This document provides information on ion exchange chromatography. It begins by describing Michael Tswett who first separated plant pigments using chromatography in 1906. It then discusses how chromatography can be used to separate mixtures with similar properties. The document outlines the basic principles of ion exchange chromatography, including that it relies on reversible exchange between ions in solution and ions bound to an insoluble stationary phase. It provides details on cation and anion exchangers, preparation of ion exchangers, choice of buffers, applications, advantages and disadvantages of ion exchange chromatography.
Ion exchange chromatography -SlideShareRIZWAN RIZWI
This ppt provide a good knowledge about ion exchange chromatography. I think this is very helpful for you .Here i have tried to explain a best way and simple method so guys you all enjoy this and gain your knowledge. And wish for me to provide more pptx for you all .at the end i want your experience give me suggestion if i made any mistake thank you .
A presentation on column efficiency parameters in chromatography.. A part of gas chromatography in pharmacutical analysis..will be helpful for all mphrm students
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
various parts of mAss spectroscopy, applications, principle, peaks, rules, typical mass spectra, various combinations, Fragmentation, rules of fragmentation and useful points which can help Chemical and analytical students and structural elucidation.
Ion exchange chromatography -SlideShareRIZWAN RIZWI
This ppt provide a good knowledge about ion exchange chromatography. I think this is very helpful for you .Here i have tried to explain a best way and simple method so guys you all enjoy this and gain your knowledge. And wish for me to provide more pptx for you all .at the end i want your experience give me suggestion if i made any mistake thank you .
A presentation on column efficiency parameters in chromatography.. A part of gas chromatography in pharmacutical analysis..will be helpful for all mphrm students
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
various parts of mAss spectroscopy, applications, principle, peaks, rules, typical mass spectra, various combinations, Fragmentation, rules of fragmentation and useful points which can help Chemical and analytical students and structural elucidation.
this slide contains all the basic about the topic ion exchange chromatography which contains all important information about topic in very easy language. it will be helpful for BSc, pharmacy and biomedical student.
INSTRUMENTAL METHODS OF ANALYSIS, B.PHARM 7TH SEM. AND FOR BSC,MSC CHEMISTRY. This is Geeta prasad kashyap (Asst. Professor), SVITS, Bilaspur (C.G) 495001
This is a type of chromatography in which similar charged ions are separated by using ion exchange resin, that exchanges ions according to their relative affinities.
Ion exchange chromatography may be defined as a reversible reaction in which free mobile ions of a solids called ion exchange are exchanged for different ions of similar charge present in solution.....................................................................
Ion pair chromatography for pharmacy studentsabhishek rai
Ion-PairChromatography
A GENERALISED OVERVIEW
Chromatography
HPLC
Reverse Phase Chromatography
Ion Pair Chromatography
Ion Pair Reagent
Mechanism of Ion Pair Chromatography
Ion Pair Wash Procedure
Principles of Ion -exchange chromatography, High performance liquid chromatography (HPLC) , chromatography generally stands for a technique which separates mixtures based on different dynamic sharing of their components between two distinct physio-chemical environments called mobile and stationary phase by repeated absorption/desorption steps. Ion chromatography (IC) is a member of large family of liquid phase
chromatographic methods (that is a mobile phase is a liquid and a stationary phase is a
solid).
Ion exchange chromatography works under the principle of reversible adsorption and this method involves the separation of ions by using different types of exchange resins based on the ions to be separated.
Theoretical background
Cont’d
Ion exchangers
There are three classes of ion exchangers , these include
Resins
Gels
Inorganic exchangers
Selectivity for ion exchange
In general , ion exchangers favour the binding of ions of
Higher charge
Decreased hydrated radius
Increased polarizability
Ion exchange resins are used for the separation of small molecules.
Ion exchange gels are used for the separation of large molecules like protiens ,nucleic acids.
Separations involving harsh chemical conditions(high temperature , high radiation levels, strongly basic solutions or powerful oxidizing agents) employ inorganic ion exchangers
Advantages
Detectability: useful for the detection of many in-organic salts and organic ions with poor uv absorptivity like alkyl amines or sulfonates.
Preparative separations: usually preferred because of the availability of volatile buffers . volatile buffers makes the removal of mobile phase easier.
