1. Asifa Bibi presented on various chromatography techniques. She discussed paper chromatography, column chromatography, gel filtration chromatography, ion exchange chromatography, and affinity chromatography.
2. The key principles of chromatography are the separation of mixtures based on differences in how components partition between a stationary and mobile phase.
3. Chromatography techniques have many applications in fields like biotechnology, pharmaceuticals, and forensic analysis.
Chromatography : A seperation techniqueSHIVANEE VYAS
Chromatography is a method of seperating mixture of components into individual components through equlibrium distribution between two phases.
Each chromatographic method essentially consists of 2 phases a staionary phase and a mobile phase.
Stationary phase : solid or liquid
Mobile phase : liquid or gas
Chromatography : A seperation techniqueSHIVANEE VYAS
Chromatography is a method of seperating mixture of components into individual components through equlibrium distribution between two phases.
Each chromatographic method essentially consists of 2 phases a staionary phase and a mobile phase.
Stationary phase : solid or liquid
Mobile phase : liquid or gas
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
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
introduction, history, principle, experimental techniques, evaluation on chromatogram, adv. & dis-adv., common problems, comparision, applications and analysis of drugs through TLC(2000-2017)
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
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
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
introduction, history, principle, experimental techniques, evaluation on chromatogram, adv. & dis-adv., common problems, comparision, applications and analysis of drugs through TLC(2000-2017)
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
• Chromatography is a method of separation in which the components to be separated are distributed between two phases, one of these is called a stationary phase and the other is a mobile phase which moves on stationary phase in a definite direction
HPLC- introduction, principle, types, working, instrumentation and operations of HPLC has been included with appropriate gifs and images for better understanding. What are all the things need to be known by a science student about HPLC (basics and working) is clearly given in this presentation.
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
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
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
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.
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Factory Supply Best Quality Pmk Oil CAS 28578–16–7 PMK Powder in Stockrebeccabio
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Hot Selling Organic intermediates
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.
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
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.
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
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
3. CHROMATOGRAPHY
• Chromatography is an analytical
technique commonly used for
separating a mixture of chemical substances into
its individual components, so that the individual
components can be thoroughly analyzed.
• It is also defined as the process of separation of
the individual components of a mixture based on
their relative affinities towards stationary and
mobile phases.
4. • The mixture is dissolved in a fluid called
the mobile phase, which carries it
through a structure holding another
material called the stationary phase.
5. • Its name is derived from two words: "chromo" meaning
colour, and "graphy" meaning writing. in other words, colour
bands are formed in the procedure, which are then measured or
analysed. these bands are due to separation of individual
compounds at different lengths on the column, as seen in
column chromatography and on paper in paper chromatography.
ORIGIN OF NAME:-
6. • The word “chromatography” was firstly coined by
the italian-born scientist Mikhail tsvet in 1903 in
russia .
• He continued to work with chromatography in the
first decade of the 20th century, primarily for the
separation of plant pigments
• Chromatography technique developed
substantially as a result of the work of archer john
porter martin and richard laurence millington
synge during the 1940s and 1950s.
HISTROY:-
7. • Chromatography is based on the principle of separation of
compounds into different bands (color graphs) and then
identification of those bands.
• Basically, the samples are subjected to flow by mobile
liquid onto or through the stable stationary phase. the
sample components are separated into fractions based on
their relative affinity towards the two phases during their
travel.
• Thefraction with a greater affinity to stationary layer
travels slower and at a shorter distance, while that with a
lesser affinity travels faster and longer
PRINCIPLE:-
8. TYPES OF CHROMATOGRAPHY
Types of chromatography to
be discussed:-
• Paper Chromatography
• Column Chromatography
• Gel Filtration
Chromatography
• Ion Exchange
Chrmoatography
• Affinity Chromatography
9. • Paper chromatography is an analytical chemistry technique for
separating and identifying color mixtures.
• Substances are distributed between stationary phase and a mobile
phase.
• Stationary phase is usually a piece of filter paper and mobile phase is
colors that travels up stationary phase.
• Components of the samples will separate readily according to how
strongly they absorb on the stationary phase vs. how readily they
dissolve in the mobile phase.
PAPER CHROMATOGRAPHY:
12. • The principle involved is partition chromatography
wherein the substances are distributed or partitioned
between liquid phases. one phase is the water, which is
held in the pores of the filter paper used; and other is the
mobile phase which moves over the paper. the compounds
in the mixture get separated due to differences in their
affinity towards water (in stationary phase) and mobile
phase solvents during the movement of mobile phase under
the capillary action of pores in the paper.
PRINCIPLE:-
14. • Ink is a solution containing a number of different molecules.
