Lcmsms

this will help to know about the advance technique to analysis the biological sample in cancer diagnosis and general separation of proteins based upon the molecular weight and helps to analysis the new drug synthesis level

1. By S.Boobash Raj 3 rd M.Tech Biotechnology Liquid Chromatography/Mass Spectroscopy/Mass Spectroscopy (LC/MS/MS)

2. CONTENT Hyphenated technique Introduction to LC/MS/MS Principles of LC/MS/MS Mass spectrometry Instrumentation of LC/MS/MS Method of working Sample preparation Ion sources Mass analyzers Applications& advantages

3. Hyphenated technique The term “hyphenation” was first adapted by “ Hirschfeld ” in 1980. The technique developed from the coupling of a separation technique and an on-line spectroscopic detection technology. Advantage of hyphenated techniques: Fast and accurate analysis Higher degree of automation Higher sample throughput Better reproducibility Reduction of contamination due to its closed system Separation and quantification achieved at same time List of Hyphenated techniques: 1. GC-MS, 2. LC/MS, 3. GC-IR, 4. LC/MS/MS, 5. GC-MS-MS

4. DEFINITION Liquid Chromatography with tandem mass spectrometry (LC-MS-MS) is a powerful analytical technique that combines the separating power of liquid chromatography with the highly sensitive and selective mass analysis capability of triple quadrupole mass spectrometry . The LC/MS/MS data may be used to provide information about the molecular weight, structure, identity and quantity of specific sample components.

5. Principles of LC/MS/MS Liquid chromatography generally utilizes very small particles and operating at relatively high pressure, and is referred to as high performanCe liquid chromatography (HPLC) In HPLC, the sample is forced by a liquid at high pressure (the mobile phase ) through a column that is packed with a stationary phase generally composed of irregularly or spherically shape particles derivatized to accomplish particular types of separations.

6. Mass spectrometry (MS) Mass spectrometry (MS) is an analytical technique that measures the mass-to-charge ratio of charged particles. MS works by ionizing chemical compounds to generate charged molecules or molecule fragments and measuring their mass-to-charge ratios.

7. Instrumentation of LC/MS/MS 1) HPLC Constitutees the LC part: Solvent system (mobile phase) Pumps Mixer Injector cOlumn 2) Mas spectrometer Ion sources electrospray ionization. Atmospheric Pressure Chemical Ionization. Atmospheric pressure photoionization

8. Sample preparation: Sample preparation consist of concentrating the analyte and removing compounds that can cause background ion or suppress ionization. Desalting to reduce the sodium and potassium adduct formation that commonly occurs in electrospray.

9. Ion sources Electrospray ionization (ESI) Atmospheric pressure chemical ionization(APCI) Atmospheric pressure photoionization(APPI)

10. Mass ANALYSER In mass analyzer the magnetic field is scanned to measure the different ions. Types of mass analyzer:- Quadra pole mass filter. Time of flight Ion trap Fourier transform ion cyclotron resonance (FT-ICR)

11. Quadra pole mass filter It is also known as a  transmission quadrupole mass spectrometer it consists of four cylindrical rods, set parallel to each other . Each opposing rod pair is connected together electrically, and a  radio frequency  (RF) voltage with a DC offset voltage is applied between one pair of rods and the other A linear series of three quadrupoles is known as a  triple quadrupole mass spectrometer . The first (Q 1 ) and third (Q 3 ) quadrupoles act as mass filters, and the middle (q 2 ) quadrupole is employed as a collision cell . .

12. Time of flight Time of flight  ( ToF ) is the measurement of the time taken by an object, particle or wave (be it acoustic, electromagnetic, etc.) to travel a distance through a medium.  Ions  are accelerated by an  electric field  of known strength . This acceleration results in an ion having the same  kinetic energy  as any other ion that has the same charge.  The time that it subsequently takes for the particle to reach a detector at a known distance is measured . this time will depend on the  mass-to-charge ratio  of the particle (heavier particles reach lower speeds). 

13. ion trap An  ion trap  is a combination of electric or magnetic fields used to capture charged particles, often in a system isolated from an external environment . Paul traps are commonly used as a components of a  mass spectrometer .

14. Work flow of lc/ ms / ms in cancer diagnostic

15. Just I wanna say Thank you