The document provides an overview of mass spectrometry. It discusses the history, principles, instrumentation, ionization techniques, mass analyzers, and applications of mass spectrometry. Mass spectrometry involves converting sample molecules to ions, separating the ions based on their mass-to-charge ratio, and detecting the ions. Key components include an ion source, mass analyzer, and ion detector. Common ionization methods include electron ionization and chemical ionization. Common mass analyzers are magnetic sector, quadrupole, time-of-flight, and ion trap. Mass spectrometry has various applications in fields like proteomics, metabolomics, and environmental analysis.
Flame photometry is a technique that uses the intensity of light emitted from a flame to determine the concentration of certain metal ions in a sample. When a sample is introduced into the flame, the metal ions are atomized and excited. As they return to the ground state, they emit light of characteristic wavelengths. The intensity of light emitted can then be measured to determine the concentration of the metal ions. Flame photometry is used to analyze samples for concentrations of ions like sodium, potassium, calcium, and lithium. It has applications in analyzing body fluids and determining metal concentrations in materials like cement.
FT-NMR uses Fourier transforms to convert time domain signals from nuclear magnetic resonance into frequency domain spectra. The sample is placed in a strong magnet and exposed to pulses of radio frequency radiation, producing a free induction decay signal that is recorded over time. This time domain signal is then digitized and analyzed using a Fourier transform program on a computer to produce the frequency domain NMR spectrum. FT-NMR provides higher sensitivity than continuous wave NMR, allowing analysis of smaller sample sizes.
Mass spectrometry(Ionization Techniques) by Ashutosh PankeAshutosh Panke
The document discusses various ionization techniques used in mass spectrometry. It describes several gas phase ionization methods including electron impact ionization, chemical ionization, and field ionization. It also discusses several desorption ionization techniques, notably fast atom bombardment, matrix assisted laser desorption/ionization, electrospray ionization, and surface enhanced laser desorption/ionization. The document provides details on the mechanisms and applications of these various ionization methods. It also categorizes mass analyzers and discusses time-of-flight mass analyzers.
Mass spectrometry deals with studying charged molecules and fragment ions produced from a sample exposed to ionizing conditions. It provides the relative intensity spectrum based on ions' mass to charge ratio, allowing identification of unknown compounds. The document discusses the basic principles, advantages, disadvantages, instrumentation, applications, and analysis techniques of mass spectrometry.
Quadrupole and Time of Flight Mass analysers.Gagangowda58
Description about important mass analysers Quadrupole and TOF: Principle, Construction and Working, Advantages and Disadvantages and their Applications.
This document provides an overview of mass spectrometry. It begins with an introduction to mass spectrometry, explaining that it is a technique used to analyze molecules by ionizing them and measuring the mass-to-charge ratios of the ions produced. It then covers the basic principles of mass spectrometry, describing how molecules are ionized by bombarding them with electrons and how the ions are separated and detected based on their mass-to-charge ratios. The remainder of the document discusses the theory behind mass spectrometry, describes common types of mass spectrometers, and outlines the ionization and fragmentation processes involved in mass spectrometry analysis.
The document discusses various ionization techniques used in mass spectrometry. It describes electron impact ionization, chemical ionization including positive and negative modes, atmospheric pressure chemical ionization, field ionization, field desorption, and electrospray ionization. Each technique is explained in terms of its construction, working principle, advantages, and limitations. Electron impact ionization is the most widely used classical method that produces extensive fragmentation, while chemical ionization and electrospray ionization are suited for high molecular weight compounds that undergo less fragmentation.
The document discusses atomic absorption spectroscopy. It begins with an introduction describing how atomic absorption spectroscopy measures the concentration of an element by measuring the amount of light absorbed at a characteristic wavelength when it passes through atoms of that element. It then describes the principle, instrumentation, applications, and sources of interference in atomic absorption spectroscopy. The key sources of interference discussed are non-spectral interferences such as matrix, chemical, and ionization interferences and spectral interferences such as background absorption.
Flame photometry is a technique that uses the intensity of light emitted from a flame to determine the concentration of certain metal ions in a sample. When a sample is introduced into the flame, the metal ions are atomized and excited. As they return to the ground state, they emit light of characteristic wavelengths. The intensity of light emitted can then be measured to determine the concentration of the metal ions. Flame photometry is used to analyze samples for concentrations of ions like sodium, potassium, calcium, and lithium. It has applications in analyzing body fluids and determining metal concentrations in materials like cement.
FT-NMR uses Fourier transforms to convert time domain signals from nuclear magnetic resonance into frequency domain spectra. The sample is placed in a strong magnet and exposed to pulses of radio frequency radiation, producing a free induction decay signal that is recorded over time. This time domain signal is then digitized and analyzed using a Fourier transform program on a computer to produce the frequency domain NMR spectrum. FT-NMR provides higher sensitivity than continuous wave NMR, allowing analysis of smaller sample sizes.
Mass spectrometry(Ionization Techniques) by Ashutosh PankeAshutosh Panke
The document discusses various ionization techniques used in mass spectrometry. It describes several gas phase ionization methods including electron impact ionization, chemical ionization, and field ionization. It also discusses several desorption ionization techniques, notably fast atom bombardment, matrix assisted laser desorption/ionization, electrospray ionization, and surface enhanced laser desorption/ionization. The document provides details on the mechanisms and applications of these various ionization methods. It also categorizes mass analyzers and discusses time-of-flight mass analyzers.
Mass spectrometry deals with studying charged molecules and fragment ions produced from a sample exposed to ionizing conditions. It provides the relative intensity spectrum based on ions' mass to charge ratio, allowing identification of unknown compounds. The document discusses the basic principles, advantages, disadvantages, instrumentation, applications, and analysis techniques of mass spectrometry.
Quadrupole and Time of Flight Mass analysers.Gagangowda58
Description about important mass analysers Quadrupole and TOF: Principle, Construction and Working, Advantages and Disadvantages and their Applications.
This document provides an overview of mass spectrometry. It begins with an introduction to mass spectrometry, explaining that it is a technique used to analyze molecules by ionizing them and measuring the mass-to-charge ratios of the ions produced. It then covers the basic principles of mass spectrometry, describing how molecules are ionized by bombarding them with electrons and how the ions are separated and detected based on their mass-to-charge ratios. The remainder of the document discusses the theory behind mass spectrometry, describes common types of mass spectrometers, and outlines the ionization and fragmentation processes involved in mass spectrometry analysis.
The document discusses various ionization techniques used in mass spectrometry. It describes electron impact ionization, chemical ionization including positive and negative modes, atmospheric pressure chemical ionization, field ionization, field desorption, and electrospray ionization. Each technique is explained in terms of its construction, working principle, advantages, and limitations. Electron impact ionization is the most widely used classical method that produces extensive fragmentation, while chemical ionization and electrospray ionization are suited for high molecular weight compounds that undergo less fragmentation.
The document discusses atomic absorption spectroscopy. It begins with an introduction describing how atomic absorption spectroscopy measures the concentration of an element by measuring the amount of light absorbed at a characteristic wavelength when it passes through atoms of that element. It then describes the principle, instrumentation, applications, and sources of interference in atomic absorption spectroscopy. The key sources of interference discussed are non-spectral interferences such as matrix, chemical, and ionization interferences and spectral interferences such as background absorption.
This document discusses the applications of mass spectrometry. It can be used for structure elucidation, detection of impurities, quantitative analysis, drug metabolism studies, and clinical, toxicological and forensic applications. Mass spectrometry has qualitative uses like determining molecular weight, molecular formula, and compound structure through fragmentation patterns. It also has quantitative uses such as determining isotope abundance, isotope ratios, heat of vaporization, heat of sublimation, ionization potentials, and detecting impurities. Mass spectrometry can also be used to study ion-molecule reactions, identify unknown compounds, and analyze proteins.
Nuclear magnetic resonance spectroscopy is an analytical technique that uses radio waves and strong magnetic fields to characterize organic molecules. The key components of an NMR spectrometer include a sample holder, permanent magnet, magnetic coil, radio frequency generator and receiver, and readout system. The magnet provides a strong and uniform magnetic field, the generator produces radio waves to excite the nuclei, and the receiver and readout system detect and display the resonance signals to identify the molecules.
Fragmentation techniques in mass spectroscopyMahendra G S
Fragmentation in mass spectrometry involves the breakdown of molecular ions into smaller daughter ions. There are several types of fragmentation including homolytic cleavage, heterolytic cleavage, and rearrangement reactions. Homolytic cleavage involves equal transfer of electrons to both atoms, forming a radical and cation. Heterolytic cleavage involves both electrons being taken by one atom, forming an even electron cation and radical. Rearrangement reactions require changes to multiple bonds and can eliminate smaller molecules. Common rearrangements include McLafferty rearrangement and elimination reactions. Fragmentation patterns provide information about functional groups present in molecules.
Mass spectroscopy, Ionization techniques and types of mass analyzers Muhammad Asif Shaheeen
Mass spectroscopy is a technique used to determine the molecular mass and elemental composition of a compound. It works by ionizing molecules using electron bombardment or chemical ionization and then separating the resulting ions based on their mass-to-charge ratio using electric and magnetic fields. The instrument consists of an ion source, a mass analyzer, and an ion detector. Common ion sources include electron impact, chemical ionization, and electrospray ionization, with each having advantages for different types of samples. The document provides detailed explanations of the basic principles and components of mass spectroscopy.
This document discusses mass spectrometry and chemical ionization. It explains that mass spectrometry can provide molecular structure information by determining molecular weight. Chemical ionization is a softer ionization technique than electron ionization, which causes less fragmentation and makes it easier to detect molecular ions. The document describes the instrumentation for chemical ionization, where a reagent gas like methane is ionized by electrons and the resulting ions collide with analyte molecules to form positively or negatively charged molecular ions depending on the mode used. The advantages of chemical ionization are also summarized.
Mass spectrometry breaks molecules into fragments using high-energy electrons. The masses of the fragments and their relative abundances provide information about molecular weight, structure, and functional groups. Key principles for interpreting mass spectra include identifying the parent ion peak from molecular weight, observing isotope effects, and examining fragmentation patterns according to the nitrogen rule and common losses of simple groups like methyl or ethyl.
Nuclear magnetic resonance (NMR) spectroscopyVK VIKRAM VARMA
SPECTROSCOPY
NMR SPECTROSCOPY
HISTORY
THEORY
PRINCIPLE
INSTRUMENTATION
SOLVENTS USED IN NMR(PROTON NMR)
CHEMICAL SHIFT
FACTORS AFFECTING CHEMICAL SHIFT
RELAXATION PROCESS
SPIN-SPIN COUPLING
푛+1 RULE
NMR SIGNALS IN VARIOUS COMPOUNDS
COUPLING CONSTANT
NUCLEAR MAGNETIC DOUBLE RESONANCE/ SPIN DECOUPLING
FT-NMR
ADVANTAGES & DISADVANTAGES
APPLICATIONS
REFERENCE
This document provides an overview of the key components and operating principles of mass spectrometry. It discusses the inlet system, ion sources, mass analyzers, detectors, and vacuum system. Common types of ion sources like electron impact and chemical ionization are described. Popular mass analyzers such as quadrupole, time-of-flight, ion trap, and double focusing are explained. The document also covers the theory behind how mass spectrometry separates ions based on their mass-to-charge ratio and discusses the need for high vacuum levels in mass spectrometers.
this ppt contain all basic information related to the mass spectrometry like introduction, principle of MS, type of ions, fragmentation processes eg. mcLafferty rearrangement, alpha clevage, sigma bond clevage, retro-diels-alder reaction
Types of Ions Produced In Mass SpectrometryAditya Sharma
Mass spectrometry produces different types of ions that can be analyzed. Aditya Sharma, an M.S. in Pharmaceutical Analysis from NIPER Guwahati, gave a presentation on the types of ions produced in mass spectrometry. The presentation covered the key topic of ions produced in mass spectrometry.
This document provides an overview of different ionization techniques used in mass spectrometry. It discusses gas phase ionization methods like electron impact and chemical impact ionization. It also covers desorptive ionization techniques such as field desorption, fast atom bombardment, and matrix-assisted laser desorption ionization. Finally, it summarizes evaporative ionization methods like atmospheric pressure chemical ionization and atmospheric pressure photoionization. The document aims to explain the basic principles, applications, advantages, and limitations of various ionization methods for mass spectrometry analysis.
Mass analyzers separate ionized molecules based on their mass-to-charge ratios. The main types are quadrupole, time-of-flight, magnetic sector, quadrupole ion trap, and ion cyclotron resonance. A quadrupole uses oscillating electric fields to selectively transmit ions through four rods. Time-of-flight separates ions by their time of flight through a field-free region, with lighter ions arriving first. Magnetic sector analyzers use magnetic and electric fields to curve ion trajectories based on m/z.
The document provides information about mass spectrometry including:
- Mass spectrometry is a powerful analytical technique that uses instruments called mass spectrometers to identify molecules by breaking them into ionized fragments and measuring their mass-to-charge ratios.
- The basic components of a mass spectrometer are the sample inlet, ionization source, mass analyzer, and ion detector. Common ionization sources are electrospray ionization, matrix-assisted laser desorption/ionization, and electron ionization. Common mass analyzers are quadrupoles, ion traps, and time-of-flight.
