SlideShare a Scribd company logo
1 of 24
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 1
A seminar as a part of curricular requirement
for I year M. Pharm. I semester.
Presented by
G. Sai Swetha, (20L81S0103)
M. Pharmacy
Department of Pharmacology
Under the guidance of
Dr. P. Ramalingam, M.Pharm., Ph. D
Research Director & Professor of Pharmaceutical and Medicinal Chemistry
Paper Electrophoresis
03-06-2021
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 2
1. Introduction
2. Paper electrophoresis
3. Practical aspects
4. Types of paper electrophoresis
5. Working & Detection
6. Factors affecting electrophoresis
7. Advantages
8. Disadvantages
9. Applications
10. Reference
Contents
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 3
• Electrophoresis refers to the migration of charged particles of
any size in liquid medium, under the influence of an electric
field.
• Depending on the kind of charge the molecule carry, they move
either towards anode or cathode.
• An ampholyte becomes positively charged in acidic condition
and migrates to cathode and in alkaline medium, they become
negatively charged and move to anode.
Introduction
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 4
Electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 5
1. Free electrophoresis
• Capillary electrophoresis
• Isotachophoresis
• Isoelectric focusing
• Immuno electrophoresis
2. Zone electrophoresis
• Paper electrophoresis
• Cellulose acetate electrophoresis
• Gel electrophoresis
• Thin layer electrophoresis
Types of electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 6
• Paper electrophoresis (PE) is a separation technique
• Mainly used to separate amino acids and proteins, peptides.
• Separation of mixture is based on charge and size of the
molecule.
• Migration of charged particles takes place through paper under
the influence of electric field.
• The paper strip used must contain at least α-cellulose at a percent
of 95% and should have a slight adsorption capacity.
• Paper does not contain charges that interfere with the separation
of compounds.
Paper electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 7
The equipment required include:
• A power pack- provides a stabilised DC and has controls for both
voltage output (0-500V) and current output (0- 150 mA).
• An electrophoretic cell- contains the electrodes, buffer reservoir, a
support for paper and a transparent insulating cover.
• Whatman filter paper, cellulose acetate filter paper or
chromatography paper are used in PE.
• Prior to use, the paper is washed with distilled water followed by
0.1M HCl or 0.01M EDTA to remove impurities.
Practical aspects of PE
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 8
Based on voltage applied, it is of two types:
• Low voltage PE
- a voltage of 100-300 V is applied across the electrodes
- more no. of samples can be separated on a single paper, at a time
- diffused sample bands may appear
• High voltage PE
- up to 1000 V is applied across the electrodes
- sharp bands are obtained
- more voltage is required so, it is dangerous to operate
Types of paper electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 9
Based on design of instrument, it is of two types:
1. Horizontal paper electrophoresis
2. Vertical paper electrophoresis
The two arrangements of the filter strips are commonly used. Both
the arrangements are equally viable and the choice usually
depends upon personal preferences.
Types of paper electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 10
Horizontal paper electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 11
Vertical paper electrophoresis
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 12
Horizontal paper electrophoresis system Vertical paper electrophoresis system
Paper electrophoresis system
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 13
Vertical paper electrophoresis system
Paper electrophoresis system
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 14
• A paper strip of suitable length is moistened with suitable buffer of
required pH and the sample is applied transversely across the central
part of paper strip.
• Ends are fixed to dip in buffer solution in two troughs, fitted with
electrodes.
• Now electric field is applied across the system.
• The charged particles of sample migrate along the strip towards
respective electrodes of opposite polarity.
• Homogenous group of particles migrate as separate band.
• The process is carried out for 14-16 hours.
Working
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 15
Some commonly used buffers include:
Intended
Separation
Buffer pH
Proteins Barbital 8.6
Phosphate 7.4
Nucleoproteins Acetate 4.5
Citrate 4.5
Amino acids Phosphate 4.6
Michaelis 8.6
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 16
Individual compounds are identified by following methods:
1. UV absorption
-Proteins, peptides and nucleic acids absorb UV radiation at a range
of 260 to 280 nm.
2. Fluorescence
- Staining with Ethidium bromide and observing the
electrophorogram under UV light makes DNA and RNA flouresce
and fascilitates detection.
- Flourescamine staining is used for detecting amino acids,
peptides, proteins.
Detection
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 17
Compound Dye
Proteins Bromophenol blue in Acetic acid,
DANSYl Chloride
Nucleic acids Methyl-green pyronine,
Ethidium bromide
Polysaccharides Iodine
Glycoproteins Alcian blue
3. Staining
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 18
Factors affecting electrophoresis:
• Properties of supporting medium
• The strength of electric field
• Temperature of the buffer
• pH of the buffer
• Molecular mass of the sample
• Net charge on the particles
• Shape and size of the particles
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721
19
Advantages
• It is simple and inexpensive process.
• It is easily available and easy to handle.
Disadvantages
• Certain compounds like proteins, hydrophilic molecules cannot be
resolved because of adsorptive and property of paper.
• It results in tailing and distortion of component bands.
• Electro osmotic flow may occur.
• Time taking process. It takes almost 14-16 hours for separation
process.
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 20
• Serum analysis for diagnostic purpose is routinely carried out by
paper electrophoresis.
• Separation of amino acids, proteins in serum.
• Separation on enzymes in blood.
• Separation of antibiotics in different samples.
• Muscle proteins, egg white proteins, and snake, insect venom
analysis is done by this technique.
• Used in the separation and identification of alkaloids.
• It can also be used to test the suitability of municipal water supplies,
the toxicity of water, and other environmental components.
Applications
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 21
Serum analysis of proteins
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 22
Paper Electrophorogram
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 23
1. V. N. Kandula & S. Gummadi. A review on electrophoresis,
capillary electrophoresis & hyphenations. IJPSR. 2020; 11(12):
6038-6056.
2. W. P. Jencks. Paper electrophoresis as a quantitative method of
serum proteins. NCBI. 1955; 60(2): 205- 215.
3. Martin N. H. The use and limitations of filter paper electrophoresis.
Journal of Clinical Pathology. 1952; 7(2): 87- 105.
4. Myer Lubran. Paper electrophoresis. JAMA. 1996; 197(5): 360-
361.
5. J. Sternberg. Paper electrophoresis. NCBI. 1953; 68(3): 284- 285.
References
RIPER
AUTONOMOUS
NAAC &
NBA (UG)
SIRO- DSIR
Raghavendra Institute of Pharmaceutical Education and Research - Autonomous
K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 24

