• 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.
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.
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.
LC/MS is a technique that combines physical separation capabilities of LC with mass analysis capability of MS.
It is a method that combines separation power of HPLC with detection power of MS.
In LC-MS we remove the detector from the column of LC and fit the column to interface of MS.
In the most of the cases the interface used in LC-MS are ionization source.
Mass Analyzers for example Magnetic Sector Mass Analyzer, Double Focusing Mass Analyzer, Quadroupole Mass Analyzer, Time of Flight Mass Analyzer and Applications of Mass Analyzer were explained
Electron Spray Ionization (ESI) and its ApplicationsNisar Ali
In this slide ,You will get to learn Electron Spray Ionization (ESI) technique used in Mass Spectroscopy and its Various Application in Pharmaceutical Drug Analysis.
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
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.
LC/MS is a technique that combines physical separation capabilities of LC with mass analysis capability of MS.
It is a method that combines separation power of HPLC with detection power of MS.
In LC-MS we remove the detector from the column of LC and fit the column to interface of MS.
In the most of the cases the interface used in LC-MS are ionization source.
Mass Analyzers for example Magnetic Sector Mass Analyzer, Double Focusing Mass Analyzer, Quadroupole Mass Analyzer, Time of Flight Mass Analyzer and Applications of Mass Analyzer were explained
Electron Spray Ionization (ESI) and its ApplicationsNisar Ali
In this slide ,You will get to learn Electron Spray Ionization (ESI) technique used in Mass Spectroscopy and its Various Application in Pharmaceutical Drug Analysis.
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
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.
Application of ICP-MS and LC-ICP-MS in Drug DevelopmentQPS Holdings, LLC
Inductively coupled mass spectroscopy plasma (ICP-MS) has big potential in preclinical and clinical studies of new drug candidates. One particular area is metallodrugs.
Chromatography is an analytical method in which compounds are physically separated and measured.
The main purpose of chromatography is to separate and quantify the target sample.
The Chromatography technique used to separate a mixture of compounds in pharmaceutical sciences , analytical analytical Chemistry with the purpose of identifying, quantifying and purifying the individual components of the mixture.
Here are the lecture notes from the presentation titled 'ICP-OES/MS Analysis: Advancements, Limitations, and Future Applications in Soil and Water Research,' delivered to a group of researchers affiliated with the Soil & Water Department, Faculty of Agriculture, Hebrew University of Jerusalem (Seagram Center) in 2023. The aim was to explore advanced technologies in elemental analysis and their application to soil and water research. The Lecture Notes Brochure (22 pages) can serve as a concise guide to ICP-OES/MS for researchers and students, assisting them in selecting the appropriate technique for their projects
Interfaces in chromatography [LC-MS, GC-MS, HPTLC, LC, GC]Shikha Popali
THE INTERFACES OF CHROMATOGRAPHY INCLUDES THE CHROMATOGRAPHY CRITEREA WHERE THE DIFFERENT CHROMATOGRAPHY ARE EXPLAINED IN DETAIL WITH PRACTICAL EXAMPLES AND THEIR IMAGES.
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
powerpoint presentation on high performance liquid chromatography which include its definition, classification, principles of seperation, instrumentation and application.
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
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These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
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Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
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micro teaching on communication m.sc nursing.pdfAnurag Sharma
Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
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Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
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MANAGEMENT OF ATRIOVENTRICULAR CONDUCTION BLOCK.pdf
Lc ms
1. SAGAR SAVALE 1
Liquid chromatography and the mass spectrometry
(LC-MS)
Mr. Sagar Kishor Savale
(Department of Pharmaceutics, North Maharashtra University, college of
R.C.Patel Institute of Pharmaceutical Education and Research, Shirpur, 425405,
Dist.Dhule, Maharashtra.)
avengersagar16@gmail.com
2. SAGAR SAVALE 2
1. Introduction
• 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.
• It has been said that over 80% of known organic species are amenable to separation
with liquid chromatography.
• Mass spectrometry is capable of providing structure, molecular weight, empirical
formula, and quantitative information about a specific analytes so, LCMS in recent
years, liquid chromatography/mass spectrometry (LC/MS) has become one of the
most powerful analytical techniques for qualitative and quantitative analysis .