- Mass spectrometry has a variety of applications and has undergone significant technological developments since its invention in the early 20th
Mass spectrometry is an analytical technique that ionizes molecules and separates the resulting ions based on their mass-to-charge ratio. It is a powerful qualitative and quantitative technique used to measure a wide range of clinically relevant analytes. Various ionization sources are used depending on the type of sample, including electron ionization, chemical ionization, electrospray ionization, and matrix-assisted laser desorption/ionization. Ions are accelerated into a mass analyzer such as a quadrupole, magnetic sector, or time-of-flight analyzer which separates the ions based on m/z. The detected ions produce a mass spectrum that provides information about molecular structure.
1) NMR spectroscopy is a technique that uses radio waves to induce transitions between magnetic energy levels of atomic nuclei, providing information about molecular structure.
2) There are two main types of NMR - 1H NMR which identifies hydrogen atoms, and 13C NMR which identifies carbon atoms.
3) An NMR instrument consists of a strong magnet to align nuclear spins, a radiofrequency transmitter to perturb the spins, and a receiver to measure the emitted radio waves as spins relax.
NMR Instrumentation
ppt
Magnet
Permanent and conventional electromagnets
The Magnetic Field Sweep
Sweep Generator
frequency sweep method
field sweep method
The Sample Holder
The Sample Probe
Radio Frequency Generator
Oscillator
Radio Frequency Receiver
Amplifier
The Signal Detector and Recording System
NMR Instrumentation
ppt
Magnet
Permanent and conventional electromagnets
The Magnetic Field Sweep
Sweep Generator
frequency sweep method
field sweep method
The Sample Holder
The Sample Probe
Radio Frequency Generator
Oscillator
Radio Frequency Receiver
Amplifier
The Signal Detector and Recording System
This document discusses flame emission spectroscopy. It begins with an introduction stating that flame emission spectroscopy uses a flame to provide energy and excite atoms introduced into the flame. It then covers the history, principle, instrumentation, applications and potential interferences of flame emission spectroscopy. The principle involves desolvation, vaporization, atomization, excitation and emission of light at characteristic wavelengths. Common instrumentation components include burners, atomizers, monochromators, detectors and readouts. Applications include analysis of chemicals, soils, plants, waters and more. Potential issues include matrix, chemical, ionization and spectral interferences.
SPECTROSCOPY
INFRARED SPECTROSCOPY
HISTORY
PRINCIPLE
MODES OF VIBRATION
INSTRUMENTATION
SAMPLE HANDLING
FTIR (FOURIER TRANSFORM INFRARED) SPECTROMETER
PRINCIPLE
INSTRUMENTATION
WORKING
DISPERSIVE VERSUS FTIR
ADVANTAGES & DISADVANTAGES OF FTIR WITH APPLICATIONS
FACTORS AFFECTING VIBRATIONAL FREQUENCIES
IR SPECTRA REGION
IR SPECTRA INTERPRETATION
EXAMPLES
ADVANTAGES AND DISADVANTAGES OF IR
APPLICATIONS OF IR
Reference
This document discusses the applications of mass spectrometry. It can be used for structure elucidation, detection of impurities, quantitative analysis, drug metabolism studies, and clinical, toxicological and forensic applications. Mass spectrometry has qualitative uses like determining molecular weight, molecular formula, and compound structure through fragmentation patterns. It also has quantitative uses such as determining isotope abundance, isotope ratios, heat of vaporization, heat of sublimation, ionization potentials, and detecting impurities. Mass spectrometry can also be used to study ion-molecule reactions, identify unknown compounds, and analyze proteins.
Nuclear magnetic resonance spectroscopy is an analytical technique that uses radio waves and strong magnetic fields to characterize organic molecules. The key components of an NMR spectrometer include a sample holder, permanent magnet, magnetic coil, radio frequency generator and receiver, and readout system. The magnet provides a strong and uniform magnetic field, the generator produces radio waves to excite the nuclei, and the receiver and readout system detect and display the resonance signals to identify the molecules.
Fragmentation techniques in mass spectroscopyMahendra G S
Fragmentation in mass spectrometry involves the breakdown of molecular ions into smaller daughter ions. There are several types of fragmentation including homolytic cleavage, heterolytic cleavage, and rearrangement reactions. Homolytic cleavage involves equal transfer of electrons to both atoms, forming a radical and cation. Heterolytic cleavage involves both electrons being taken by one atom, forming an even electron cation and radical. Rearrangement reactions require changes to multiple bonds and can eliminate smaller molecules. Common rearrangements include McLafferty rearrangement and elimination reactions. Fragmentation patterns provide information about functional groups present in molecules.
Mass spectroscopy, Ionization techniques and types of mass analyzers Muhammad Asif Shaheeen
Mass spectroscopy is a technique used to determine the molecular mass and elemental composition of a compound. It works by ionizing molecules using electron bombardment or chemical ionization and then separating the resulting ions based on their mass-to-charge ratio using electric and magnetic fields. The instrument consists of an ion source, a mass analyzer, and an ion detector. Common ion sources include electron impact, chemical ionization, and electrospray ionization, with each having advantages for different types of samples. The document provides detailed explanations of the basic principles and components of mass spectroscopy.
This document discusses mass spectrometry and chemical ionization. It explains that mass spectrometry can provide molecular structure information by determining molecular weight. Chemical ionization is a softer ionization technique than electron ionization, which causes less fragmentation and makes it easier to detect molecular ions. The document describes the instrumentation for chemical ionization, where a reagent gas like methane is ionized by electrons and the resulting ions collide with analyte molecules to form positively or negatively charged molecular ions depending on the mode used. The advantages of chemical ionization are also summarized.
Mass spectrometry breaks molecules into fragments using high-energy electrons. The masses of the fragments and their relative abundances provide information about molecular weight, structure, and functional groups. Key principles for interpreting mass spectra include identifying the parent ion peak from molecular weight, observing isotope effects, and examining fragmentation patterns according to the nitrogen rule and common losses of simple groups like methyl or ethyl.
Nuclear magnetic resonance (NMR) spectroscopyVK VIKRAM VARMA
SPECTROSCOPY
NMR SPECTROSCOPY
HISTORY
THEORY
PRINCIPLE
INSTRUMENTATION
SOLVENTS USED IN NMR(PROTON NMR)
CHEMICAL SHIFT
FACTORS AFFECTING CHEMICAL SHIFT
RELAXATION PROCESS
SPIN-SPIN COUPLING
푛+1 RULE
NMR SIGNALS IN VARIOUS COMPOUNDS
COUPLING CONSTANT
NUCLEAR MAGNETIC DOUBLE RESONANCE/ SPIN DECOUPLING
FT-NMR
ADVANTAGES & DISADVANTAGES
APPLICATIONS
REFERENCE
This document provides an overview of the key components and operating principles of mass spectrometry. It discusses the inlet system, ion sources, mass analyzers, detectors, and vacuum system. Common types of ion sources like electron impact and chemical ionization are described. Popular mass analyzers such as quadrupole, time-of-flight, ion trap, and double focusing are explained. The document also covers the theory behind how mass spectrometry separates ions based on their mass-to-charge ratio and discusses the need for high vacuum levels in mass spectrometers.
this ppt contain all basic information related to the mass spectrometry like introduction, principle of MS, type of ions, fragmentation processes eg. mcLafferty rearrangement, alpha clevage, sigma bond clevage, retro-diels-alder reaction
Types of Ions Produced In Mass SpectrometryAditya Sharma
Mass spectrometry produces different types of ions that can be analyzed. Aditya Sharma, an M.S. in Pharmaceutical Analysis from NIPER Guwahati, gave a presentation on the types of ions produced in mass spectrometry. The presentation covered the key topic of ions produced in mass spectrometry.
This document provides an overview of different ionization techniques used in mass spectrometry. It discusses gas phase ionization methods like electron impact and chemical impact ionization. It also covers desorptive ionization techniques such as field desorption, fast atom bombardment, and matrix-assisted laser desorption ionization. Finally, it summarizes evaporative ionization methods like atmospheric pressure chemical ionization and atmospheric pressure photoionization. The document aims to explain the basic principles, applications, advantages, and limitations of various ionization methods for mass spectrometry analysis.
Mass analyzers separate ionized molecules based on their mass-to-charge ratios. The main types are quadrupole, time-of-flight, magnetic sector, quadrupole ion trap, and ion cyclotron resonance. A quadrupole uses oscillating electric fields to selectively transmit ions through four rods. Time-of-flight separates ions by their time of flight through a field-free region, with lighter ions arriving first. Magnetic sector analyzers use magnetic and electric fields to curve ion trajectories based on m/z.
The document provides information about mass spectrometry including:
- Mass spectrometry is a powerful analytical technique that uses instruments called mass spectrometers to identify molecules by breaking them into ionized fragments and measuring their mass-to-charge ratios.
- The basic components of a mass spectrometer are the sample inlet, ionization source, mass analyzer, and ion detector. Common ionization sources are electrospray ionization, matrix-assisted laser desorption/ionization, and electron ionization. Common mass analyzers are quadrupoles, ion traps, and time-of-flight.
- Mass spectrometry has a variety of applications and has undergone significant technological developments since its invention in the early 20th
Mass spectrometry is an analytical technique that ionizes molecules and separates the resulting ions based on their mass-to-charge ratio. It is a powerful qualitative and quantitative technique used to measure a wide range of clinically relevant analytes. Various ionization sources are used depending on the type of sample, including electron ionization, chemical ionization, electrospray ionization, and matrix-assisted laser desorption/ionization. Ions are accelerated into a mass analyzer such as a quadrupole, magnetic sector, or time-of-flight analyzer which separates the ions based on m/z. The detected ions produce a mass spectrum that provides information about molecular structure.
1) NMR spectroscopy is a technique that uses radio waves to induce transitions between magnetic energy levels of atomic nuclei, providing information about molecular structure.
2) There are two main types of NMR - 1H NMR which identifies hydrogen atoms, and 13C NMR which identifies carbon atoms.
3) An NMR instrument consists of a strong magnet to align nuclear spins, a radiofrequency transmitter to perturb the spins, and a receiver to measure the emitted radio waves as spins relax.
NMR Instrumentation
ppt
Magnet
Permanent and conventional electromagnets
The Magnetic Field Sweep
Sweep Generator
frequency sweep method
field sweep method
The Sample Holder
The Sample Probe
Radio Frequency Generator
Oscillator
Radio Frequency Receiver
Amplifier
The Signal Detector and Recording System
NMR Instrumentation
ppt
Magnet
Permanent and conventional electromagnets
The Magnetic Field Sweep
Sweep Generator
frequency sweep method
field sweep method
The Sample Holder
The Sample Probe
Radio Frequency Generator
Oscillator
Radio Frequency Receiver
Amplifier
The Signal Detector and Recording System
This document discusses flame emission spectroscopy. It begins with an introduction stating that flame emission spectroscopy uses a flame to provide energy and excite atoms introduced into the flame. It then covers the history, principle, instrumentation, applications and potential interferences of flame emission spectroscopy. The principle involves desolvation, vaporization, atomization, excitation and emission of light at characteristic wavelengths. Common instrumentation components include burners, atomizers, monochromators, detectors and readouts. Applications include analysis of chemicals, soils, plants, waters and more. Potential issues include matrix, chemical, ionization and spectral interferences.
SPECTROSCOPY
INFRARED SPECTROSCOPY
HISTORY
PRINCIPLE
MODES OF VIBRATION
INSTRUMENTATION
SAMPLE HANDLING
FTIR (FOURIER TRANSFORM INFRARED) SPECTROMETER
PRINCIPLE
INSTRUMENTATION
WORKING
DISPERSIVE VERSUS FTIR
ADVANTAGES & DISADVANTAGES OF FTIR WITH APPLICATIONS
FACTORS AFFECTING VIBRATIONAL FREQUENCIES
IR SPECTRA REGION
IR SPECTRA INTERPRETATION
EXAMPLES
ADVANTAGES AND DISADVANTAGES OF IR
APPLICATIONS OF IR
Reference
Mass spectrometry is a powerful analytical technique used to identify unknown compounds based on their mass. It works by converting analyte molecules to ions, separating the ions based on their mass-to-charge ratio, and detecting the ions. Common applications include determining the structures of biomolecules like proteins, identifying drugs and metabolites in biological samples, and quantifying elemental compositions. The document provides details on the principles, instrumentation components like ion sources and mass analyzers, and various applications of mass spectrometry.
SPECTROSCOPY
UV-VISIBLE SPECTROSCOPY
PRINCIPLE
THEORY
LAWS
LAMBERT LAW
BEER'S LAW
LAMBERT BEER LAW
ABSORPTION & Intensity SHIFTS
WOODWARD-FIESCER RULE WITH EXAMPLES
INSTRUMENTATION (DETAILED)
CHOICE OF SOLVENT
SOLVENT EFFECT
EFFECT OF CONCENTRATION, TEMPERATURE & 푝퐻
ADVANTAGES & DISADVANTAGES
APPLICATIONS
REFERENCE
An opto-electric nuclear battery is a device that converts nuclear energy into light, which it then uses to generate electrical energy. A beta-emitter such as technetium-99 or strontium-90 is suspended in a gas or liquid containing luminescent gas molecules of the excimer type, constituting a "dust plasma." This permits a nearly lossless emission of beta electrons from the emitting dust particles
Introduction to Semiconductor elect (1).pdfmansi21bphn002
Semiconductor electrochemistry is the study of electrochemical processes involving semiconductors. It involves the interaction of light, electrons, and chemical reactions at semiconductor surfaces. When light is absorbed by a semiconductor, electrons are excited from the valence to conduction band, creating electron-hole pairs that can undergo electrochemical reactions. Applications of semiconductor electrochemistry include solar energy conversion using photoelectrochemical cells, water splitting to produce hydrogen fuel, environmental remediation of pollutants, and chemical sensing.