More Related Content

What's hot (20)

Capillary electrophoresis
Capillary electrophoresisCapillary electrophoresis
Capillary electrophoresis
 
AFFINITY CHROMATOGRAPHY
AFFINITY CHROMATOGRAPHYAFFINITY CHROMATOGRAPHY
AFFINITY CHROMATOGRAPHY
 
Techniques of electrophoresis
Techniques of electrophoresisTechniques of electrophoresis
Techniques of electrophoresis
 
Radio Immuno Assay
Radio Immuno AssayRadio Immuno Assay
Radio Immuno Assay
 
Ion exchange chromatography
Ion exchange chromatography Ion exchange chromatography
Ion exchange chromatography
 
Moving Boundary Electrophoresis by Anamika Dey
Moving Boundary Electrophoresis by Anamika DeyMoving Boundary Electrophoresis by Anamika Dey
Moving Boundary Electrophoresis by Anamika Dey
 
Electrophoresis & its types
Electrophoresis & its typesElectrophoresis & its types
Electrophoresis & its types
 
Radioimmunoassay
RadioimmunoassayRadioimmunoassay
Radioimmunoassay
 
ZONE ELECTROPHORESIS
ZONE ELECTROPHORESISZONE ELECTROPHORESIS
ZONE ELECTROPHORESIS
 
Column chromatography
Column chromatographyColumn chromatography
Column chromatography
 
Ion Exchange Chromatography ppt
Ion Exchange Chromatography pptIon Exchange Chromatography ppt
Ion Exchange Chromatography ppt
 
Radio immunoassay (RIA)
Radio immunoassay (RIA)Radio immunoassay (RIA)
Radio immunoassay (RIA)
 
Mass spectroscopy
Mass spectroscopyMass spectroscopy
Mass spectroscopy
 
Gel electrophoresis
Gel electrophoresisGel electrophoresis
Gel electrophoresis
 
TLC, thin layer chromatography
TLC, thin layer chromatographyTLC, thin layer chromatography
TLC, thin layer chromatography
 
