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MASS SPECTROSCOPY
-: Presented By :-
M Asif shaheen
Lecturer
KEMU Lahore
1
Spectroscopy
Spectroscopy
Spectroscopy
Spectroscopy
Spectroscopy
Spectroscopy
Basic principle
Spectroscopy
8
Spectroscopy Principle and Instrumentation
9
Ionisation
Spectroscopy
The atom is ionised by knocking one or more electrons
off to give a positive ion. (Mass spectrometers always
work with positive ions).
These positive ions are persuaded out into the rest of the
machine by the ion repeller which is another metal plate
carrying a slight positive charge.
11
Acceleration
Spectroscopy
The ions are accelerated so that they all have the
same kinetic energy.
12
Spectroscopy
13
Spectroscopy
Deflection
The ions are then deflected by a magnetic field
according to their masses. The lighter they are, the
more they are deflected.
14
Spectroscopy
Different ions are deflected by the magnetic field by
different amounts. The amount of deflection depends
on:
15
Detection
Spectroscopy
The beam of ions passing through the machine is detected
electrically.
The other ions collide with the walls where they will
pick up electrons and be neutralised.
17
Spectroscopy
Spectroscopy
Mass Spectrometer Block Diagram
Turbo molecular
pumps
High Vacuum System
HPLC
Flow injection
Sample plate
Spectroscopy
Ion Source
MALDI
ESI
FAB
LSIMS
EI
CI
Spectroscopy
Mass Analyzer
Time of flight
(TOF)
Quadrupole
Ion Trap
Magnetic Sector
Spectroscopy
Spectroscopy
The Sample Inlet System
Batch Inlets
The batch inlet system is considered the most
common and simplest inlet system. Normally, the
inside of the system is lined with glass to elude losses
of polar analyte by adsorption.
This system externally volatizes the sample which
leaks into an empty ionization region. Boiling points up
to 500 degrees C of gaseous and liquid samples can
be used on typical systems.
The system's vacuum contains a sample pressure of
10-4 to 10-5 Torr. Liquids are introduced using a
microliter syringe into a reservoir; gases are enclosed
in a metering area that is confined between two valves
before being expanded into a reservoir container.
Spectroscopy
ADVANTAGES
Gives molecular mass and also the fragmentation
pattern of the sample.
Extensive fragmentation and consequent large number
of peaks gives structural information.
Gives reproducible mass spectra.
DISADVANTAGES
Sample must be thermally stable and volatile.
A small amount of sample is ionized (1 in 1000
molecules).
Unstable molecular ion fragments are formed so readily
that are absent from mass spectrum.
Spectroscopy
Spectroscopy
Spectroscopy
For example,
Ammonia is used as reagent gas.
First ammonia radical cations are generated by electron
impact and this react with neutral ammonia to form
ammonium cation (reactive species of ammonia CI).
ADVANTAGES
Used for high molecular weight compounds.
Used for samples which undergo rapid fragmentation in
EI.
LIMITATIONS
Not suitable for thermally unstable and non-volatile
samples.
Relative less sensitive then EI ionization.
Samples must be diluted with large excess of reagent gas
to prevent primary interaction between the electrons and
sample molecules.
Spectroscopy
Spectroscopy
Field Ionization
FI is used to produce ions from volatile compounds
that do not give molecular ions by EI.
It produces molecular ions with little or no
fragmentation.
Application of very strong electric field induces
emission of electrons.
FI utilizes 10-micron diameter tungsten emitter wires
on which carbon whiskers, or dendrites, have been
grown.
A high electric field gradient (1010 V/cm) at the tips of
the whiskers produces ionization
Spectroscopy
ADVANTAGES
As fragmentation is less, abundance of molecular ions
(M+) is enhanced, hence this method is useful for
relative molecular mass and empirical formula
determination.
DISADVANTAGES
Not suitable for thermally unstable and non volatile
samples.
Sensitivity is les than EI ion source.
No structural information is produced as very little
fragmentation occurs.
Spectroscopy
Spectroscopy
Spectroscopy
Spectroscopy
Field Desorption
Also known as offspring of field ionization.
In field desorption method, a multitipped emitter
(made up of tungsten wire with carbon or silicon
whiskers grown on its surface) similar to that used in FI
is used.
The sample solution is deposited on the tip of the
emitter whiskers either by
dipping the emitter into analyte solution or
using a microsyringe.
Then the sample is ionized by applying a high voltage
to the emitter.
Spectroscopy
ADVANTAGES
Works well for small organic molecules, low
molecular weight polymers and petrochemical fractions.
DISADVANTAGES
Sensitive to alkali metal contamination.
Sample must be soluble in a solvent.
Not suitable for thermally unstable and non volatile
samples.
Structural information is not obtained as very little
fragmentation occurs.
Spectroscopy
Spectroscopy
Spectroscopy
Electrospray ionization
Electrospray ionization is a technique used in mass
spectrometry to produce ions from macromolecules
such as proteins, polypeptides and oligonucleotides
having molecular weights of 10,000 Da or more.
The method generates ions from solution of a sample by
creating fine spray of charged droplets.
• A solution of sample is pumped through a fine,
charged stainless steel capillary needle at a rate of few
microlitres/minute.
The needle is maintained at a high electric field
(several kilovolts) with respect to cylindrical electrode.
• The liquid pushes itself out of the capillary as a mist
or aerosol of fine charged droplets.
Spectroscopy
These charged droplets are then passed through
desolvating capillary where the solvent is evaporated in
the vacuum and attachment of charge to the analyte
molecules takes place.
Desolvating capillary uses warm nitrogen as nebulising
gas.
The desolvating capillary is maintained under high
pressure.
• As the droplets evaporate the analyte molecules comes
closer together.
Spectroscopy
ADVANTAGES
Most important techniques for analysis of high
molecular weight biomolecules such as polypeptides,
proteins, oligonucleotides and synthetic polymers.
Can be used along with LC and capillary
electrophoresis.
Spectroscopy
Spectroscopy
Partial
Sample Inlet Nozzle
Pressure = 1 atm vacuum
(Lower Voltage)
Inner tube diam. = 100 um
MH+
N2
++
+ + + + + +
+ + ++ ++ +
++
+ ++
+
+ +
+ MH2+
++ +
Sample in solution ++
+ +
+ + +
++ ++ +
++ +
N2 gas ++ + +
MH3+
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