Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
Expt. 7 Bioassay of acetylcholine using rat ileum by four point bioassayVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
Expt. 5 Bioassay of oxytocin using rat uterine horn by interpolation methodVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of oxytocin standard solution
Preparation of De Jalon solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Graphical presentation of DRC
Calculation
Result and interpretation
Expt. 6 Bioassay of histamine using guinea pig ileum by matching methodVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of histamine standard solution
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
Expt. 9 Effect of atropine on DRC of acetylcholine using rat ileumVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh and Atropine stock and std. solutions
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Graphical presentation of CRC/ DRC
Result and interpretation
Expt. 10 effect of spasmogens and spasmolytics using rabbit jejunumVISHALJADHAV100
Overview of Discussion
Objective
Principle
Requirements
Experimental specifications (conditions)
Drugs and solutions used in rabbit intestine experiment
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Result and interpretation
Expt. 7 Bioassay of acetylcholine using rat ileum by four point bioassayVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
Expt. 5 Bioassay of oxytocin using rat uterine horn by interpolation methodVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of oxytocin standard solution
Preparation of De Jalon solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Graphical presentation of DRC
Calculation
Result and interpretation
Expt. 6 Bioassay of histamine using guinea pig ileum by matching methodVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of histamine standard solution
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Result and interpretation
Expt. 9 Effect of atropine on DRC of acetylcholine using rat ileumVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh and Atropine stock and std. solutions
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation
Graphical presentation of CRC/ DRC
Result and interpretation
Expt. 10 effect of spasmogens and spasmolytics using rabbit jejunumVISHALJADHAV100
Overview of Discussion
Objective
Principle
Requirements
Experimental specifications (conditions)
Drugs and solutions used in rabbit intestine experiment
Preparation of Tyrode solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Result and interpretation
Expt. 3 Introduction to principles of bioassay, its types including advantage...VISHALJADHAV100
Biological standardization (Bioassay)- Definition
Bioassays are employed- BUT WHEN???
Applications of bioassay methods
Principles of bioassay of drugs
Standard preparation (Reference Standard)
Types of bioassays-
1) Quantal response bioassay
2) Graded response bioassays
a) Matching bioassay
b) Interpolation bioassay
c) Bracketing bioassay
d) Multiple point bioassays
Three-point bioassay
Four-point bioassay
Six-point bioassay
Latin square designs- Multiple point bioassays
Intact animal studies
Expt. 4 DRC of acetylcholine using frog rectus abdominis muscleVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation of magnification value (Mf)
Graphical presentation of CRC/ DRC
Result and interpretation
Expt. 6 Study of effect of drugs on gastrointestinal motilityVISHALJADHAV100
Objective
Principle
Requirements
Preparation of Tyrode solution
Procedure
Kymograph recording of contractions
Observation table
Result and Interpretation
Expt. 8 Effect of physostigmine on DRC of acetylcholine using frog rectus abd...VISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh and Physostigmine stock and std. solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation of magnification value (Mf)
Graphical presentation of CRC/ DRC
Result and interpretation
Expt. 1 Introduction to in vitro pharmacology and physiological salt solutionsVISHALJADHAV100
Definitions of pharmacology & drug
Aims of experimental pharmacology
Pre-clinical pharmacology
Clinical pharmacology
Types of experiments in pharmacology
Assembly for isolated organ/ tissue related experiments
Recording (writing) levers
Physiological salt solution (PSS)
Introduction
Examples
Composition
Role of ingredients
Precautions in preparation of PSS
Selection of PSS
Expt. 3 Introduction to principles of bioassay, its types including advantage...VISHALJADHAV100
Biological standardization (Bioassay)- Definition
Bioassays are employed- BUT WHEN???
