Tandem mass spectrometry is a technique that uses multiple stages of mass spectrometry to analyze molecular ions and their fragmentation patterns. It works by first analyzing parent ions (MS1), then inducing fragmentation through collision with gas molecules to produce daughter ions, which are then analyzed (MS2). This allows for more detailed structural information to be obtained compared to single-stage mass spectrometry. Common applications include protein sequencing, DNA/RNA sequencing, and newborn screening for metabolic diseases.

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2. 
Presented by:
Jameel Ahmed Buledi
 Assigned by: Prof. Dr. Shuhabdin Memon
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Tandem Mass

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 INTRODUTION
 PRINCIPLE
 INSTRUMENTATION
 APPLICATION
 REFRENCES
 ACKNOLEDMENT
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CONTENTS

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5. 
 Mass spectroscopy in an analytical technique used to
ionize the chemical species and sorts them according
to Mass-to-Charge ratio
 This technique is used in different fields of science
 Medicine
 Geology
 Forensic
 Pharmaceutical
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Mass spectroscopy

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It provides the information
about.
 Molecular formula
 Molecular weight
 Structural formula
 Protein sequence
 Isotopic abundances of different elements
 And much more…
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Cont.

7.  Tandem mass is an spectroscopic
technique
has same principle as mass
spectroscopy but in multiple
stages
 This technique is also known as
MS/MS or 𝑴𝑺 𝟐
 It has mostly two portion of
analysis
1.MS1
2.MS2 7
Tandem mass

8.  Tandem mass was introduced by W.M.A Neissen in 1999
 This technique currently is an important tool while in fundamental
studies concerning the behavior and structure of gas phase ions
 It analyze the parent and its daughter ions in the cell called Collision
Induced Dissociation cell
 The major breakthrough of Tandem mass was introduction of triple-
quadrupole mass analyzers
 World wide it is most probably use in determining the structures of
 Protein
 Peptides
 Nucleic acids and other biomolecules 8
Cont…

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a
Tandem
MS/MS
It separate the
ions according
to Mass-to-
Charge ratio
MS1 analyze
the parent
ions
MS2 analyze
the daughter
ions

11.  Tandem mass has same instrumentation as MS but differ
to very small extent
1. Sample inlet
2. Ion source
3. Mass analyzer
4. CID
5. Triple quadrupole
6. Detectors
7. Data analysis 11
INSTRUMENTATION

12.  Firstly the ion mixture is allowed to inter in ionization chamber in MS1
and also known as Q1
 The mixture is then ionized with different ionization sources like
ESI . CI. MALDI. ESI
 This process takes up time where larger or huge molecule breaks up into
fragments but these fragments are not enough to characterize the sample
because of its huge mass , although some fragments are formed.
 Thereafter we have two option with our fragmentized sample
 1. To move with the selected fragment into Q2
 2. To move up with all types of fragments into Q2
 This Q2 involves the Collision Induced Dissociation (CID)
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Cont.

13.  Now in CID the high pressure inert molecules are in contact
with the parent ion fragments
 Because of high kinetic energy is induced in the fragments and
now they became the Metastable ions
 The ions are further fragmentized into a Daughter one
 These daughter ions are accelerated towards the triple-
quadrupole analyzer
 The ions are now in MS2 which is called Q3
 Total three analyses steps are involved Q1 Q2 Q3
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cont.

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15.  The molecular ion are accelerated in CID
where most
often they interact with neutral
molecule e.g helium
nitrogen, organ.
 In collision kinetic energy converted
into internal energy which
results in the bond breakage and
fragmentation of molecular ion
into the smaller fragments.
 CID provides partial as well complete
structural information 15
COLLISION INDUCED
DISSOCIATION

16.  Quadrupole mass analyzer is made up of four metallic rods
connected with AC as well DC current which helps to resonate
the ions in direct trajectory .
 DC current is applied to keep the ions within the rods
 While AC is varied to resonate the ion towards the detector
 There are four rods in quadrupole among of them two are
negatively charged and other two are positively charged.
 The trajectory of the ions towards the detector depends on the
frequency of ions as well as on applied AC strength . 16
QUADURPOLE MASS
ANALYZER

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 Tandem Mass can be used for protein sequencing
Protein is identified in tandem mass spectrometer in two
ways
 Top-down proteomic
 Bottom-down proteomics
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Protein sequence

20. 
 This technique is also used in DNA and RNA
sequencing
 DNA sequencing is the method of determining the
precise order of nucleotides within a DNA molecule
 RNA sequencing is also called Whole Shotgun
Sequencing to reveal the presence and quantity of
RNA in biological sample at a given moment
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DNA and RNA

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 The development of Tandem Mas spectrometry
screening in early 1990s led to large expansion of
potentially congenital metabolic diseases that affect
blood level of organic acids.
 Newborn screening is the process testing newborn
babies for treatable genetic , metabolic and
hematologic diseases.
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NEWBORN
SCREENING

22. 
Introduction to spectroscopy by PAVIA Fifth edition
P#120 , 122 ,123
http://en.wikipedia.org/wiki/tandem_mass_spectrosc
opy.
http://www.intechopen.com.
http://www.sciencedirect.com.
Silver Stein Book
And many more….
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References

23. 
 I am thankful to Allah Almighty who had always been
taken at the edge of good and bad circumstances
 I am also keenly grateful to Prof. Dr. Shuahbdin Memon
for his tremendous support all the time whenever we
need him
 And nevertheless I am so much thankful to my colleagues
for critically finding holes to do better things and also for
their soulful encouragement.
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Acknowlegdment

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