- 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.
- GC is used to separate and analyze compounds that can be vaporized without decomposition, while MS is used to measure the mass-to-charge ratios of ions to determine the molecular mass and structure of molecules eluting from the GC.
- The combination of GC and MS provides a powerful tool for analyzing complex mixtures by separating compounds and identifying their chemical structures.
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 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
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 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.
This ppt consist of basic principle of GC-MS, instrumentation of GC-MS, components of GC-MS ,Advantages and disadvantages of GC-MS and application of GC-MS
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
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 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.
This ppt consist of basic principle of GC-MS, instrumentation of GC-MS, components of GC-MS ,Advantages and disadvantages of GC-MS and application of GC-MS
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
Gas chromatography–mass spectrometry (GC-MS) is an analytical method that combines the features of gas-chromatography and mass spectrometry to identify different substances within a test sample.[1] Applications of GC-MS include drug detection, fire investigation, environmental analysis, explosives investigation, food and flavor analysis, and identification of unknown samples, including that of material samples obtained from planet Mars during probe missions as early as the 1970s. GC-MS can also be used in airport security to detect substances in luggage or on human beings. Additionally, it can identify trace elements in materials that were previously thought to have disintegrated beyond identification. Like liquid chromatography–mass spectrometry, it allows analysis and detection even of tiny amounts of a substance.[2]
GC-MS has been regarded as a "gold standard" for forensic substance identification because it is used to perform a 100% specific test, which positively identifies the presence of a particular substance. A nonspecific test merely indicates that any of several in a category of substances is present. Although a nonspecific test could statistically suggest the identity of the substance, this could lead to false positive identification. However, the high temperatures (300°C) used in the GC-MS injection port (and oven) can result in thermal degradation of injected molecules,[3] thus resulting in the measurement of degradation products instead of the actual molecule(s) of interest.The first on-line coupling of gas chromatography to a mass spectrometer was reported in the late 1950s.[4][5] An interest in coupling the methods had been suggested as early as December 1954.
Introduction to gas Chromatography
,Principle of gas chromatography
Instrumentation of gas Chromatography
Type of detectors of gas chromatography
Advantages of gas chromatography
Disadvantages of gas chromatography
Applications of gas chromatography
mass spectrometry for pesticides residue analysis- L1sherif Taha
This is the first lecture in series of lectures on mass spectrometry for pesticides residue analysis. This lecture (1) include Pesticides classification, introduction to mass spectrometry, vacuum system for Agilent GC MS/ MS and AB SCIEX LC MS/ MS
Peptide vaccine containing only epitopes capable of inducing positive, desirable T cell and B cell mediated immune response.
Peptides‖ used in these vaccines are 20–30 amino acid sequences that are synthesized to form an immunogenic peptide molecule representing the specific epitope of an antigen.
sufficient for activation of the appropriate cellular and humoral responses
Eliminating allergenic and/or reactogenic responses.
The different types of external stresses that influence the plant growth and development.
These stresses are grouped based on their characters
Biotic
Abiotic
Almost all the stresses, either directly or indirectly, lead to the production of reactive oxygen species (ROS) that create oxidative stress in plants.
This damages the cellular constituents of plants which are associated with a reduction in plant yield.
Bioreactors are devices in which biological or biochemical processes develop under a closely monitored and tightly controlled environment. Bioreactors have been used in animal cell culture since the 1980s in order to produce vaccines and other drugs and to culture large cell populations. Bioreactors for use in tissue engineering have progressed from such devices.
A tissue engineering bioreactor can be defined as a device that uses mechanical means to influence biological processes. In tissue engineering, this generally means that bioreactors are used to stimulate cells and encourage them to produce extracellular matrix (ECM). There are numerous types of bioreactor which can be classified by the means they use to stimulate cells.
Microgravity is the condition in which people or objects appear to be weightless (In space). Astronauts and cosmonauts returning from long-term space missions exhibited various health problems, among them changes of the immune system, bone loss, muscle atrophy, ocular problems, and cardiovascular changes. Space biologists investigated various cell types in space to find the molecular mechanisms responsible for the observed immune disorders. Experimental cell research studying three-dimensional (3D) tissues in space and on Earth using new techniques to simulate microgravity is currently a hot topic in Gravitational Biology and Biomedicine.
An idea was considered as to producing an entire organ in vivo by bypassing many of the steps like cell isolation and expansion, culturing in bioreactors, scaffolds and growth factor delivery ect. involved in traditional tissue engineering. This concept was called the in vivo bioreactor (IVB).
