1. It is one of the type of Hyphenated technique.
2. It is a combination of gas chromatographic technique and spectroscopic technique.
3. It is having a high resolution capacity.
4. It is used has volatile and Non-volatile compounds.
5. It is used for qualitative and quantitative analysis.
GAS CHROMATOGRAPHY-MASS SPECTROSCOPY [GC-MS]Shikha Popali
THIS PRESENTATION GIVES A DETAIL ACCOUNT ON THE GC-MS WITH ITS INTRODUCTION, BASIC PRINCIPLE OF BOTH COMBINED AND INDIVIDUALLY WITH ITS INSTRUMENTATION, APPLICATION AND EXAMPLES, MAKES EASY TO COLLECT ALL THE DATA AT A PLACE ACCORDING TO THE M.PHARM SYLLABUS S PER PCI
1. It is one of the type of Hyphenated technique.
2. It is a combination of gas chromatographic technique and spectroscopic technique.
3. It is having a high resolution capacity.
4. It is used has volatile and Non-volatile compounds.
5. It is used for qualitative and quantitative analysis.
GAS CHROMATOGRAPHY-MASS SPECTROSCOPY [GC-MS]Shikha Popali
THIS PRESENTATION GIVES A DETAIL ACCOUNT ON THE GC-MS WITH ITS INTRODUCTION, BASIC PRINCIPLE OF BOTH COMBINED AND INDIVIDUALLY WITH ITS INSTRUMENTATION, APPLICATION AND EXAMPLES, MAKES EASY TO COLLECT ALL THE DATA AT A PLACE ACCORDING TO THE M.PHARM SYLLABUS S PER PCI
GCMS & LCMS
htps://youtube.com/vishalshelke99
https://instagram.com/vishal_stagram
Sub :- Advanced Analytical Techniques
M.Pharmacy Sem1
Savitribai Phule Pune University
Contents :-
GC-MS
Introduction
Principle
Instrumentation
Application
LC-MS
Introduction
Principle
Instrumentation
Application
Introduction to Gas chromatography-Mass spectroscopy
Gas chromatography-Mass spectroscopy is one of the so-called hyphenated analytical techniques. It is actually two techniques that are combined to form a single method of analyzing mixtures of chemicals
GC-MS is an instrumental technique, comprising a gas chromatograph coupled to a mass spectrometer by which complex mixtures of chemicals may be separated, identified & quantified. In order to a compound to be analysed by GC-MS it must be sufficiently volatile & thermally stable.
Principle :-
The Sample solution is injected into the GC inlet where it is vapourized & swept onto a chromatographic column by the carrier gas ( usually helium). The sample flows through the column & compounds comprising the mixture of interest are separated by virtue of their relative interaction with the coating of the column (stationery phase) & the carrier gas (mobile phase). The later part of the column passes through a heated transfer line & ends at the entrance to ion source where compounds eluting from the column are converted to ions
Gas chromatography-mass spectrometry (GC-MS) is the synergistic combination of two analytical method to separate and identify different substances within a test sample.
Gas chromatography separates the components of a mixture in time.
Mass spectrometer provides information that aids in the identification and structural elucidation of each component.
HPLC Principle,Instrumentation and ApplicationAlakesh Pradhan
HPLC Chromatography and its principle
Liquid chromatography
High Performance Liquid Chromatography ( HPLC )
The components of the high performance liquid chromatograph (HPLC).
The separation process.
The chromatogram
various parts of mAss spectroscopy, applications, principle, peaks, rules, typical mass spectra, various combinations, Fragmentation, rules of fragmentation and useful points which can help Chemical and analytical students and structural elucidation.
Gas chromatography-Mass spectrometry (GC-MS)Saira Fatima
PRESENTED BY
SAIRA FATIMA
SABAHAT MEHMOOD
SANA USMAN
MSc 4 (2018-2020)
Department of MicroBiology & Molecular Genetics
University of the Punjab
Lahore, Pakistan
ION EXCHANGE CHROMATOGRAPHY
ByM.Vharshini
B.Sc. Bio Medical Science
Sri Ramachandra University
ION EXCHANGE CHROMATOGRAPHY
Ion-exchange chromatography is a process that allows the separation of ions and polar molecules based on their affinity to the ion exchanger.
