This presentation provides an overview of electrophoresis. It begins by defining electrophoresis as the differential migration of ionized molecules in an electric field based on their charge-to-mass ratio. It then discusses the basic principles, factors affecting mobility, supporting media like paper and gels, common techniques including low voltage and capillary electrophoresis, and applications for analyzing proteins, DNA, antibiotics and more. The presentation concludes that electrophoresis is a useful method for separating charged substances, from small ions to large molecules, and is widely used despite some limitations.
It would be use full to All Needy People. It involve information about NMR Spectroscopy ( a spectroscopic techniques), factors influencing , proton NMR and their applications of NMR as well as Nuclear magnetic imaging.
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
It would be use full to All Needy People. It involve information about NMR Spectroscopy ( a spectroscopic techniques), factors influencing , proton NMR and their applications of NMR as well as Nuclear magnetic imaging.
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
In this slide contains types, working principle, factors affecting, advantage and disadvantage of paper electrophoresis.
Presented by: G.Sai Swetha. (Department of pharmacology),
RIPER, anantapur.
The technique of paper electrophoresis is simple and inexpensive and requires only micro quantities of plasma for separation.
The support medium is a filter paper
The electrophoresis apparatus in its simplest form consists of two troughs to contain buffer solution, through which electric current is passed.
Frequently used in isolating proteins, amino acids and oligopeptides.
This presentation gives you thorough knowledge about the IR Spectroscopy. This include basic principle, type of vibrations, factors influencing vibrational frequency, instrumentation and applications of IR Spectroscopy. This is the most widely used technique for identifying unknown functional group depending on the vibrational frequency.
Ultraviolet-visible (UV-Vis) spectrophotometry is a technique used to measure light absorbance across the ultraviolet and visible ranges of the electromagnetic spectrum. When incident light strikes matter it can either be absorbed, reflected, or transmitted. The absorbance of radiation in the UV-Vis range causes atomic excitation, which refers to the transition of molecules from a low-energy ground state to an excited state.
principle, application and instrumentation of UV- visible Spectrophotometer Ayetenew Abita Desa
This Presentation powerpoint includes the principle, application, and instrumentation of UV- Visible Spectrophotometer. It covers beer-lambert low and its quantitative applications. It also includes the qualitative applications in different fields of study. Presented at Addis Ababa University, School of medicine, department of medical biochemistry.
Electrophoresis principle and types by Dr. Anurag YadavDr Anurag Yadav
the general principle on how the electrophoresis performs.
the different types of electrophoresis and the mechanism of separation based on different character of the medium and type of electrophoresis.
In this slide contains types, working principle, factors affecting, advantage and disadvantage of paper electrophoresis.
Presented by: G.Sai Swetha. (Department of pharmacology),
RIPER, anantapur.
The technique of paper electrophoresis is simple and inexpensive and requires only micro quantities of plasma for separation.
The support medium is a filter paper
The electrophoresis apparatus in its simplest form consists of two troughs to contain buffer solution, through which electric current is passed.
Frequently used in isolating proteins, amino acids and oligopeptides.
This presentation gives you thorough knowledge about the IR Spectroscopy. This include basic principle, type of vibrations, factors influencing vibrational frequency, instrumentation and applications of IR Spectroscopy. This is the most widely used technique for identifying unknown functional group depending on the vibrational frequency.
Ultraviolet-visible (UV-Vis) spectrophotometry is a technique used to measure light absorbance across the ultraviolet and visible ranges of the electromagnetic spectrum. When incident light strikes matter it can either be absorbed, reflected, or transmitted. The absorbance of radiation in the UV-Vis range causes atomic excitation, which refers to the transition of molecules from a low-energy ground state to an excited state.
principle, application and instrumentation of UV- visible Spectrophotometer Ayetenew Abita Desa
This Presentation powerpoint includes the principle, application, and instrumentation of UV- Visible Spectrophotometer. It covers beer-lambert low and its quantitative applications. It also includes the qualitative applications in different fields of study. Presented at Addis Ababa University, School of medicine, department of medical biochemistry.
Electrophoresis principle and types by Dr. Anurag YadavDr Anurag Yadav
the general principle on how the electrophoresis performs.
the different types of electrophoresis and the mechanism of separation based on different character of the medium and type of electrophoresis.
Electrophoresis:
Electrophoresis is separation technique based on movement of charge particle in an electric field.