Applications of bioassay methods
Principles of bioassay of drugs
Standard preparation (Reference Standard)
Types of bioassays-
1) Quantal response bioassay
2) Graded response bioassays
a) Matching bioassay
b) Interpolation bioassay
c) Bracketing bioassay
d) Multiple point bioassays
Three-point bioassay
Four-point bioassay
Six-point bioassay
Latin square designs- Multiple point bioassays
Intact animal studies
Expt. 4 DRC of acetylcholine using frog rectus abdominis muscleVISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh stock and standard solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation of magnification value (Mf)
Graphical presentation of CRC/ DRC
Result and interpretation
Expt. 6 Study of effect of drugs on gastrointestinal motilityVISHALJADHAV100
Objective
Principle
Requirements
Preparation of Tyrode solution
Procedure
Kymograph recording of contractions
Observation table
Result and Interpretation
Expt. 8 Effect of physostigmine on DRC of acetylcholine using frog rectus abd...VISHALJADHAV100
Objective
Principle
Requirements
Experimental specifications (conditions)
Preparation of ACh and Physostigmine stock and std. solutions
Preparation of frog ringer solution (PSS)
Procedure
Kymograph recording of contractions
Observation table
Calculation of magnification value (Mf)
Graphical presentation of CRC/ DRC
Result and interpretation
Expt. 1 Introduction to in vitro pharmacology and physiological salt solutionsVISHALJADHAV100
Definitions of pharmacology & drug
Aims of experimental pharmacology
Pre-clinical pharmacology
Clinical pharmacology
Types of experiments in pharmacology
Assembly for isolated organ/ tissue related experiments
Recording (writing) levers
Physiological salt solution (PSS)
Introduction
Examples
Composition
Role of ingredients
Precautions in preparation of PSS
Selection of PSS
Lecture includes definition of bioassay, Types of Assay and Bioassay , Indications, principles, advantages of bioassay. Example of a Bioassay with calculations. This lecture will be of help for postgraduate pharmacology students as well as undergraduates
For HPLC, sample solvents that adequately dissolve target compounds are required. Therefore, sample solvents that contain a high concentration of organic solvent are often used for reversed phase chromatography. The problem is that these solvents sometimes cause peak broadening.
This presentation discusses techniques for reducing the effects of sample solvents on UHPLC analyses.
Microbiological inspection of mineral water by redox-potential measurement Olivér Reichart
MicroTester as a validated method is suitable for rapid microbiological testing of mineral water, carbonated water, tank and running drinking water and other types of water. The time needed for a reliable detection of microorganisms is of key importance: in water industry the real-time (or at least as fast as possible) monitoring of the microbiological properties of the production is indispensable; in public water supply the essential basis of the epidemiological and public health measures is the fast and reliable result of the microbiological inspection. Beside the most important and most widely inspected microbiological contaminants the most relevant disturbing flora was involved to the validation process as well.
Analysis of Phenolic Antioxidants in Edible Oil/Shortening Using the PerkinEl...PerkinElmer, Inc.
Phenolic antioxidants are commonly used in food to prevent the oxidation of oils. Oxidized oil and fats cause foul odor and rancidity in food products, which is a major cause for concern to the food industry. Globally, regulations vary, but current maximum allowable levels are as low as 100 μg/g (100 ppm). This application note presents a UHPLC method for the analysis of the ten most common phenolic antioxidants that may be found in such products.
Using THGA and Zeeman Background Correction for Blood-Lead Determination in C...PerkinElmer, Inc.
Validated applications determining whole blood levels are generally performed using graphite furnace atomic absorption spectroscopy (GFAAS). GFAAS is cost effective, allows for detection limits well under the blood-lead level action guideline, and requires less operator training than more advanced elemental techniques.2 In this study, we will demonstrate the applicability of the PerkinElmer® PinAAcle™ 900T atomic absorption spectrometer (Figure 1) using the stabilized temperature platform furnace (STPF) and transversely-heated graphite atomizer (THGA), for use in customer-validated applications to determine lead amounts in blood samples.
Learn more about our solutions: http://bit.ly/IG2kI1
Microbial assays or microbiological assays could be a sort of bioassays designed to analyse the compounds or substances that have impact on micro-organisms. They help to estimate concentration and efficiency of antibiotics. Also facilitate in determination of the simplest anti-biotic appropriate for patient recovery.