These different molecules have different characteristics such
as size and solubility. Solubility is a molecule's ability to
dissolve in a particular solvent such as alcohol, water or nail
polish remover. Because of their different characteristics,
each molecule travels at a different speed when pulled along
the piece of paper toweling by the solvent. The lightest
particles, which are not necessarily the lightest coloured
particles, move more quickly and a greater distance than the
heavier particles. Thus, all of the pigments that make up an
ink sample are separated out.
WHAT HAPPEN IN PAPER
CHROMATOGRAPHY:-
15. • Paper chromatography is specially used for the
separation of a mixture having polar and non-polar
compounds.
• separation of amino acids.
• Determine organic compounds, biochemical in urine
• In the pharma sector it is used for the determination of
hormones, drugs, etc.
• Sometimes it is used for evaluation of inorganic
compounds like salts and complexes
USES AND APPLICATIONS:-
17. Column chromatography is a preparative technique used to
purify compounds depending on their polarity or hydrophobicity.
In column chromatography, a mixture of molecules is separated
based on their differentials partitioning between a mobile phase
and a stationary phase.
Stationary phase:- is confined to a glass or plastic tube
and is mostly silica gel
Mobile phase:- (a solvent or buffer) is allowed to flow
through the solid adsorbent.
Sample:- to be analyzed is layered on top of the
column.
“2.COLUMN CHROMATOGRAPHY”
18. • Russian-italian botanist Mikhail Tsvet.
• Tsvet applied his observations with filter paper extraction to the
new methods of column fractionation for separating the
components of petroleum.
“HISTORY”
19. • The rate at which the components of a mixture are
separated depends on the
• activity of the adsorbent
• polarity of the solvent.
• If the activity of the adsorbent is very high and polarity of
the solvent is very low, then the separation is very slow but
gives a good separation. On the other hand, if the activity
of adsorbent is low and polarity of the solvent is high the
separation is rapid but gives only a poor separation,
“PRINCIPLE”
20.
21. “PROCEDURE”
• Packing the column
• Loading the column
• Eluting The column
• Collecting the Eluent
• Detection of Eluting Components
26. • Analytes larger than the
pores cannot enter the
interior of the gel beads,
so they are limited to the
space between the beads.
As a result, they are not
slowed in their progress
through the column and elute rapidly in a single zone.
• Small molecules capable of diffusing in and out of the beads have a
much larger volume available to them. Therefore, they are delayed in
their journey through the column bed. Molecules of intermediate size
migrate through the column at a rate somewhere between those for
large and small molecules.
“PRINCIPLE”
27. 1. Select columns not greater
than 100 cm in length.
2. Selection the proper gel,
which act as stationary phase
3.Layer sample on the column.
4.Add buffer to wash proteins
through the column.
5. Eluting the sample
6. At th end collect the fraction.
“HOW DOES IT WORKS?”
28. • Fractionation of molecules and complexes within a predetermined
size range
• Size analysis and determination
• Removal of large proteins and complexes
• Desalting
• Removal of small molecules
• Assessment of sample purity
• purification.
• molecular weight determination and quantitative analysis of
molecular interactions.
“APPLICATIONS”:-
31. • Allows the separation of ions and polar molecules based on the
charge properties of the molecules.
• A type of column chromatography
• Useful in the separation of charge compounds like proteins
differing by only one charged amino acid.
(ion exchanger components)
(buffered sollution)
( positive/negative)
“INTRODUCTION”:-
32. Polymers that are capable of exchanging particular ions within the polymer with ions in a
solution that is passed through them
A-polystyrene: polymerisation reaction
Of styrene and divinylbenzene, useful
For separating small molecular weight
Compounds
B-cellulose: readily obtained in a high
Pure state,greater permeability to
Macromolecules
“COMPONENT OF ION EXCHANGE”:-
33. The choice of ion exchangers depends upon the stability,
molecular weight, and ionic strength of the sample components
Volume of exchanger 2.5 fold greater than to exchange with the
ion in the sample
Anion exchangers
Retains positively charged cations
Because the stationary phase displays
A negatively charged group
Cation exchangers
Retains anions using positively charged group
“COMPONENT OF ION EXCHANGE”:-
34. Ion exchange chromatography relies on the attraction between oppositely charged stationary
phase, known as an ion exchanger, and analyze.
To these covalently bound functional groups the oppositely charged ions are bounded (mobile
counter ion), which will be exchanged with like charge ions in the sample having charge
magnitude more than the ions bounded to the matrix.
Thus if anion exchange chromatography is performed, negatively charged sample
components will interact more with the stationary phase and will be exchanged for like
charged ions already bounded to the matrix.