Mass spectrometry is a technique that involves (1) creating gas phase ions from analyte atoms or molecules, (2) separating the ions based on their mass-to-charge ratio, and (3) measuring ion abundance. It provides qualitative and quantitative analysis, molecular structure information, and is used for identification and characterization. The basic components include a sample introduction system, ion source, mass analyzer, detector, and vacuum system. Common applications are in drug discovery, clinical testing, genomics, environmental analysis, and geology.
In situ hydrodynamic spectroscopy was used to characterize porous energy storage electrodes during lithium ion insertion/extraction. The technique uses electrochemical quartz crystal microbalance (EQCM) which monitors changes in resonance frequency and width during the electrochemical process. Three experiments were conducted:
1) Validation of the technique by measuring model surfaces with known properties.
2) Characterization of spray-pyrolyzed LiMn2O4 coatings of different mass loadings, allowing determination of specific capacity.
3) In situ measurements during charging/discharging of a LiMn2O4 electrode, showing swelling and shrinking of the porous layer associated with solvent insertion and extraction. The technique provides detailed information on changes
This document discusses quantum levitation using superconductors. It begins by defining superconductivity as zero electrical resistance and magnetic field expulsion below a critical temperature. It then discusses Meissner effect and different types of superconductors. The document introduces the concept of quantum locking, where magnetic flux lines do not move inside an ultra-thin type-2 superconductor. Applications of quantum levitation include frictionless bearings, gravity manipulation, and lossless electrical machines. Future prospects include quantum levitation trains with greater stability than maglev trains.
The document summarizes research into using plasmonic silver nanowires to harvest hot electrons for chemical reactions. Nanowires were synthesized inside anodic aluminum oxide nanopores using electrodeposition. Scanning electron microscopy images showed the nanowires had a unique stacked ring structure, and absorption spectroscopy indicated absorption in the visible spectrum. Removing the alumina template left just the silver nanowire structures. Future work will refine the nanostructures and integrate them into a device for energy storage applications.
This document discusses radioactivity and atomic structure. It begins by explaining the need to study radioactivity for diagnosis, therapy, and medical research. It then defines atoms and their structure, including atomic number and mass number. The document discusses isotopes and radioactive decay, including different types of decay and half-life. It also covers radiation properties, detection and measurement of radioactivity, and radiation safety quantities and units.
Mass spec and thermal methods of analysis - By France ChavangwaneFranceChavangwane
Heat flux Differential Scanning Calorimetry (DSC) diagram
Working principles of Heat Flux DSC
Advantages and disadvantages of HEAT FLUX DSC
labelled diagram showing Matrix-Assisted Laser Desorption-Ionization (MALDI)
Working principle of MALDI
Advantages and disadvantages of MALDI
Labelled Diagram of FAB
.Working principle of FAB (Fast Atom Bombardment)
Advantages and Disadvantages of FAB
Conclusion
Labelled Diagram Of continuous- dynode electron multiplier diagram
.Working principle Of continuous- dynode electron multiplier continued
Advantages and disadvantages of continuous- dynode electron multiplier
Labelled diagram of Faraday Cup Collector
Working principle of Faraday Cup Collector
Advantages and disadvantages of Faraday Cup Collector
References
This document discusses the cooking mechanism in microwave ovens. It begins by explaining what microwave radiation is and how it is used to cook food by causing water molecules in the food to vibrate, generating heat. It then discusses the history and development of microwave ovens, how they work, and the key components involved like the magnetron. It also covers safety considerations regarding microwave radiation exposure and standards.
Analytical Spectroscopic systems
Mass Spectrometry
Atomic mass to charge ratio
Laser Raman
Spectroscopy
Molecular vibrational modes
Laser Induced
Breakdown
Spectroscopy
Atomic emission
Visible Reflectance
Spectroscopy
Reflected color
This document discusses nuclear battery technology. It begins with objectives like developing small, reliable power sources. It outlines the report's phases and literature review. It introduces nuclear batteries, which use radioactive isotope decay rather than a chain reaction. Conversion techniques are thermal (based on temperature differences) or non-thermal. Thermal examples include thermionic and radioisotope thermoelectric generators. Non-thermal include direct charging and betavoltaics. Advantages are long life, high energy density, and use of nuclear waste. Applications include spacecraft and pacemakers.
This paper analyzes the reliability of MOSFETs that use indium-tin oxide as the gate oxide instead of silicon dioxide. Interface trap charges at the oxide-silicon interface can degrade MOSFET performance by changing the threshold voltage over time. The paper finds that MOSFETs using indium-tin oxide exhibit improved immunity to the effects of interface trap charges compared to those using silicon dioxide. Specifically, indium-tin oxide MOSFETs show enhanced static, linearity, and intermodulation performance metrics when subjected to both positive and negative interface trap charges. Thus, indium-tin oxide has potential to improve MOSFET reliability by reducing sensitivity to interface trap charge effects.
Characterization of the electrical properties of interfaces by impedance spec...Edmund Mills
Summary of major research accomplishments during PhD. Emphasizes my most recent work: the development of a new method for the determination of Peukert's constant, and the demonstration of the utility of Peukert's equation for the analysis of supercapacitor performance.
Similar to Mass spectrometry (Analytical Technique) (20)
MICROBIAL CONTAMINATION IN HERBS AND THEIR FORMULATIONSVK VIKRAM VARMA
INTRODUCTION
SOURCES OF CONTAMINATION
RAW MATERIALS
PACKAGING MATERIALS
LIMITS FOR MICROBIAL CONTAMINATION
LIMITS AS PER WHO
TYPES OF CONTAMINATION
DIRECT CONTAMINATION
CROSS CONTAMINATION
DETERMINATION OF CONTAMINATION
TOTAL VIABLE COUNT
PRETREATMENT OF HERBAL MATERIALS
REFRENCES
Micropropagation
steps of micropropagation
system used to regenerate plantlets by micropropagation
methods of micropropagation
embrogensis
organogenesis
bud culture
How does micropropogation work?
Examples with flow diagrams
Advantages & Disadvantages
Applications
Reference
CCRAS (central council for reasearch in ayurvedic sciences)VK VIKRAM VARMA
Introduction
Sowa-Rigpa
CCRAS Website
CCRAS Vision & Mission
Organisation chart
Seniority List
Institutes
Research Activities
The Broad Areas of Research Comprise
Clinical Research
Fundamental Research
Pharmacology Research
Medicinal Plant Research, Drug Standardisation Research
Literature Research
AYUSH Research Portal
Outreach Activities
Publications
E-books
Reference
Introduction
Classification with examples
Regulatory provisions relating to manufacturing the cosmetics
Requirements of the factory premises for manufacture of cosmetics
Requirements of plant and equipment
Manufacturing record
Prohibition of the manufacture
Offences and penalties
Export and import of potential herbal cosmetics
Some Indian importers of herbal cosmetics
WHAT IS PHARMACOPOEIA?
HISTORY OF PHARMACOPOEIA
CONTENT OF PHARMACOPOEIA
WHAT IS MONOGRAPH?
PURPOSE OF MONOGRAPH
TYPES OF MONOGRAPH
IMPORTANCE OF CONTENT OF MONOGRAPH
MONOGRAPH DEVELOPMENT PROCESS
INDIAN PHARMACOPOEIA
format of monograph
AYURVEDIC PHARMACOPOEIA
format of monograph
UNNANI PHARMACOPOEIA
format of monograph
SIDDHA PHARMACOPOEIA
format of monograph
GERMAN HOMOEPATHIC PHARMACOPOEIA
format of monograph
US PHARMACOPOEIA
format of monograph
JAPANESE PHARMACOPOEIA
format of monograph
BRITISH PHARMACOPOEIA
format of monograph
EUROPEAN PHARMACOPOEIA
format of monograph
COMPARATIVE STUDY
CONCLUSION
Reference
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2. CONTENTS
• INTRODUCTION
• HISTORY
• PRINCIPLE
• THEORY
• TERMINOLOGY
• INSTRUMENTATION
• IONISATION TECHNIQUES
• MASS ANALYSERS & TYPES
• MASS DETECTORS & TYPES
• TYPES OF IONS
• MASS FRAGMENTATION RULES
• MASS FRAGMENTATION PATTERNS
• MASS FRAGMENTATION TYPES
• TYPES OF PEAKS
• TANDEM MASS SPECTROMETRY
• ADVANTAGES & DISADVANTAGES OF
MS
• APPLICATIONS OF MS
• REFERENCES
21-12-2019V.K. VIKRAM VARMA 2
3. INTRODUCTION
•MASS SPECTROMETRY(MS) IS THE MOST ACCURATE
METHOD AMONGST OTHER SPECTROSCOPIC METHODS.
•IN WHICH SAMPLE IS CONVERTED TO RAPIDLY MOVING
POSITIVE IONS BY ELECTRON BOMBARDMENT &
CHARGED PARTICLES ARE SEPARATED ACCORDING TO
THEIR MASSES.
•MASS SPECTRA: IT IS A PLOT OF RELATIVE ABUNDANCE
AGAINST THE RATIO OF MASS/CHARGE.
21-12-2019V.K. VIKRAM VARMA 3
4. HISTORY
1897
•J.J. Thomson. Discovered electrons by cathode rays experiment.
Nobel prize in 1906.
1919
•Francis Aston recognized 1st mass spectrometer and measure z/m
of ionic compounds.
1934
•First double focusing magnetic analyser was invented by Johnson
& Neil.
1966
•Munson & field described chemical ionisation.
21-12-2019V.K. VIKRAM VARMA 4
5. CONTD.
1968
•Electrospray ionisation was invented by Dole, Mack & friends.
1975
•Atmospheric pressure chemical ionisation(APCI) was developed
by Caroll & others.
1985
•F.Hillenkamo, M.Karas & co-workers describe & coin the term
matrix assisted laser desorption ionisation(MALDI)
1989
•Paul discovered the ion trap technique.
21-12-2019V.K. VIKRAM VARMA 5
8. PRINCIPLE
Organic molecules
are bombarded with
electrons
Converted into highly
energetic positively
charged ions
(Molecular/Parent
ions )
Further breakup into
smaller ions
(Fragment/Daughter
ions)
Formed ions are
separated by
deflection of magnetic
field according to
their mass & charge.
Mass spectrum
21-12-2019V.K. VIKRAM VARMA 8
11. THEORY
•IT IS AN INSTRUMENT WHICH HELP IN SEPARATING THE
INDIVIDUAL ATOMS OR MOLECULES BECAUSE OF THE
DIFFERENCE IN THE MASSES.
•THREE BASIC FUNCTIONS:
TO VAPORISE COMPOUNDS OF VARYING VOLATILITY.
PRODUCE IONS FROM THE NEUTRAL COMPOUNDS IN THE
VAPORISE PHASE.
TO SEPARATE IONS ACCORDING TO THEIR MASS/CHARGE
RATIO & TO RECORD THEM.
21-12-2019V.K. VIKRAM VARMA 11
12. TERMINOLOGY
•MASS NUMBER (A): TOTAL NUMBER OF PROTONS AND
NEUTRONS IN AN ATOMIC NUCLEUS.
•MOLECULAR ION: IONS OBTAINED BY THE LOSS OF
AN ELECTRONS FROM THE MOLECULE.
•BASE PEAK: MOST INTENSE PEAK IN THE MASS
SPECTROMETRY, ASSIGNED 100% INTENSITY.
21-12-2019V.K. VIKRAM VARMA 12
13. CONTD.
• 𝐌+
: SYMBOL OFTEN GIVEN TO THE MOLECULAR ION.
•RADICAL ION: POSITIVE CHARGED SPECIES WITH AN
ODD NUMBER OF ELECTRONS.
•FRAGMENT ION: LIGHTER CATIONS FORMED BY THE
DECOMPOSITION OF THE MOLECULAR ION. THESE
OFTEN CORRESPOND TO STABLE CARBOCATIONS.
21-12-2019V.K. VIKRAM VARMA 13
15. COMPONENTS OF MASS
SPECTROMETER
•INLET SYSTEM
•ION SOURCE
oIONISATION METHODS
•MASS ANALYSERS
•ION DETECTORS
•VACCUM SYSTEM
21-12-2019V.K. VIKRAM VARMA 15
16. INLET SYSTEM
For Solids, Liquids, Gases
IONISATION SOURCE
EI, CI, FD, FAB, MALDI, ESI, APCI, APPI.