Useful to resolve very complex samples, i.e in the case of multi step separation
Useful for separation of mixtures of biological origin, in organic salts and some organo- metallics
Applications
Conversion from one salt to other e.g we can prepare tetra propyl ammonium hydroxide from a tetra propyl salt of some other anion.
household (laundry detergents and water filters) to produce soft water
Ion exchange is used to prepare de-ionized water
separate and purify metals
Dealkalization
analysis and purification of immunoglobulins
Separation of inorganic ions
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
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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.
Richard's aventures in two entangled wonderlandsRichard 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.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
2. • The first detailed description of chromatography is generally
credited to Michael Tswett, a Russian biochemist, who separated
chlorophyll from a mixture of plant pigments in 1906.
• Because of the nature of the pigments in the sample, each band had
a distinctive color.
• Thus the name of the process was coined from the Greek words for
color (chromo) and to write (graphy).
• If the individual components of a mixture have widely dissimilar
physical and chemical properties, it is very easy to separate one
from another.
CHROMATOGRAPHY
3. • But as the individual components of a mixture get more and more
similar in physical and chemical properties, it becomes increasingly
difficult to separate them from one another.
• However it can be readily achieved using chromatography.
• The feature common to them all is that two mutually immiscible
phases are brought into contact with each other.
• One of these phases is stationary, while the other is mobile.
• The mobile phase either moves over the surface or percolates
through the interstices of the stationary phase.
4. PARTITION
COEFFICIENT• Partition Coefficient (also known as distribution coefficient) is a
definitive term normally used to describe the way in which a given
compound distributes or partitions itself between two immiscible
phases, the stationary and the mobile phase.
K = Cs
Cm
where Cs and Cm are the concentrations of the compound in the
stationary and the mobile phases respectively.
• This concept of Partition Coefficient is the basic principle of all
chromatographic methods.
5. TECHNIQUES OF
CHROMATOGRAPHY
• There are two basic techniques of chromatography : plane
chromatography and column chromatography.
PLANE CHROMATOGRAPHY
• There are two variations of plane chromatography: paper
chromatography and thin layer chromatography.
• In paper chromatography the stationary phase is supported by
cellulose fibres of the paper sheet.
• In thin layer chromatography the stationary phase is coated onto a
glass or plastic surface.
6. COLUMN CHROMATOGRAPHY
• In column chromatography the stationary phase is packed into a
glass or metal column.
• The stationary phase is either coated onto discrete small particles
(the matrix) and packed into the column or applied as a thin film to
the inside wall of the column.
• As the eluent flows through the column the analytes separate on the
basis of their distribution coefficients and emerge individually in
the eluate as it leaves the column.
9. INTRODUCTION
• This technique has been developed since 19th century which was
firstly used for purifying the drinking water.
• Ion exchange chromatography is a distinct principle of
chromatography performed in the column.
• Ion exchange chromatography may be defined as the reversible
exchange of ions in the solution with ions electrostatically bound to
some sort of insoluble matrix or a stationary phase.
• This technique is extremely useful in the separation of charge
compound like proteins differing by only one charged amino acid.
11. PRINCIPLE
• Ion exchange chromatography relies on the attraction between
oppositely charged stationary phase, known as an ion exchanger and
analyte.
• It is frequently chosen for the separation and purification of
proteins, peptides, nucleic acids, polynucleotides and other charged
molecules, mainly because of its high resolving power and high
capacity.
• There are two types of ion exchanger, namely cation and anion
exchangers.
• Cation exchangers possess negatively charged groups and these will
attract positively charged cations. These exchangers are also called
acidic ion exchangers.
12. PRINCIPLEContinued…
• Anion exchangers have positively charged groups that will attract
negatively charged anions. The term basic ion exchangers is also
used to describe these exchangers.
• The ion exchanger consists of an inert support medium coupled to
positive (anion exchanger) or negative (cation exchanger)
functional groups.
• 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.
13. WORKING
• Consider a column having E- Y+ cation exchanger in which E- is
negative charged exchanger and Y+ is the mobile counter ion.
• Let X+ be the cation in the sample having charge greater than Y+ .
• The X+ ion can exchange sites with the counter ion Y+ with
satisfying the following relationship :
E- Y+ + X+ E- X + + Y+
15. Continued…
• Bounded interest of ion (X +) can now be eluted by either of the two
ways;
1. By adding the component Y+ having magnitude of charge more
than that of X + so that Y+ will replace X + and X + will be eluting
out.
2. By changing pH of the solvent (mobile phase) so that X + have no
charge and is then unbounded from the matrix and can be eluted
out.
