes in eimass spectrometry by torben lund. Timeofflight mass spectrometry tofms is used in tandem with both our supersonic jet and electrospray ion sources.
Mass spectrometry fragmentation patterns chemistry. These reactions are well documented over the decades and fragmentation pattern is useful to determine the molar weight and structural. A mass spectrometer a mass spectrometer is designed to do three things 1. The infrared photofragmentation experiments performed in the johnson lab rely on mass spectrometric methods for both the selective interrogation of molecular ions and the acquisition of their infrared spectra. Be able to predict the fragmentation patterns expected to arise in the mass spectrum of alkanes, alkyl halides, ethers, alcohols, and ketones. Mass spectrometry the mass spectrometer bombards the sample with electrons this knocks one electron off the molecule and forms a cation with the same mass as the molecular mass. Understanding mass spectrometry in a mass spectrometer, the same thing is happening, except its atoms and molecules that are being deflected, and its electric or magnetic fields causing the deflection. Mass spectrometry ms is one of the key analytical technology on which the emerging omics approaches are based.
Evaluation of the mass spectrometric fragmentation of codeine. Mass spectrometry is essentially a technique for weighing molecules. That is covered in three other pages which you can get at via the mass spectrometry menu. When molecules go through a mass spectrometer, they break into pieces in a variety of different ways. The delay for the timelagfocusing ion source was 500 ns and the pulse voltage was 3. From the time of the second edition published in 2001 until now, much progress has.
Fragment screening by native state mass spectrometry. The mass spectra of simple hydrocarbons have peaks at mz values corresponding. Zenk a,jurgen schmidt b, a biozentrum universitat halle, weinbergweg 22, d06120 halles. Application of fragmentation mechanisms for the assignment of some of the higher mz fragment peaks in appearing in ei mass spectra are shown in figures 11, 12, and for a ketone, and amine and an ester, respectively. General mass table elemental mass values, amino acid residues and immonium ions. It may provide detection and quantization of thousands of proteins and biologically active metabolites from a tissue, body fluid or cell culture working in a global or targeted manner, down to ultratrace levels. Evaluation of the mass spectrometric fragmentation of codeine and morphine after cisotope biosynthetic labeling chotima poeaknapo a, ursula fisinger a, meinhart h. For example, lines with mz values 1 or 2 less than one of the easy lines are often due to loss of one or more hydrogen atoms during the fragmentation process. Mass spectrometry is a gas phase technique the sample must be vaporized. Monoisotopic mass is typically expressed in unified atomic. These reactions are well documented over the decades and fragmentation pattern is useful to. Use the text to clarify whats covered on the slides. As a result, chemical ionization would need to be utilized to.
Mass spectrometry ms is a very powerful instrument that can be used to analyze a wide range of materials such as proteins, peptides, dna, drugs, and polymers. Etd and ecd mass spectrometry fragmentation for the. Fragments lost as neutral molecules in positive ionization mode, a trace of formic acid is often added to. A number of tables can be downloaded through this page.
The fragmentation pattern is characterized by peaks separated by 14 mass units a ch 2 group. The trimethylsilyl ethers of alditols with 3, 4, 5 and 6 carbon atoms were investigated. Etd and ecd mass spectrometry fragmentation for the characterization of protein post translational modifications, tandem mass spectrometry applications and principles, jeevan k. Department of engineering chemistry, chalmers university of technology, goteborg, sweden. Maldi mass spectra were recorded with a micromass tofspec 2e mass spectrometer micromass, wythenshawe, manchester, uk from 2,5dhb. Practice questions for mass spectrometry home chemistry. Among these methods, mass spectrometry represents the most advanced technique because of its high sensitivity and specificity and the possibility to determine the desired substance in not highly purified samples by using lcmsms. General not a spectroscopic technique no absorption of radiation spectroscopy. Rearrangements are common but usually do not give intense peaks. Chapter 2 fragmentation and interpretation of spectra 2. When students ask what books to read on mass spectrometry, there are various excellent monographs, but the ideal textbook still seemed to be missing at least in my opinion. The leading scientist in this field was fred mclafferty who together with frantisek turecek has written a very famous text book. The mass spectrum of the product molecule contains intense signals at mz 71, 59, and 43 all of which can arise from the ucleaveage of the most likely molecular ion, draw a reaction mechanism for the reaction pathways that leads to these species.
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