IONISATION MECHANISMS
Protonation, Deprotonation, Cationization,
Electron ejection, Electron capture
MASS ANALYSER
Magnetic field deflection, Double
focusing, Quadrupole, Time of
flight, Ion trap analyser, FT-ICR
DETECTOR
Electron multiplier, Faraday cup,
Photomultiplier conversion
dynode, Array
Mass
Spectrometer
21-12-2019V.K. VIKRAM VARMA 16
17. SAMPLE INLET•THREE TYPES OF SAMPLES, IF SOLID OR LIQUID SAMPLE IS
PRESENT CONVERTED INTO GAS UNDER SUITABLE ATMOSPHERIC
PRESSURE.
21-12-2019V.K. VIKRAM VARMA 17
SAMPLE
SOLID
Solids with lower vapour
pressure directly inserted
into ionisation chamber.
Volatilisation is
controlled by heating the
probe.
LIQUID
Handled by hypodermic
needles.
Injected by silicon rubber
dam.
GAS
Directly introduced into
ionisation chamber by
mercury manometer.
18. IONISATION CHAMBER
•IONISES THE MATERIAL UNDER ANALYSIS.
•MOLECULAR IONS ARE FORMED WHEN ENERGY OF
THE ELECTRON BEAM IS 10 − 15eV.
•FRAGMENT IONS ARE FORMED WHEN ENERGY OF
THE ELECTRON BEAM IS 70eV.
•ELECTRONS ARE RELEASED BY M ARE COLLECTED
BY ION COLLECTOR.
21-12-2019V.K. VIKRAM VARMA 18
19. ION ACCELERATION
CHAMBER
•ELECTRODE POTENTIAL OF ION ACCELERATION
CHAMBER IS 8 − 10KV.
•THE ACCELERATED IONS FURTHER GOES TO MAGNETIC
FIELD & DEFLECTION OF IONS.
•IONS HAVING HIGH MOLECULAR WEIGHT WILL REACH
DETECTOR FIRST.
21-12-2019V.K. VIKRAM VARMA 19
20. CONTD.
ENERGY OF ACCELERATED IONS
𝟏
𝟐
𝑴𝒗 𝟐
= ⅇ𝑽 (EQUATION 1)
ENTERED INTO MAGNETIC FIELD
𝑯ⅇ𝑽 =
𝑴𝒗 𝟐
𝒓
(EQUATION 2)
SQUARING BOTH THE SIDES IN EQUATION 2
𝑯 𝟐ⅇ 𝟐 𝑽 𝟐 =
𝑴 𝟐 𝒗 𝟒
𝒓 𝟐 (EQUATION 3)
𝑯 𝟐ⅇ 𝟐 =
𝑴 𝟐 𝒗 𝟐
𝒓 𝟐 (EQUATION 4)
21-12-2019V.K. VIKRAM VARMA 20
Where,
M= mass of the ion.
e= charge on the ion.
v= velocity of the ion.
V= potential difference.
Where,
H= Applied magnetic field.
r= radius of path.
21. CONTD.
REARRANGING EQUATION 1
𝑴𝒗 𝟐 = 𝟐ⅇ𝑽 (EQUATION 5)
PUT THE VALUE OF 𝑀𝑣2 FROM EQUATION 5 TO 4
𝑯 𝟐
ⅇ 𝟐
=
𝑴.𝟐ⅇ𝑽
𝒓 𝟐
𝑯 𝟐ⅇ =
𝑴𝑽
𝒓 𝟐
𝑴
ⅇ =
𝑯 𝟐 𝒓 𝟐
𝟐𝑽
(EQUATION 6)
21-12-2019V.K. VIKRAM VARMA 21
22. MASS ANALYSER
•IONS WILL BE SEPARATED ON
THE BASIS OF MOLECULAR
WEIGHT.
•TYPES OF ANALYSER
oMAGNETIC FIELD
DEFLECTION
oDOUBLE FOCUSING
oQUADRUPOLE
oTIME OF FLIGHT
oION TRAP ANALYSER
oFT-ICR
21-12-2019V.K. VIKRAM VARMA 22
AMPLIFIER•DIRECT ELECTRIC AMPLIFIER IS USED TO AMPLIFY THE
SIGNAL.
23. RECORDER
•RECORDS THE SPECTRA.
21-12-2019V.K. VIKRAM VARMA 23
VACCUM SYSTEM
•ALL MASS SPECTROMETERS NEED A VACCUM TO ALLOW
IONS TO REACH THE DETECTOR WITHOUT COLLIDING
WITH OTHER GASEOUS MOLECULES OR ATOMS.
•IF SUCH COLLISION OCCUR, THE INSTRUMENT WOULD
SUFFER FROM REDUCED RESOLUTION & SENSITIVITY
10−2 𝑡𝑜10−5OR 10−4 𝑡𝑜10−7 𝑡𝑜𝑟𝑟.
24. IONISATION TECHNIQUES
•MASS SPECTRUM IS SIGNIFICANTLY DEPENDED UPON THE
IONISATION METHOD.
•VARIATION IN THE SPECTRUM IS INTRODUCED IN TERMS OF
NUMBER OF PEAKS.
INTENSITY OF PEAKS(SPECIALLY MOLECULAR ION).
•IONISATION TECHNIQUE CAN BE CATEGORIES INTO 2 PARTS:
HARD IONISATION TECHNIQUE:
–HIGH ENERGY, INCREASED FRAGMENTATION.
SOFT IONISATION TECHNIQUE:
–LOW ENERGY, DECREASED FRAGMENTATION.
21-12-2019V.K. VIKRAM VARMA 24
25. 21-12-2019V.K. VIKRAM VARMA 25
TYPES or METHODS
GAS PHASE
Electron Ionisation (EI)
Chemical Ionisation
(CI)
DESORPTION
Field Desorption (FD)
Fast Atom
Bombardment (FAB)
Matrix Assisted Laser
Desorption Ionisation
(MALDI)
EVARPORATIVE
Electron Spray
Ionisation (ESI)
Atomic Pressure
Chemical Ionisation
(APCI)
Atomic Pressure Photo
Ionisation (APPI)
26. GAS PHASE IONISATION
•OLDEST & MOST POPULAR METHOD.
•SAMPLE IS VAPORISED BEFORE IONISATION.
•CLASSIFIED INTO:
– ELECTRON IONISATION (EI)
–CHEMICAL IONISATION (CI)
–FIELD IONISATION (FI)
21-12-2019V.K. VIKRAM VARMA 26
28. CONTD.
•DIRECT IONISATION THROUGH ELECTRON BEAM.
•IT IS THE MOST WIDELY USED & HIGHLY DEVELOPED METHOD. IT
IS ALSO KNOW AS ELECTRON IMPACT IONISATION OR ELECTRON
BOMBARDMENT.
•IONISATION TECHNIQUE IS USED TO CONVERT THE GASEOUS
SAMPLE INTO MOLECULAR IONS.
•HARD IONISATION TECHNIQUE (70𝑒𝑉).
𝐌 + ⅇ− → 𝐌⦁+ + 𝟐ⅇ−
21-12-2019V.K. VIKRAM VARMA 28
29. CONTD.
•OPERATING PRESSURE 10−5 𝑡𝑜10−6 𝑡𝑜𝑟𝑟.
•POTENTIAL DIFFERENCE FROM 𝐺3 𝑡𝑜𝐺5 IS UP TO 8000V OR 8KV.
21-12-2019V.K. VIKRAM VARMA 29
Gaseous sample
is entered from
slit 1.
Electrons emitted
from the filament
(tungsten) by
thermic emission.
Gaseous sample
& electrons
collides in the
ionisation
chamber.
𝐌⦁+Radical is
converted into
𝑴 𝟏
+
, 𝑴 𝟐
+
, 𝑴 𝟑
+
&
electrons
collected at
anode.
30. CONTD.
21-12-2019V.K. VIKRAM VARMA 30
Positive ions will
move to
acceleration
chamber.
Potential difference
b/w the repller
plate(+vely charged
G3) & first
accelerating plate (-
vely charged G4)
Move to second
accelerating plate (-
vely charged G5)
Accelerated ions
are separated to
their masses to
their final
velocities.
The ions emerge from the final accelerating slit as a collimated
ribbon of ions. The energy and velocity of ions are given by :
𝒁𝑽 =
𝟏
𝟐
𝑴 𝟏 𝒗 𝟏 =
𝟏
𝟐
𝑴 𝟐 𝒗 𝟐 =
𝟏
𝟐
𝑴 𝟑 𝒗 𝟑
Where, z = charge of the ion V
= accelerating potential v =
velocity of ion.
31. CONTD.
ADVANTAGES
• CAN BE USED AS GC/MS INTERFACE.
• GIVES REPRODUCIBLE MASS
SPECTRA.
• GIVES MOLECULAR MASS & ALSO
THE FRAGMENTATION PATTERN OF
THE SAMPLE.
• EXTENSIVE FRAGMENTATION &
LARGE NUMBER OF PEAKS GIVES
STRUCTURAL INFORMATION.
DISADVANTAGES
• SAMPLE MUST BE THERMALLY
STABLE & VOLATILE.
• UNSTABLE MOLECULAR ION
FRAGMENTS ARE FORMED SO
READILY THAT ARE ABSENT FROM
MASS SPECTRUM.
• A SMALL AMOUNT OF SAMPLE IS
IONISED (1 IN 1000 MOLECULES).
21-12-2019V.K. VIKRAM VARMA 31
33. CONTD.
•CARRIER GAS WILL BE IONISED → SECONDARY IONS WILL BE
PRODUCED & IONS WILL BE TRANSFERRED TO THE ANALYTES.
•IT IS VERY IMPORTANT SOFT IONISATION TECHNIQUE.
•FRAGMENTATION IS LESS & GIVES INTENSE PEAK OF
MOLECULAR ION.
•SOME MOLECULES LIKE ALCOHOLS, ETHERS, AMINES,
ESTERS, AMINO ACIDS ARE HIGHLY FRAGMENTED IN
ELECTRON IONISATION (EI), SO MOLECULAR ION PEAKS WILL
NOT BE DETECTED, TO GET PROPER ION PEAK WE ARE USING
THIS TECHNIQUE.
21-12-2019V.K. VIKRAM VARMA 33
34. CONTD.
•STEPS OF CHEMICAL IONISATION:
A CARRIER / REAGENT GAS IS INTRODUCED INTO
IONISATION SOURCE AT SLIGHTLY HIGHER PRESSURE(1
TORR) (METHANE, AMMONIA & ISOBUTANE GASES ARE
USED)
CARRIER GAS WILL BE IONISED DUE TO ELECTRON
IMPACT FROM THE IONISATION SOURCE.
𝑪𝑯 𝟒 + ⅇ−
→ 𝑪𝑯 𝟒
+⦁ + 𝟐ⅇ−
𝑪𝑯 𝟒
+⦁ → 𝑪𝑯 𝟑
+
+ 𝑯∎
21-12-2019V.K. VIKRAM VARMA 34
Where,
𝑪𝑯 𝟒
+⦁ &𝑪𝑯 𝟑
+
:Primary
ions
36. CONTD.
ADVANTAGES
•USED FOR SAMPLES WHICH
UNDERGO RAPID
FRAGMENTATION IN EI.
•USED FOR HIGH MOLECULAR
WEIGHT COMPOUNDS.
DISADVANTAGES
• RELATIVE LESS SENSITIVE THAN
EI IONISATION.
• NOT SUITABLE FOR THERMALLY
UNSTABLE & NON VOLATILE
SAMPLES.
• SAMPLES MUST BE DILUTED
WITH LARGE EXCESS OF
REAGENT GAS TO PREVENT
PRIMARY INTERACTION BETWEEN
THE ELECTRONS & SAMPLE
MOLECULES.
21-12-2019V.K. VIKRAM VARMA 36
37. DESORPTION IONISATION
TECHNIQUE
•LIQUID/ SOLID SAMPLES WILL BE DIRECTLY
CONVERTED INTO GASEOUS IONS.
•CLASSIFIED INTO:
–FIELD DESORPTION (FD).
–FAST ATOM BOMBARDMENT (FAB).
–MATRIX ASSISTED LASER DESORPTION IONISATION
(MALDI).
21-12-2019V.K. VIKRAM VARMA 37
39. CONTD.
•DESORPTION OF ELECTRONS FROM THE ANALYTE THROUGH
ANODE.
•LOW VOLATILE SAMPLES ARE USED TO PRODUCE STABLE
MOLECULAR IONS.
•WORKING:
SAMPLES ARE LOADED ON THE SURFACE OF THE CARBON
MICRONEEDLE BY DIPPING IN THE SAMPLE SOLUTION.
CARBON NEEDLES WILL PRODUCE HIGH GRADIENT VOLTAGE
ON THEIR TIPS, THAT IS WHY SHARP TIPS ARE USED.
21-12-2019V.K. VIKRAM VARMA 39
40. CONTD.
ION FORMATION TAKES PLACE MAINLY BY 2 MECHANISMS:
FIELD IONISATION: ELECTRONS ARE REMOVED FROM THE SPECIES/
ANALYTE IN A HIGH ELECTRIC FIELD.
𝑴 → 𝑴+ + ⅇ−
CATIONS ATTACHED: CATIONS WILL BE ATTACHED WITH ANALYTE
MOLECULE 𝑖. 𝑒. 𝐻+
𝑜𝑟 𝑁𝑎+
𝑒𝑡𝑐.