It can be used for almost any kind of charged molecule including large proteins, small nucleotides and amino acids.
Cations or Anions can be separated using this method.
PRINCIPLE
It is based on the reversible electrostatic interaction of ions with the separation matrix (i.e.)
The separation occurs by reversible exchange of ions between the ions present in the solution and those present in the ion exchange resin.
CLASSIFICATION OF RESINS
According to the chemical nature they classified as-
1. Strong cation exchange resin
2. Weak cation exchange resin
3. Strong anion exchange resin
4. Weak anion exchange resin
According to the Source they can -
Natural resins : Cation - Zeolytes, Clay
Anion - Dolomite
Synthetic resins: Inorganic & Organic resins
◘Organic resins are polymeric resin matrix.
The resin composed of –
Polystyrene (sites for exchangeable functional groups)
Divinyl benzene(Cross linking agent)-offers stability.
Ion exchange resin should have following requirements
»It must be chemically stable.
»It should be insoluble in common solvents.
» It should have a sufficient degree of cross linking.
»The swollen resin must be denser than water.
»It must contain sufficient no. of ion exchange groups.
Physical properties of ion exchange resins
Cross linking:
It affects swelling & strength & solubility
Swelling:
When resin swells, polymer chain spreads apart
Polar solvents → swelling
Non-polar solvents → contraction
Swelling also affected electrolyte concentration.
Particle size and porosity
Increase in surface area & decrease in particle size will increase the rate of ion exchange.
Regeneration
Cation exchange resin are regenerated by treatment with acid, then washing with water.
Anion exchange resin are regenerated by treatment with NaOH, then washing with water until neutral.
EXPERIMENTAL SETUP OF ION EXCHANGE CHROMATOGRAPHY
Metrohm 850 Ion chromatography system
Instrumentation of ion exchange chromatography
PRACTICAL REQUIREMENTS
1.Column
» glass, stainless steel or polymers
2.Packing the column
» Wet packing method:
A slurry is prepared of the eluent with the stationary phase powder and then carefully poured into the column. Care must be taken to avoid air bubbles.
3.Application of the sample
After packing, sample is added to the top of the stationary phase, use syringe or pipette.
This layer is usually topped with a small layer of sand or with cotton or glass wool to protect the shape of the organic layer from the velocity of newly added eluent.
4.Mobile phase
Acids, alkalis, buffers…
6.Stationary phase
The ionic
GCMS & LCMS
htps://youtube.com/vishalshelke99
https://instagram.com/vishal_stagram
Sub :- Advanced Analytical Techniques
M.Pharmacy Sem1
Savitribai Phule Pune University
Contents :-
GC-MS
Introduction
Principle
Instrumentation
Application
LC-MS
Introduction
Principle
Instrumentation
Application
Introduction to Gas chromatography-Mass spectroscopy
Gas chromatography-Mass spectroscopy is one of the so-called hyphenated analytical techniques. It is actually two techniques that are combined to form a single method of analyzing mixtures of chemicals
GC-MS is an instrumental technique, comprising a gas chromatograph coupled to a mass spectrometer by which complex mixtures of chemicals may be separated, identified & quantified. In order to a compound to be analysed by GC-MS it must be sufficiently volatile & thermally stable.
Principle :-
The Sample solution is injected into the GC inlet where it is vapourized & swept onto a chromatographic column by the carrier gas ( usually helium). The sample flows through the column & compounds comprising the mixture of interest are separated by virtue of their relative interaction with the coating of the column (stationery phase) & the carrier gas (mobile phase). The later part of the column passes through a heated transfer line & ends at the entrance to ion source where compounds eluting from the column are converted to ions
Gas chromatography-mass spectrometry (GC-MS) is the synergistic combination of two analytical method to separate and identify different substances within a test sample.
