66218 - Mass Spectrometry with Exercises

Learning outcomes

At the end of the course, the student is able to explain electron ionization mass spectra of the principal classes of organic molecules and to recognize spectra obtained by other ionization techniques. About the instrumental part of mass spectrometry, he/she knows: (a) the main parts of the instrument (ion sources and analyzers, detectors, vacuum pumps); (b) fundamentals of mass spectrometer functioning; (c) applications of mass spectrometry to analytical problems. He/she can use, at a basic level, (a) a gas chromatograph/mass spectrometer (GC/MS); (b) softwares for instrument control, qualitative and quantitative analysis, and mass spectra search in databases; (c) some sample purification techniques. In conclusion, the student (a) learns how to plan a simple experiment to identify and quantify pollutants in environmental samples; (b) refreshes his ability to perform simple calculations using the concepts of mole, concentration, and dilution; (c) strengthens how to properly conduct himself/herself and follow the safety rules in a chemistry laboratory; (d) learns how to write a report about chemical instrumental analyses.

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:
a) Lectures (32 hours).

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): i.e. bromobenzene.
2. How explain an electron ionization (EI) mass spectrum. Odd and even electron ions. Examples of mass spectra: saturated and unsaturated hydrocarbons, aromatic, piridinic and pirrolic compounds, carboxylic acids, esters, alcohols, compounds containing chlorine or bromine. Identification of the molecular ion. Factors influencing fragmentation rates. Stevenson rule and its exceptions. Gamma-hydrogen rearrangement. Tropylium and azatropylium ions. Even-electron rule. H and McLafferty rearrangements. Multiple rearrangements (i.e. ftalates, tributylphosphate).
3. 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.
4. 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.
5. Hyphenated techniques: gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS).

b) Laboratory experiments (24 hours).

1. Analysis of volatile organic compounds in water samples by Purge and Trap/GC/MS and Solid Phase Micro Extraction-GC/MS.
2. Analysis of nicotine in cigarette smoke by Solid Phase Extraction-GC/MS.
3. Identification of unknown compounds by searching mass spectra in NIST library database (parameters used to compare the unknown spectra with the library ones).

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 laboratory activity. 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 laboratory classes, the students use a gas chromatograph/mass spectrometer (GC/MS) along with techniques of sample purification to identify pollutants in environmental samples.

Assessment methods

The examination is divided into three parts: (a) to solve a problem of stoichiometry of the solutions (written test); (b) to intepret EI mass spectra (written test); and (c) the students who pass the above tests will orally answer to questions regarding the content of the above program and of the oral lectures. The students are not allowed to use personal notes or books.

Teaching tools

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

Teaching materials published on AMS Campus web site.

Office hours

See the website of Guido Galletti