Overview of Discussion-
Renin-Angiotensin system (RAS)
a) Circulating renin-angiotensin system
b)Tissue (local) renin-angiotensin systems
c)Alternative (ACE-independent) pathway
Other angiotensin peptides
Angiotensin receptors and transducer mechanisms
Actions of angiotensins
Pathophysiological roles of angiotensins
Inhibition of renin-angiotensin system
Overview of Discussion-
Anti-rheumatoid drugs
Classification of anti-rheumatoid drugs
Pharmacology of disease modifying anti-rheumatic drugs (DMARDs)
Pharmacology of adjuvant drugs
Overview of Discussion
Introduction
Which are the features of inflammation…?
Functional importance of eicosanoids and other chemical mediators
Pharmacological/physiological effects of inflammatory mediators
How PGs produce PAIN?
How PGs produces FEVER?
How PGs produces INFLAMMATION?
About NSAIDs...
Classification of NSAIDs
Mechanism of Action: NSAIDs
Pharmacology of Individual Class of NSAIDs
Choice of NSAIDs
Analgesic combinations
Overview of Discussion-
About Substance P (SP)
Discovery of SP
SP Receptor
Functions mediated by SP
Clinical significance of the SP-NK1R
NK1 receptor antagonists
Overview of Discussion
About Plasma Kinins
Generation and metabolism
Kinin receptors
Actions of kinins
Pathophysiological roles of kinins
Bradykinin antagonists
Overview of Discussion-
What is Serotonin?
Physiologic Distribution of Serotonin
Synthesis, Storage and Destruction
Biosynthesis of 5HT compared to CAs
Serotonin Uptake
5-HT Receptors
Actions
Pathophysiological Roles
Use
Drugs Affecting 5-HT System
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
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.
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
Synthetic Fiber Construction in lab .pptxPavel ( NSTU)
Synthetic fiber production is a fascinating and complex field that blends chemistry, engineering, and environmental science. By understanding these aspects, students can gain a comprehensive view of synthetic fiber production, its impact on society and the environment, and the potential for future innovations. Synthetic fibers play a crucial role in modern society, impacting various aspects of daily life, industry, and the environment. ynthetic fibers are integral to modern life, offering a range of benefits from cost-effectiveness and versatility to innovative applications and performance characteristics. While they pose environmental challenges, ongoing research and development aim to create more sustainable and eco-friendly alternatives. Understanding the importance of synthetic fibers helps in appreciating their role in the economy, industry, and daily life, while also emphasizing the need for sustainable practices and innovation.
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.
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.
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.
Expt. 2 Bioassay of acetylcholine using rat ileum by four point bioassay
1. ExperimentNo. 2
Bioassay of acetylcholine using rat ileum
by four point bioassay
Mr. Vishal Balakrushna Jadhav
Assistant Professor (Pharmacology)
GES’s Sir Dr. M. S. Gosavi COPER, Nashik-5
2. Overview of Discussion
• Objective
• Principle
• Requirements
• Experimental specifications (conditions)
• Preparation of ACh stock and standard solutions
• Preparation of Tyrode solution (PSS)
• Procedure
• Kymographrecording of contractions
• Observation table
• Calculation
• Result and interpretation
2
3. Objective
To determine unknown concentration of acetylcholine
(ACh) using isolated rat ileum preparation by four
point bioassay.
3
4. Four point bioassay
4
In such a type of multiple point bioassay, two responses of
standard drug and two responses of test drug are recorded.
The selection of two responses of standard should be such
that they lie on the linear portion of the concentration
response curve and also the ratio between the doses
should be 1:2. The selection of test response is determined
by hit and trial method so that the responses fall on the
linear part of the curve.
The precision, reliability and reproducibility of this bioassay are
very high. It is most commonly used for estimating the
concentration of the unknown sample.
