Maharishi University of

information technology

Name - Nitya Verma

Class - MSc. Biotechnology 1st year 1st semester
Submitted to - Dr. Himani Kulshrestha
● Topic: mass spectrometry
● Content -
1. Introduction
2. Basic principle
3. Instruction and working
4. Ionization technique
5. Types of ions
6. Applications
 Introduction of spectroscopy
● Spectroscopy is the study of the absorption and emission of light and other radiation by
matter. It involves the splitting of light (or more precisely electromagnetic radiation) into its
constituent wavelengths (a spectrum), which is done in much the same way as a prism
splits light into a rainbow of colours.

● Spectroscopy is based on the ability of atoms and molecules to absorb or emit

electromagnetic (EM) radiation. The absorption or emission of different forms of EM
radiation is related to different types of transitions.

● Spectroscopy is used in virtually all technical fields of science and

technology.
Mass spectrometry
● Mass spectroscopy, also known as mass spectrometry, is an analytical technique that identifies
chemical substances by sorting gaseous ions in electric and magnetic fields according to their
mass-to-charge ratios. The results are presented as a mass spectrum, which is a plot of
intensity as a function of the mass-to-charge ratio.
● Mass spectrometry can be used to:
● Separate ionized particles such as atoms, molecules, and clusters
● Determine the molecular weight of the particles
● Calculate the mass-to-charge ratio (m/z) of one or more molecules in a sample
● Basic principle
1. MS) is to generate ions from a sample, separate them by their
mass-to-charge ratio (m/z), and record the relative abundance
of each ion type. This produces a mass spectrum of the molecule.
2. The steps in mass spectrometry are:
3. Generate ions from the sample
4. Separate the ions by their m/z
5. Record the relative abundance of each ion type
6. Deflect the ions by a magnetic field
Move the ions with the correct charge and mass to the detector
● Mass spectrometry works by:
● Sorting gaseous ions in electric and magnetic fields according to
their mass-to-charge ratios
● Using differences in the ratios of their charges to their respective
masses (mass/charge; m/z)
● By the 1980s, small organic molecules were routinely being
analyzed by mass spectrometry. However, proteins, especially large
ones, and other macromolecules such as nucleic acids and complex
carbohydrates provided more of a challenge.
● Instrumation and working
● The four main parts of mass spectrometry are discussed below:
●
● Ionizer – The bombarding of the sample is done by the electrons.
These electrons move between cathode and anode. When the
sample passes through the electron stream between the cathode
and anode, electrons with high energy knock electrons out of the
sample and form ions.
● Accelerator – The ions placed between a set of charged parallel
plates get attracted to one plate and repel from the other plate. The
acceleration speed can be controlled by adjusting the charge on the
plates.
 ● Deflector – Magnetic field deflects ions based on its charge and
mass. If an ion is heavy or has two or more positive charges, then
it is least deflected. If an ion is light or has one positive charge,
then it is deflected the most.
● Detector – The ions with correct charge and mass move to the
detector. the ratio of mass to charge is analyzed through the ion
that hits the detector.
● Working
● In a regular mass spectrometer, we initially have the
material to be analyzed, but we need it to be ionized to pass
through the spectrometer with enough energy. Thus, the
sample is bombarded by electrons to ionize it.
●
● This ionized beam is now passed through a series of electric
or magnetic fields depending on the type of the sample and
its properties.
● The ions are deflected by the field through which they are passed
through in such a way that the ions with the same mass to signal
ratio will follow the same path to the detector.
●
● These charged and deflected ions are now incident onto a detector
which is capable of distinguishing the charged particles falling on
it. Based on the mass spectrum produced by the charged ions, we
can identify the atoms or molecules constituting the sample by
comparing them with known masses or through a characteristic
fragmentation pattern
Mass spectrometry
● Ionization technique
● There are many ionization techniques used in mass
spectrometry, including:
● Electron impact (EI): One of the first ionization techniques
developed for mass spectrometry. EI uses a beam of
high-energy electrons to bombard a sample, producing
multiple ions. EI is best suited to relatively nonpolar, volatile
samples.
● Fast-atom bombardment (FAB): Uses a stream of inert gas
atoms (argon or xenon) to sputter a sample, generating
individualized ions.
● Secondary ion mass spectrometry (SIMS): Uses ions such as Cs+
to sputter a sample, generating individualized ions.
● Other ionization techniques include:
● Chemical ionization (CI)
● Plasma and glow discharge
● Electrospray ionization (ESI)
● Field ionization
● Laser ionization (LIMS)
● Matrix-assisted laser desorption ionization (MALDI)
● Matrix-assisted laser desorption/ionization (MALDI) is a mass spectrometry
technique that uses a laser to create ions from large molecules. MALDI is a "soft"
ionization technique that minimizes fragmentation of the molecules. It's used to
analyze molecules that are non-volatile or thermally unstable.
● MALDI works by:
● Using a laser to strike a matrix of small molecules
● Making the analyte molecules into the gas phase without fragmenting or
decomposing them
● Measuring the mass of molecules from a sample that has been embedded in a
matrix
● MALDI is one of the most successful "soft" ionization methods in mass
spectrometry. It enables the analysis of a broad range of molecules, including
lipid.
● Types of ions
● Mass spectrometry produces many types of ions, including:
● Molecular ion: The parent ion
● Fragment ions: These can be even-electron cations or odd-electron radical cations
● Rearrangement ions
● Metastable ions
● Multiple charged ions
● Other types of ions produced in mass spectrometry include:
● [AB]-H+ - deprotonated molecule
● [AB]- - molecular anion
● [B]- - anion
● [AB-N]- - neutral loss fragment ion
● [A]+ - cation
● [AB]+H+ - protonated molecule
● The most commonly used ions are:
● Cs+ and O2+ for positively charged ion beams
● O- for negatively charged beams
● Applications
● Mass spectrometry (MS) is an analytical technique that identifies chemical
substances by sorting gaseous ions in electric and magnetic fields according
to their mass-to-charge ratios. MS is used in many applications, including:
● Drug testing and discovery
● Food contamination detection
● Pesticide residue analysis
● Isotope ratio determination
● Protein identification
● Carbon dating
● Analysis of amino acid sequences in proteins and peptides
● Determination of elemental concentrations of surface contaminants
● Forensic science
● Planetary exploration
● Metabolomics