Gas chromatography separates the components of a mixture in time.
Mass spectrometer provides information that aids in the identification and structural elucidation of each component.
HPLC Principle,Instrumentation and ApplicationAlakesh Pradhan
HPLC Chromatography and its principle
Liquid chromatography
High Performance Liquid Chromatography ( HPLC )
The components of the high performance liquid chromatograph (HPLC).
The separation process.
The chromatogram
various parts of mAss spectroscopy, applications, principle, peaks, rules, typical mass spectra, various combinations, Fragmentation, rules of fragmentation and useful points which can help Chemical and analytical students and structural elucidation.
Gas chromatography-Mass spectrometry (GC-MS)Saira Fatima
PRESENTED BY
SAIRA FATIMA
SABAHAT MEHMOOD
SANA USMAN
MSc 4 (2018-2020)
Department of MicroBiology & Molecular Genetics
University of the Punjab
Lahore, Pakistan
ION EXCHANGE CHROMATOGRAPHY
ByM.Vharshini
B.Sc. Bio Medical Science
Sri Ramachandra University
ION EXCHANGE CHROMATOGRAPHY
Ion-exchange chromatography is a process that allows the separation of ions and polar molecules based on their affinity to the ion exchanger.
It can be used for almost any kind of charged molecule including large proteins, small nucleotides and amino acids.
Cations or Anions can be separated using this method.
PRINCIPLE
It is based on the reversible electrostatic interaction of ions with the separation matrix (i.e.)
The separation occurs by reversible exchange of ions between the ions present in the solution and those present in the ion exchange resin.
CLASSIFICATION OF RESINS
According to the chemical nature they classified as-
1. Strong cation exchange resin
2. Weak cation exchange resin
3. Strong anion exchange resin
4. Weak anion exchange resin
According to the Source they can -
Natural resins : Cation - Zeolytes, Clay
Anion - Dolomite
Synthetic resins: Inorganic & Organic resins
◘Organic resins are polymeric resin matrix.
The resin composed of –
Polystyrene (sites for exchangeable functional groups)
Divinyl benzene(Cross linking agent)-offers stability.
Ion exchange resin should have following requirements
»It must be chemically stable.
»It should be insoluble in common solvents.
» It should have a sufficient degree of cross linking.
»The swollen resin must be denser than water.
»It must contain sufficient no. of ion exchange groups.
Physical properties of ion exchange resins
Cross linking:
It affects swelling & strength & solubility
Swelling:
When resin swells, polymer chain spreads apart
Polar solvents → swelling
Non-polar solvents → contraction
Swelling also affected electrolyte concentration.
Particle size and porosity
Increase in surface area & decrease in particle size will increase the rate of ion exchange.
Regeneration
Cation exchange resin are regenerated by treatment with acid, then washing with water.
Anion exchange resin are regenerated by treatment with NaOH, then washing with water until neutral.
EXPERIMENTAL SETUP OF ION EXCHANGE CHROMATOGRAPHY
Metrohm 850 Ion chromatography system
Instrumentation of ion exchange chromatography
PRACTICAL REQUIREMENTS
1.Column
» glass, stainless steel or polymers
2.Packing the column
» Wet packing method:
A slurry is prepared of the eluent with the stationary phase powder and then carefully poured into the column. Care must be taken to avoid air bubbles.
3.Application of the sample
After packing, sample is added to the top of the stationary phase, use syringe or pipette.
This layer is usually topped with a small layer of sand or with cotton or glass wool to protect the shape of the organic layer from the velocity of newly added eluent.