Biomaterials were defined as “any substance, other than a drug, or a combination of substances, synthetic or natural in origin, which can be used for any period of time, as a whole or as a part of a system, which treats, augments or replaces any tissue, organ or function of the body”
Hematopoiesis is the process through which the body manufactures blood cells. It begins early in the development of an embryo, well before birth, and continues for the life of an individual. Hematopoiesis begins during the first weeks of embryonic development. All blood cells and plasma develop from a stem cell that can develop into any other cell.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
Comparative structure of adrenal gland in vertebrates
Chromatography(gc ms & lc ms)
1.
2. • Chromatography is the science which is studies
the seperation of molecules based on
differences in their structure and/or
composition.
• Chromatography was first developed and
defined by the Russion Botonist Mikhail
Tswett in 1903. He produced a colourful
seperation of plant pigments using a column of
calcium carbonate(chalk).
3. Charged molecules or molecular fragments are
generated in a high vacuum or immediately prior to a
sample entering a high vacuum using a variety of
methods for ion production.
magnetic fields to enable the determination of their
molecular weight (m/z).The history of mass
spectroscopy (MS) started in 1912 when Thomson obtd.
4. Combining the two processes reduces the possibility of error, as it is extremely
unlikely that two different molecules will behave in the same way in both a gas
chromatograph and a mass spectrometer.
Therefore, when an identifying mass spectrum appears at a characteristic retention
time in a GC-MS analysis, it typically lends to increased certainty that the analyte
of interest is in the sample.
5. Gas Chromatography was first described in 1952 by James
and martin with the separation of a mixture of small
carboxylic acids.
The power of GC was substantially enlarged by the
introduction of open capillary columns in 1958 by Golay.
The introduction of the fused-silica capillary column in
1976 by Dandeneau and Zerner can be considered as a
breakthrough in the development of GC.
The use of a mass spectrometer as the detector in gas
chromatography was developed during the 1950s by
Roland Gohlke and Fred McLafferty.
6.
7. • It is an instrumental method for the seperation and
identification of chemical compounds.
• Gas chromatography is a chromatographic
technique that can be used to separate volatile
organic compounds.
• The organic compounds are seperates due to
differences in their partitioning behaviour between
the mobile phase and the stationary phase in the
column.
8. The sample solution is injected into the GC inlet
where it is vaporized and swept onto a
chromatographic column by the carrier gas (usually
helium).
The sample flows through the column and the
compounds comprising the mixture of interest are
separated by interaction with the coating of the
column (stationary phase) and the carrier gas
(mobile phase).
The latter part of the column passes through a
heated transfer line and ends at the entrance to ion
source where compounds eluting from the column
are converted to ions.
9. The GC-MS is composed of two
major building blocks: the gas
chromatograph and the mass
spectrometer.
Carrier Gas, N2 or He, 1-2 mL/min
Injector
Oven
Column
Detector
10. Inert
Helium (hydrogen/ nitrogen).
Choice dictated by detector, cost, availability
Pressure regulated for constant inlet pressure(
below 0.3MPa).
Flow controlled for constant flow rate
~20ml/min for packed column
~1ml/min for open capillary column
Chromatographic grade gases (high purity)
11. State
Organic compounds must be in solution for injection into the
gas chromatograph.
The solvent must be volatile and organic.
Amount
Depending on the ionization method, analytical sensitivities of
1 to 100 pg per component are routine.
Preparation
Sample preparation can range from simply dissolving some of
the sample in a suitable solvent .
12. Purge and Trap
(Aqueous and Soils / Volatiles Preparation)
Courtesy of Environmental Conservation
18. Made of the highest purity fused silica obtained
with an external polyimide coating
Length-10 to 100 m -depends on application.
For fast analysis shorter column are applied e.g.-
for heat sensitive and for high boiling compound.
Large columns are required for high resolution
seperation.
19. A more polar stationary phase is applied for the analysis
of more polar compounds.
So least polar column- CP-Sil 5 and CP-Sil 8 applied.
For high resolution more polar stationary phase have to
be applied.
In GC-MS low bleed columns are applied.
20.
21. These columns, less commonly used today, have
diameter of 1.6 to 9.5mm and a length of
between 1–3m.
Manufactured from steel or glass, the internal
wall of the tube is treated to avoid catalytic
effects with the sample.
They can withstand a carrier gas flow rate
within the range 10–40 mL/min.
