66218 - Mass Spectrometry with Exercises

Learning outcomes

When positively evaluated at the end of the course, the student can interpret the mass spectrum af the main families of organic compounds as obtained by electron ionization and can recognize the characteristics of those mass spectra obtained by other ionization techniques.

As to the mass spectrometer, the student knows how sources, ion analyzers, detectors, vacuum pumps operate and their application to analytical problems.

Course contents

Prerequisites:
Students are required to have a good knowledge about math, stoichiometry, fundamentals of chemistry, and organic chemistry. Foreign students should master Italian (writing, speaking and listening) at a C1-level.

Program:

The course starts with practice (24 hours - 2 CFU) and carries on with lectures (32 hours - 4 CFU), both held in the classroom.

a) Practice (24 hours - 2 CFU).

1. EI-MS interpretation.
2. Interpretation of EI mass spectra. Even- and odd-electron ions. Molecular ion identification. Ion abundances. Stevenson rule with exceptions. Gamma-hydrogen rearrangement. Tropylium and aza-tropylium ions. Even-electron rule. H- and McLafferty rearrangement. Multiple rearrangements (e.g., phthalates, tributylphosphate). Examples of mass spectra of the main classes of compounds: saturated and unsaturated hydrocarbons, alcohols, carboxylic acids, esters, amines. aromatics, pyridines, pyrroles, fluorinated, chlorinated and brominated cpds. Identification of unknown compounds.
3. Identification of unknown compounds by searching mass spectra in NIST library database (parameters used to compare the unknown spectra with the library ones).
4. Examples of practical solutions of analytical problems through GC/MS.

b) Lectures (32 hours - 4 CFU).

1. Students are motivated to study mass spectrometry. Application and fundamentals of mass spectrometry. Diagram of a mass spectrometer. Definition of average mass, nominal mass, monoisotopic mass, and exact mass. Isotopic ions (A+2 rule): e.g. bromobenzene.
2. Ion sources. Electron Ionization (EI). Chemical Ionization (CI). Field Ionization (FI). Field Desorption (FD). Fast Atom Bombardment (FAB). Continuous Flow FAB. Matrix Assisted Laser Desorption Ionization (MALDI): chemical structure of principal matrixes, calibration compounds spectra, mass spectra resolution, monoisotopic masses and average masses, adduct ions, cationized ions, multicharged ions. ThermoSpray Ionization (TSP). ElectroSpray Ionization (ESI) (multicharged ions recognize and methods to calculate the number of charges). Atmospheric Pressure Chemical Ionization (APCI). Particle Beam (PB). Inductively Coupled Plasma Ionization (ICP). Examples of mass spectra are shown when necessary.
3. Ion analyzers. Magnetic analyzer (B): equation which describes the action of a magnetic field on a moving ion, resolution, metastable ions. Electrostatic analyzer (E): equation which describes the action of an electrostatic field on a moving ion. Energy and angular dispersions. Double focusing instruments EB and BE. IKE and MIKE spectra. Resolution: definitions, high and low resolution. Linked scan B/E, B2/E, [B2(1-E)]/E2. Quadrupole analyzer (Q): equation of ion motion in a quadrupolar field, Mathieu diagram for a bi-dimensional quadrupole. Multipole radiofrequency-only guides (Multipole Rf-only guides). Tandem Mass Spectrometry (MS2) using a triple quadrupole analyzer (QQQ). Ion Trap analyzer (ITMS): equation of ion movtion, Mathieu diagram for an ion trap, ion analysis by resonant ejection i and instability ejection. MS2 and MSn in an ion trap by resonant ejection. Ion trap as a single device for both ion production and analysis. Automatic control of the filament current emission in electron ionization (automatic gain control). Time of Flight analyzer (TOF): equation of ion motion, delayed extraction (DE), reflectron, post source decay (PSD). Fundamentals of Ion Cyclotrone Resonance (ICR) and Orbitrap.
4. Hyphenated techniques: gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS).
   

Readings/Bibliography

E. de Hoffmann, V. Stroobant, Mass Spectrometry – Principles and Applications, Third Edition, Wiley, 2007. (*)

T.A. Lee, A Beginner's Guide to Mass Spectral Interpretation, Wiley, 1998. (*)

F. W. McLafferty, F. Turecek, Interpretation of Mass Spectra, University Science Books, Fourth Edition, 1993. (°)

J. H. Gross, Mass Spectrometry – A textbook, Springer, Second Edition, 2011. (^)

J. T. Watson, O. D. Sparkman, Introduction to Mass Spectrometry – Instrumentation, Applications and Strategies for Data Interpretation, Wiley, Fourth Edition, 2007. (^)

R. E. March, J. F. J. Todd, Practical Aspects of Trapped IonMass Spectrometry, Volume V, Applications of Ion Trapping Devices, pp 491-507, CRC Press, 2010. (^)

O. D. Sparkman, Z. E. Penton, F. G. Kitson, Gas Chromatography and Mass Spectrometry – A Practical Guide, Academic Press, Second Edition, 2011. (^)

M. C. McMaster, LC/MS – A Practical User's Guide, Wiley, 2005. (^)

R. B. Cole, Electrospray and MALDI Mass Spectrometry – Fundamentals, Instrumentation, Practicalities, and Biological Applications, Wiley, Second Edition, 2010. (^)

(*) The text contains sections that students have to study to pass the exam.

(°) The text contains sections useful to pass the exam.

(^) The text is not necessary to pass the exam, but contains sections which may be useful for future focusing.

 

Teaching methods

The course consists of 32 hours of lectures and 24 hours of practice. The lectures provide explanations about the following topics (a) theory, instrumentation and applications of mass spectrometry (basics, mass spectrum representation, techniques for ion production and separation, detectors); (b) methods for the interpretation of mass spectra of organic compounds as obtained by electron ionization and other ionization techniques. During the practice, the students learn how to interpret EI spectra and the features of the EI spectra of the main classes of substances. They also learn the characteristics of the spectra generated by other ionization methods, such as CI, MALDI and ESI.

Assessment methods

As a consequence of the recent pandemia, lectures are delivered both in lecture room and on-line. Changes may occur and will be announced in due time. 

Exams.

The exam regards (A) EI spectra interpretation and (B) open and/or multiple-choice questions regarding instrumental and theoretical aspects of mass spectrometry. Part (A) is propedeutical to part (B). Students must pass both parts to pass the exam.

Final score.

The final score is the average of the scores obtained in parts (A) and (B), weighed by the respective credits (i.e. 2 CFU of part A and 4 CFU of part B). The this score, one point may be summed for a short presentation given during the lecture period, when occurred. Student is failed when part (A) is failed or part (B) is failed. Students may reject a sufficient score one time only. Unibo didactic regulations apply for all matters not expressly specified above.

Teaching tools

Blackboard; transparencies; power-point and other similar softwares; instruments which can be studied and/or used in laboratory.

Teaching materials published on virtuale.unibo.it web site.

The above teaching material is not enough per se to pass the exam and is not to be considered as a substitute of the recommended books, which are the only and true "teaching material".

Office hours

See the website of Guido Galletti