Capillary electrophoresis
Capillary electrophoresisCapillary electrophoresis
Capillary electrophoresis
 
LCMS
LCMS LCMS
LCMS
 
UV-Visible spectroscopy
UV-Visible spectroscopyUV-Visible spectroscopy
UV-Visible spectroscopy
 
FLUORIMETRY
FLUORIMETRYFLUORIMETRY
FLUORIMETRY
 
Mass Spectroscopy
Mass SpectroscopyMass Spectroscopy
Mass Spectroscopy
 

Similar to PAPER ELECTROPHORESIS.

DETECTORS USED IN HPLC
DETECTORS USED IN HPLCDETECTORS USED IN HPLC
DETECTORS USED IN HPLCTarunk23
 

Similar to PAPER ELECTROPHORESIS. (20)

Electrophoresis Principle and its Types
Electrophoresis Principle and its Types Electrophoresis Principle and its Types
Electrophoresis Principle and its Types
 
TYPES OF ELECTROPHORESIS
TYPES OF ELECTROPHORESISTYPES OF ELECTROPHORESIS
TYPES OF ELECTROPHORESIS
 
GEL ELECTROPHORESIS
GEL ELECTROPHORESISGEL ELECTROPHORESIS
GEL ELECTROPHORESIS
 
Horizontal Electrophoresis.
Horizontal Electrophoresis.Horizontal Electrophoresis.
Horizontal Electrophoresis.
 
ANALYTICAL TECHNIQUES IN PHASE III
ANALYTICAL TECHNIQUES IN PHASE IIIANALYTICAL TECHNIQUES IN PHASE III
ANALYTICAL TECHNIQUES IN PHASE III
 
ANALYTICAL TECHNIQUES IN PHASE -III
ANALYTICAL TECHNIQUES IN PHASE -IIIANALYTICAL TECHNIQUES IN PHASE -III
ANALYTICAL TECHNIQUES IN PHASE -III
 
Vertical Gel Electrophoresis
Vertical Gel ElectrophoresisVertical Gel Electrophoresis
Vertical Gel Electrophoresis
 
Sampling Techniques in IR Spectroscopy
Sampling Techniques in IR SpectroscopySampling Techniques in IR Spectroscopy
Sampling Techniques in IR Spectroscopy
 
Ultra Performance Liquid Chromatography (UPLC).
Ultra Performance Liquid Chromatography (UPLC).Ultra Performance Liquid Chromatography (UPLC).
Ultra Performance Liquid Chromatography (UPLC).
 
JOURNAL CLUB PRESENTATION (20L81S0714-PA )
JOURNAL CLUB PRESENTATION (20L81S0714-PA )JOURNAL CLUB PRESENTATION (20L81S0714-PA )
JOURNAL CLUB PRESENTATION (20L81S0714-PA )
 
JOURNAL CLUB PRESENTATION (20L81S0707-PA)
JOURNAL CLUB PRESENTATION (20L81S0707-PA)JOURNAL CLUB PRESENTATION (20L81S0707-PA)
JOURNAL CLUB PRESENTATION (20L81S0707-PA)
 
DETECTORS USED IN HPLC
DETECTORS USED IN HPLCDETECTORS USED IN HPLC
DETECTORS USED IN HPLC
 
DETECTORS IN HPLC
DETECTORS IN HPLCDETECTORS IN HPLC
DETECTORS IN HPLC
 
JOURNAL CLUB PRESENTATION (20L81S0709-PA)
JOURNAL CLUB PRESENTATION (20L81S0709-PA)JOURNAL CLUB PRESENTATION (20L81S0709-PA)
JOURNAL CLUB PRESENTATION (20L81S0709-PA)
 
Use of XRD in Preformulations
Use of XRD in Preformulations Use of XRD in Preformulations
Use of XRD in Preformulations
 
Qualification of Weights and Measures
Qualification of Weights and MeasuresQualification of Weights and Measures
Qualification of Weights and Measures
 
Ionization techniques in lcms.ppt
Ionization techniques in lcms.pptIonization techniques in lcms.ppt
Ionization techniques in lcms.ppt
 
Qualification Of HPLC
Qualification Of HPLCQualification Of HPLC
Qualification Of HPLC
 
Interference In Atomic Absorption Spectroscopy.
Interference In Atomic Absorption Spectroscopy.Interference In Atomic Absorption Spectroscopy.
Interference In Atomic Absorption Spectroscopy.
 
JOURNAL CLUB PRESENTATION (20L81S0711-PA)
JOURNAL CLUB PRESENTATION (20L81S0711-PA)JOURNAL CLUB PRESENTATION (20L81S0711-PA)
JOURNAL CLUB PRESENTATION (20L81S0711-PA)
 

More from Raghavendra institute of pharmaceutical education and research .

More from Raghavendra institute of pharmaceutical education and research . (20)

JOURNAL CLUB PRESENTATION (20L81S0710-PA)
JOURNAL CLUB PRESENTATION (20L81S0710-PA)JOURNAL CLUB PRESENTATION (20L81S0710-PA)
JOURNAL CLUB PRESENTATION (20L81S0710-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0706-PA)
JOURNAL CLUB PRESENTATION (20L81S0706-PA)JOURNAL CLUB PRESENTATION (20L81S0706-PA)
JOURNAL CLUB PRESENTATION (20L81S0706-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0712-PA)
JOURNAL CLUB PRESENTATION (20L81S0712-PA)JOURNAL CLUB PRESENTATION (20L81S0712-PA)
JOURNAL CLUB PRESENTATION (20L81S0712-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0713-PA)
JOURNAL CLUB PRESENTATION (20L81S0713-PA)JOURNAL CLUB PRESENTATION (20L81S0713-PA)
JOURNAL CLUB PRESENTATION (20L81S0713-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0702-PA)
JOURNAL CLUB PRESENTATION (20L81S0702-PA)JOURNAL CLUB PRESENTATION (20L81S0702-PA)
JOURNAL CLUB PRESENTATION (20L81S0702-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0705-PA)
JOURNAL CLUB PRESENTATION (20L81S0705-PA)JOURNAL CLUB PRESENTATION (20L81S0705-PA)
JOURNAL CLUB PRESENTATION (20L81S0705-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0402-PA & QA)
JOURNAL CLUB PRESENTATION (20L81S0402-PA & QA)JOURNAL CLUB PRESENTATION (20L81S0402-PA & QA)
JOURNAL CLUB PRESENTATION (20L81S0402-PA & QA)
 
JOURNAL CLUB PRESENTATION (20L81S0401-PA & QA)
JOURNAL CLUB PRESENTATION (20L81S0401-PA & QA)JOURNAL CLUB PRESENTATION (20L81S0401-PA & QA)
JOURNAL CLUB PRESENTATION (20L81S0401-PA & QA)
 
JOURNAL CLUB PRESENTATION (20L81S0703-PA)
JOURNAL CLUB PRESENTATION (20L81S0703-PA)JOURNAL CLUB PRESENTATION (20L81S0703-PA)
JOURNAL CLUB PRESENTATION (20L81S0703-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0701-PA)
JOURNAL CLUB PRESENTATION (20L81S0701-PA)JOURNAL CLUB PRESENTATION (20L81S0701-PA)
JOURNAL CLUB PRESENTATION (20L81S0701-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0704-PA )
JOURNAL CLUB PRESENTATION (20L81S0704-PA )JOURNAL CLUB PRESENTATION (20L81S0704-PA )
JOURNAL CLUB PRESENTATION (20L81S0704-PA )
 
JOURNAL CLUB PRESENTATION (20L81S0708-PA)
JOURNAL CLUB PRESENTATION (20L81S0708-PA)JOURNAL CLUB PRESENTATION (20L81S0708-PA)
JOURNAL CLUB PRESENTATION (20L81S0708-PA)
 
JOURNAL CLUB PRESENTATION (20L81S0403-PA & QA)
JOURNAL CLUB PRESENTATION (20L81S0403-PA & QA)JOURNAL CLUB PRESENTATION (20L81S0403-PA & QA)
JOURNAL CLUB PRESENTATION (20L81S0403-PA & QA)
 
Validation master plan
Validation master planValidation master plan
Validation master plan
 
Physical Evaluation of Crude Drugs
Physical Evaluation of Crude Drugs Physical Evaluation of Crude Drugs
Physical Evaluation of Crude Drugs
 
QUALIFICATION OF FT-IR
QUALIFICATION OF FT-IRQUALIFICATION OF FT-IR
QUALIFICATION OF FT-IR
 
Qualification Of HPTLC
Qualification Of HPTLCQualification Of HPTLC
Qualification Of HPTLC
 
Hair products
Hair productsHair products
Hair products
 
Study of Quality of Raw Materials and General methods of analysis of Raw mate...
Study of Quality of Raw Materials and General methods of analysis of Raw mate...Study of Quality of Raw Materials and General methods of analysis of Raw mate...
Study of Quality of Raw Materials and General methods of analysis of Raw mate...
 
Determination of Acid value, Saponification value and Ester value
Determination of Acid value, Saponification value and Ester value Determination of Acid value, Saponification value and Ester value
Determination of Acid value, Saponification value and Ester value
 

Recently uploaded

Topic 9- General Principles of International Law.pptx
Topic 9- General Principles of International Law.pptxTopic 9- General Principles of International Law.pptx
Topic 9- General Principles of International Law.pptxJorenAcuavera1
 
Biological classification of plants with detail
Biological classification of plants with detailBiological classification of plants with detail
Biological classification of plants with detailhaiderbaloch3
 
CHROMATOGRAPHY PALLAVI RAWAT.pptx
CHROMATOGRAPHY  PALLAVI RAWAT.pptxCHROMATOGRAPHY  PALLAVI RAWAT.pptx
CHROMATOGRAPHY PALLAVI RAWAT.pptxpallavirawat456
 
Dubai Calls Girl Lisa O525547819 Lexi Call Girls In Dubai
Dubai Calls Girl Lisa O525547819 Lexi Call Girls In DubaiDubai Calls Girl Lisa O525547819 Lexi Call Girls In Dubai
Dubai Calls Girl Lisa O525547819 Lexi Call Girls In Dubaikojalkojal131
 
GenAI talk for Young at Wageningen University & Research (WUR) March 2024
GenAI talk for Young at Wageningen University & Research (WUR) March 2024GenAI talk for Young at Wageningen University & Research (WUR) March 2024
GenAI talk for Young at Wageningen University & Research (WUR) March 2024Jene van der Heide
 
Harmful and Useful Microorganisms Presentation
Harmful and Useful Microorganisms PresentationHarmful and Useful Microorganisms Presentation
Harmful and Useful Microorganisms Presentationtahreemzahra82
 
Base editing, prime editing, Cas13 & RNA editing and organelle base editing
Base editing, prime editing, Cas13 & RNA editing and organelle base editingBase editing, prime editing, Cas13 & RNA editing and organelle base editing
Base editing, prime editing, Cas13 & RNA editing and organelle base editingNetHelix
 
User Guide: Pulsar™ Weather Station (Columbia Weather Systems)
User Guide: Pulsar™ Weather Station (Columbia Weather Systems)User Guide: Pulsar™ Weather Station (Columbia Weather Systems)
User Guide: Pulsar™ Weather Station (Columbia Weather Systems)Columbia Weather Systems
 
Organic farming with special reference to vermiculture
Organic farming with special reference to vermicultureOrganic farming with special reference to vermiculture
Organic farming with special reference to vermicultureTakeleZike1
 
Ai in communication electronicss[1].pptx
Ai in communication electronicss[1].pptxAi in communication electronicss[1].pptx
Ai in communication electronicss[1].pptxsubscribeus100
 
Microteaching on terms used in filtration .Pharmaceutical Engineering
Microteaching on terms used in filtration .Pharmaceutical EngineeringMicroteaching on terms used in filtration .Pharmaceutical Engineering
Microteaching on terms used in filtration .Pharmaceutical EngineeringPrajakta Shinde
 
Pests of soyabean_Binomics_IdentificationDr.UPR.pdf
Pests of soyabean_Binomics_IdentificationDr.UPR.pdfPests of soyabean_Binomics_IdentificationDr.UPR.pdf
Pests of soyabean_Binomics_IdentificationDr.UPR.pdfPirithiRaju
 
Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...
Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...
Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...D. B. S. College Kanpur
 
Environmental Biotechnology Topic:- Microbial Biosensor
Environmental Biotechnology Topic:- Microbial BiosensorEnvironmental Biotechnology Topic:- Microbial Biosensor
Environmental Biotechnology Topic:- Microbial Biosensorsonawaneprad
 
User Guide: Orion™ Weather Station (Columbia Weather Systems)
User Guide: Orion™ Weather Station (Columbia Weather Systems)User Guide: Orion™ Weather Station (Columbia Weather Systems)
User Guide: Orion™ Weather Station (Columbia Weather Systems)Columbia Weather Systems
 
REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...
REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...
REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...Universidade Federal de Sergipe - UFS
 
trihybrid cross , test cross chi squares
trihybrid cross , test cross chi squarestrihybrid cross , test cross chi squares
trihybrid cross , test cross chi squaresusmanzain586
 
Pests of Blackgram, greengram, cowpea_Dr.UPR.pdf
Pests of Blackgram, greengram, cowpea_Dr.UPR.pdfPests of Blackgram, greengram, cowpea_Dr.UPR.pdf
Pests of Blackgram, greengram, cowpea_Dr.UPR.pdfPirithiRaju
 

Recently uploaded (20)

Topic 9- General Principles of International Law.pptx
Topic 9- General Principles of International Law.pptxTopic 9- General Principles of International Law.pptx
Topic 9- General Principles of International Law.pptx
 
Biological classification of plants with detail
Biological classification of plants with detailBiological classification of plants with detail
Biological classification of plants with detail
 
CHROMATOGRAPHY PALLAVI RAWAT.pptx
CHROMATOGRAPHY  PALLAVI RAWAT.pptxCHROMATOGRAPHY  PALLAVI RAWAT.pptx
CHROMATOGRAPHY PALLAVI RAWAT.pptx
 
Dubai Calls Girl Lisa O525547819 Lexi Call Girls In Dubai
Dubai Calls Girl Lisa O525547819 Lexi Call Girls In DubaiDubai Calls Girl Lisa O525547819 Lexi Call Girls In Dubai
Dubai Calls Girl Lisa O525547819 Lexi Call Girls In Dubai
 
GenAI talk for Young at Wageningen University & Research (WUR) March 2024
GenAI talk for Young at Wageningen University & Research (WUR) March 2024GenAI talk for Young at Wageningen University & Research (WUR) March 2024
GenAI talk for Young at Wageningen University & Research (WUR) March 2024
 
Harmful and Useful Microorganisms Presentation
Harmful and Useful Microorganisms PresentationHarmful and Useful Microorganisms Presentation
Harmful and Useful Microorganisms Presentation
 
Base editing, prime editing, Cas13 & RNA editing and organelle base editing
Base editing, prime editing, Cas13 & RNA editing and organelle base editingBase editing, prime editing, Cas13 & RNA editing and organelle base editing
Base editing, prime editing, Cas13 & RNA editing and organelle base editing
 
User Guide: Pulsar™ Weather Station (Columbia Weather Systems)
User Guide: Pulsar™ Weather Station (Columbia Weather Systems)User Guide: Pulsar™ Weather Station (Columbia Weather Systems)
User Guide: Pulsar™ Weather Station (Columbia Weather Systems)
 
Organic farming with special reference to vermiculture
Organic farming with special reference to vermicultureOrganic farming with special reference to vermiculture
Organic farming with special reference to vermiculture
 
Ai in communication electronicss[1].pptx
Ai in communication electronicss[1].pptxAi in communication electronicss[1].pptx
Ai in communication electronicss[1].pptx
 
Microteaching on terms used in filtration .Pharmaceutical Engineering
Microteaching on terms used in filtration .Pharmaceutical EngineeringMicroteaching on terms used in filtration .Pharmaceutical Engineering
Microteaching on terms used in filtration .Pharmaceutical Engineering
 
Pests of soyabean_Binomics_IdentificationDr.UPR.pdf
Pests of soyabean_Binomics_IdentificationDr.UPR.pdfPests of soyabean_Binomics_IdentificationDr.UPR.pdf
Pests of soyabean_Binomics_IdentificationDr.UPR.pdf
 
AZOTOBACTER AS BIOFERILIZER.PPTX
AZOTOBACTER AS BIOFERILIZER.PPTXAZOTOBACTER AS BIOFERILIZER.PPTX
AZOTOBACTER AS BIOFERILIZER.PPTX
 
Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...
Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...
Fertilization: Sperm and the egg—collectively called the gametes—fuse togethe...
 