Movement of charge particles can be determined by following formula--
V= Eq/f
Where,
V= Velocity of the charged particle;
E= electric field of the molecule;
q= Net charge of the molecule; and
f= Frictional co-efficient of the molecule
Types of electrophoresis:
1. Agarose gel electrophoresis ;
2. Poly-acryl amide gel electrophoresis [PAGE];
3. Sodium do-decyl sulphate Poly- acrylamide gel electrophoresis [SDS-PAGE] ;
4. Two dimensional –Poly-acrylamide gel electrophoresis [2D-PAGE];
5. Pulse field gel electrophoresis [PFGE];
6. Capillary gel electrophoresis [CGE]; and
7. Disc electrophoresis for Protein.
Application of electrophoresis:
1. Estimation of the DNA molecule.[ Agarose , PAGE ]
2. Analysis of PCR product. [ Agarose ]
3. Separation of restricted genomic DNA and RNA. [Agarose and PAGE respectively]
4. Conformation of newly isolated DNA .[Agarose]
5. Separation of most small fragments of DNA. [PAGE]
6. In forensic science.[Agarose , PAGE, SDS-PAGE, 2D PAGE ,Capillary gel electrophoresis , PFGE]
8. In determining molecular wt. of protein.[SDS-PAGE].etc
A Business Idea upto the Feasibility Analysis: EntrepreneurshipSultan Islam
This task we have prepared for our Entrepreneurship course, a business idea along with its feasibility analysis. The we developed is based on the sylhet region particularly to some extent.
Mobile banking is a term used for performing balance checks, account transactions, payments, credit applications, pay bills, authorize fund transfers, and other banking transactions through a mobile device such as a mobile phone or Personal Digital Assistant (PDA)
Electrophoresis is a scientific laboratory technique that is used to separate DNA, RNA, or protein molecules based on their size and electrical charge. An electric current is passed through the molecules to move them so that they can be separated via a gel. The pores present in the gel work like a sieve, allowing smaller molecules to pass through more quickly and easily than the larger molecules. According to the way conditions are adjusted during electrophoresis, the molecules can be separated in the desired size range.
What is electrophoresis and what are its uses?
Electrophoresis is a very broadly used technique that, fundamentally, applies electric current to biological molecules – they’re usually DNA, but they can be protein or RNA, too – and separates these fragments into pieces that are larger or smaller in size.
The phenomenon of electrophoresis was first observed by Russian professors Peter Ivanovich Strakhov and Ferdinand Frederic Reuss in 1807 at Moscow University. A constant application of electric field caused the particles of clay dispersed in water to migrate, showing an electrokinetic phenomenon.
Electrophoresis can be defined as an electrokinetic process that separates charged particles in a fluid using an electrical field of charge. Electrophoresis of cations or positively charged ions is sometimes referred to as cataphoresis (or cataphoretic electrophoresis). In contrast, sometimes, the electrophoresis of anions or negatively charged ions is referred to as anaphoresis (or anaphoric electrophoresis).
It’s used in a variety of applications. Though it is most often used in life sciences to separate protein molecules or DNA, it can be achieved through several different techniques and methods depending upon the type and size of the molecules.
The methods differ in some ways, but all we need is a source for the electrical charge, a support medium and a buffer solution. Electrophoresis is also used in laboratories for the separation of molecules based on their size, density and purity.
The method used to separate macromolecules such as DNA, RNA, or protein molecules is known as gel electrophoresis.
It is used in forensics for –
Nucleic acid molecule sizing
DNA fragmentation for southern blotting
RNA fragmentation for northern blotting
Protein fragmentation for western blotting
Separation of PCR products analysis
Detection and analysis of variations or mutations in the sequence
Its clinical applications involve –
Serum protein electrophoresis
Lipoprotein analysis
Diagnosis of haemoglobinopathies and hemoglobin A1c.
The fundamental principle of electrophoresis is the existence of charge separation between the surface of a particle and the fluid immediately surrounding it. An applied electric field acts on the resulting charge density, causing the particle to migrate and the fluid around the particle to flow.
It is the process of separation or purification of protein molecules, DNA, or RNA that differ in charge, size.
Capillary electrophoresis is an analytical technique that separates charged particles using electricity and a very small tube called “capillary”.
Popularized by Jorgenson and Lukacs in the late 1980’s.
Capillary electrophoresis is used most predominately because it gives faster results and provides high resolution separation.
The rate at which the particle moves is directly proportional to the applied electric field--the greater the field strength, the faster the mobility.
Neutral species are not affected, only ions move with the electric field. If two ions are the same size, the one with greater charge will move the fastest.
For ions of the same charge, the smaller particle has less friction and overall faster migration rate.
Dielectric Spectroscopy in Time and Frequency DomainGirish Gupta
This presentation describes the basics and technicalities of Dielectric Spectroscopy in both time and frequency domain. IT also includes the procedure and results involved in Dielectric Spectroscopy on different dielectrics.