They contain an inert and stable porous support
on which the stationary phase can be
impregnated or bounded (between 3 and 20 per
cent).
22.
23. Two capillary tubes aligned with a small
space between them. (1 mm)
A vacuum is created between the two tubes
using a rotary pump.
The GC effluent enters the vacuum region,
those molecules which continue in the same
direction enter the second capillary tube and
continue to the ion source.
24. The carrier gas molecules are more easily
diverted from the linear path by collisions.
The analyte molecules are much larger and
carry more momentum.
The surface of the separator must be inactive
and a reasonably even temperature.
26. The insides of the GC-MS, with the column of the
gas chromatograph in the oven on the right.
27. • The gas chromatograph utilizes a capillary column
which depends on the column's dimensions (length,
diameter, film thickness) as well as the phase properties.
• The difference in the chemical properties between
different molecules in a mixture will separate the
molecules as the sample travels the length of the column.
• The molecules take different amounts of time (called the
retention time) to come out of (elute from) the gas
chromatograph, and this allows the mass spectrometer
downstream to capture, ionize, accelerate, deflect, and
detect the ionized molecules separately.
• The mass spectrometer does this by breaking each
molecule into ionized fragments and detecting these
fragments using their mass to charge ratio.
29. The physics behind mass spectrometry is that a charged
particle passing through a magnetic field is deflected along a
circular path on a radius that is proportional to the mass to
charge ratio, m/e.
30. • Used to confirm molecular weight.
• Known as a “soft” ionisation technique.
• Differs from EI in that molecules are ionised
by interaction or collision with ions of a
reagent gas rather that with electrons.
• Common reagent gases used are Methane ,
Isobutane and Ammonia.
• Reagent gas is pumped directly into ionisation
chamber and electrons from Filament ionise
the reagent gas.
31. The most common type of mass spectrometer (MS)
associated with a gas chromatograph (GC) is the
quadrupole mass spectrometer, sometimes referred
to by the Hewlett-Packard (now Agilent) trade name
"Mass Selective Detector" (MSD).
Another relatively common detector is the ion trap
mass spectrometer.
Other detectors may be encountered such as time of
flight (TOF), tandem quadrupoles (MS-MS).
32.
33.
34. • A mass spectrometer is typically utilized in one of two ways:
• Full Scan or Selective Ion Monitoring (SIM).
• The typical GC/MS instrument is capable of performing both
functions either individually or concomitantly, depending on
the setup the particular instrument.
36. Environmental Monitoring and
Cleanup
GC-MS is becoming the tool of choice for
tracking organic pollutants in the
environment. The cost of GC-MS
equipment has decreased significantly, and
the reliability has increased at the same
time, which has contributed to its increased
adoption in environmental studies. There
are some compounds for which GC-MS is
not sufficiently sensitive, including certain
pesticides and herbicides, but for most
organic analysis of environmental samples,
including many major classes of pesticides,
it is very sensitive and effective.
37. • GC-MS can analyze the particles from a human body in order
to help link a criminal to a crime. The analysis of fire debris
using GC-MS is well established, and there is even an
established American Society for Testing Materials (ASTM)
standard for fire debris analysis.
• GCMS/MS is especially useful here as samples often contain
very complex matrices and results, used in court, need to be
highly accurate.
38. Law Enforcement
GC-MS is increasingly used for detection of illegal narcotics, and may
eventually supplant drug-sniffing dogs. It is also commonly used in forensic
toxicology to find drugs and/or poisons in biological specimens of suspects,
victims, or the deceased.
Food, Beverage and Perfume Analysis
Foods and beverages contain numerous aromatic compounds, some naturally
present in the raw materials and some forming during processing. GC-MS is
extensively used for the analysis of these compounds which include esters, fatty
acids, alcohols, aldehydes, terpenes etc. It is also used to detect and measure
contaminants from spoilage or adulteration which may be harmful and which is
often controlled by governmental agencies, for example pesticides.
39. Astrochemistry
• Several GC-MS have left earth. Two
were brought to Mars by the Viking
program.Venera 11 and 12 and Pioneer
Venus analysed the atmosphere of Venus
with GC-MS.
• The Huygens probe of the Cassini-
Huygens mission landed one GC-MS on
Saturn's largest moon, Titan.The material
in the comet 67P/Churyumov-
Gerasimenko will be analysed by the
Rosetta mission with a chiral GC-MS in
2014.
40. • It is the combination of liquid
chromatography and the mass
spectrometry.
• In LC-MS we are removing the detector
from the column of LC and fitting the
column to interface of MS.
