Mass Spectrometry

Group members:
o Arshia Bakshi- 1132220175
o Danish Shaikh- 1132220143
o Firdaus Sayyed- 1132220510
o Tushara Jayaprasad- 1132220153
 Introduction
• Analytical technique where molecules in a test
sample are converted to gaseous ions that are
subsequently separated in a mass spectrometer
according to their mass-to-charge (m/z) ratio and
detected.
• To identify unknown compounds within a sample
and to elucidate the structure ,position of functional
groups and chemical properties of different
molecules. To test the presence of isotopes.
• This technique basically studies the effect of
ionizing energy on molecules.
• The mass spectrum is a plot of the (relative)
abundance of the ions at each m/z ratio. • The essential features of all mass spectrometers are therefore:
 production of ions in the gas phase;
• The development of electrospray (ESI) and matrix-  acceleration of the ions to a specific velocity in an electric field;
assisted laser desorption/ionization (MALDI), has
 separation of the ions in a mass analyser; and
enabled the accurate mass determination.
 detection of each species of a particular m/z ratio.
 History
• First performed at the Cambridge university, in 1912 by J.J Thomson (1856-1940) when he
obtained the mass spectra of O2, N2, CO.
• Modern techniques of mass spectrometry were devised by Arthur Jeffrey Dempster and
F.W. Aston in 1918 and 1919 respectively.
• 1989: half of the Nobel Prize in Physics to Hans Dehmelt and Wolfgang Paul for the
development of the ion trap technique in the 1950s and 1960s.
• 2002: Nobel Prize in Chemistry to John Bennett Fenn for the development of electrospray
ionization (ESI).
• 1934: First double focusing magnetic analyzer was invented by Johnson and Neil.
• 1966: Munson and Field described Chemical Ionization.
• 1968: Electrospray Ionization was invented by Dole, Mack and friends.
• 1985: F. Hillenkamp, M. Karas and co-workers describe and coined the term Matrix
Assisted Laser Desorption Ionization (MALDI).
• 1989: W. Paul discovered the Ion Trap Technique.
Fragmentation and interpretation
Fragmentation and interpretation
System Assembly
Ionisers and types
Electron Impact
Chemical Ionization and FAB
MALDI
Analyser And Types
Magnetic Sector
Time Of Flight (TOF) Quadropole

Time Of Flight (TOF)

 Detectors
• Detectors function by recording the induced charge or
current generated by an ion hitting or passing through a
surface.
• The objective of ion current detectors used with mass
spectrometers is to provide a measure of the intensity of
ions with different m/z values that have been separated in
a mass analyzer.
• A key element to all mass spectrometry systems is the type
of detector used to convert a current of mass separated
ions into measurable signal. Different types of detectors
are used depending upon factors including dynamic range,
spatial information retention, noise and suitability to
the mass analyzer.
FARADAY CUP
A metal or carbon cup with a small orifice serves to capture ions and store the charge
Principle: measurement of direct charge current that is produced when an ion hits a surface
and is neutralized. Ions reach the inside of the cylinder and are neutralized by either accepting
or donating electrons as they strike the walls. This leads to a current through the resistor. The
discharge current is then amplified and detected.
The total number of ions hitting the cup per unit time is:-
N/t=i/e

Advantages:
Cup shape decreases loss of electrons.
Least expensive detector

Disadvantages:
Secondary electrons that are emitted when an ion strikes
the wall of the detector are an important source of errors.
The cup is coated with carbon because it produces few secondary ions
ELECTRON MULTIPLIERS
The basic physical process that allows an electron multiplier to operate is called secondary electron emission.
When a charged particle strikes a surface, it causes secondary electrons to be released from atoms in the
surface layer.
A Faraday cup uses one dynode and as a result produces one level of signal amplification .

There are two types of electron multipliers:-

1. Discrete type 2. Channel Electron Multiplier
Discrete type of electron multiplier has series This type has a curved horn shape, where
of dynodes maintained at increasing potentials amplifications occur through repeated collisions
resulting in a series of amplifications. with the dynode surface.
ARRAY DETECTORS
The first detector on a mass spectrometer was an array detector – photographic film. Since
that time, array detectors have evolved into a variety of types using different principles. Array
detectors can cover a broad range of detector types and systems but can be generally broken down
into two categories:
• Detectors that can measure many ions of differing mass-to-charge ratio (m/z) values
simultaneously.
• Detectors that are position sensitive.