4.Mobile phase
Acids, alkalis, buffers…
6.Stationary phase
The ionic
Gas Chromatography-Mass Spectrometry (GC-MS) is an analytical method that combines the features of gas-liquid chromatography and mass spectrometry to identify different substances within a test sample.
in this presentation we learn about the mass spectrometery principal and its mass to charge ratio.
components of mass spectrometers .
sample inoculation and its processing. i feel these are very good slides.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
ANTI - INFLAMMATORY ACTIVITY OF LEAVES OF JATROPHA GOSSYPIFOLIA L.Raju Sanghvi
BY HRBC MEMBRANE STABILIZATION METHOD, Introduction, Preparation of extracts, Chemicals and instruments, In vitro anti – inflammatory activity, Acute oral toxicity study, Discussion
Introduction & Definition, Theory, instrumentation, Continuous – wave (CW) instrument, The pulsed Fourier Transform [FT] instrument, Solvents, Chemical shift
i. Shielding and de-shielding
ii. Factors affecting chemical shift
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
How to Split Bills in the Odoo 17 POS ModuleCeline George
Bills have a main role in point of sale procedure. It will help to track sales, handling payments and giving receipts to customers. Bill splitting also has an important role in POS. For example, If some friends come together for dinner and if they want to divide the bill then it is possible by POS bill splitting. This slide will show how to split bills in odoo 17 POS.
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
GIÁO ÁN DẠY THÊM (KẾ HOẠCH BÀI BUỔI 2) - TIẾNG ANH 8 GLOBAL SUCCESS (2 CỘT) N...
GCMS
1. GGAASS CCHHRROOMMAATTOOGGRRAAPPHHYY--
MMAASSSS SSPPEECCTTRROOMMEETTRRYY ((GGCC--MMSS))
Presented by
APARNA.T UNDER THE GUIDANCE OF
Mr. Ch. DEVADASU M.Pharm.,
Assistant professor
Department of PA & QA
VIGNAN PHARMACY COLLEGE
(Approved by AICTE, PCI &
Affiliated to JNTU-K)
VADLAMUDI, 522213.
VIGNAN PHARMACY COLLEGE 13/9/2014
2. IUPAC: chromatography is a physical method of separation in which the
components to be separated are distributed between two phases. One of
which is stationary (stationary phase) while the other (the mobile phase)
moves in a definite direction.
Elution chromatography is a procedure in which the mobile phase is
continuously passed through or along the chromatographic bed and the
sample is fed into the system as a finite slug. EX: Gas Liquid
Chromatography(GC)
Gas chromatography is a separation method in which the components
of a sample partition between two phases one of these phases is a
stationary bed with a large surface area, and the other is a gas which
percolates through the stationary bed.
VIGNAN PHARMACY COLLEGE 13/9/2014 2
3. The father of modern gas chromatography is Nobel Prize
winner John Porter Martin, who also developed the first
liquid-gas chromatograph. (1950)
VIGNAN PHARMACY COLLEGE 13/9/2014 3
6. In GC the main principle of separation is partition.
1. A gaseous mobile phase flows continuously through
the column which is coated with a liquid stationary
phase.
2. The sample is introduced into the heated injection
port where it is vaporized and carried in to the column.
3. The sample partition between the mobile phase and
stationary phase, and is separated in to individual
components based on relative solubility in liquid
stationary phase at the given temperature.
4. The components of the sample separate from one
another based on their relative vapor pressures and
affinities for the stationary bed.
VIGNAN PHARMACY COLLEGE 13/9/2014 6
7. THE CHROMATOGRAPHIC PPRROOCCEESSSS -- PPAARRTTIITTIIOONNIINNGG
(gas or liquid)
MMOOBBIILLEE PPHHAASSEE
SSTTAATTIIOONNAARRYY PPHHAASSEE
Sample
out
Sample
in
(solid or heavy liquid coated onto a solid or support system)
VIGNAN PHARMACY COLLEGE 13/9/2014 7
8. In the animation below the red molecules are more soluble in the
liquid (or less volatile) than are the green molecules.
VIGNAN PHARMACY COLLEGE 13/9/2014 8
9. Distribution Coefficient
Definition:
Concentration of component A in stationary phase
Concentration of component A in mobile phase
Different affinity of any 2 components to
stationary phase causes the separation.