Introduction to Electrophoresis and its ClassificationsRAMEG
VIDEO WITH EXPLANATION IN THE LINK BELOW 👇👇
https://www.youtube.com/watch?v=s1dF0BuW64E&t=9s
👇👇👇 (Reference Books 📚 Available below) 👇👇👇
References📚
🔑 PASSWORD - RAMEG123
📚 Clinical Biochemistry An Illustrated Colour Text, 5e by Allan Gaw https://tii.la/l8zwft4u297
📚 Clinical Chemistry Principles Techniques Correlations by Michael Bishop
https://tii.la/h7aqB52kT
📚 Naik Essentials of Biochemistry
https://tii.la/Q65
📚 Satyanarayana-biochemistry
https://tii.la/038zfK7dd5v
📚 Tietz Fundamentals of Clinical Chemistry and Molecular Diagnostic
https://tii.la/wJRgNhF
This presentation will give you idea about one of the mostly used Electrophoresis technique which is Capillary Electrophoresis. It covers one part of syllabus. It will help you to learn about the new aspects in CE and also to upgrade your knowledge.
Introduction
History
Elecrophoresis
Principle
Types of electrophoresis
Application
Conclusion
Reference
When a potential difference is applied between the two electrodes in a colloidal solution, It has been observed that the colloidal particles are carried to either the positive or negative electrode.
In other words , they behave as if they are electrically charged w.r.t. the dispersion medium. This phenomenon is known as electrophoresis.
Many important biological molecules, such as amino acids, peptides, proteins, nucleotides and nucleic acids, possess ionisable groups and, therefore, at any given pH, exist in solution as electrically charged species either as cations or anions.
Under the influence of an electric field these charged particles will migrate either to the cathode or to the anode, depending on the nature of their net charge.
A Study on Liquid Dielectric Breakdown in Micro-EDM Discharge - Cognitio paperSantosh Verma
The research work carried out in this paper, aims at
understanding the breakdown phenomenon of liquid dielectric by the low energy ultra-short pulsed electric discharge produced between tiny electrodes (~ 100µm diameter electrode) through experimental studies In literature not many studies are reported on liquid dielectric breakdown mechanism, and in micro-EDM no published literature discusses about this. Therefore, a detailed study on literature has been performed and preliminary
experiments have conducted on micro-EDM to understand the glow discharge and its breakdown phenomenon better, towards validation of scientific analogies for micro-EDM process conditions.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
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.
Richard's entangled aventures in wonderlandRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Multi-source connectivity as the driver of solar wind variability in the heli...Sérgio Sacani
The ambient solar wind that flls the heliosphere originates from multiple
sources in the solar corona and is highly structured. It is often described
as high-speed, relatively homogeneous, plasma streams from coronal
holes and slow-speed, highly variable, streams whose source regions are
under debate. A key goal of ESA/NASA’s Solar Orbiter mission is to identify
solar wind sources and understand what drives the complexity seen in the
heliosphere. By combining magnetic feld modelling and spectroscopic
techniques with high-resolution observations and measurements, we show
that the solar wind variability detected in situ by Solar Orbiter in March
2022 is driven by spatio-temporal changes in the magnetic connectivity to
multiple sources in the solar atmosphere. The magnetic feld footpoints
connected to the spacecraft moved from the boundaries of a coronal hole
to one active region (12961) and then across to another region (12957). This
is refected in the in situ measurements, which show the transition from fast
to highly Alfvénic then to slow solar wind that is disrupted by the arrival of
a coronal mass ejection. Our results describe solar wind variability at 0.5 au
but are applicable to near-Earth observatories.
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.
2. Shahjalal University of Science
&
Technology, Sylhet
Department of Chemistry
Course No. : CHE 300
Course title : Seminar and Oral
A presentation on Electrophoresis
Presented by,
Tanjila Islam
Reg. No. : 2010131019
Semester : 3/2
4. Basic of electrophoresis :
Differential rate of
migration of ion molecule
in an electrolyte solution
under the influence of an
applied electric field in a
support medium (e.g.
paper, gel or capillary
tube)
Figure 1: Motion of a charged particle by electrophoresis
*
* A useful method to separate
substances based on their
charge – to – mass ratios
5. Principle :
* Charged ion or molecule migrates when placed in an electric field
Rate of migration depends on its net charge, size, shape and the
applied electric current
v = μeE
where, v = velocity of an ion
E = electric field strength (Vcm-1)
μe = electrophoretic mobility
= distance migrated in a certain time period
The electrophoretic mobility is given by
μe =
q
6πηr
(when electric force = frictional drag)
showing that small highly charged species have high mobility and vice versa.