Environmental Biotechnology Topic:- Microbial Biosensor
Environmental Biotechnology Topic:- Microbial BiosensorEnvironmental Biotechnology Topic:- Microbial Biosensor
Environmental Biotechnology Topic:- Microbial Biosensor
 
User Guide: Orion™ Weather Station (Columbia Weather Systems)
User Guide: Orion™ Weather Station (Columbia Weather Systems)User Guide: Orion™ Weather Station (Columbia Weather Systems)
User Guide: Orion™ Weather Station (Columbia Weather Systems)
 
Let’s Say Someone Did Drop the Bomb. Then What?
Let’s Say Someone Did Drop the Bomb. Then What?Let’s Say Someone Did Drop the Bomb. Then What?
Let’s Say Someone Did Drop the Bomb. Then What?
 
REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...
REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...
REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 - Artigo_Bioterra_V24_...
 
trihybrid cross , test cross chi squares
trihybrid cross , test cross chi squarestrihybrid cross , test cross chi squares
trihybrid cross , test cross chi squares
 
Pests of Blackgram, greengram, cowpea_Dr.UPR.pdf
Pests of Blackgram, greengram, cowpea_Dr.UPR.pdfPests of Blackgram, greengram, cowpea_Dr.UPR.pdf
Pests of Blackgram, greengram, cowpea_Dr.UPR.pdf
 

PAPER ELECTROPHORESIS.