VIGNAN PHARMACY COLLEGE 13/9/2014 9
11. G. C is a separation technique.
Small amounts of sample for example 1 ml of air,1 micro
lit. of the solution either liquids and solids in solution are
injected in to an Instrument .
The machine is called a Gas chromatograph
this machine by using injection port, column and detector
generates a written record of analysis .. a series of peaks .
Series of peaks are called a chromatogram.
Chromatogram is simply a written record of the
analysis performed by the gas chromatograph.
VIGNAN PHARMACY COLLEGE 13/9/2014 11
13. The concept of mass spectrometry was first put forth by
Sir J.J Thomson, English Physicist Who discovered the
electron in 1887.
He got 1906 Nobel Laureate in Physics.
DEMPSTER
Sir J.J Thomson
VIGNAN PHARMACY COLLEGE 13/9/2014 13
14. What does a mass spectrometer do?
Mass –spec or simply MS is a super important technique
Mass spec is easy technique to give you Molecular weight
(from molecular ion (M+)
You can get Molecular formula (Elements present).
Nearly ALL ELEMENTS in the periodic table can be determined by
mass spectrometry.
MS is incredibly valuable in getting structure (from fragments) of Bio
molecules such as peptides and proteins and also
natural products and also organic structures.
It can give information about chemical structures.
VIGNAN PHARMACY COLLEGE 13/9/2014 14
17. THEORY
The mass spectrometer is an instrument which help in separating the individual
atoms or molecules because of difference in their masses.
17
Consider a molecule M, Which is bombarded with a beam of electrons
M + e- M+. +2e-where,
M+. is molecular ion or radical ion
2e- is electron
Now voltage “v” is applied in an electric field then ions are accelerated. In this
condition the energy given to each particle is zV and this is equal to kinetic
energy which is equal to 1/2mv2 .
VIGNAN PHARMACY COLLEGE 13/9/2014
18. i.e. potential energy=kinetic energy
zV = 1/2mv2
2zV = mv2
2zV/m= v2
= v
Where V = Velocity of particle
m = mass
z = charge of an electron
V = Acceleration voltage
All the particles posses some energy zV with some kinetic energy
1/2mv2, but m value changes from molecule to molecule with respect velocity
‘v’ also changes. i.e. ½ mv=zV
VIGNAN PHARMACY COLLEGE 13/9/2014 18
19. When all charged particles have been accelerated by an applied voltage, they
enter into a magnetic field “H”. Then attractive force is HzV. And balancing
force of particle is mv2 /r.
Centripetal = Centrifugal
HzV=mv2/r
Hz=mv/r
From the above equation v=
Hz=m / r
By squaring on both sides
H2 z2 = m2 (2zV/m) / r2
H2 z = 2v m/ r2
m/z = H2 r2 / 2v
VIGNAN PHARMACY COLLEGE 13/9/2014 19
20. MASS SPECTRUM
The mass spectrum is the plot of mass to charge ratio of positively charged
ions against their relative abundance. The m/z ratio are taken along the
abscissa, while relative abundance is taken on ordinate.
BASE PEAK:
The most intense peak in the mass spectrum is called the base peak. Base
peak is the highest peak it is assigned a relative intensity of 100%.
VIGNAN PHARMACY COLLEGE 13/9/2014 20
21. MOLECULAR ION PEAK:
The ion formed from a molecule by removal of one electron of lowest
ionization potential is known as molecular ion.
The molecular ion is detected as mass to charge ratio that corresponds to
molecular weight of molecule. The molecular ion peak gives the molecular
weight of compound . The molecular ion peak is highest mass number except
isotope peak. Base peak Molecular ion peak
Fragment ions
VIGNAN PHARMACY COLLEGE 13/9/2014 21
22. FRAGMENT IONS:
The ions produced from the molecular ion by cleavage of bonds are called
fragment ions
They have lower masses and used as building blocks to reconstruct the
molecular structure. Fragmentation of molecular ion cleavage bond occurs in
heterolytic and homolytic cleavage.