• In the most of the cases the interface used
in LC-MS are ionization source.
42. To reduce separation time to achieve fast analysis. (
From hours to minutes)
HOW?
The solutes must move faster through stationary phase MEANS
By increasing the mobility of the liquid mobile phase.
43. The MIGRATION VELOCITY of the liquid mobile phase should be
HIGH.
We can achieve faster migration velocities of liquid mobile
phase by-
1.Applying vacuum at the other end of the chromatographic column
2.Applying high pressure on the liquid mobile phase.
Application of higher pressure on liquid mobile phase
seems to be an easier and more practical way for achieving
fast analysis.
44. Sample preparation generally consists of concentrating
the analyte and removing compounds that can cause
background ion or suppress ionization.
Example of sample preparation include:-
(1) on –column concentration to increase analyte
concentration.
(2) desalting to reduce the sodium and potassium
adduct formation that commonly occurs in electro
spray.
(3) filtration to separate a low molecular-weight
drug from proteins in plasma, milk, or tissue.
45. • Column type:-
• Specialized mode:-
• The use of di-functional or tri-functional silanes
to create bonded groups with two or three
attachement points leading to phases with higher
stability in low or higher pH and lower bleed for
LCMS
• Most widely used columns for LCMS are:-
(1) fast LC column.
the use of short column. (15-50mm)
(2) Micro LC column.
the use of large column. ( 20-150mm)
46. • Glass is normally the choice for chromatographic
column material due to its chemical inertness towards
stationary phase, mobile phase and solutes being
separated.
• We could not increase the pressure of a glass column?
• Hence glass will not be able to withstand high
pressures of liquid mobile phase because glass has
relatively small mechanical strength.
47. • Which IDEAL MATERIAL should be selected in place of
glass?
• STAINLESS STEEL due to its chemical inertness &
mechanical strength.
48. 48
Previous condition :- Liquid mobile phase moving
across the stationary phase in a glass column under
atmospheric pressure.
Present condition :- Liquid mobile phase with high
migration velocity moving across the stationary
phase in a stainless steel column under high
pressure..
Will the column separation efficiency of the
latter be good?
NO
49. With the increase in the liquid mobile
phase velocity under high pressures,
solute molecules will not get enough
time to interact with the stationary phase
and separation efficiency will be reduced
and we will not be able to achieve good
separation of solutes.
49
50. How can we achieve good column separation
efficiency?
50
1. By reducing the Particle size of the stationary phase
support.
2. By reducing the width of chromatographic column.
51. We use Narrow bore (3mm, id) SS columns filled
with small particles of stationary phase support (5µ)
will give us high separation efficiency with low
analytical time when liquid mobile phase moves
through the column under high pressures of the
order of 500 –5000 psi.
51
52. Can the physically coated stationary phases used
in GC be used in narrow bore SS columns through
which liquid mobile phase moves under high
pressures?
52
Obviously… NO
53. Because under high pressures of liquid mobile phase,
physically coated liquid stationary phases will be
removed physically or by dissolution in liquid mobile
phase.
We need physically&chemically stable stationary phase.
53
54. Chemically treated inert (not strictly) particles have
free silinol group at their surface.
54
Silica
particle
CCC
CCC
Si - OH
Si - OH
Si - OH
Free silinol
groups
55. Early attempts were then made to chemically bonded long chain
aliphatic alcohols with the free silinol groups present on the
surface of silica particles.
55
Si - OH + HO - R
- H2O
Si - O - R
56. However these chemically bonded phases with
Si - O - C bonds are stable only in acidic media.
56
57. We cannot restrict our separation work always in acidic
media
57
In basic media Si– O- C bond undergoes hydrolysis.
Si - O - R basic medium Si - OH + R - OH
58. We therefore would like to use chemical bonding
with ‘ Si ’ of free silinol groups which will be
hydrolytically stable over a wide acidic to basic
pH range.
58
59. Attempts were then made to carry out chemical
reactions with substituted silane ( Si H4) having
suitable long chain of hydrocarbons attached to it.
59
60. 60
Si - OH
1 2 18
+ Cl – Si – C – C C - H
CH3
CH3
H
H
H
H
H
H
Substituted siliane
1 2 18
– Si – C – C C - H
CH3
CH3
H
H
H
H
H
H
Si - O
-H Cl
61. This chemically bonded phase ( Si-O-Si bond )
is therefore mechanically strong and hydrolytically
stable over a wide pH range (2-9).