𝑴 + 𝑯+
→ [𝑴 − 𝑯]+
POSITIVE IONS WILL BE REPELLED BY THE ANODE & THEY WILL
GO TOWARDS THE MASS ANALYSER.
IONS DOESN’T HAVE SUFFICIENT INTERNAL ENERGY FOR
FRAGMENTATION, DUE TO THIS STABLE IONS FORMED.
21-12-2019V.K. VIKRAM VARMA 40
41. CONTD.
ADVANTAGES
•WORKS WELL FOR SMALL
ORGANIC MOLECULES, LOW
MOLECULAR WEIGHT
POLYMERS & PETROCHEMICAL
FUNCTIONS.
DISADVANTAGES
• SAMPLE MUST BE IN A SOLVENT.
• SENSITIVE TO ALKALI METAL
CONTAMINATION.
• NOT SUITABLE FOR THERMALLY
UNSTABLE & NON VOLATILE
SAMPLES.
• STRUCTURAL INFORMATION IS
NOT OBTAINED AS VERY LITTLE
FRAGMENTATION OCCURS.
21-12-2019V.K. VIKRAM VARMA 41
42. FAST ATOM BOMBARDMENT (FAB)
21-12-2019V.K. VIKRAM VARMA 42
43. CONTD.
•SAMPLE IS MIXED WITH MATRIX & NEUTRAL ATOM BEAM
IS BOMBARDED.
•SOFT IONISATION METHOD.
•DETERMINE THE MOLECULAR WEIGHT OF THE
COMPOUNDS HAVING THE SIZE FROM 300-6OOODALTONS.
GENERALLY USED TO DETERMINE MOLECULAR WEIGHTS
OF PEPTIDES.
21-12-2019V.K. VIKRAM VARMA 43
44. CONTD.
•METHODOLOGY:
CHARACTERISTICS OF THE MATRIX:
NON VOLATILE
LOW VAPOUR PRESSURE LIQUID
EXAMPLES: GLYCEROL, THIOGLYCEROL, DIMETHANOLAMINE, TRIETHANOLAMINE.
𝑋𝑒 OR 𝐴𝑟 (ACCELERATED NEUTRAL ATOMS)WILL BE BOMBARDED TO THE SAMPLE
MATRIX MIXTURE & IONISE THE SAMPLE DUE TO TRANSLATIONAL ENERGY. E.G.:
𝑿ⅇ + ⅇ− → 𝑿ⅇ⦁ + 𝟐ⅇ−
𝑿ⅇ + 𝑿ⅇ+⦁ → 𝑿ⅇ + 𝑿ⅇ+⦁
𝑮𝒍𝒚𝒄ⅇ𝒓𝒐𝒍 − 𝑯+
→ [𝑴𝑯]+
•IF 𝐶𝑠+
ION IS USED THAN IT IS KNOWN AS SIMS (SECONDARY
IONISATION MASS SPECTROMETRY).
21-12-2019V.K. VIKRAM VARMA 44
45. CONTD.
ADVANTAGES
• FAB SPECTRA USUALLY PROVIDE
RELATIVELY ABUNDANT MOLECULAR
OR QUASIMOLECULAR IONS & SHOW
SOME STRUCTURALLY IMPORTANT
FRAGMENT IONS.
• USED FOR IONISATION OF HIGH
MOLECULAR WEIGHT SAMPLES OF
BIOLOGICAL ORIGIN.
• EXTENSIVELY USED FOR OBTAINING
MASS SPECTRA OF SALTS DEPENDING
UPON THE NATURE OF ITS CATION &
ANION.
DISADVANTAGES
• FAB SAMPLES THE SURFACE
RATHER THAN THE BULK
CONCENTRATION OF THE SOLUTE
PRESENT & HENCE LIMITS
QUANTITATIVE MEASUREMENTS.
• THE MATRIX ALSO FORMS IONS
ON BOMBARDMENT, IN ADDITION
TO THOSE FORMED BY THE
SAMPLE WHICH COMPLICATES
THE SPECTRUM.
21-12-2019V.K. VIKRAM VARMA 45
46. CONTD.
•APPLICATIONS
THE SEPARATION & MS ANALYSIS OF PEPTIDES
ARISING FROM PROTEIN ENZYMATIC DIGESTION.
ELUCIDATION OF THE AMINO ACID SEQUENCE OF
THE OLIGOPEPTIDE EFRAPEPTIN D. THIS IS THE
POTENT INHIBITOR OF MITOCHONDRIAL 𝑨𝑻𝑷𝒂𝒔ⅇ
ACTIVITY.
21-12-2019V.K. VIKRAM VARMA 46
48. CONTD.
21-12-2019V.K. VIKRAM VARMA 48
Laser beams:
337nm- nitrogen laser of UV
range.
355nm- Frequency tripled
Nd:YAG
(Neodynium:Yterium:Alumini
um:Garnet)
326nm- Frequency
Quadrupole ND:YAG
294nm- IR laser
GENERALLY TIME OF FLIGHT(TOF) ANALYSER IS
USED.
49. CONTD.
• MIXED WITH POLAR MATRIX & LASER LIGHT IS USED FOR IONISATION.
• SOFT IONISATION METHOD, WHICH USES PULSED LASER BEAM.
• DETERMINE THE MOLECULAR WEIGHT OF PEPTIDES, ANTIBODIES,
PROTEIN, MOLECULES 𝑒𝑡𝑐 UP TO THE SIZE OF 300𝐾𝐷𝑎.
• LASER BEAM WILL HIT THE SAMPLE: MATRIX MIXTURE & ANALYTE/
SAMPLE WILL CONVERT INTO THE FORM OF GAS.
• ANALYTE/ SAMPLE & MATRIX WILL ALSO CONVERTS INTO THE IONS
DUE TO TRANSITIONAL ENERGY.
PROTONATION: 𝑴 + 𝑯+ → [𝑴𝑯]+
DEPROTONATION: 𝑴 → [𝑴 − 𝑯]−+𝑯+
21-12-2019V.K. VIKRAM VARMA 49
50. CONTD.
ADVANTAGES
• HIGH MOLECULAR WEIGHT ANALYTE
CAN BE IONISED.
• GENTLE IONISATION TECHNIQUE.
• MOLECULE NEED TO BE VOLATILE.
• WIDE ARRAY OF MATRIXES.
• PRIOR SEPARATION BY
CHROMATOGRAPHY IS NOT REQUIRED.
• PRODUCES SINGLY CHARGED IONS
THUS INTERPRETATION BECOMES EASY.
DISADVANTAGES
• ANALYTE MUST HAVE VERY LOW
VAPOUR PRESSURE.
• MALDI MATRIX CLUSTER IONS NOT
DISCOVERED LOW 𝑚
𝑧 SPECIES (<6OO).
• PULSED NATURE OF SOURCE LIMITS
COMPATIBILITY WITH MANY MASS
ANALYSERS.
• ANALYTES THAT ABSORB THE LASER
CAN BE PROBLEMATIC.
21-12-2019V.K. VIKRAM VARMA 50
51. CONTD.
•APPLICATIONS:
PHARMACEUTICAL ANALYSIS
a.DRUG METABOLISM STUDIES, PHARMACOKINETICS.
b.BIOAVAILABILITY STUDIES.
c. CHARACTERISATION OF POTENTIAL DRUGS.
d.IDENTIFYING DRUG TARGETS.
e. SCREENING OF DRUG CANDIDATES.
f. DRUG DEGRADATION PRODUCT ANALYSIS.
21-12-2019V.K. VIKRAM VARMA 51
52. CONTD.
MICROBIOLOGY
a. IDENTIFICATION OF MICROORGANISMS.
b. SPECIES DIAGNOSIS BY THIS PROCEDURE IS MUCH FASTER, MORE
ACCURATE & CHEAPER THAN OTHER PROCEDURES BASED ON
BIOCHEMICAL TESTS.
FORENSIC & ENVIRONMENTAL ANALYSIS
a. PESTICIDES ON FOOD.
b. SOIL & GROUND WATER CONTAMINATION.
PROTEOMICS
a. TO IDENTIFY, VERIFY & QUANTITATE: METABOLITES, RECOMBINANT
PROTEINS, PROTEINS ISOLATED FROM NATURAL SOURCE, PEPTIDES &
THEIR AMINO ACIDS SEQUENCES.
21-12-2019V.K. VIKRAM VARMA 52
53. EVARPORATIVE IONISATION
METHOD
•THESE TECHNIQUES ARE USED IN
CHROMATOGRAPHY.
•CLASSIFIED INTO:
–ELECTRON SPRAY IONISATION (ESI).
–ATMOSPHERIC PRESSURE CHEMICAL IONISATION
(APCI).
–ATMOSPHERIC PRESSURE PHOTO IONISATION
(APPI).
21-12-2019V.K. VIKRAM VARMA 53
54. ELECTRON SPRAY IONISATION (ESI)
21-12-2019V.K. VIKRAM VARMA 54
55. CONTD.
•SOFT IONISATION TECHNIQUE.
•USED TO ANALYSE THE HIGH MOLECULAR WEIGHT
BIOMOLECULES, LIABLE & NON VOLATILE COMPOUNDS.
•USED TO IONISE PROTEINS, PEPTIDES, LIPIDS,
OLIGOSACCHARIDE, OLIGONUCLEOTIDE & SYNTHETIC
POLYMER.
21-12-2019V.K. VIKRAM VARMA 55
56. CONTD.
Solution
containing the
sample through
the high voltage
potential
capillary by the
help of
Nebulisation gas.
Sprayed droplets
are ionised due to
high voltage
potential at
capillary.
Heated
desolvation gas
will evaporate the
solvent & it will
produce the
molecular ion.
Moves towards
ion accelerator
chamber.
21-12-2019V.K. VIKRAM VARMA 56
Working:
• It can also produce multiply charged ions along with singly
charged.
57. CONTD.
ADVANTAGES
• GOOD SENSITIVITY & THEREFORE,
USEFUL IN ACCURATE QUANTITATIVE
& QUALITATIVE MEASUREMENTS.
• HAS THE ABILITY TO HANDLE
SAMPLES WITH LARGE MASSES.
• ONE OF THE SOFT IONISATION
TECHNIQUE AVAILABLE & HAS THE
ABILITY TO ANALYSE BIOLOGICAL
SAMPLES WITH NON COVALENT
INTERACTIONS.
DISADVANTAGES
• CANNOT ANALYSE MIXTURES VERY WELL &
WHEN FORCED TO DO SO, RESULTS ARE
UNRELIABLE.
• PRIOR SEPARATION OF CHROMATOGRAPHY
IS REQUIRED
• APPARATUS IS VERY DIFFICULT TO CLEAN &
HAS A TENDENCY TO BECOME OVERLY
CONTAMINATED WITH RESIDUES FROM
PREVIOUS EXPERIMENTS.
• MULTIPLE CHARGES THAT ARE ATTACHED TO
THE MOLECULAR IONS CAN MAKE FOR
CONFUSING SPECTRAL DATA.
21-12-2019V.K. VIKRAM VARMA 57
58. CONTD.
•APPLICATIONS:
STUDYING NON COVALENT INTERACTION.
IDENTIFICATION & QUANTIFICATION OF HAEMOGLOBIN
VARIANTS.
PROTEIN IDENTIFICATION & CHARACTERISATION.
PROBING MOLECULAR DYNAMICS.
CHEMICAL IMAGING.
MONITORING CHEMICAL REACTIONS & STUDYING REACTIVE
INTERMEDIATES.
SCREENING FOR INBORN ERRORS OF METABOLISM.
21-12-2019V.K. VIKRAM VARMA 58
60. CONTD.
•SOFT IONISATION TECHNIQUE, BASED ON THE
MECHANISM OF EVAPORATION & CARRIED OUT
ATMOSPHERIC PRESSURE.
•APCI IS A COMBINATION OF CI & ESI WITH
DEVIATION(ADVANCED VERSION OF CHEMICAL
IONISATION).
•GENERALLY APCI IS COUPLED WITH CHROMATOGRAPHIC
INSTRUMENT LIKE HPLC.
21-12-2019V.K. VIKRAM VARMA 60
61. CONTD.
•WORKING:
SAMPLE WILL BE INJECTED THROUGH THE CAPILLARY THEN IT
WILL BE CONVERTED INTO SPRAYED DROPLET & FINALLY
ANALYTE.
SOLVENT VAPOUR DUE TO HEATING BY 𝑵 𝟐.
CORONA DISCHARGE ELECTRODE WILL IONISE THE SOLVENT
VAPOUR MOLECULE JUST LIKE PRODUCTION OF PRIMARY IONS
IN CHEMICAL IONISATION.
SOMETIMES ANALYTE VAPOURS MAY ALSO IONISED BY THE
ELECTRODE.
21-12-2019V.K. VIKRAM VARMA 61
62. CONTD.
CORONA DISCHARGE ELECTRODE 𝒐𝒓 𝜷 −PARTICLE EMITTER IS
USED FOR IONISATION.