PHOTON MULTIPLIER
In the photon multiplier (or scintillation counter) the ions initially strike a dynode which results
in electron emission.
These electrons then strike a phosphorous screen which in turn releases a burst of photons.
The photons then pass into the multiplier where amplification occurs in a cascade fashion -
much like with the electron multiplier.
The main advantage of using photons is that the multiplier can be kept sealed in a vacuum
preventing contamination and greatly extending the lifetime of the detector.
 Electrospray Ionization(ESI)
• Electrospray ionization is an evaporative ionization technique.
• It is used to analyze the high molecular weight biomolecules, labile and non volatile compounds.
• ESI is generally coupled with chromatography.
• A solution containing sample is sprayed
through the high voltage capillary by the
help of nebulization gas.
• Sprayed droplets are ionized due to high
voltage potential at capillary and solvent
is evaporated by desolvation gas.
The reaction taking place during ionization is:-
M-SH+ MH+ (S evaporated)
This technique can be used to ionize –
Proteins, peptides, lipids, oligosaccharides and
Oligonucleotides, synthetic polymerases etc.
 Atmospheric Pressure Chemical Ionization (APCI)
• Based on the mechanism of evaporation and carried out
at atmospheric pressure. It is generally
coupled with a chromatographic instrument like HPLC.
• Sample solution enters through the heated capillary
and sprayed droplets are formed due to nebulizing gas.
• Desolvation gas is supplied which converts the
sprayed droplets into vapours of analyte and solvent.
• Corona discharge electrode or beta particle
emitter is used for ionization.
• Mostly solvent molecules are ionized but
sometimes the analyte may also get ionized.

APCI is used to analyze polar, thermostable compounds with

molecular weight less than 1500Da.
 Atmospheric Pressure Photo Ionization (APPI)
• APPI is based on the technique of evaporation and carried out at atmospheric pressure.
• The process of APPI is very similar to APCI, but here the ionization is carried out by photons generated by UV light of
Krypton lamp.
• Photons emitted by the Krypton lamp have a specific energy i.e. 10eV, which is sufficient to ionize the target molecule
i.e. analyte and solvent.
 MALDI TOF
● Matrix-assisted laser desorption/ionization (MALDI) is a soft
ionization technique used in mass spectroscopy, allowing the
analysis of biomolecules (biopolymers such as DNA,
Proteins, peptide and sugars) and organic molecules (such as
polymers, dendrimers and other macromolecules), which tend
to be fragile and fragment when ionized by more
conventional ionization methods.

● Method where a laser is used to generate ions of high

molecular weight samples, such as proteins and polymers.

● Time of flight (TOF), separates ions by their mass to charge

ratio and determines that mass to charge ratio by time it takes
for ions to reach a detector.
Components of MALDI TOF:

Sample plate- Laser

Matrix
Analyte added

Time mass Flight Vacuum

detector tube system
Sample plate
 How it works?
One inside spectrometer, the target is brought into a vacuum and struck by a UV laser, wavelength at 337nm.

The laser energy is absorbed by matrix molecule which energetically desorbed or ablated from the surface of the
sample into gas phase.

During the ablation process, the analyte molecule is ionized usually by proton transfer with the nearby matrix
molecules.
Time Mass Detector
The typical detector used with MALDI is the time of flight mass detector
(TOF-MD)-
I. TOF is a method where the ions are accelerated by an electric field,
resulting in ions of the same strength to have the same kinetic energy.
II. Kinetic energy = ½ mv2
III.The time it takes for each ion to traverse the flight tube and arrive at
the detector is based on its mass-to-charge ratio; therefore the heavier
ions have shorter arrival times compared to lighter ions.
Application https://youtu.be/0jeFpXHZ8W0
Application
• Mass spectrometers provides a specific method for determining or
confirming the structure or the identity of dugs and raw materials.
• Mass spectrometers are sensitive detectors of isotopes bases on their masses.
• They are used in carbon dating and other radioactive dating processes .
• The combination of a mass spectrometer and gas chromatograph makes it
helpful for the detection of trace quantities of contaminants or toxins.
• Its used as important tool in proteomics, which is currently the major tools in
drug discovery.