In this slide ,You will get to learn Electron Spray Ionization (ESI) technique used in Mass Spectroscopy and its Various Application in Pharmaceutical Drug Analysis.

1. ELECTRONSPRAY
IONIZATION(ESI)
AND ITS APPLICATION
Presented by:-
Shaikh Nisar Ali (M.Pharm)
Dept. Of Pharmacology
Guided by:- Dr. PSS

2. Mass Spectroscopy:-
• Mass spectrometry is an analytical tool useful for measuring
the mass-to-charge ratio of one or more molecules present in a
sample.
• Every Mass Spectrometer consists of at least these three
components:
1. Ionization Source
2. Mass Analyzer
3. Ion Detection System

3. Basics...
• Organic Molecules are bombarded with beam of Energetic electrons
(70 ev) in gaseous state.
• Converted into highly energetic positively charged ions.(Molecular ion
or Parent ion)
• Further breakup into smaller ions.(Fragmented or daughter ions).
• The formed ions are separated by deflection in Magnetic field
according to their MASS and CHARGE.

4. MASS SPECTROSCOPY

5. Different Ionization Method

6. • Electrospray Ionization
• Electrospray ionization (ESI) is a technique to generate ions for
mass spectrometry using electrospray by applying a high voltage
to a liquid to produce an aerosol.
• Is a type of evaporative Ionization technique used to analyze the high
Molecular weight biomolecules Labile and Non Volatile compound.
• Due to Electron Spray Ionization (ESI) technique mass Spectroscopy
becomes very popular and generally coupled with Chromatography.

7. ESI Method

8. • A solution containing the sample molecule is sprayed through the
high voltage potential capillary by the help of nebulization gas.
• The sample comes to capillary tube and due to the nebulizing gas the
sample will be converted into sprayed droplets.
• And here we are supplying power.
• Due to the power supply here high voltage potential will be generated
at the tip of the capillary.
• Due to the high voltage potential that droplets becomes ionized.
• In next step the droplets comes to ionisation chamber and this
chamber is operated at normal atmospheric pressure.
• In this ionization chamber we are supplying heated dissolution gas
that is heated Nitrogen gas and due to that temperature of the
chamber becomes high.

9. • And at high temperature the sample is removed from the analyte
due to due to evaporation and it will converted in the form of
Molecular Ion.
• Heated desolvation gas will evaporate the solvent and it will produce
the molecular ion.
• For example --- If we have the molecule M and it is surrounded by
the Solvent S and this Solvent is Ionised like MSH+
• Now due to this heated dissolvation gas – Temp∆ Rises – And
evaporation takes place and it will converted into MH+
• Now this molecular Ion will go further towards ION accelerator
chamber and then to mass analyser.

10. Power is supplied.
Positive side is attached to capillary tube.
Negative side is attached to Ion accelerator chamber and it will
produce negative potential.
That negative potential will help this positive Ion to get the velocity
and ultimately it will move towards the mass analyzer.
It can also produce multiple charged ions along with the single
charged ions.

11. Uses :-
• Mass spectra is used in two general ways.
• 1. To prove the identity of two compounds.
• 2. To establish the structure of a new compound.
• It deals with the examination of the characteristics fragments (ions)
arising from the breakdown of organic molecules.
• Used to Ionised Proteins , Lipids , Olegosacchrides , Olegonucleotides,
Synthetic Polymer etc.

12. • It helps to establish the structure of a new compound in several ways
1. It can give the exact molecular mass.
2. It can give a molecular formula or it can reveal the presence of
certain structural units in a molecule.
• Trace gas analysis, respired gas monitors .
• Determining the isotopic compounds of elements within a sample .
• Pharmacokinetics, protein characterization.
• Glycan analysis and space exploration.

13. Applications:-
• It has both quantitative and qualitative uses.
• Helps in identification of unknown compounds.
• Determining the isotopic composition of elements in a molecule.
Quantifying the amount of compound in the sample.
• Determining the structure of a compound by observing fragmentation.
• Mass spectroscopy is now in very common use in analytical laboratories
that study physical, chemical or biological properties of a great variety of
compounds.

14. Thank you