DUE TO COLLISION & ION MOLECULAR CHARGE TRANSFER
BETWEEN SOLVENT & ANALYTE TAKES PLACE & IT WILL
PRODUCE
𝑴𝑯+
𝒊𝒐𝒏 𝑨 + 𝑺+
→ 𝑴𝑯+
+ 𝑺−
𝒑𝒐𝒔𝒊𝒕𝒊𝒗ⅇ
[𝑴 − 𝑯]− 𝒊𝒐𝒏 𝑨+ + 𝑺 → [𝑴 − 𝑯]−+𝑺𝑯+ 𝒏ⅇ𝒈𝒊𝒕𝒊𝒗ⅇ
•APCI IS USED TO ANALYSE POLAR, THERMOSTABLE SUBSTANCE
WITH MOLECULAR WEIGHT LESS THAN 1500DALTONS.
21-12-2019V.K. VIKRAM VARMA 62
63. CONTD.
ADVANTAGES
• MULTIPLE CHARGING IS TYPICALLY NOT
OBSERVED AS THE IONISATION PROCESS IS
MORE ENERGETIC THAN ESI.
• ELECTRON TRANSFER OR PROTON LOSS,
([𝑀 − 𝐻]−
) OCCURS IN THE NEGATIVE MODE.
• PROTON TRANSFER OCCURS IN THE POSITIVE
MODE.
• AT ATMOSPHERIC PRESSURE ANALYTE
MOLECULES COLLIDE WITH THE REAGENT
IONS FREQUENTLY & HENCE IONISATION IS
VERY DIFFICULT.
DISADVANTAGES
• RELATIVELY LOW ION CURRENTS.
• VERY SENSITIVE TO CONTAMINANTS
SUCH AS ALKALI METALS OR BASIC
COMPOUNDS.
• RELATIVELY COMPLEX HARDWARE
COMPARED TO OTHER ION SOURCES.
21-12-2019V.K. VIKRAM VARMA 63
64. CONTD.
• APPLICATIONS:
DETERMINATION OF VITAMIN D3 IN POULTRY FEED SUPPLEMENTS.
CAN BE USED AS LC/MS INTERFACE.
ANALYSIS OF ORGANIC COMPOUNDS WITH MEDIUM-HIGH POLARITY.
ANALYSIS OF PESTICIDES.
SINCE POSITIVE IONISATION IS DEPENDENT ON PROTONATION, MOLECULES
CONTAINING BASIC FUNCTIONAL GROUPS SUCH AS AMINO, AMIDE ESTERS,
ALDEHYDE/ KETONE & HYDROXYL CAN BE ANALYSED.
NEGATIVE IONISATION DEPENDS UPON DEPROTONATION, MOLECULES
CONTAINING ACIDIC FUNCTIONAL GROUPS ARE ANALYSED BY THIS METHOD.
ANALYSIS OF TRIAZINES, PHENYLUREAS, CARBAMATES, &
ORGANOPHOSPHORUS COMPOUNDS.
21-12-2019V.K. VIKRAM VARMA 64
66. CONTD.
•SOFT IONISATION TECHNIQUE.
•APPI SIMILAR TO APCI, BUT IONISATION IN APPI IS DUE TO
PHOTONS GENERATED BY UV LIGHT OF KRYPTON LAMP.
•SAMPLE SOLUTION WILL COME THROUGH THE HEATED
CAPILLARY & SPRAYED DROPLETS WILL BE FORMED DUE TO
NEBULISING GAS(𝑁2).
•DESOLVATION GAS(𝑁2) WILL BE SUPPLIED WHICH WILL
CONVERT THE SPRAYED DROPLETS INTO THE FORM OF
VAPOURS OF ANALYTE & SOLVENT.
21-12-2019V.K. VIKRAM VARMA 66
67. CONTD.
•PHOTONS EMITTED BY THE KRYPTON LAMP HAVE A
SPECIFIC ENERGY 𝑖. 𝑒. 10𝑒𝑉; WHICH IS SUFFICIENT TO
IONISE THE TARGET MOLECULE 𝑖. 𝑒.ANALYTE & SOLVENT.
•PHOTONS OF 10𝑒𝑉 WILL NOT IONISE OTHER ATMOSPHERIC
GAS PRESENT, DUE TO LOW ENERGY.
•PHOTONS WILL IONISE THE ANALYTE BY 3 MECHANISMS
DIRECT APPI
𝑴 + 𝒉𝒗 → 𝑴⦁+ + ⅇ−
21-12-2019V.K. VIKRAM VARMA 67
68. CONTD.
INDIRECT APPI
𝑺 + 𝒉𝒗 → 𝑺⦁+ + ⅇ−
𝑴 + 𝑺+
→ 𝑴+
+ 𝑺⦁
DOPANT ASSISTED APPI
TOLUENE IS USED AS DOPANT AGENT TO INCREASE THE
PERCENTAGE OF MOLECULAR ION.
𝑫 + 𝒉𝒗 → 𝑫⦁+ + ⅇ−
𝑺 + 𝑫⦁+ → 𝑴+ + 𝑫⦁
21-12-2019V.K. VIKRAM VARMA 68
69. CONTD.
ADVANTAGES
•ADVANTAGE OVER APCI:
APPLICABLE TO HIGHLY NON
POLAR COMPOUNDS & LOW
FLOW RATED(<100𝜇𝑙/𝑚𝑖𝑛).
DISADVANTAGES
•IT CAN GENERATE
BACKGROUND IONS FROM
SOLVENTS.
•IT REQUIRES VAPORISATION
TEMPERATURES RANGING
FROM 350-500℃, WHICH CAN
CAUSE THERMAL
DEGRADATION.
21-12-2019V.K. VIKRAM VARMA 69
70. CONTD.
•APPLICATIONS:
IT HAS THE CAPABILITY TO IONISE COMPOUNDS WITH A
WIDE RANGE OF POLARITIES WHILE BEING
REMARKABLY TOLERANT OF MATRIX COMPONENTS OF
HPLC ADDITIVES.
APPI HAS BEEN PROVED TO BE A VALUABLE TOOL FOR
ANALYTES WHICH ARE POORLY IONISED OR NOT
IONISED BY ESI & APCI, IN PARTICULAR.
21-12-2019V.K. VIKRAM VARMA 70
71. CONTD.
APPI WAS SHOWN TO BE ABLE TO DETECT STEROID
HORMONES & HAD BEEN PROVEN TO HAVE MUCH
HIGHER SENSITIVITY THAN ESI.
RESULTS INDICATE THAT APPI USING TOLUENE AS
DOPANT PROVIDES EXCEPTIONAL IONISATION
CAPABILITIES FOR A BROAD RANGE OF COMPOUNDS,
IN PARTICULAR FOR HORMONES & STEROLS
COMPARED TO APCI & HESI.
21-12-2019V.K. VIKRAM VARMA 71
72. MASS ANALYSER
•IONS AFTER LEAVING ION SOURCE, THE IONS ARE SEPARATED
ACCORDING TO THEIR 𝑚
𝑒 RATIO.
•IN THIS AREA, THE IONS ARE ACCELERATED BY BOTH
ELECTROSTATIC & MAGNETICALLY
•TYPES OF ANALYSERS:
MAGNETIC SECTOR ANALYSER.
DOUBLE FOCUSING ANALYSER.
QUADRUPOLE ANALYSER.
TIME OF FLIGHT ANALYSER(TOF).
ION TRAP ANALYSER.
21-12-2019V.K. VIKRAM VARMA 72
74. CONTD.
•POSITIVELY CHARGED PARTICLES ARE SEPARATED BY
APPLICATION OF MAGNETIC FIELD, THEY TRAVEL I THE
CURVED PATH & MOLECULAR IONS ARE SEPARATED
ACCORDING TO THEIR MASSES & COLLECTED.
•GIVEN BY
𝒎
𝒛
=
𝑯 𝟐 𝒓 𝟐
𝟐𝑽
21-12-2019V.K. VIKRAM VARMA 74
Where,
•
𝒎
𝒛
= mass
• H=magnetic
field
• r= Radius of
curvature
• V= Applied
voltage
76. CONTD.
•DOUBLE FOCUSING MAGNETIC SECTOR MASS ANALYSER
ARE THE CLASSICAL MODEL AGAINST WHICH OTHER MASS
ANALYSERS ARE COMPARED.
•THE LIMITATION IN THE SINGLE FOCUSING INSTRUMENT
IS THAT THE RESOLVING POWER IS LIMITED BY INITIAL
SPREAD OF TRANSLATIONAL ENERGY OF ION LEAVING
THE SOURCE.
•MAGNETIC SECTOR ANALYSER + ELECTROSTATIC
SECTOR→RESOLUTION INCREASED.
21-12-2019V.K. VIKRAM VARMA 76
77. CONTD.
ADVANTAGES
•CLASSICAL MASS SPECTRA.
•HIGH RESOLUTION.
•HIGH SENSITIVITY.
•BEST QUANTITATIVE
PERFORMANCE OF ALL MS
ANALYSERS.
•VERY HIGH REPRODUCIBILITY.
DISADVANTAGES
•REQUIRED SKILLED OPERATOR
•DIFFICULT TO INTERFACE TO
ESI.
•USUALLY LARGER & HIGHER
COST THAN OTHER MASS
ANALYSERS.
21-12-2019V.K. VIKRAM VARMA 77
80. CONTD.•QUADRUPOLE ANALYSER CONSISTS OF TWO PAIRS OF
RODS WITH A HYPERBOLIC CROSS SECTION THAT ARE
ACCURATELY POSITIONED PARALLEL IN RADIAL ARRAY.
•APPLIED DC & RF VOLTAGE.
•IF RF>DC:- LARGER IONS WILL HIT THE DETECTOR FIRST.
•IF RF<DC:- SMALLER IONS WILL HIT THE DETECTOR
FIRST.
•UNSTABLE OR NON TRANSMITTED IONS WILL HIT THE
RODS & WILL NOT BE DETECTED.
21-12-2019V.K. VIKRAM VARMA 80
81. CONTD.
ADVANTAGES
• RELATIVELY SMALL & COST
EFFECTIVE SYSTEMS.
• GOOD REPEATABILITY.
• CLASSICAL MASS SPECTRA.
• LOW ENERGY COLLISION INDUCED
DISSOCIATION (CID) MS/MS SPECTRA
LEADS TO EFFICIENT CONVERSION OF
PRECURSOR TO PRODUCT.
DISADVANTAGES
• LIMITED RESOLUTION.
• PEAK HEIGHT 𝑉𝑆 MASS RESPONSE
SHOULD BE TUNED.
• PEAK HEIGHTS ARE VARIABLE AS A
FUNCTION OF MASS DISCRIMINATION.
• LOW ENERGY COLLISION INDUCED
DISSOCIATION (CID) MS/MS SPECTRA
RELY MOST PROBABLY ON ENERGY,
COLLISION GAS, PRESSURE, &
ALTERNATIVE FACTORS.
21-12-2019V.K. VIKRAM VARMA 81
83. TIME OF FLIGHT ANALYSER (TOF)
21-12-2019V.K. VIKRAM VARMA 83
84. CONTD.
•TOF ANALYSER- IONS OF DIFFERENT MASS/CHARGE RATIO ARE
SEPARATED BY THE DIFFERENCE IN TIME THEY TAKE TO TRAVEL
OVER AN IDENTICAL PATH FROM THE ION SOURCE TO THE
COLLECTOR AT THE DETECTOR.
•SORTING OF IONS IS DONE IN ABSENCE OF MAGNETIC FIELD.
•IONS PRODUCED HAVE DIFFERENT VELOCITIES DEPENDS ON THEIR
MASSES.
•LIGHTER IONS HAVE HIGHER VELOCITY COMPARED TO HEAVIER
IONS.
•LIGHTER IONS WILL STRIKE THE DETECTOR FIRST DUE TO HIGHER
VELOCITY.
21-12-2019V.K. VIKRAM VARMA 84
85. CONTD.
ADVANTAGES
• FASTEST MS ANALYSER.
• HIGH ION TRANSMISSION.
• WELL SUITED FOR PULSED
IONISATION METHODS(METHOD OF
CHOICE FOR MAJORITY OF MALDI
MS SYSTEMS).
• HIGHEST PRACTICAL MASS RANGE
OF ALL MS ANALYSERS.
• MS/MS INFORMATION FROM POST
SOURCE DECAY.
DISADVANTAGES
•LOW RESOLUTION
•LIMITED PRECURSOR ION
SELECTIVITY FOR MOST MS/MS
EXPERIMENTS.
•REQUIRES PULSED IONISATION
METHOD OR ION BEAM
SWITCHING (DUTY CYCLE IS A
FACTOR).
21-12-2019V.K. VIKRAM VARMA 85
88. CONTD.
•QUADRUPOLE ION TRAP CONSISTS OF A RING ELECTRODE & 2
HYPERBOLIC END CAP ELECTRODES.
•AS THE RF VOLTAGE IS INCREASED, THE ORBITS OF HEAVIER IONS
BECOME STABILISED, & PASSED INTO THE DETECTOR.
•IONS ARE INJECTED INTO THE TRAP & ALL IONS ARE TRAPPED.
•RF & DC ARE SCANNED TO SEQUENTIALLY EJECT IONS FOR
DETECTION.
•SPECIFIC IONS CAN BE TRAPPED WHILE OTHERS ARE EJECTED.
•ION VELOCITY CAN BE INCREASED TO INDUCED FRAGMENTATION.
21-12-2019V.K. VIKRAM VARMA 88
89. CONTD.
ADVANTAGES
•INEXPENSIVE.