METASTABLE IONS:
Mass spectrum of molecule shows sharp peaks at m/z integrals. But some show
diffuse, broad low intensity peaks at non integral m/z values these are called
metastable ions
m1
+ m2
++ neutral fragment
VIGNAN PHARMACY COLLEGE 13/9/2014 22
23. If in the reaction m1
+--------->m2
++ takes place in source then the
daughter ion may be m2
+. But m1
+----->m2
++ if occurs after the source and
before arrival at collector at lower mass than m2+ and is said to be
metastable ion m*.The peak (m*) due to such fragmentation therefore
occurs at lower mass than m2+ and generally broad. The relation between
the m* with that of m1
+ & m2+ can be written as
m*=(m2)2/m1
VIGNAN PHARMACY COLLEGE 13/9/2014 23
25. Gas chromatography–mass spectrometry (GC-MS, or
alternatively HPLC-MS) is an ADVANCED ANALYTICAL
INSTRUMNTAL technique that combines the physical
separation capabilities of GGAASS CCHHRROOMMAATTOOGGRRAAPPHHYY with
the mass analysis capabilities of MASS SPECTROMETER
27. 27
Gas
chromatograp
hy
Mass
spectrometry
GC-MS
Separates
mixture of
components
into individual
Identifies
molecules
based on their
mass
A chemical
analysis technique
combining two
instruments to
provide for
powerful
separation &
identification.
VIGNAN PHARMACY COLLEGE 13/9/2014
28. Coupling of GC to
GCM S:
Atmospheric
density
heated (200-
300 ∘C)
Interfaces
MS
High vacuum
(10-6 torr)
heated
The interface b/w the GC&MS is an important
role to play in the overall efficiency of the
instrument.
Both system are heated (200 -300 ∘C) both deal
Ownitlhy coonme pporuonbdlesm in i st hteh avta tphoer asttamtoes. pheric pressure
output of the GC must be reduced to vacuum of
10-5 – 10-6 torr for the MS inlet
30. Capillary direct interface:
Today most GC-MS systems use capillary columns &
Capillary direct interface:
Today most GC-MS systems use capillary columns &
fused silica tubing permits an inert,high efficiency,direct
transfer between the 2 systems.
fused silica tubing permits an inert,high efficiency,direct
transfer between the 2 systems.
Flow rates is 5ml/min.
Flow rates is 5ml/min.
VIGNAN PHARMACY COLLEGE 13/9/2014 30
31. Jet separator (packed column):
•The separator consist of two glass tubes aligned with a
Small distance between them.
•Carrier gas entering from the GC column is pumped away
by a separate vacuumed system.
•The larger sample molecules maintain their momentum
&pass preferentially in to the second capillary.
•Sample enrichment occurs & the initial atmospheric
pressure is reduced.
VIGNAN PHARMACY COLLEGE 13/9/2014 31
32. Bothe T & P surfaces activity of the glass jet separator must be controlled.
VIGNAN PHARMACY COLLEGE 13/9/2014 32
33. Watson & Biermann effusion
separator:
• It consists of a sintered glass tube .
• The carrier usually Helium, passes preferentially
through the sintered glass tube & the effluent in
concentrated by a factor of up to 100.
• The gas flow rates in the order of 20-60ml/min.
VIGNAN PHARMACY COLLEGE 13/9/2014 33
34. It converts the components of a sample into ions by
bombardment with electrons, ions, molecules.
IONIZER;
CH3OH + 1e CH3OH+ + 2e
molecular ion or radical ion
The gas molecules exiting the GC are bombarded by a
high energy electron beam.
VIGNAN PHARMACY COLLEGE 13/9/2014 34
35. An electron which sticks a molecule may impart enough
energy to remove another electron from that molecule.
The charged molecule is known as molecular ion.
The molecular ion can causes that ion to break into
smaller pieces.
