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Mass spectrometry:
Mass spectrometer measures molecular
masses.
Molecular mass and formula and structure
information
Ionization: - generate similar ions from all
molecules in the mixture.
Different molecule same charge but
different mass.
Basis of separation: mass / charge ratio
(m/z) =mass of ion.
Mass spectrometry:
Mass Spectrometer
A mass spectrometer needs to perform three
functions:
1. Creation of ions sample molecules are
subjected to a high energy beam of electrons,
converting them in to ions
2. Separation of ions as they are accelerated in
an electric field, the ions are separated according
to mass-to-charge ratio (m/z)
3. Detection of ions as each separated
population of ions is generated, the spectrometer
needs to qualify and quantify them
Principle:
Instrument is mass spectrometer
Separates gas phase ionized atoms, molecules,
and fragments of molecules
Separation is based on the difference in mass-tocharge ratio (m/z)
m = unified atomic mass units (u)
1 dalton (Da) = 1 u = 1.665402 x 10-27 kg
z = charge on the ion (may be positive or
negative)
Process:
A small quantity of sample is injected and vaporized
under high vacuum
The sample is then bombarded with electrons having
25-80 eV of energy
A valence electron is punched off of the molecule,
and an ion is formed
Process:
ions are accelerated to a constant velocity in
vacuum by a series of electrostatic plates.
Then subjected to strong magnetic field
Deflection of ions from its path-least mass more
reflection.
This will lead to separates of ions based on mass
Detector measures the ion flow.
If there is a mixture of different molecules in a
sample, all the masses are measured
simultaneously. So you get a mass spectrum.
INSTRUMENTATION
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1. Vacuum system
Vacuum minimises the collisions between ions
and air molecules. Without a high vacuum, the
ions produced in the source will not reach the
detector
Two vacuum pump types are used:
Rotary vane pump
Turbo molecular pump or Diffusion pump.
Rotary vane pump can be an oil pump to provide
initial vacuum (approximately1torr)
Turbo molecular pump provides working high
vacuum(1mtorr to1 ntorr).
Sample introduction:
Two things must be
achieved here
1) Sample must be
introduced into
vacuum
2) Vaporization of
sample prior to
ionization
IONIZATION SOURCE
Electron Ionization (EI) :
Mass analyzer
DETECTORS
DETECTORS
Electron
Multiplier (EM)
Faraday Cup
Photomultiplier
Conversion
Dynode
Applications
Determine protein structure, function, folding
and interactions
Identify a protein from the mass of its peptide
fragments
Monitor enzyme reactions, chemical
modifications and protein digestion
Applications
Genomics
Sequence oligonucleotides
Environment
Geology
Measure petroleum
composition
Perform carbon dating
Source: https://www.thermofisher.com
Some Pictures
MALDI-R
FT-ICR
Q-Tof Micro
LTQ-Orbitrap
Reference: