This document provides information about radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) techniques. It discusses the principles, requirements, methodology, and applications of RIA. Key steps in RIA include radio label production, conjugate preparation, antibody production and characterization, and separation techniques. RIA has applications in pharmaceutical analysis and pharmacokinetic studies. ELISA can be used to detect antigens or antibodies and has advantages of sensitivity and accurate measurement of low analyte levels. Both techniques have widespread uses in fields like immunoassay, drug analysis, and HIV testing.
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.
In this slide contains principle, instrumentation, methodology, and application of gel chromatography.
Presented by: SATHEES CHANDRA (Department of pharmaceutical analysis).
RIPER, anantapur
This document describes detailed information about Radio immuno assay (RIA) including its principle, procedure, advantages, disadvantages, application etc
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.
In this slide contains principle, instrumentation, methodology, and application of gel chromatography.
Presented by: SATHEES CHANDRA (Department of pharmaceutical analysis).
RIPER, anantapur
This document describes detailed information about Radio immuno assay (RIA) including its principle, procedure, advantages, disadvantages, application etc
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
Introduction
working principle
fragmentation process
general rules for fragmentation
general modes of fragmentation
metastable ions
isotopic peaks
applications
The following presentation contains helpful information regarding Radioimmunoassay (RIA) and Enzyme-Linked Immunosorbent Assay (ELISA), including their history, introduction, advantages, procedures and applications.
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
Introduction
working principle
fragmentation process
general rules for fragmentation
general modes of fragmentation
metastable ions
isotopic peaks
applications
The following presentation contains helpful information regarding Radioimmunoassay (RIA) and Enzyme-Linked Immunosorbent Assay (ELISA), including their history, introduction, advantages, procedures and applications.
Enzyme linked immunosorbent assay (elisa) and its clinical significancerohini sane
A comprehensive presentation on Enzyme Linked Immunosorbent Assay (ELISA) and its clinical significance for MBBS, BDS, B Pharm & Biotechnology students to facilitate self- study.
ELISA use an enzyme to detect the binding of antigen (Ag) antibody (Ab). • The enzyme converts a colorless substrate (chromogen) to a colored product, indicating the presence of Ag:Ab binding. • An ELISA can be used to detect either the presence of antigens or antibodies in a sample depending how the test is designed
immunoassay of digoxin.pptx (analytical methods of immunoassay)NikitaBankoti2
Immunoassays are bioanalytical methods in which the quantitation of the analyte depends on the reaction of an antigen (analyte) and an antibody.
Principle - Immuno assay methods are based on a competitive binding between a fixed amount of labelled form of an analyte and available amount of unlabeled sample analyte for a limited amount of binding sites on a highly specific anti-analyte antibody.
002. Cephalosporins for students 2023 Prof. P. Ravisankar.pdfDr. Ravi Sankar
Cephalosporins, Why Cephalosporins? Advantages of cephalosporins over penicillin, Mechanism of action of cephalosporins, Classification of cephalosporins, Structures of some important cephalosporins and cephamycins, Oximinocephalosporins, SAR of cephalosporins,Hydrolytic reactions, degradation and stability of cephalosporins, Uses of cephalosporins, Comparison between 6-APA and 7-ACA and penam and cepham.
1a. Prof. P. Ravisankar, Vignan Pharmacy College, Vadlamudi, Guntur, AP., Ind...Dr. Ravi Sankar
Antibiotics, characteristics of an antibiotic, Brief historical background of antibiotics, Modern History of antibiotics, Classification of antibiotics, four groups of beta-lactam antibiotics, Classification of Penicillins, Penicillinase resistant Penicillins, Narrow spectrum anti-staphylococcal Penicillins, Amino Penicillins, Nomenclature of Penicillins, Stereochemistry of Penicillins, Hydrolysis of Penicillins, SAR of Penicillins, Mechanism of Penicillins, Beta-lactamase inhibitors, Therapeutic uses of Penicillins, Penicillin advantages, and disadvantages, Hypersensitivity or allergic reactions of Penicillins.
Tetracyclines are Octahydro napthacene derivatives which are bacteriostatic potent broad spectrum antibiotics and are the most widely prescribed form of antibiotic after penicillins.
TETRA means = four
CYCL means = hydrocarbon rings
INE means = derivation.
Tetracyclines are introduced 50 years ago as potent broad spectrum antibiotics.
They are biosynthesized form acetic acid and propionic acid units in microorganisms.
Finally, one of the best ways of reducing cancer is …..
Cancer prevention is an essential component of all cancer control plans because about 40 % of cancers are preventable.
Firstly…Public education campaigns are important in highlighting the dangers of smoking because possibly as many as 22 % (in the UK 30%) of cancers are caused by smoking.
Secondly, another 30% of cancers are diet-related. Decades of research have clearly shown that by living a healthy life, people can reduce the risk of developing the disease.
Screening tests help in finding Cancer earlier before developing any symptoms.
The benefits of eating high-fiber foods, fruit, and vegetables.
Epigallocatechin gallate, an antioxidant present in green tea, is another potential protective agent.
Finally, The new research funded by the UK and USA which aims to show the number of cancer cases could be prevented by known lifestyle and environmental factors,
like being a non-smoker, keeping a healthy weight, drinking less alcohol, eating a healthy balanced diet, and avoiding being exposed to certain infections or radiation.
Every country, regardless of resource level, can take steps to curb the cancer epidemic, save lives and prevent unnecessary suffering.
Finally, cancer prevention efforts should be preceded by a systematic planning process on cancer control.
Antibiotics definition, Early and modern history, classification of antibiotics, Mechanism of antibacterial action, bacterial cell and drug targets, penicillins nomenclature, degradation reactions of penicillins, medical classification of penicillins, SAR of penicillins, Mechanism of penicillins, Betalactamase inhibitors, Therapeutic uses of penicillins, toxicity of penicillins, Side effects of penicillins.
NOVEL SEPARATION AND QUANTITATIVE DETERMINATION OF LEVOFLOXACIN, PRULIFLOXACI...Dr. Ravi Sankar
The core AIM of the present study is to develop a novel, rapid, precise and accurate RP-HPLC method for simultaneous separation and quantification of six fluoroquinolones OF LEVOFLOXACIN (LEVO), PRULIFLOXACIN (PRFX), GATIFLOXACIN (GATI), SPARFLOXACIN (SPAR), MOXIFLOXAXIN (MOXI) AND BALOFLOXACIN (BALO) for the the day to day analysis.
The author felt that a novel single method for separation and quantification of all the above said drugs on single chromatographic system without any minor changes in detection wavelength and mobile phase composition.
To develop rapid, sensitive and economical analytical method based on HPLC for separation and estimation of six fluoroquinolones pharmaceutical dosage forms.
To develop method with shorter run time and better sensitivity.
Reducing the solvent consumption to make it more eco-friendly.
Avoid the column damage by minimizing the buffer strength and pH of mobile phase than reported methods.
To validate the method for different parameters like Accuracy, Precision, Linearity, specificity, Robustness International Conference on Harmonization ICH Q2(R1) guidelines..
To apply the developed RP-HPLC method in the analysis of pharmaceutical formulations.
Analytical method development and validation of novel Levofloxacin, Pruliflox...Dr. Ravi Sankar
For the first time simple, selective, sensitive RP-HPLC method was developed for the separation and quantitative development of LEVO, PRFX, GATI, SPAR, MOXI and BALO relating to fluoroquinolone anti bacterials in pharmaceutical dosage forms. The most important advantage of developed method was that the 6 separate drugs can be determined on a single chromatographic system without modifications in detection wavelength and mobile phase by RP-HPLC. The chromatographic separation of the selected drugs was carried out on Welchrom C18 column consisting of 250 mm X 4.6 mm, 5 µm particle size utilizing mixture of 10 mM phosphate buffer (pH 3.1): Acetonitrile in the ratio of 70:30,v/v as mobile phase at the flow rate of 1mL/min with detection wave length at 293 nm by using UV spectrophotometric detector with total run time of 10 minutes and 3.613, 4.230, 4.707, 5.497, 5.880 and 6.253 minutes of retention time obtained for LEVO, PRFX, GATI, SPAR, MOXI and BALO respectively. All calibration curves for six drugs showed indicated linearity over a concentration range of 2-10 µg/mL. The results regarding to limit of detection (LOD) and limit of quantitation (LOQ) for LEVO, PRFX, GATI, SPAR, MOXI and BALO were found to be 0.116 µg/mL and 0.348 µg/mL; 0.152 µg/mL and 0.460 µg/mL; 0.084 µg/mL and 0.255 µg/mL; 0.186 µg/mL and 0.558 µg/mL, 0.162 and 0.493, 0.112 and 0.390 respectively. These results clearly show low values of LOD and LOQ. The said developed method was ultimately utilized for quantification of marketed formulation.