Principle
5. 5
Rat ileum or colon are the smooth muscles which are sensitive
to ACh and shows contraction.
Mechanism of smooth muscle contraction
ACh acts on muscarinic M3- subtype of receptor which
function through Gq protein and activate membrane bound
phospholipase C (PLc) → generating inositol trisphosphate
(IP3) and diacylglycerol (DAG), the secondary messengers
release Ca2+ intracellularly → cause depolarization and
thereby smooth muscle contraction.
Overnight fasted rats are used to record better response of
AChon intestinal smooth muscles such as ileum and colon.
6. Requirements
Animal: Albino rats (150-200 g, overnight fasted)
Physiological solution: Tyrode solution.
Drug- ACh (Stock solution: 1 mg/ml), ACh std. (1 or 10 or 100
μg/ml)
Chemical- Fixing solution.
Instruments: Sherrington recording drum , Student organ
bath, Aerator, Insulin or tuberculin syringe to inject drugs
in small fractions, Dissecting board and various dissecting
instruments. Frontal writing lever and stand, Pipette,
Stop watch etc.
Miscellaneous: Kymograph paper, plasticin, clips, and
thread.
6
7. Experimentalspecifications(conditions)
Isolated tissue- Isolated rat ileum preparation
Drug- ACh (Stock solution: 1 mg/ml), ACh std. (1/ 10/ 100
μg/ml)
Physiological salt solution (PSS)- Tyrode solution
Time cycle- Total- 5 minutes, Base line- 30 seconds, Contact
time- 90 seconds, Washing period- 3 minutes
Applied load/ tension- 0.5 g
Bath capacity- 40 ml
Bath temperature- 32- 35°C
Speed of rotation of drum- 0.25 mm/ second
Magnification value (Mf) = d (F-W)/ d (F-T)
Aeration- Normal air (1- 2 bubbles/ second)
7
8. Preparationof ACh stock and standardsolutions
Stock solution- 1 mg/ml or 1000 μg/ml
Dissolve1 mg of ACh in 1 ml of distilled water → 1000 μg/ml
Standard solutions
Dilute 1 ml of stock solution up to 10 ml with distilled water
→ 100 μg/ml
Dilute 1 ml of 100 μg/ml solution up to 10 ml with distilled water
→ 10 μg/ml
Dilute 1 ml of 10 μg/ml solution up to 10 ml with distilled water
→ 1 μg/ml
8
9. Preparationof Tyrode solution (PSS)
Prepare 1 litre of Tyrode solution by dissolving NaCl (8.0
g), KCl (0.2 g), MgCl2 (0.1 g), NaHCO3 (1.0 g), NaH2PO4
(0.05 g) and glucose (2.0 g) in distilled water.
MgCl2 should be added at last.
CaCl2 (0.2 g) should be dissolved separately in distilled
water to avoid chances of precipitation of salt.
Mix CaCl2 solution to the higher volume of PSS.
9
10. Procedure
Albino rat is sacrificed by a blow on the head and carotid
bleeding.
Cut open the abdomen and lift the caecum to trace the ileocecal
junction. Cut and remove a few centimeter long of the ileal
portion and immediately place it in the watch glass containing
Tyrode solution. Trim the mesentery and with gentle care clean
the contents of the ileum by pushing the Tyrode solution into
the lumen of the ileum. Utmost care should be taken to avoid
any damage to the gut muscle. Cut the ileum into small
segments of 2-3 cm long.
Take a piece of ileum of 2-3 cm long and tie the thread to the top
and the bottom ends without closing the lumen, and mount the
tissue in the organ bath containing Tyrode solution maintained
at 32- 35°C and bubbled with O2 or air. A tension of 0.5 g is
applied and the tissue is allowed to equilibrate for 30 minutes
before adding the drugs to the organ bath.
10
11. Record graded responses (CRC/ DRC) with standard solution of
ACh until the peak effect is obtained.
Select two concentrations (A, B) of the standard ACh eliciting
submaximal responses (S1, S2) and bearing a dose ratio 1:2
preferentially.
