The aim of the coupling is to obtain an information-rich detection for both identification and quantification compared to that with a single analytical technique.
An Infrared spectrum represents a fingerprint of a sample with absorption peaks which correspond to the frequencies of vibrations between the bonds of the atoms making up the material-Because each different material is a unique combination of atoms, no two compounds produce the exact same spectrum, therefore IR can result in a unique identification of every different kind of material!
The aim of the coupling is to obtain an information-rich detection for both identification and quantification compared to that with a single analytical technique.
An Infrared spectrum represents a fingerprint of a sample with absorption peaks which correspond to the frequencies of vibrations between the bonds of the atoms making up the material-Because each different material is a unique combination of atoms, no two compounds produce the exact same spectrum, therefore IR can result in a unique identification of every different kind of material!
Detectors are the brain of any chromatograhic system. It help us to record the chromatogram based on certain characteristics of the analyte and help us in identifying that compound both qualitatively and quantitatively.
a type of an analyzer used in mass spectrometer. separates the ions based on mass to charge ratios. useful for the detection of ions present in the sample
Fourier transform infrared spectroscopy: advantage and disadvantage of conventional infrared spectroscopy, introduction to FTIR ,principle of FTIR, working, advantage, disadvantage and application of FTIR.
• It is the combination of liquid chromatography and the mass spectrometry.
• Liquid chromatography-mass spectrometry (LC-MS) is an analytical chemistry
technique that combines the physical separation capabilities of liquid
chromatography with the mass analysis capabilities of mass spectrometry.
• The combination of these two powerful techniques gives the chemical analyst the
ability to analyze virtually any molecular species; including, thermally labile, non
volatile, and high molecular weight species.
Detectors are the brain of any chromatograhic system. It help us to record the chromatogram based on certain characteristics of the analyte and help us in identifying that compound both qualitatively and quantitatively.
a type of an analyzer used in mass spectrometer. separates the ions based on mass to charge ratios. useful for the detection of ions present in the sample
Fourier transform infrared spectroscopy: advantage and disadvantage of conventional infrared spectroscopy, introduction to FTIR ,principle of FTIR, working, advantage, disadvantage and application of FTIR.
• It is the combination of liquid chromatography and the mass spectrometry.
• Liquid chromatography-mass spectrometry (LC-MS) is an analytical chemistry
technique that combines the physical separation capabilities of liquid
chromatography with the mass analysis capabilities of mass spectrometry.
• The combination of these two powerful techniques gives the chemical analyst the
ability to analyze virtually any molecular species; including, thermally labile, non
volatile, and high molecular weight species.
an overview of lcms and gcms and its applications....
LC-MS:
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
INTERFACE
Apart from being an inlet system for the
MS, an LC–MS interface is also the coupling
of a detector (MS) to a chromatograph.
The choice of LC–MS interface strongly
influences the characteristics of the MS as
a detector for LC. Therefore, we should
keep in mind what characteristics are ideal
for an LC detector
1. Direct liquid introduction (DLI):
The DLI interface was developed in order to solve the problems with in-capillary evaporation in the capillary inlet.
In a DLI interface, the column effluent is nebulized by the disintegration into small droplets of a liquid jet formed at a small diaphragm After desolvation of the droplets in a desolvation chamber, the analytes can be analysed using solvent-mediated CI with the LC solvents as the reagent gas.
Advantages:
• No heat is applied to the interface and it is therefore able to deal with thermally labile materials better than the moving-belt interface.
• The interface contains no moving parts and is cheap and simple to construct and operate and is inherently more reliable than the moving-belt interface.
• Both positive- and negative-ion CI spectra can be generated and the interface provides molecular weight information, plus it can also be used for sensitive quantitative and semi-quantitative procedures.
Disadvantages:
• Involatile compounds are not usually ionized with good efficiency.
• The pinhole is prone to blockage and therefore the system must be kept completely free of solid materials.
• Only a small proportion of the flow from a conventional HPLC column is able to enter the source of the mass spectrometer and sensitivity is consequently low
2. Moving belt/wire:
In a moving-belt interface (MBI), the column effluent is deposited onto an endless moving belt from which the solvent is evaporated by means of gentle heating and efficiently exhausting the solvent vapours. After removal of the solvents, the analyte molecules are (thermally) desorbed from the belt into the ion source and mass analysed.
The MBI for LC.MS was used in a wide variety of applications, including the analysis of drugs and their metabolites, pesticides, steroids, alkaloids, polycyclic, aromatic hydrocarbons and others.
Advantages:
• The interface can be used with a wide range of HPLC conditions.
• The analyst does have some choice of the ionization method to be used; EI, CI and FAB are available, subject to certain limitations, and thus both molecular weight and structural information may be obtained from the analyte(s) under investigation.
this will help to know about the advance technique to analysis the biological sample in cancer diagnosis and general separation of proteins based upon the molecular weight and helps to analysis the new drug synthesis level
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.
Mass spectrometer converts molecules to ions under vacuum so that they can be moved about and manipulated by external electric and magnetic fields.
These ions are then separated and determined. Separation is achieved on different trajectories of moving ions in electrical and/or magnetic fields.
*Electrospray Ionization (ESI)
*Matrix-Assisted Laser Desorption/Ionization (MALDI)
*Time-of-Flight (TOF) Mass Analyzer
Recent advances in the application of mass spectrometry in food-related analysis
*LC-MS coupling techniques
*HPLC-MS coupling techniques
*MALDI-TOF-MS
*ESI-MS
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The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
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Liquid chromatography and mass spectrometry.(LCMS)
1.
2. Chromatography is the collective term for a set of
laboratory techniques for the separation of mixtures. The
mixture is dissolved in a fluid called the mobile phase,
which carries it through a structure holding another
material called the stationary phase. The various
constituents of the mixture travel at different speeds,
causing them to separate. The separation is based on
differential partitioning between the mobile and
stationary phases. Subtle differences in a compound's
partition coefficient result in differential retention on the
stationary phase and thus changing the separation.
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. Modern techniques of mass spectrometry were
devised by Arthur Jeffrey Dempster and F.W. Aston
in 1918 and 1919 respectively.
Mass spectrometry (MS) is an analytical chemistry
technique that measures the mass-to-charge ratio
(m/z) and abundance of gas-phase ions. A mass
spectrum (plural spectra) is a plot of the ion signal as
a function of the mass-to-charge ratio. The spectra are
used to determine the elemental or isotopic signature
of a sample, the masses of particles and of molecules,
and to elucidate the chemical structures of molecules,
such as peptides and other chemical compounds.
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 liquid chromatograph and a mass spectrometer.
Therefore, when an identifying mass spectrum appears at a
characteristic retention time in a LC-MS analysis, it typically lends
to increased certainty that the analyte of interest is in the sample.
9. 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.
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.
11. • 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)
13. • 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.
14.
15.
16. 16
As soon as the solutes are eluted they should be detected and
quantitated with sensitive and specific or universal detectors.
With specific and universal detectors we have achieved the
following attributes:
High sensitivity.
High reliability and accuracy
20. 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)
28. 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.
29. 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.