*
*
6. Driving force of migration :
* Resultant of the electrostatic
force of attraction between the
electric field and the charged
molecule, and the retarding
forces due to friction and
electrostatic repulsion from
molecules of the transport
medium.
Figure 2: Illustration of electrophoresis retardation
7. Supporting media for electrophoresis :
* Paper
- filter paper such as Whatman no.1 and no.3MM
- Used to good effect
* Cellulose acetate
- containing 2 to 3 acetyl groups
- to give sharper bands
- more easily rendered transparent
- low solvent capacity
- enhancing the resolution
Gels
- 3 dimensional semisolid colloids
- resolving power enhanced due to sieve effect operating
- prepared from starch, agar, or polyacrylamide
*
8. General procedure for electrophoresis :
Immersion of two electrodes in two separate
buffer chambers but not fully isolated from
each other
Migration of charged particles from one
chamber to the other by using an electric
field
Separation of different ions migrating at
different speeds Figure 3: Fundamental of electrophoresis
9. Factors affecting electrophoretic mobility :
*
*
*
Charge – higher the charge greater the mobility
Size – bigger the molecule greater the frictional and electrostatic
forces exerted on it by the medium i.e. larger particles have smaller
electrophoretic mobility compared to smaller particles
Electric field – increase of migration with the
increase of voltage gradient
* Buffer – dependence of migration on pH of the buffer
* Ionic strength – greater the ionic strength of the buffer solution higher
proportion of the current hence electrophoretic mobility
10. Types of electrophoresis :
Electrophoresis
Frontal
Electrophoresis
Micro
electrophoresis
Moving
boundary
Zone electrophoresis
Paper
electrophoresis
Cellulose acetate
electrophoresis
Gel
electrophoresis
Figure 4: Types of electrophoresis
11. Techniques of electrophoresis:
Techniques
Low voltage (LVE)
High voltage (HVE)
SDS polyacrylamaide gel (SDS-PAGE)
Isoelectric focusing
Immunoelectrophoresis
Discontinuous electrophoresis
Figure 5: Different techniques of electrophoresis
12. Low voltage electrophoresis :
* Two compartments to hold
the buffer and electrodes
Figure 6: Apparatus for low voltage electrophoresis
*A suitable carrier for support
medium
ending in contact with the buffer
medium
*To provide voltage
gradient ̴ 5 Vcm-1, a
power pack supplying up
to 500 V or even 1000 V
and 0 – 150 mA
13. Application of LVE :
*
*
*
To separate any ionic substances
The examination of biological
and clinical specimens for
amino acids and proteins
Separation of sugars
Figure 7: Electrophoretogram of plasma proteins
on cellulose acetate at pH 8.6
14. High voltage electrophoresis :
To obtain voltage gradients
up to 100 Vcm-1, high voltage
and current supplying
*
*
*
Using cooling plates for
heat dissipation generated
by high voltage
Less than of 1h analysis time
* Working best with small ions
deriving from small peptides
and amino acids
Figure 8: HVE apparatus
15. Capillary electrophoresis (CE) :
* Separation of
analyte species
achieved on the
basis of
differential
migration in an
electric field
through narrow
bore fused silica
capillary columns
(25 – 100 μm).
Capillary electrophoresis
Capillary
zone
electrophoresis
(CZE)
Capillary gel
electrophoresis
(CGE)
Isoelectric
focusing
Isotachophoresis
(ITP)
Miscellar
electrokinetic
capillary
chromatography
(MECC)
Figure 9: Separation modes of capillary electrophoresis
16. Overview of instrumentation of CE :
* A fused capillary column
dipping into two electrolyte
buffers containing Pt foil
cathode or anode across 15
– 60 kV voltage applied
* Introducing a small volume of
sample at one end of capillary
* Migration of sample through the
capillary under the force of
applied electric field
Figure 10: Schematic of a capillary zone electrophoresis
17. Advantages of CE :
Power dissipation minimized by high electrical resistance*
* Having voltage gradients up to 100 – 500 Vcm-1 necessary
for rapid separations
*
No Joule–Thompson effect
*
No band broadening
*
Most prominently used because of its faster results and
high resolution separation
*
Large range of detection methods available
18. Applications of electrophoresis :
*
* DNA analysis
Protein analysis
* Antibiotic analysis
*Vaccine analysis
*
Detection of damaged
genes by gel
electrophoresis
*
To use in forensic research Figure 11: A simple view of protein separation
19. Conclusion :
* Although not in principle a chromatographic method,
electrophoresis used in conjunction with paper chromatography
and gel materials, proves an extremely useful method for
separation of charged substances, ranging from small ions to
large charged macromolecules, of biological and biochemical
interest.
* It is widely used yet it has some limitations.