•EASILY INTERFACED TO
MANY IONISATION
METHODS.
•MS/MS IN ONE ANALYSER.
DISADVANTAGES
•LOW ACCURACY (>100𝑝𝑝𝑚).
•LOW RESOLUTION (<4000).
•SLOW SCANNING.
•SPACE CHARGING CAUSES
MASS SHIFTS.
•LOW MASS RANGE (<4000).
21-12-2019V.K. VIKRAM VARMA 89
91. MASS DETECTORS
•ONCE THE IONS ARE SEPARATED BY THE MASS ANALYSER,
THEY REACH THE ION DETECTOR, WHICH GENERATES A
CURRENT SIGNAL FROM THE INCIDENT IONS.
•TYPES OF DETECTORS:
FARADAY CUP DETECTOR.
ELECTRON MULTIPLIER DETECTOR.
PHOTOMULTIPLIER DYNODE DETECTOR.
ARRAY DETECTOR.
21-12-2019V.K. VIKRAM VARMA 91
92. FARADAY CUP MASS DETECTOR
21-12-2019V.K. VIKRAM VARMA 92
93. CONTD.
• BASIC PRINCIPLE: THE INCIDENT ION STRIKES THE DYNODE SURFACE WHICH
EMITS ELECTRONS & INDUCES A CURRENT WHICH IS AMPLIFIED & RECORDED.
• THE DYNODE ELECTRODE IS MADE OF A SECONDARY EMITTING MATERIALS LIKE
𝐶𝑠𝑆𝑏, 𝐺𝑎𝑃 𝑜𝑟 𝐵𝑒𝑂.
• IT IS IDEALLY SUITED TO ISOTOPE ANALYSIS.
• ADVANTAGES:
GOOD FOR CHECKING ION TRANSMISSION & LOW SENSITIVITY
MEASUREMENTS.
• DISADVANTAGES:
LOW AMPLIFICATION.
21-12-2019V.K. VIKRAM VARMA 93
95. CONTD.
• ELECTRON MULTIPLIER ARE THE MOST COMMON ESPECIALLY WHEN POSITIVE &
NEGATIVE IONS NEED TO BE DETECTED ON THE SAME INSTRUMENT.
• DYNODES MADE UP OF COPPER-BERYLLIUM WHICH TRANSDUCES THE INITIAL
ION CURRENT & ELECTRON EMITTED BY FIRST DYNODE ARE FOCUSED
MAGNETICALLY FROM DYNODE TO THE NEXT.
• FINAL CASCADE CURRENT IS AMPLIFIED MORE THAN MILLION TIMES.
• ADVANTAGES:
FAST RESPONSE
SENSITIVE
• DISADVANTAGES:
SHORTER LIFETIME THAN SCINTILLATION COUNTING(~3 𝑌𝑒𝑎𝑟𝑠).
21-12-2019V.K. VIKRAM VARMA 95
97. CONTD.
• THE DYNODE CONSISTS OF A SUBSTANCE (A SCINTILLATOR) WHICH EMITS
PHOTONS.
• THE EMITTED LIGHT IS DETECTED BY PHOTO MULTIPLIER TUBE & IS
CONVERTED INTO ELECTRIC CURRENT.
• USEFUL IN STUDIES ON METASTABLE IONS.
• ADVANTAGES:
SENSITIVE
LONG LIFETIME(>5YEARS)
• DISADVANTAGES:
CANNOT BE EXPOSED TO LIGHT WHILE IN OPERATION.
21-12-2019V.K. VIKRAM VARMA 97
99. CONTD.
• AN ARRAY DETECTOR IS A GROUP OF INDIVIDUAL DETECTORS ALIGNED IN AN ARRAY
FORMAT.
• ARRAY DETECTOR, WHICH SPATIALLY DETECTS IONS ACCORDING TO THEIR DIFFERENT
𝑚
𝑧 , HAS BEEN TYPICALLY USED ON MAGNETIC SECTOR MASS ANALYSERS.
• SPATIALLY DIFFERENTIATED IONS CAN BE DETECTED SIMULTANEOUSLY BY AN ARRAY
DETECTOR.
• ADVANTAGES:
FAST & SENSITIVE.
• DISADVANTAGES:
REDUCES RESOLUTION
EXPENSIVE
21-12-2019V.K. VIKRAM VARMA 99
101. MOLECULAR / PARENT
IONS:
•MOLECULE IS BOMBARDMENT WITH ELECTRONS IN
HIGH VACCUM IT IS CONVERTED TO POSITIVE IONS BY
LOSS OF ELECTRONS.
𝐌 + ⅇ−
→ 𝐌⦁+
+ 𝟐ⅇ−
21-12-2019V.K. VIKRAM VARMA 101
102. •GENERATED BY THE FRAGMENTATION OF THE
MOLECULAR ION IN THE IONISATION CHAMBER.
𝐌⦁+
→ 𝐌 𝟏
+
+ 𝐌 𝟐
+
DUE TO UNSTABILITY OF 𝐌⦁+
& ENERGY OF
IONISATION POTENTIAL.
21-12-2019V.K. VIKRAM VARMA 102
FRAGMENT IONS / DAUGHTER
IONS:
103. REARRANGEMENT IONS:
•IONS RESULTS FROM THE INTRAMOLECULAR
ATOMIC REARRANGEMENT DURING
FRAGMENTATION.
21-12-2019V.K. VIKRAM VARMA 103
METASTABLE IONS:
•FRAGMENT OF PARENT ION WILL GIVE RISE TO A NEW
ION(DAUGHTER ION) + NEUTRAL MOLECULE /
RADICAL.
𝐌 𝟏
+
→ 𝐌 𝟐
+
+ 𝐧𝐨𝐧 𝐜𝐡𝐚𝐫𝐠ⅇ𝐝 𝐩𝐚𝐫𝐭𝐢𝐜𝐥ⅇ𝐬
104. MULTICHARGED IONS:
•IONS MAY EXIST AS 2 OR 3 CHARGES INSTEAD OF
USUAL SINGLE CHARGE.
𝐌⦁+
+ ⅇ−
→ 𝐌++ + 𝟑ⅇ−
•RARELY FORMED UNDER NORMAL CONDITIONS.
•COMMON IN INORGANIC MASS SPECTRA.
21-12-2019V.K. VIKRAM VARMA 104
105. NEGATIVE IONS:
•POSITIVE IONS PREDOMINATE IN ELECTRONIC IMPACT IONISATION
BECAUSE OF GREATER STABILITY.
•BUT NEGATIVE IONS ARE NOT VERY USEFUL IN STRUCTURE
DETERMINATION, FORMATION OF NEGATIVE IONS ARE VERY RARE.
𝐀𝐁 + ⅇ−
→ 𝐀𝐁−
𝐀𝐁 + ⅇ−
→ 𝐀+
+ 𝐁−
𝐀𝐁 + ⅇ−
→ 𝐀+
+ 𝐁−
+ ⅇ−
21-12-2019V.K. VIKRAM VARMA 105
106. QUASI-MOLECULAR IONS:
•A PROTONATED MOLECULAR ION “OR” AN ION
FORMED BY REMOVAL OF ONE HYDROGEN ATOM
FROM MOLECULAR ION.
𝐌 + 𝐇+
→ 𝐌𝐇 +
(M+1)
𝐌+
→ 𝐌 − 𝐇 +
+ H (M+2)
21-12-2019V.K. VIKRAM VARMA 106
108. MASS FRAGMENTATION RULES
۞HEIGHT OF M+⦁ PEAK DECREASES WITH INCREASING
DEGREE OF BRANCHING.
۞HEIGHT OF M+⦁ PEAK DECREASES WITH INCREASING
MOLECULAR WEIGHT.
۞CLEAVAGE IS FORMED AT ALKYL SUBSTITUTED
CARBONS WHICH LEADS TO FORMATION OF
CARBOCATION.
21-12-2019V.K. VIKRAM VARMA 108
109. CONTD.
۞DOUBLE BONDS, CYCLIC STRUCTURES, AROMATIC RINGS
STABILISE M+⦁ & INCREASE THE PROBABILITY OF ITS
APPEARANCE.
۞DOUBLE BONDS FAVOUR ALLYLIC CLEAVAGE TO GIVE
THE RESONANCE STABILISED CATION.
CH
+
𝟐
− 𝐂𝐇 = 𝐂𝐇 𝟐 ↔ 𝐂𝐇 𝟐 = 𝐂𝐇 − 𝐂𝐇
+
𝟐
21-12-2019V.K. VIKRAM VARMA 109
110. CONTD.
۞SATURATED RINGS TENDS TO LOOSE ALKYL SIDE
CHAINS AT THE 𝛼 – BOND & UNSATURATED RINGS
UNDERGO RETRO-DIELS ALDER REACTION.
21-12-2019V.K. VIKRAM VARMA 110
111. CONTD.
۞ALKYL SUBSTITUTED AROMATIC COMPOUNDS ARE
CLEAVED PREFERABLY AT 𝛽- BOND TO THE RING, GIVING
RESONANCE STABILISED “BENZYL ION” OR “TROPYLIUM
ION”.
21-12-2019V.K. VIKRAM VARMA 111
112. CONTD.
۞CLEAVGE IS OFTEN ASSOCIATED WITH ELIMINATION OF
SMALL STABLE, NEUTRAL MOLECULES, SUCH AS CO,𝐇 𝟐 𝐎,
N𝑯 𝟑, 𝑯 𝟐 𝐒, KETONES. E.G.: MCLAFFERTY REARRANGEMENT.
۞C-C BONDS NEXT TO HETEROATOM ARE FREQUENTLY
CLEAVED, LEAVING THE CHARGE ON THE HETEROATOM.
21-12-2019V.K. VIKRAM VARMA 112
113. CONTD.
۞NITROGEN RULE:
•NONE/EVEN NUMBER OF N ATOMS: EVEN NOMINAL
MASS.
•ODD NUMBER OF N ATOMS: ODD NOMINAL MASS.
•IHD (INDEX OF HYDROGEN DEFICIENCY INDEX OR
DEGREE OF UNSATURATION)
FOR CXHY 𝐈𝐇𝐃 =
𝟐𝐱+𝟐𝐲
𝟐
DOUBLE BONDS/RING= IHD=1 ; TRIPLE BOND= IHD=2
21-12-2019V.K. VIKRAM VARMA 113
114. CONTD.
۞STEVENSON RULE: WHEN AN ION FRAGMENTS, THE POSITIVE
CHARGE REMAIN ON THE FRAGMENT OF LOWEST IONISATION
POTENTIAL.
21-12-2019V.K. VIKRAM VARMA 114
۞RING RULE: CAN CALCULATE THE NUMBER OF UNSATURATED
SITES IN THE COMPOUND FROM RING RULE. 𝐑 = 𝐂 + 𝟏 +
𝐍−𝐇
𝟐
FOR HALOGEN 𝐑 = 𝐂 + 𝟏 +
𝐗−𝐍
𝟐
−
𝐱
𝟐
Where, R= no. of unsaturated sites.
C= no. of carbons
N= no. of nitrogen X= halogen
116. CONTD.
•HETEROLYTIC CLEAVAGE: CLEAVAGE OF C-X (X=O, N, S,
CHLORINE) BOND IS MORE DIFFICULT THAN C-C BOND.
IN SUCH CLEAVAGE, THE POSITIVE CHARGE IS CARRIED
BY THE CARBON ATOM & BY THE HETEROATOM.
21-12-2019V.K. VIKRAM VARMA 116
117. CONTD.
•MCLAFFERTY REARRANGEMENT: OCCUR IN
KETONE, ALDEHYDES, CARBOXYLIC ACIDS &
ESTER.
21-12-2019V.K. VIKRAM VARMA 117
SEPERATION OF 𝛾- HYDROGEN FOLLOWED BY 𝛽-
BOND CLEAVAGE TO FORM FRAGMENTS.
118. MASS FRAGMENTATION
•TYPES OF FRAGMENTATION:
COLLISION INDUCED DISSOCIATION (CID)
ELECTRON CAPTURE DISSOCIATION (ECD)
ELECTRON TRANSFER DISSOCIATION (ETD)
ELECTRON DETACHMENT DISSOCIATION (EDD)
PHOTO DISSOCIATION
SURFACE INDUCED DISSOCIATION (SID)
CHARGE REMOTE FRAGMENTATION
HIGH ENERGY C-TRAP DISSOCIATION (HCD)
21-12-2019V.K. VIKRAM VARMA 118
119. COLLISION INDUCED DISSOCIATION (CID):
•MOLECULAR IONS ARE ACCELERATED BY ELECTRICAL
POTENTIAL TO HIGH KINETIC ENERGY AND THEN ALLOWED
TO COLLIDE WITH NEUTRAL MOLECULES LIKE HELIUM,
NITROGEN OR ARGON.
•COLLISION BETWEEN THESE MOLECULES LEADS TO
FORMATION OF FRAGMENT IONS WHICH ARE ANALYZED BY
MASS SPECTROMETER.
•EXAMPLE:- TRIPLE QUADRUPOLE SPECTROMETER
PRODUCES CID FRAGMENTS
21-12-2019V.K. VIKRAM VARMA 119
120. ELECTRON CAPTURE
DISSOCIATION(ECD):
•IT IS A METHOD OF FRAGMENTING GAS PHASE IONS
FOR TANDEM MASS SPECTROMETRIC ANALYSIS
(STRUCTURAL ELUCIDATION).