NOVEL SIMULTANEOUS SEPARATION AND QUANTITATIVE DETERMINATION OF FOUR SARTANS...Dr. Ravi Sankar
The core AIM of the present study is to develop a novel, rapid, precise and accurate RP-HPLC method for simultaneous separation and quantification of Hydrochlorothiazide along with four sartans Telmisartan(TELM), Losartan(LOSA), Olmesartan(OLME) and Valsartan(VALS).
To develop Novel methods for separation and quantification of all the above said drugs on single chromatographic system without any minor changes in detection wavelength and mobile phase composition.
Total quality management (TQM), and current Good Manufacturing Practice (cGMP...Dr. Ravi Sankar
TQM, cGMP, Introduction, Definition, Importance, TQM frame work, Key concepts (Principles) of TQM, specific steps in the cycle, Benefits of TQM, cGMP, principles of GMP, Improtance of GMP, why GMP established?, difference between GMP and cGMP, GMP and cGMP regulations, code of federal regulations.
Tetracyclines BY Dr. P. Ravisankar M. Pharm., Ph.D.Dr. Ravi Sankar
Tetracyclines by Dr. P. Ravisankar M. Pharm., Ph.D.
Definition
Introduction
Classification
Historical background
Sources
Chemistry
SAR of tetracyclines
Mechanism of action of tetracyclines
Spectrum of activity
Uses of tetracyclines
Side effects of tetracyclines
Dr. P. Ravisankar M. Pharm., Ph.D.
Vitamin A
Vitamin D
Vitamin E
Vitamin K
Definition
Introduction
Classification
Structures,Functions,Deficiency,Diseases,Toxicity and uses.
Introduction to diuretics.
Therapeutic approaches.
Normal physiology of urine formation.
Classification of drugs .
Mechanism of action of Acetazolamide.
Mechanism of action of Thiazides.
Mechanism of action of Loop diuretics.
Mechanism of action of potassium sparing diuretics &aldosterone antagonists.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
How to Create Map Views in the Odoo 17 ERPCeline George
The map views are useful for providing a geographical representation of data. They allow users to visualize and analyze the data in a more intuitive manner.
This is a presentation by Dada Robert in a Your Skill Boost masterclass organised by the Excellence Foundation for South Sudan (EFSS) on Saturday, the 25th and Sunday, the 26th of May 2024.
He discussed the concept of quality improvement, emphasizing its applicability to various aspects of life, including personal, project, and program improvements. He defined quality as doing the right thing at the right time in the right way to achieve the best possible results and discussed the concept of the "gap" between what we know and what we do, and how this gap represents the areas we need to improve. He explained the scientific approach to quality improvement, which involves systematic performance analysis, testing and learning, and implementing change ideas. He also highlighted the importance of client focus and a team approach to quality improvement.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
Andreas Schleicher presents at the OECD webinar ‘Digital devices in schools: detrimental distraction or secret to success?’ on 27 May 2024. The presentation was based on findings from PISA 2022 results and the webinar helped launch the PISA in Focus ‘Managing screen time: How to protect and equip students against distraction’ https://www.oecd-ilibrary.org/education/managing-screen-time_7c225af4-en and the OECD Education Policy Perspective ‘Students, digital devices and success’ can be found here - https://oe.cd/il/5yV
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
2. Introduction
principle
Schematic representation of cometetive binding in
RIA
Objective of RIA
Instrumentation
Requirements for RIA
Methodology of the assay
Applications of RIA in pharmaceutical analysis
Novel applications of RIA-techniques
Conclusion
references
3. INTRODUCTION:
Ria was primarili developed by
berson & yalow(1959) for the quantitative measurement of
insulin in human plasma.