• AIMS - To identify the different proteins, peptides drugs in various samples also to
study the bioequivalence in future
2. Advantages of LC-MS
2.1 Provides compound identity.
2.2 Provides sensitive response to most analytes.
2.3 Provides compound class information.
2.4 Provides compound structure.
2.5 Provides sequence information.
2.6 Provides molecular weight information.
3. SAGAR SAVALE 3
2.7 Provides the five 5 Important Criteria
Speed, Selectivity, Specificity, Sensitivity, Low Cost
3. Sample Consideration for LC-MS
3.1 The analytes must have ionizable groups such as Amines, Carboxylic Acids, Ketones
and Aldehydes.
3.2 For best sensitivity, work at a pH where the analytes is ionized
3.3 i.e. for acid, Neutral to basic pH (7-9) and Acidic pH (3-4) for bases.
4. Parts of LC-MS
5. Flow Chart of LC-MS
Sample Column Detector Eluent
Port
Collector
Ionization Source Vacuum
Mass analyzer
Detector
Read out device
4. SAGAR SAVALE 4
6. Liquid Chromatography
6.1 Liquid chromatography is type of chromatography in which analytes molecule get
partitioned between moving mobile phase and stationary phase
6.2 In liquid chromatography mobile phase is always liquid while stationary phase is either
liquid or solid.
6.3 Liquid chromatography includes following chromatographic techniques:
6.3.1 Paper chromatography
6.3.2 Thin layer chromatography
6.3.3 Adsorption column chromatography
6.3.4 High performance liquid chromatography
6.3.5 Ion exchange chromatography
6.3.6 Liquid-liquid chromatography
7. Instrumentation of
1.1 Detectors
1.2 Pumps
1.3 Column heaters
1.4 Auto samplers
8. Applications of LC:
8.1 Separation
8.2 Purification
8.3 Quantification
9. Mass Spectroscopy
5. SAGAR SAVALE 5
9.1 Mass spectroscopy is an analytical technique used to measure the mass-to-charge-
ratio of ions it is most generally used to find the composition of a physical sample by
generating a mass spectrum representing the masses of sample components.
Sample Inlet Ionization source Mass Analyzer Detector Read out
Vacuum
9.2 The stages within the mass spectrometer are:
9.2.1 Producing ions from the sample.
9.2.2 Separating ions of differing masses.
9.2.3 Detecting the number of ions of each mass produced.
9.2.4 Collecting the data and generating the mass spectrum.
9.3 Application of MS
9.3.1 Identifying unknown compounds by the mass of the compound molecules or their
fragments. Determining the isotopic composition of elements in a compound.
9.3.2 Determining the structure of a compound by observing its fragmentation.
9.3.3 Quantifying the amount of a compound in a sample using carefully designed methods.
9.3.4 Determining other physical, chemical, or even biological properties of compounds with
a variety of other approaches.
6. SAGAR SAVALE 6
10. Interfaces Used In LC-MS
10.1. Direct Liquid Introduction
The first attempts to introduce a liquid into an MS using the classic electron impact
ionization (EI)/chemical ionization (CI) source were based on the simple principle that by
minimizing the amount of liquid, the vacuum system would remove the solvent leaving the
analytes in the gas phase for ionization.
Figure 1: Scheme of the DLI interface. 1 _ connection to LC column, 2 _ diaphragm 5 μm
Opening to MS, 3 _ needle valve, 4 _ cooling region, 5 _ to UV detector or waste.
10.2 Moving belt/wire interface
The moving-belt interface separates the condensed liquid-phase side of the LC from the high
vacuum of the MS and uses a belt to transport the analytes from one to the other. The mobile
phase of the LC is deposited on a band and evaporated. The analytes remain on the continuously
cycling belt and are transported from atmospheric pressure into the vacuum of the ion source
through two differentially pumped vacuum locks. A heater in the ion source evaporates the
sample from the belt allowing MS analysis.