“PRINCIPLE”:-
35. Positively charged molecules are attracted to a negatively charged
solid support. Commonly used cation exchange resins are s-resin,
sulfate derivatives; and CM resins, carboxylate derived ions
“TYPES OF ION EXCHANGE
CHROMATOGRAPHY”:-
36. Negatively charged molecules is attracted to a
positively charged solid support. Commonly used anion
exchange resins are q-resin and DEAE resin
“TYPES OF ION EXCHANGE
CHROMATOGRAPHY”:-
37. Step1
A sample introduced into a sample
Loop of known volume
Step 2 “Set the column (equlibrium phase)”
The mobile phase carries the sample
From the loop onto a column that contains
Some form of stationary phase material
Stationary phase material is a resin or gel matrix consisting of
cellulose beads with covalently bonded charged functional groups.
“PROCEDUE”:-
38. Step3 “sample application into the column”
The target analytes (anions or cations) are retained on the stationary phase
Step4 “elution”
When we increased the concentration of a similarly charged species it will displace
the analyte ions from the stationary phase.
The analytes of interest must then be detected by some means, typically by
conductivity or uv/visible light absorbance.
Step5 “regeneration”
Cation exchanger = by acid then washed with water
Anion exchanger =by alkali then washed with water
39.
40. 1. Separating the proteins
Separates proteins according to their net charge, which is
dependent on the composition of the mobile phase(by
adjusting the ph or the ionic concentration)
Proteins are charged molecules. At specific ph, it can exist in
anionic (-), cationic (+) or zwitterion (no net charge) stage.
cationic pH =pI anionic
pH increase
“APPLICATIONS”:-
41. Hardness due to the presence of ca2+.Mg2+
Removed by passing hard water to cation exchanger changed
with na+
nar+nca2+ car + nna+
Resin hard water resin solution
Canr2 + 2nna+ nca2+ +2nanr
Resin solution resin regenerated
“APPLICATIONS”:-
42. Separation of similar ions from one another
antibody purification monoclonal antibodies
Hemoglobins seperation
Separation of vitamins
Separation of inorganic cations and anions
“APPLICATIONS”:-
43.
44. • Also known as bioselective adsorption
• It is a protein purification technique developed thirty
years ago.
• It is the isolation of a particular protein on a bioligand
that is attached to an inert matrix by a spacer arm.
• It relies on the specificity of a protein binding site for a
particular ligand
“INTRODUCTION”:-
46. • Specificity is based on three aspects:
Matix: for ligand attachment
Spacer arm: bind ligand to matrix
Ligand: bind with protein to purify it
“SPECIFICITY”:-
47. • Principle of affinity chromatography
is that the matrix combines with the
specific ligand due to specific
functional groups present on matrix .
• as the mixture of proteins is passed
through the chromatography
column, those proteins that have a
binding site for the immobilised
ligand will bind to the matrix,
while all other proteins will be
eluted from the column.
“PRINCIPLE”:-
48. •Step 1
Attachment of ligand to matrix
•Step 2
Attachment of proteins to complex
•Step 3
After washing proteins that are
unable to bind separated
•Step 4
Finally elution occurs and protein is
purified
“PROCEDURE”:-
50. Advantages
• Purify and concentrate protein
• Used in genetic engineering
• Production of vaccines
• The binding sites of biological molecules can be investigated
“ADVANTAGES”:-
Separating proteins Proteins have numerous functional groups that can have both positive and negative charges. Ion exchange chromatography separates proteins according to their net charge, which is dependent on the composition of the mobile phase. 15
Affect of pH in the separation of proteins By adjusting the pH or the ionic concentration of the mobile phase, various protein molecules can be separated. For example, if a protein has a net positive charge at pH 7, then it will bind to a column of negatively-charged beads, whereas a negatively charged protein would not. 16
.Softening of hard water
We are aware of the fact that the hardenes of water is due to the presence of Ca2+.Mg2+ and other devalent ions these ions may be removed by passing hard water to cation exchanger changed with Na+ when the following exchange reaction takes place
NaR+nCa+2 CaR+ nNa+
resin hard water resin solution
the ca+ and Mg+ ions retained in the column wherease the sodium ions passes through the solution these na ions are harmless for washing purpose .after using ion exchange for a long time it becomes inactive its activity can be revived by perculating sodium chloride solution through the reaction
CanR2+ 2nNa+ nCa2+ +2NanR
Resin solution resin regenerated
Monoclonal antibodies produced in mouse ascites can be separated from other components of the ascites fluid.
Immunoglobulins can be separated from albumin, transferrins and proteases.
Hemoglobins can be separated by high-performance cation-exchange chromatography using the Vydac strong cation-exchange column.