•DIRECT INTRODUCTION OF LOW ENERGY
ELECTRONS TO TRAPPED GAS PHASE IONS.
•ECD TYPICALLY INVOLVES A MULTIPLY PROTONATED
MOLECULE M INTERACTING WITH A FREE
ELECTRON TO FORM AN ODD-ELECTRON ION.
21-12-2019V.K. VIKRAM VARMA 120
121. •ETD INDUCES FRAGMENTATION OF CATIONS BY
TRANSFERRING ELECTRONS TO THEM.
• EXAMPLE:-PEPTIDES OR PROTEINS.
21-12-2019V.K. VIKRAM VARMA 121
ELECTRON TRANSFER DISSOCIATION(ECD):
ELECTRON DETACHMENT DISSOCIATION(EDD):
• METHOD FOR FRAGMENTING ANIONIC SPECIES.
122. PHOTO DISSOCIATION:
•PHOTODISSOCIATION IS A CHEMICAL REACTION IN
WHICH A CHEMICAL COMPOUND IS BROKEN DOWN BY
PHOTONS.
•IRMPD:- ABSORPTION OF MULTIPLE INFRA RED PHOTONS
BY A MOLECULE AND LEADS TO DISSOCIATION.
•BIRD:- LONG INTERACTION OF MOLECULE WITH
RADIATION FIELD LIKE CARBON DIOXIDE LASER.
21-12-2019V.K. VIKRAM VARMA 122
123. HIGH ENERGY C-TRAP
DISSOCIATION (HCD):
• IONS PASS THROUGH C-TRAP & INTO HCD CELL, WHERE
DISSOCIATION TAKES PLACE.
•IT IS A FRAGMENTATION TECHNIQUE, USED FOR PEPTIDE
MODIFICATION ANALYSIS.
• IMMONIUM IONS GENERATED VIA HCD PINPOINT MODIFICATIONS
SUCH AS PHOSPHO-TYROSINE.
•AN ADDED OCTOPOLE COLLISION CELL FACILITATES DE NOVO
SEQUENCING.
21-12-2019V.K. VIKRAM VARMA 123
124. •IT IS A TYPE OF COVALENT BOND BREAKING THAT OCCURS IN A GAS
PHASE ION IN WHICH THE CLEAVED BOND IS NOT ADJACENT TO THE
LOCATION OF THE CHARGE. THIS FRAGMENTATION CAN BE OBSERVED
USING TANDEM MASS SPECTROMETRY.
21-12-2019V.K. VIKRAM VARMA 124
SURFACE INDUCED DISSOCIATION(SID):
•IT IS A TECHNIQUE USED IN MS TO FRAGMENT MOLECULAR IONS IN
THE GAS PHASE BY COLLISION OF AN ION WITH A SURFACE UNDER
HIGH VACCUM
CHARGE REMOTE FRAGMENTATION:
125. FACTORS INFLUENCING
FRAGMENTATION
• THERMAL DECOMPOSITION: UNDERGO THERMAL DECOMPOSITION IN THE ION
SOURCE BEFORE IONISATION.
DUE TO THIS FRAGMENTATION OF COMPOUND WILL BE AFFECTED & PROBLEM
WILL OCCUR DURING INTERPRETATION OF SPECTRA.
• BOMBARDMENT ENERGIES: MORE NUMBER OF FRAGMENTS –HIGH
BOMBARDMENT ENERGY REQUIRED.
LESS FRAGMENTATION-LESS BOMBARDMENT ENERGY REQUIRED.
• FUNCTIONAL GROUPS.
21-12-2019V.K. VIKRAM VARMA 125
126. TYPES OF PEAKS
•MOLECULAR ION PEAK
•FRAGMENT ION PEAK
•REARRANGEMENT ION PEAK
•METASTABLE ION PEAK
•MULTICHARGED ION PEAK
•BASE PEAK
•NEGATIVE ION PEAK
21-12-2019V.K. VIKRAM VARMA 126
127. CONTD.
•MOLECULAR ION PEAK: SAMPLE IS BOMBARDED WITH
ELECTRONS OF 9-15EV THE MOLECULAR ION IS
PRODUCED BY THE LOSS OF SIMPLE ELECTRON.
•FRAGMENT ION PEAK: ENERGY IS GIVEN FURTHER
MORE UP TO 70EV. FRAGMENTS WHICH HAS LOWER
MASS NUMBER.
•REARRANGEMENT ION PEAK: RECOMBINATION OF
FRAGMENT ION.
21-12-2019V.K. VIKRAM VARMA 127
128. CONTD.
•METASTABLE ION PEAK: IONS RESULTING FROM
DECOMPOSITION BETWEEN THE SOURCE & MAGNETIC
ANALYSER.(BROAD PEAKS)
•MULTICHARGED ION PEAK: IONS EXIST AS 2 OR 3
CHARGES INSTEAD OF SINGLE CHARGE.
•BASE PEAK: MOST INTENSE PEAK IN THE MS, ASSIGNED
100% INTENSITY.
•NEGATIVE ION PEAK: NEGATIVE IONS FORMED FROM
ELECTRON BOMBARDMENT OF SAMPLE.
21-12-2019V.K. VIKRAM VARMA 128
129. ISOTOPIC PEAK•EACH ISOTOPE WILL SHOW UP AS A SEPARATE LINE IN MS.
•THE PRESENCE OF ISOTOPES READILY PRODUCE THE ISOTOPE
IONS IN THE SPECTRUM ACCOMPANIED BY A MAIN MOLECULAR
& FRAGMENT ION PEAK.
•.12
C – 98.9% NATURAL ABUNDANCE- VERY HIGH PEAK- M+
PEAK
(BASE PEAK).
•.13 C – 1.1% NATURAL ABUNDANCE- VERY LOW PEAK-
[M + 1]+PEAK.
•BASE PEAK IS THE LARGEST PEAK IN THE SPECTRUM &
INTENSITY OF EVERY OTHER PEAK IS REPORTED IN
COMPARISON TO BASE PEAK.
21-12-2019V.K. VIKRAM VARMA 129
130. TANDEM MASS
SPECTROMETRY
• PURPOSE IS TO FRAGMENT IONS FROM PARENT ION TO PROVIDE STRUCTURAL
INFORMATION ABOUT A MOLECULE
• ALSO ALLOWS MASS SEPARATION AND AA IDENTIFICATION OF COMPOUNDS IN
COMPLEX MIXTURES
• USES TWO OR MORE MASS ANALYZERS/FILTERS SEPARATED BY A COLLISION CELL
FILLED WITH ARGON OR XENON
• COLLISION CELL IS WHERE SELECTED IONS ARE SENT FOR FURTHER
FRAGMENTATION
21-12-2019V.K. VIKRAM VARMA 130
131. CONTD.
• IN TANDEM MASS SPECTROMETRY (MS/MS), DISTINCT IONS OF INTEREST ARE
SELECTED BASED ON THEIR M/Z FROM THE FIRST ROUND OF MS AND ARE
FRAGMENTED BY A NUMBER OF METHODS OF DISSOCIATION.
• ONE SUCH METHOD INVOLVES COLLIDING THE IONS WITH A STREAM OF INERT
GAS, WHICH IS KNOWN AS COLLISION-INDUCED DISSOCIATION (CID) OR HIGHER
ENERGY COLLISION DISSOCIATION (HCD). OTHER METHODS OF ION
FRAGMENTATION INCLUDE ELECTRON-TRANSFER DISSOCIATION (ETD) AND
ELECTRON-CAPTURE DISSOCIATION (ECD).
21-12-2019V.K. VIKRAM VARMA 131
132. DIAGRAM OF TANDEM MASS
SPECTROMETRY (MS/MS)
21-12-2019V.K. VIKRAM VARMA 132
133. CONTD.
21-12-2019V.K. VIKRAM VARMA 133
• THESE FRAGMENTS ARE THEN SEPARATED BASED ON THEIR INDIVIDUAL M/Z RATIOS IN A
SECOND ROUND OF MS. MS/MS (I.E., TANDEM MASS SPECTROMETRY) IS COMMONLY USED TO
SEQUENCE PROTEINS & OLIGONUCLEOTIDES AND THESE CAN BE MATCH WITH DATABASES
SUCH AS IPI, REFSEQ & UNIPROTKB/SWISS-PROT.
• THESE SEQUENCE FRAGMENTS CAN THEN BE ORGANIZED IN SILICO INTO FULL- LENGTH
SEQUENCE PREDICTIONS.
• A SAMPLE IS INJECTED INTO THE MASS SPECTROMETER, IONIZED, ACCELERATED &
ANALYZED BY MASS SPECTROMETRY (MS1).
• IONS FROM THE MS1 SPECTRA ARE THEN SELECTIVELY FRAGMENTED & ANALYZED BY A
SECOND STAGE OF MASS SPECTROMETRY (MS2) TO GENERATE THE SPECTRA FOR THE ION
FRAGMENTS.
134. HOW TANDEM MS SEQUENCING
WORKS• USE TANDEM MS: TWO MASS ANALYZERS IN SERIES WITH A COLLISION
CELL IN BETWEEN
• COLLISION CELL: A REGION WHERE THE IONS COLLIDE WITH A GAS (HE,
NE, AR) RESULTING IN FRAGMENTATION OF THE ION
• FRAGMENTATION OF THE PEPTIDES OCCUR IN A PREDICTABLE FASHION,
MAINLY AT THE PEPTIDE BONDS.
• THE RESULTING DAUGHTER IONS HAVE MASSES THAT ARE CONSISTENT
WITH KNOWN MOLECULAR WEIGHTS OF DIPEPTIDES, TRIPEPTIDES,
TETRAPEPTIDES… SER-GLU-LEU-ILE-ARG-TRP COLLISION CELL SER-GLU-
LEU-ILE-ARG SER-GLU-LEU SER-GLU-LEU-ILE ETC.…
21-12-2019V.K. VIKRAM VARMA 134
135. CONTD.
ADVANTAGES
• FAST
• NO GELS
• DETERMINES MW AND AA SEQUENCE
• CAN BE USED ON COMPLEX
MIXTURES-INCLUDING LOW COPY
• CAN DETECT POST-TRANSLATIONAL
MODIFICATION.
DISADVANTAGES
• VERY EXPENSIVE-CAMPUS
• REQUIRES SEQUENCE DATABASES
FOR ANALYSIS
21-12-2019V.K. VIKRAM VARMA 135
136. ADVANTAGES & DISADVANTAGES
OF MASS SPECTROMETRY:
ADVANTAGES
• MOLECULAR WEIGHT & FORMULA
DETERMINATION.
• QUANTITATIVE & QUALITATIVE
ANALYSIS.
• LESS AMOUNT OF SAMPLE REQUIRED.
• LESS THAN 1 MINUTE REQUIRED FOR
ANALYSIS.
DISADVANTAGES
• SAMPLE DESTRUCTION.
• SAMPLE SHOULD BE IN GASEOUS
FORM.
• COMPLEX & HIGH COST.
• SHOULD MAINTAIN VACCUM
THROUGHOUT THE PROCESS
21-12-2019V.K. VIKRAM VARMA 136
137. APPLICATIONS OF MASS
SPECTROMETRY
•STRUCTURE ELUCIDATION.
•PHARMACEUTICAL ANALYSIS:
oBIOAVAILABILITY STUDIES.
oDRUG METABOLISM STUDIES, PHARMACOKINETICS.
oCHARACTERISATION OF POTENTIAL DRUGS.
oDRUG DEGRADATION OF PRODUCT ANALYSIS.
oSCREENING OF DRUG CANDIDATES.
oIDENTIFYING DRUG TARGETS.
21-12-2019V.K. VIKRAM VARMA 137
138. CONTD.
•BIOMOLECULE CHARACTERIZATION:
oPROTEINS & PEPTIDES.
oOLIGO NUCLEOTIDES.
•ENVIRONMENTAL STUDIES:
oPESTICIDES IN FOOD.
oSOIL & GROUND WATER CONTAMINATION.
•FORENSIC ANALYSIS/ CLINICAL STUDIES:
oINVESTIGATE USE OF ILLEGAL DRUGS THROUGH ANALYZING BODY FLUIDS &
TISSUES. THE SAMPLE FOR FORENSICS IN THE CASE OF DRUG ABUSE IS
MAINLY URINE, HAIR & BLOOD.
21-12-2019V.K. VIKRAM VARMA 138
139. REFERENCE
• INTRODUCTION TO SPECTROSCOPY BY PAVIA.
• A TEXTBOOK OF ORGANIC CHEMISTRY BY BAHL ARUN & BAHL B.S.
• HTTP://WWW.CHEM.UCALGARY.CA/COURSES/350/CAREY5TH/CH13/CH13-0.HTML
• HTTP://PREMIERBIOSOFT.COM/TECH_NOTES/MASS-SPECTROMETRY.HTML
• HTTPS://EN.WIKIPEDIA.ORG/WIKI/MASS_SPECTROMETRY
• WWW.YOUTUBE.COM
• WWW.SLIDESHARE.COM
• WWW.GOOGLE.COM
21-12-2019V.K. VIKRAM VARMA 139