61
62. 62
When the liquid mobile phase is moving
under high pressures through
chromatographic columns, introduction of
a sample on to the column will require
some specially designed injection system.
63. 63
The most popular and commonly used
injector system, is the syringe –loop
injector type.
It is a fixed volume universal injector
which allows introduction of micro
litres of samples on to the
chromatographic column.
64. 64
Thus so far with we have achieved the
following attributes with LC
Fast Analysis
Small Sample Size
Versatility
Efficiency
Non-destructive
65. • It is difficult to interface a liquid
chromatography to a mass-spectrometer
cause of the necessity to remove the solvent.
• The commnly used interface are:-
(1) Electrospray ionization (ESI)
(2) Thermospray ionization (TSI)
(3) Atmospheric pressure chemical ionization
(APCI)
(4) Atmospheric pressure
photoionization(APPI)
(5) Partical beam ionization.
66.
67.
68.
69. 69
As soon as the solutes are eluted they should be
detected and quantitated with sensitive and
specific or universal detectors.
70. 70
With specific and universal detectors we have achieved the
following attributes:
High sensitivity
High reliability and accuracy
71. 71
Thus in place of glass column liquid
chromatography now we have
1 4 5
2
3
1- Pumping system
2- Universal injector
3- Column
4- Detector
5- Recorder
72. 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.
Types of mass analyzer:-
(1) Quadrapole mass filter.
(2) time of flight
(3) Ion trap
(4) Fourier transform ion cyclotron
resonance (FT-ICR or FT-MS)
73.
74. 74
1.Pumping system capable of giving
pulseless flow of liquid mobile phase under
high pressures of the order of 500 to
5000psi.
*This makes fast analysis.
76. 76
3.Narrow bore SS columns with suitable chemically
bonded phases with small particle size (3-5µ), desired
polarity and pore size and hydrolytically stable over a
wide acidic to basic pH range.
*This gives high separation efficiency , high
versatility in separating solutes with diverse
nature.
77. 77
4.Sensitive and Specific or Universal detector
*This gives high sensitivity , reliability and
accuracy of detection and quantitation of
solutes.
79. 79
All the above factors add
“High Performance” to normal Liquid
Chromatography.
It is therefore known as
“High Performance Liquid Chromatography”
abbreviated as HPLC.
80. Molecular weight determination
Determining the molecular weight of green
fluorescent proteins
Structural determination e.g. structural
determination of ginsenoside.
Pharmaceutical application e.g. identification
of bile acids metabolites.
Biochemical application e.g. rapid protein
identification using capillary lc/ms/ms.
81. Food application e.g. identification of aflatoxin
in food determination of vitamin D3 in poultry
feed supplement using MS3
Environmental application e.g. detection of
phenyl urea herbicides, detection of low level
of carbaryl in food.
84. GC-MS LC-MS
Volatile or stable
compounds can be used
as samples.
All gases are
determined.
Mobile phase used is
gas.
inner wall of the column
is lined with a thin layer
of support material such
as diatomaceous earth.
Non-volatile compounds
can be used as samples.
Can be used for all salts &
macromolecules.
Mobile phase used is
liquid.
a column that is packed
with a stationary phase
composed of irregularly or
spherically shaped
particles, a porous
monolithic layer, or a
porous membrane
85. 1. Principles and Intrumentation of gas chromatography-mass
spectroscopy by W. M. A. Niessen, hyphen MassSpec
Counsultancy,Leiden,The Netherlands.
2. gas chromatography-mass spectroscopy from Wikipedia ,The free
encyclopedia.
3. Gas chromatography by U. A. Devkate Sir.
4. Encyclo pedia of Chromatography – Jack Cazes
5. Gas chromatography by Ian A[1]. Fowlis 2nd Edition
6. hollas_J.M._ modern _Spectroscopy
7. Handbook of Instrumental Technique for Analytical Chemistry by
Frank Settle.
8. Moderm Instrumentation Methods and Techniques by Francis Rouessac
and Annick Rouessac.
9. Instrumental methods of Analysis by Willard,Merritt,Dean,Settle,7th
86. 10. W. Paul & H. Steinwedel; Zeitschrift für
Naturforschung, 8A; 1953, p448.
11. W. Paul; Agewandte Chemie - International
Edition, 29; 1990, p739.
12. G. C. Stafford et al.; International Journal of
Mass Spectrometry and Ion Processes, 60; 1984,
p85 and Analytical Chemistry, 59; 1987, p1677.