Ria principles have found wide application in the field of
drug analysis,pharmacokinetic studies,immunodiagnosis.
Ria is specific,sensitive &rapid.
The major disadvantage of ria is health&safety risks by the
use of radiation &maintaining licensed radiation &disposal
program is difficult.
4. ria is competative binding assay.
The antibody &labelled antigen are always
present as limiting factors&the concentrations
of unlabelled antigen(sample) under
examination is increased continually.
It has been observed that the % of antibody-
bound labelled antigen declines progressively.
5.
6.
7. The most 2 vital equipments essentiall required for
RIA are:
i)centrifuge
ii)radioactive counters
centrifuge
↓
↓ ↓
swing- bucketrotor fixedangleheadrotor
↓ ↓
capable of generating capable of generating
1200-2500rpm 3500-4000rpm
↓ ↓
8. the pellect is formed pellet is performed at
at bottom of test tube an angle
RADIOACTIVE COUNTERS
↓
↓ ↓
gamma counters scintillation
counters
↓ ↓
Used for counting gamma- used for counting
beta-
Energy emitting isotopes energy emmiting
isotopes
Such as i125 such as ,3H,14c.
9. The following steps are involved in RIA:
1)Radio label production
2)Conjugate preparation
3)Antibody production & characterization
4)Separation techniques
10. Eg:the most commonly used radiolabels in RIA
are 3H , I.125
I125 -> for iodination drugs are coupled with
trimethylester,tryamine,tyrosine to serve
as sites for iodination.
-> iodination is performed by enzymatic
iodination,monochloride exchange etc.
11. The combined hapten(such as drug) and carrier(a
protein,polypeptide)is called conjugate.
→common carriers are human γ-
globulin,albumin,syntheticpeptides etc.
→several reactive groups on protein can be used for
purpose of conjugating the standard drug.
eg:terminal amino,carbovyl groups,phenolic
groups of tyrosine&SH group of cysteine etc.
→conjugate is prepared by various conjugation methods.
eg:diazotization,gluteraldehyde reactions etc.
12. →the conc.of antibody(called its liter)is important for the antigenbeing
assayed.
→general method of inducing antibody formation is to inject 0.2 to 2mg of
pure antigen mixed with “freunds adjuvant”
(a mixture of mineral oil,waves,killed bacilli which enhances&prolongs
the antigenicresponse).
→animals used include rabbits,sheep,guinea pigs depending on vol.of
antiserum desired.
→once an animal has been imminized,it can be injected several times to
obtain different lots of antisera.
→antiserum collected is stored in liquid-nitrogen at -196 c.after thawing
the antiserum is stored at 4 c.
13. To separate free from bound labelled antigen
separation techniques are:
a) physical methods:-
filtration,chromatography,electrophoreses,charcoal
dextran adsorption etc.
b) Chemical methods:-
organic solvents such as
ethanol,dioxane,PEG,ammonium sulfate for
precipitating antibody- boundhapten.
14. In RIA the sequential steps followed are:
1)Mix a fixed concentration of antiserum containing
specific antibody with a constant of radiolabelled
antigen.
2)Incubate it for specified duration&at an appropriate
temperature usually +4°c.
3)A definite volume of sample containing unlabelled
antigen to be measured is added to the reaction
test tube.
15. 4)The antibody reacts with both radioactive&unlabelled antigen
forming an ab-radiolabelled antigen complexes.
5)Both radioactive&unlabelled antigens are more or less same
immunochemically,they will compete for limited number of
antibody sites available.
6)The radio activity falls because the unlabelled antigen dilutes
it.
i.e)reducing the no of labelled antigen combining with
antibody.
7)The counts obtained from the radioactivity are used to
determine the unlabelled antigen concentration in the
sample,the interpretation being done on the standard curve.
16. RIA is used for estimation of pharmaceutical drugs like:
1) morphine - narcotic analgesic
2) Hydromorphone & hydrocodone in humanplasma -
narcoticanalgesic,antitussive,antipyretic.