  • 1. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 1 A seminar as a part of curricular requirement for I year M. Pharm. I semester. Presented by G. Sai Swetha, (20L81S0103) M. Pharmacy Department of Pharmacology Under the guidance of Dr. P. Ramalingam, M.Pharm., Ph. D Research Director & Professor of Pharmaceutical and Medicinal Chemistry Paper Electrophoresis 03-06-2021
  • 2. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 2 1. Introduction 2. Paper electrophoresis 3. Practical aspects 4. Types of paper electrophoresis 5. Working & Detection 6. Factors affecting electrophoresis 7. Advantages 8. Disadvantages 9. Applications 10. Reference Contents
  • 3. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 3 • Electrophoresis refers to the migration of charged particles of any size in liquid medium, under the influence of an electric field. • Depending on the kind of charge the molecule carry, they move either towards anode or cathode. • An ampholyte becomes positively charged in acidic condition and migrates to cathode and in alkaline medium, they become negatively charged and move to anode. Introduction
  • 4. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 4 Electrophoresis
  • 5. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 5 1. Free electrophoresis • Capillary electrophoresis • Isotachophoresis • Isoelectric focusing • Immuno electrophoresis 2. Zone electrophoresis • Paper electrophoresis • Cellulose acetate electrophoresis • Gel electrophoresis • Thin layer electrophoresis Types of electrophoresis
  • 6. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 6 • Paper electrophoresis (PE) is a separation technique • Mainly used to separate amino acids and proteins, peptides. • Separation of mixture is based on charge and size of the molecule. • Migration of charged particles takes place through paper under the influence of electric field. • The paper strip used must contain at least α-cellulose at a percent of 95% and should have a slight adsorption capacity. • Paper does not contain charges that interfere with the separation of compounds. Paper electrophoresis
  • 7. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 7 The equipment required include: • A power pack- provides a stabilised DC and has controls for both voltage output (0-500V) and current output (0- 150 mA). • An electrophoretic cell- contains the electrodes, buffer reservoir, a support for paper and a transparent insulating cover. • Whatman filter paper, cellulose acetate filter paper or chromatography paper are used in PE. • Prior to use, the paper is washed with distilled water followed by 0.1M HCl or 0.01M EDTA to remove impurities. Practical aspects of PE
  • 8. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 8 Based on voltage applied, it is of two types: • Low voltage PE - a voltage of 100-300 V is applied across the electrodes - more no. of samples can be separated on a single paper, at a time - diffused sample bands may appear • High voltage PE - up to 1000 V is applied across the electrodes - sharp bands are obtained - more voltage is required so, it is dangerous to operate Types of paper electrophoresis
  • 9. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 9 Based on design of instrument, it is of two types: 1. Horizontal paper electrophoresis 2. Vertical paper electrophoresis The two arrangements of the filter strips are commonly used. Both the arrangements are equally viable and the choice usually depends upon personal preferences. Types of paper electrophoresis
  • 10. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 10 Horizontal paper electrophoresis
  • 11. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 11 Vertical paper electrophoresis
  • 12. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 12 Horizontal paper electrophoresis system Vertical paper electrophoresis system Paper electrophoresis system
  • 13. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 13 Vertical paper electrophoresis system Paper electrophoresis system
  • 14. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 14 • A paper strip of suitable length is moistened with suitable buffer of required pH and the sample is applied transversely across the central part of paper strip. • Ends are fixed to dip in buffer solution in two troughs, fitted with electrodes. • Now electric field is applied across the system. • The charged particles of sample migrate along the strip towards respective electrodes of opposite polarity. • Homogenous group of particles migrate as separate band. • The process is carried out for 14-16 hours. Working
  • 15. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 15 Some commonly used buffers include: Intended Separation Buffer pH Proteins Barbital 8.6 Phosphate 7.4 Nucleoproteins Acetate 4.5 Citrate 4.5 Amino acids Phosphate 4.6 Michaelis 8.6
  • 16. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 16 Individual compounds are identified by following methods: 1. UV absorption -Proteins, peptides and nucleic acids absorb UV radiation at a range of 260 to 280 nm. 2. Fluorescence - Staining with Ethidium bromide and observing the electrophorogram under UV light makes DNA and RNA flouresce and fascilitates detection. - Flourescamine staining is used for detecting amino acids, peptides, proteins. Detection
  • 17. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 17 Compound Dye Proteins Bromophenol blue in Acetic acid, DANSYl Chloride Nucleic acids Methyl-green pyronine, Ethidium bromide Polysaccharides Iodine Glycoproteins Alcian blue 3. Staining
  • 18. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 18 Factors affecting electrophoresis: • Properties of supporting medium • The strength of electric field • Temperature of the buffer • pH of the buffer • Molecular mass of the sample • Net charge on the particles • Shape and size of the particles
  • 19. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 19 Advantages • It is simple and inexpensive process. • It is easily available and easy to handle. Disadvantages • Certain compounds like proteins, hydrophilic molecules cannot be resolved because of adsorptive and property of paper. • It results in tailing and distortion of component bands. • Electro osmotic flow may occur. • Time taking process. It takes almost 14-16 hours for separation process.
  • 20. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 20 • Serum analysis for diagnostic purpose is routinely carried out by paper electrophoresis. • Separation of amino acids, proteins in serum. • Separation on enzymes in blood. • Separation of antibiotics in different samples. • Muscle proteins, egg white proteins, and snake, insect venom analysis is done by this technique. • Used in the separation and identification of alkaloids. • It can also be used to test the suitability of municipal water supplies, the toxicity of water, and other environmental components. Applications
  • 21. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 21 Serum analysis of proteins
  • 22. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 22 Paper Electrophorogram
  • 23. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 23 1. V. N. Kandula & S. Gummadi. A review on electrophoresis, capillary electrophoresis & hyphenations. IJPSR. 2020; 11(12): 6038-6056. 2. W. P. Jencks. Paper electrophoresis as a quantitative method of serum proteins. NCBI. 1955; 60(2): 205- 215. 3. Martin N. H. The use and limitations of filter paper electrophoresis. Journal of Clinical Pathology. 1952; 7(2): 87- 105. 4. Myer Lubran. Paper electrophoresis. JAMA. 1996; 197(5): 360- 361. 5. J. Sternberg. Paper electrophoresis. NCBI. 1953; 68(3): 284- 285. References
  • 24. RIPER AUTONOMOUS NAAC & NBA (UG) SIRO- DSIR Raghavendra Institute of Pharmaceutical Education and Research - Autonomous K.R.Palli Cross, Chiyyedu, Anantapuramu, A. P- 515721 24