CH3OH+ CH2OH+ + H-VIGNAN
PHARMACY COLLEGE 13/9/2014 35
37. The most common form of ionization is EI.
Electrons are produced by tungsten filament.
These electrons accelerated towards the ion source chamber.
The electrons require an energy equal to the voltage B/W the
filament & ion source chamber.
70 ev is commonly used.
A proportion of electron beam will strike the electron trap
producing trap current.
VIGNAN PHARMACY COLLEGE 13/9/2014 37
39. A permanent magnet is positioned across the ion chamber to
produce a magnetic flux in parallel to the electron beam.
A (+)ve ion repelle voltage & (-)ve ion excitation voltage
works to gather to produce an electric field in the source
chamber.
Such that ions leaves through ion exit slit.
The ions are directed through the various focusing &
centering lenses are focused on to the source exit slit.
VIGNAN PHARMACY COLLEGE 13/9/2014 39
40. In CI a reagent gas methane or ammonia or isobutene
are introduced into the mass spectrometer.
The reagent gas will interact with the electron to produce
radical electrons.
EG;
R + e R+ + 2e
VIGNAN PHARMACY COLLEGE 13/9/2014 40
42. It is a good for organic compounds.
Usually produces (M + H)+ (M+ CH3)+ adducts.
Adducts are not always abundant.
ISOBUTANE
Usually produces (M + H)+ ,(M+C4H9)+ adducts & some
fragmentation.
Adducts are more abundant than for methane CI .
VIGNAN PHARMACY COLLEGE 13/9/2014 42
43. Fragmentation is absent.
Polar compounds produces ( M+NH4)+ adducts.
Basic compounds produces (M+H)+ adducts.
Non polar, non basic compounds are not ionized.
VIGNAN PHARMACY COLLEGE 13/9/2014 43
44. 44
Ionization
method
electron
impact
Chemical
ionization
Typical analyses Relatively small
volatile
Relatively small
volatile
Sample
introduction
GC (or) liquid/solid
probe
GC/Liquid /solid
probe
Mass range 1-1000 Dolton's 1-1000 Daltons
Method highlights Hard method
versatile
provides structure
information
Soft method
molecular ion peak
(M+H)+
VIGNAN PHARMACY COLLEGE 13/9/2014
45. 45
Negative ion chemical ionization(NICI):
In NICI a r eagent gas is used & the electrons collide with it so
that their energies are reduced to 10Ev.
Molecules with a high affinity for electrons are able to
capture these low energy thermal electrons.
This is known as NICI but it does not involved in the
formation of a chemical adduct.
VIGNAN PHARMACY COLLEGE 13/9/2014
46. 46
AB+ e
AB-Resonance
electron capture
A- +B
Dissociative
electron capture.
VIGNAN PHARMACY COLLEGE 13/9/2014
47. 47
They deflects ions down a curved tubes in a magnetic fields based on
their kinetic energy determined by the mass, charge and velocity. The
magnetic field is scanned to measure different ions.
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48. In a quadrupole mass analyser a set of four rods are arranged parallel to
the direction. Here a DC current and radio frequency RF is applied to
generate oscillating electrostatic field in between the rods. Based on this
only m/z is been determined and stable oscillation takes place. And ion
travels in quadrupole axis with cork screw type of trajectory.
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49. TOF mass analyser is based on simple idea that the velocities of
two ions are created by uniform electromagnetic force applied to
all the ions at same time, causing them to accelerate down a flight
tube.
Lighter ions travels faster and strike the detector first so that the
m/z ratio of ions is detected.
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51. The ion trap mass analyser operates by similar principles where it
consists of circular ring electrode
Plus two end caps that form a chamber. Here AC or DC power
along RF potential is applied between the cups and the ring
electrode.
There the ions entering into the chamber are trapped by
electromagnetic fields and they oscillates in concentric
trajectories. This process is called resonant ejection.
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53. DATA HANDLING
All the mass spectrometers now employ computer control of same functions
and also use a computerised display and output.