3) Clonazepam - sedative&anticonvulsant
4) Flurazepam - hypnotic&anticonvulsant
5) Barbiturates - hypnotic&anticonvulsant
6) Flunisolide - a steroid having marked anti-inflammatory activity
20. Various dilutions of antiserum are incubated
inpresence of fixed concentration of tritium labelled
morphine.then standard unlabelled antigen is added.
incubated&saturated ammonium sulphate solution
added
↓
The precipitate is sedimented by centrifugation at
5000rpm
↓
21. The washed with 50% ammonium sulphate
solution
↓
the precipitate contains antibodybound
morphine&radioactivity counted with help of
packrd- iri- card liquid scintillation
spectrometer.
22. → combined RIA-technique &isotope dilution has
been successfully developed to estimate
SULINDAC along with its 2 prominent
metabolites,namely:sulindac-sulphone
&sulindac –sulphide,present in plasma-level.
23. Prepranolol is a racemic mixture.it contains
equimolecular portion of D-and L-isomers.
→two antisera have been developed
experimentally
a) antisera against the DL- racemicmixture&
b) antisera against the L-isomers.
→the DL- propranotol antiserum exhibits almost
equal affintity for both D-and L-isomers
24. →by the application of two RIA- techniques,dl-
and l- propronolol is quantified.
→thus the concentrations of d- propranol is
known by subtracting the concentration of l-
isomer from the dl-mixture.
25. → RIA have been successfully applied to a wide
variety of pharmacological agents.
→ RIA is an important method in the quantitative
analysis of drugs.
→the methods for preparing immunogenic drug -
protien conjugates have improved during the
past few years & high-affinity antisera are
becoming more &more commonly available.
26. INTRODUCTION:-
→ELISA is a biochemical technique used in
immunology to detect the presence of an
antibody or antigen in a sample.
→here,the tag employed is an enzyme.
→ELISA is so named because the technique
involves
the use of an immunosorbent.
27. (immunosorbent is an absorbing material specific
for one of the components of the reaction,the
antigen or antibody)
→eg.of immunosorbents :cellulose or agarose
→ELISA is actually done using 96-well microlitre
plates suitable for automation.
28. ELISA are devided into 3 types:-
1)competitive binding ELISA
2)sandwich assays
3)indirect ELISA
29. Competitive binding ELISA:-
{here analyse antigen&taggedantigen compete for
sites on the absorbed antibody}
the antibody to the antigen analyte is absorbed on to
the solid phase by hydrophobic interactions
↓
Then a known amount of enzyme-labeled
antigen&sample containing unlabeled antigen is
added
30. ↓
Incubation is done & the wells are washed &
enzyme substrate at 37°c for 30 min is added to
produce coloured product via enzyme
catalysed reaction
↓
Maximum coloration occurs if there is no
antigen in sample to compete with binding of
labelled antigen.
31. SANDWICH ASSAYS:-[these are non
competetive]
Sample is added to the wells containing
absorbed antibody
↓
incubation done
↓
Then enzyme labelled antibody added &it will
bound to antigen
32. ↓
the unbound labeled antibody is washed away
↓
The solid phase then contains antigen
sandwiched b/w labeled&unlabeled antibody
↓
The colour produced upon enzymatic reaction is
directly preportional to the amount of antigo
33. sample antigen first adsorbed the solid phase
↓
Then unlabeled primary antibody is added
↓
Then incubated &washed
↓
Then a secondary labeled antibody is added
↓
Then incubated &washed
↓
the enzyme activity is proportional to the antigen
35. ELISA can be performed to evaluate the
presence of antigen or antibody in a sample.it
is useful for determining serum antibody
concentrations in HIV test.
ELISA is highly sensitive &allows accurate
measurement of very low levels of IGH
subclass.
36. 1)Analytical chemistry (sixth edition)
→by GARY D.CHRISTIAN
2)Practical pharmaceutical chemistry(fourth edition)
→by A.H.BECKETT J.B.STENLAKE
3)Pharmaceutical drug analysis(second edition)
→by ASHUTOSHKAR
4)Pharmaceutical analysis modern methods (partA)
→by JAMES W.MUNSON