7. SAGAR SAVALE 7
Figure 2: Schematic showing the principal components of a moving-belt interface
10.3 Thermo spray Interface
As the name thermo spray implies, heating the liquid flow leaving an LC system creates a
spray of superheated mist containing small liquid droplets.
The most successful method involves directing the liquid flow through an electrically heated
capillary, which can be directly introduced into the MS ion source. The droplets are further
vaporized as they collide against the walls of the heated ion source.
Figure 3: Thermo spray interface.
8. SAGAR SAVALE 8
10.4 Particle Beam Interface (MAGIC)
MAGIC, an acronym for monodisperse aerosol generation interface for chromatography. The
LC eluent is forced through a small nebulizer using a He gas flow to form a stream of uniform
droplets. These droplets move through a desolvation chamber and evaporate to a solid particle.
These particles are separated from the gas and transported into the MS source using a
differentially pumped momentum separator.
10.5 Atmospheric pressure chemical ionization
10.5.1 In APCI, the LC eluent is sprayed through a heated (typically 250°C – 400°C)
vaporizer at atmospheric pressure. The heat vaporizes the liquid.
10.5.2 The resulting gas-phase solvent molecules are ionized by electrons discharged from
a corona needle.
10.5.3 The solvent ions then transfer charge to the analyze molecules through chemical
reactions (chemical ionization).
10.5.4 The analytes ions pass through a capillary sampling orifice into the mass analyzer.
10.5.5 APCI is applicable to a wide range of polar and non polar molecules.
9. SAGAR SAVALE 9
10.5.6 It rarely results in multiple charging so it is typically used for molecules less than
1,500 u.
10.5.7 Because it involves high temperatures, APCI is less well-suited than electrospray for
analysis of large biomolecules that may be thermally unstable.
10.5.8 APCI is used with normal-phase chromatography more often than electrospray is
because the analytes are usually non polar.
Figure 5 Atmospheric pressure chemical ionization
10.6. Atmospheric pressure photoionization
10.6.1 As in APCI, a vaporizer converts the LC eluent to the gas phase.
10.6.2 A discharge lamp generates photons in a narrow range of ionization energies.
10.6.3 The range of energies is carefully chosen to ionize as many analyte molecules as
possible while minimizing the ionization of solvent molecules.
10.6.4 The resulting ions pass through a capillary sampling orifice into the mass analyzer.
10. SAGAR SAVALE 10
Figure 6 Atmospheric pressure photoionization
10.7 Electrospray Ionization (ESI)
The analytes solution flow passes through the electrospray needle that has a high potential
difference (with respect to the counter electrode) applied to it. This forces the spraying of
charged droplets from the needle with a surface charge of the same polarity to the charge on
the needle. The droplets are repelled from the needle towards the source sampling cone on the
counter electrode. As the droplets traverse the space between the needle tip and the cone and
solvent evaporation occurs.
Figure 7 Electrospray Ionization (ESI)
11. SAGAR SAVALE 11
11. Applications
LC-MS USED IN FOLLOWING AREAS
11.1 Drug Discovery
11.2 Clinical Analysis.
11.3 Proteomics
11.4 Forensic Chemistry
11.5 Drug Metabolism study
11.6 Environmental chemistry
11.7 Diagnostic studies.
11.8 Molecular Weight Determination
11.9 Determination Of Molecular Weight Of Green Florescent Protein
11.10 Structural Determination
11.11 Pharmaceutical Applications
11.12 Identification Of Bile Acid Metabolites
11.13 Clinical Applications
11.14 Biochemical Genetics
11.15 Therapeutic Drug Monitoring
12. Conclusion
The study has been shown that the LC-MS is the valuable tool for analysis of various
biological samples.
The LCMS provides accuracy, specificity, selectivity & rapid less time consuming.
12. SAGAR SAVALE 12
Instrumentation of LC-MS is quite complicated but very efficient for their accuracy,
sensitivity.
It is mostly useful in pharmaceutical industries finds application in pharmacokinetic
study, proteomics, drug development, radio pharmaceutics and clinical analysis & in
toxicological studies.
13. Future Scope
LC-MS is the technique which have application in future in bioequivalence study.
It can be use for blood analysis, for detection of drugs whose identity is not known till
now.