Select two suitable volumes of test solution by trial and error
method in such a way that the response T1 due to lower dose of
test (C) lies preferentially between S1 and S2. The higher volume
of test solution selected would be D such that the dose ratio
B/A= D/C and the response of D is T2. All the four responses (S1,
S2, T1 , T2) due to the doses thus selected (A, B, C, D) must lie on
the linear part of the standard (sigmoid) curve.
Record four sets of responses due to A, B, C and D adding them
to the organ bath in a random fashion (Latin square design) as
shown.
Properly label and fix the kymograph recording of responses
with the help of fixing solution.
11
12. Measure various responses to calculate mean of each response
(Mean of S1, S2, T1 and T2).
Calculate the potency ratio (M) using the following formula-
Where, A= Lower volume of the standard drug,
B= Higher volume of the standard drug,
C= Lower volume of the test drug, and
= Mean standard and test responses.
Determine the strength of unknown solution of ACh using the
concentration of the standard ACh solution and potency ratio
(M). (Simply multiply potency ratio, M with standard
concentration of ACh, Cs).
12
14. Kymograph recording of contractions- continue
14
Part II- Recording of responses of A, B, C & D in four point bioassay.
A- Lower volume of the standard drug (0.4ml)
B- Higher volume of the standard drug (0.8 ml)
C- Lower volume of the test drug (0.6ml)
D- Highervolume of the test drug (1.2ml)
A B C D
B C D A
C D A B
D A B C
Latin Square Design
Fourpoint bioassay
15. Observation table- Example
15
Sr.
No.
Drug
(Conc.)
Dose
(ml)
Dose
(μg)
log
(dose)=
log (dose
in μg )
Response in Conc. of ACh in 40 ml
organ bath=
Dose (μg)/ Bath
Capacity (ml)
Height
(cm/mm)
%
1
ACh
(100 μg/ml )
0.1 10 1.0 0.3 16.67 0.25
2 0.2 20 1.3010 0.5 27.78 0.5
3 0.4 40 1.6021 1.4 77.78 1
4 0.8 80 1.9031 1.8 100.00 2
5 1.6 160 2.2041 1.8 100.00 4
6 3.2 320 2.5051 1.8 100.00 8
Part I- Dose Response Curve (DRC) of ACh Std.
0.00
20.00
40.00
60.00
80.00
100.00
120.00
0 0.5 1 1.5 2 2.5 3
%
RESPONSE
LOG(DOSE)
Log (DRC)of ACh Standard (100μg/ml)
16. Observation table- Example
16
Sr.
No.
Drug
(Conc.)
Dose
(ml)
Response in
Height
(cm/mm)
1 A 0.4
2 B 0.8
3 C 0.6
4 D 1.2
5 B 0.8
6 C 0.6
7 D 1.2
8 A 0.4
Part II- Recording of responses of A, B, C & D in four point bioassay.
Sr.
No.
Drug
(Conc.)
Dose
(ml)
Response in
Height
(cm/mm)
9 C 0.6
10 D 1.2
11 A 0.4
12 B 0.8
13 D 1.2
14 A 0.4
15 B 0.8
16 C 0.6
17. Calculation
17
Magnificationvalue (Mf) = d (F-W)/ d (F-T)
Where-
d (F-W) → distance between fulcrumand stylus (writing tip)
d (F-T) → distance between fulcrumand point of attachment of tissue
Unknown concentration of ACh T (μg/ml)
Calculate mean of heightsof standard and test responses due to A, B, C and D
Unknown concentration of ACh T (μg/ml)= PotencyRatio(M) X Cs
18. Result and interpretation
The concentration response curve (CRC) or dose response
curve (DRC) of standard concentration of ACh (100 μg/ml)
by using isolated rat ileum preparation was recorded and
interpreted graphically as sigmoid or S-shape curve.
The unknown concentration of ACh (T) using isolated rat
ileum preparation by four point bioassay was found to be
.........μg/ml.
The magnification value was found to be-.........
18