17. ION EXCHANGERS
• An ion exchanger consists of two basic components :
1. Ion exchange resin
2. Exchange medium (Cation exchanger and Anion exchanger)
- Charged functional group.
- Mobile counter ion.
18. ION EXCHANGE RESIN
Two main groups of materials are used to prepare ion exchange
resins :
• Polystyrene
• Cellulose
Resin made from both of these materials differ in their flow
properties, ion accessibility and chemical and mechanical stability.
Selection of one or the other type of resin is done on the basis of
compounds being separated.
19. 1. POLYSTYRENE
• Polystyrene resins are prepared by polymerisation reaction of
styrene and divinyl benzene.
• Higher concentrations of divinyl benzene produces higher cross
linkages.
• Polystyrene resins are very useful for separating small molecular
weight compounds, however, unsatisfactory for the separation of
macromolecules.
20. 2. CELLULOSE
• Cellulose is a high molecular weight compound which can
be readily obtained in a high pure state.
• Cellulose has much greater permeability to
macromolecules.
21. EXCHANGE MEDIUM
• The choice of ion exchangers depends upon the stability, molecular
weight and ionic strength of the sample components.
• The ion exchanger are packed in column having suitable buffer.
• The ion exchangers are of two types :
1. Anion Exchangers
2. Cation Exchangers
22. 1. Anion Exchangers
• The anion exchangers have positively charged exchanger with
negatively charged mobile counter ion available for exchange.
• If the basic functional groups are introduced, the resin becomes
anion exchanger.
• Tertiary amines Strong anion exchangers
• Secondary amines Weak anion exchangers
23. Examples : Anion Exchangers
Source : Wilson K & Walker J (2010). Principles and Techniques of Biochemistry and
Molecular biology.
24. 2. Cation Exchangers
• The cation exchangers have negatively charged exchanger with
positively charged mobile counter ion available for exchange.
• If acidic functional group are introduced, then the resin becomes
cation exchangers.
• Sulphonic acid Strong cation exchangers
• Carboxylic acid Weak cation exchangers
25. Examples : Cation Exchangers
Source : Wilson K & Walker J (2010). Principles and Techniques of Biochemistry and
Molecular biology.
26. PREPARATION OF ION
EXCHANGERS
• Preparation of the exchange medium is essential for satisfactory
performance of ion exchange chromatography.
• Apart from removing impurities, there are three major steps that are
absolutely important in ion exchanger preparation.
1. SWELLING: For anion exchangers it is done by treating it first
with an acid (0.5N HCl) and then with base (0.5N NaOH).
Reverse is true for preparation of cation exchangers.
27. 2. REMOVAL OF FINE PARTICLES: Large number of such
particle can result in decreased flow rate and improper resolution.
For this the exchangers are repeatedly suspended in large volume of
water.
3. ADDITION OF COUNTER IONS: Accomplished by washing
the exchanger with suitable reagent depending upon which counter
ion to be introduced.
For e.g : NaOH (Na+), HCl (H+) etc…
28. CHOICE OF BUFFER
• The choice of buffers which maintain the pH of the column is
dictated by the compounds to be separated and the type of ion
exchange being carried out (anionic or cationic).
• Anion exchange chromatography should be carried out with
cationic buffers. Reverse is true for cation exchange
chromatography.
• If anion exchange or cation exchange is carried out with anionic or
cationic buffers respectively, the buffer ions will indulge in ion
exchange and hamper sample component exchange.
• The pH of the buffer should impart the same charge to the sample
ions as is present on the counterion.
29. Some Volatile Buffers Used in Ion-Exchange
Chromatography
Avinash Upadhyay - Biophysical Chemistry-Principles & Techniques
30. APPLICATIONS
• Softening of hard water.
• Demineralization of water.
• To analyze base composition of nucleic acid.
• To concentrate the metal ions in the sample.
• To measure the additives in food and drug sample.
• To separate protein mixtures.
31. ADVANTAGES
• Detectability: useful for the detection of many inorganic salts and
also for the detection of organic ions with poor UV absorptivity
like alkyl amines or sulfonates.
• Preparative separations: usually preferred because of the
availability of volatile buffers. Volatile buffers makes the removal
of mobile phase easier.
• Useful to resolve very complex samples, i.e in the case of multi
step separation.
• Useful for separation of mixtures of biological origin, in organic
salts and some organo – metallics.
33. DISADVANTAGES
• Column efficiency is less.
• It is difficult to achieve control over selectivity and resolution.
• Stability and reproducibility of the columns become questionable
after repeated use.