Tandem mass

it is a hyphenated technique used to separated and identify the compounds.

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

3. INTRODUTION PRINCIPLE INSTRUMENTATION APPLICATION REFRENCES ACKNOLEDMENT 3 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 5 Mass spectroscopy

6. It provides the information about. Molecular formula Molecular weight Structural formula Protein sequence Isotopic abundances of different elements And much more… 6 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 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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11. Tandem mass has same instrumentation as MS but differ to very small extent Sample inlet Ion source Mass analyzer CID Triple quadrupole Detectors 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) 12 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 13 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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19. Tandem Mass can be used for protein sequencing Protein is identified in tandem mass spectrometer in two ways Top-down proteomic Bottom-down proteomics 19 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 20 DNA and RNA

21. 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. 21 NEWBORN SCREENING

22. Introduction to spectroscopy by PAVIA Fifth edition P#120 , 122 ,123 http://en.wikipedia.org/wiki/tandem_mass_spectroscopy . http://www.intechopen.com . http://www.sciencedirect.com . Silver Stein Book And many more…. 22 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. 23 Acknowlegdment

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