The amount of data generated even by a fairly modest mass spectrometer is
very large indeed, a single run may store data for upto 100 fragments from
each type of molecule and if, GCMS analyses is being performed, a complete
mass spectrum is generated and stored every sec for upto 90 min
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55. •Only Compounds with Vapour Pressure exceeding about 1010 torr
can be analyzed b as chromatography –mass spectrometry.
•Certain isomeric compounds cannot be distinguished by mass
spectrometry (EG : naphthalene vs. azulene).
57. Elucidation of the structure of organic & biological
molecules.
Impurity profiling of pharmaceuticals.
Identification of components in thin layer & paper
chromatograms.
Identification of drugs of abuse & metabolites of drugs of abuse
in blood, urine & saliva.
Testing for the presence of the drugs in blood in race horses &
in Olympic athletic (in forensic GC-MS).
Analyzer of aerosol particles.
Determination of pesticide residues in food.
Polymer characterization (pyrolysis methods combined GCMS).
Drug monitoring & toxicology studies.
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62. Impurities can arise from
•Manufacturing process
•Degradation of drug substance
If impurities are greater than 0.1% concentration in drug substance
the name and structure of impurity is to be submitted to regulatory agencies.
USFDA CONSIDERATIONS:
IMPURITIES SHOULD BE LESS THAN 1.0%
Presence of impurities must be illustrated with GC-MS/LC-MS chromatograms
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69. GC-MS is becoming the tool of choice for tracking organic
pollutants in the environment.
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70. GC-MS can analyze the particles from a human body in order
to help link a criminal to a crime.
GC-MS especially useful here as samples often contain very
complex matrices &results used in court.
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71. GC-MS used for detection of illegal narcotics & may
eventually supplant drug-sniffing dogs.
It’s also commonly used in forensic toxicology to find drugs
&poisons in biological specimens of victims .
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72. GC-Ms is main tool used in sports anti doping
laboratories to test athletes urine samples for prohibited
performance enhancing drugs.
EG : anabolic steroids.
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73. Food & beverage contain numerous aromatic compounds ,
some naturally present in the raw materials &some forming
during process.
GC-MS is extensively used for the analysis of these
compounds which include ester, fatty acids , alcohols,
aldehydes, terpenes etc……
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74. GC-MS 2 were brought to mars by the Viking program.
Venera11&12 pioneer Venus analyzed the atmosphere of
Venus with GC-MS.
The material in the comet 67p will be analyzed by the rosetla
mission with a chiral GC-MS in 2014.
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75. In born errors of metabolism are now detectable by new born
screening tests, especially the testing using GC-MS .
It can determine compounds in urine even in minor
concentration.
The measurement of c13-c12 ratio with an isotope ratio mass
spectrometer.
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76. GC-MS is one of the best analytical tool iinn aannaallyyssiinngg tthhee
mmoosstt ooff tthhee ccoommppoouunnddss. GGCC--MMSS aass aann aannaallyyttiiccaall mmeetthhoodd
ooff ooppttiioonn iiss iittss iinnccrreeaasseedd sseennssiittiivviittyy && rreelliiaabbiilliittyy eevveenn iinn
vveerryy ssmmaallll qquuaannttiittiieess ((nngg)).
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78. Gurdeep R.Chatwal, K.Anand, Instrumental methods of
chemical analysis (Pg.No 2.272,2.673)
Pavia,Lampman, Kriz, (Pg.No 401-415).
B.K Sharma, Instrumental methods of chemical analysis
(Pg.No 844-938,180-224)
Dr.S.Ravi Sankar text book of pharmaceutical analysis
3rd Edition (Pg.No 8.1,17.1)
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79. AACCKKNNOOWWLLEEDDGGEEMMEENNTTS
I sincerely thank my guide
Mr. Ch. DEVADASU sir
for his constant guidance & support.
I thank our respected Principal
Dr. P.SRINIVASA BABU
& seminar Committee for giving me
this opportunity.
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