Practical Manual of Pharmacology

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Practical Manual of Pharmacology
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CMH INSTITUTE OF MEDICAL SCEINCES BAHAWALPUR

PAKISTAN
Practical Manual of Pharmacology i

Authors
Dr. Syed Asif Jahanzeb Kazmi: MBBS., M.Phil., Ph.D. Scholar
Professor and Head of the Department, Pharmacology and Therapeutics, CMH Institute of
Medical Sciences, Bahawalpur
Fayyaz Anjum (R.ph): Pharm.D., M.Phil. Pharmacology
Lecturer, Department of Pharmacology and Therapeutics, CMH Institute of Medical Sciences
Bahawalpur
Co-Authors
Dr. Hafiz Muhammad Ahsan: Ph.D., Post Doctorate Pharmacology
Assistant Professor, Department of Pharmacology and Therapeutics, CMH Institute of
Medical Sciences Bahawalpur
Syeda Memoona Gillani: Pharm.D., M.Phil. Pharmacology
Bahawalpur
Irsah Maqbool: Pharm.D., M.Phil.
Bahawalpur
Dr. Rafeeq Alam Khan: Ph.D. Pharmacology
Meritorious Professor and Dean Faculty of Pharmacy, Ziauddin University, Karachi
Dr. Qaiser Jabeen: Ph.D., Post Doctorate Pharmacology
Professor of Pharmacology, Faculty of Pharmacy and Alternative Medicine, The Islamia
University Bahawalpur
Reviewers
Dr. Syed Asif Jahanzeb Kazmi: MBBS., M.Phil., Ph.D. Scholar
Professor and Head of the Department, Pharmacology and Therapeutics, CMH Institute of
Medical Sciences, Bahawalpur
Dr. Hafiz Muhammad Ahsan: Ph.D., Post Doctorate Pharmacology
Assistant Professor, Department of Pharmacology and Therapeutics, CMH Institute of
Medical Sciences Bahawalpur
Practical Manual of Pharmacology ii

Preface
Pharmacology is a very important and highly growing subject of medical science. A sound
understanding of basic and clinical applications of Pharmacology in different clinical
scenarios is a key for clinicians and specialists. Laboratory practicals have been proved an
important tool to learn the applications of any theoretical information. Pakistan Medical and
Dental Council has recommended two third of assigned teaching time for on-job training. A
curriculum is a vision and road map to meet the academic objectives. The undergraduate
pharmacology curriculum has always been a topic of intense debate. It has been generally felt
that pharmacology practical course in medical institutions is not coping with the rapid
changes and requirements of clinical practice. Traditionally, it has focused more on factual
information with little or no emphasis on clinical and applied aspects. Dispensing pharmacy
and experimental pharmacology has remained the cornerstone of conventional pharmacology
practical exercises. Clinical utility and relevance of these practical exercises have always
been questioned and criticized. Keeping in view the deficiencies in available practical
manuals and need of growing applied and clinical pharmacology, this practical manual is
designed to meet the needs of the hour. The pharmaceutical calculations are correlated with
clinical scenarios and practical about dose calculation in special groups is added. First time,
the practicals of behavioral studies are included for better understanding of psychotic drugs.
This effort is being made to include practical application of maximum theoretical syllabus.
Another important purpose of the compilation of this manual was to reduce the unnecessary
dictation and writing time of facilitator and students and better understanding of theoretical
knowledge.
Hopefully, second edition of this manual will be worthwhile for students and teachers. The
authors would be pleased and welcome any suggestion and positive criticism for further
improvement of this manual.
1st Edition 2018
2nd Edition 2019
Authors
Practical Manual of Pharmacology iii

Acknowledgements
In the name of ALLAH, the most Merciful and the most Beneficial. Alhamdulillah, all
praises to ALLAH, Who conferred us with His countless blessings and empowered us to
complete this practical manual successfully.
Countless salutations to the HOLY PROPHET HAZRAT MUHAMMAD (PBUH) a
symbol of humanity, perfection and a book of guidance for the whole universe.
We are highly indebted to Brig. CH ALTAF HUSSAIN (R) SI (M) (Principal, CMH
Institute of Medical Sciences Bahawalpur) and Brig. KHALID MEHMOOD (Vice
Principal, CMH Institute of Medical Sciences Bahawalpur) for their full support and co-
operation in writing and compilation of the manual.
We would like to say special thanks to our mentors, Dr. Rafeeq Alam Khan
(Meritorious Professor and Dean, Faculty of Pharmacy, Ziauddin University Karachi) and
Dr. Qaiser Jabeen (Professor of Pharmacology, Department of Pharmacy, The Islamia
University of Bahawalpur) for their passion and patience to guide us all the way in
completion of this practical manual.
Last but no least special thanks to Co-Authors, Reviewers and Faculty members of the
department of Pharmacology and Therapeutics for their valuable inputs and suggestions in
making the practical manual diverse, comprehensive and according to the needs of the hour.
Authors
Practical Manual of Pharmacology iv

Table of Contents
Practical Page
Title Signatures
No No
Pharmacy Practical
Understanding and Introduction to Equipment used in

1 Pharmacology laboratory 1
Understanding of different systems of measurements and their

2 interconversion used in medical profession 6
Study of different biological Salt Solutions used in vitro studies

3 and their functions and composition 9
Understanding and calculations of Serial and Stock Dilutions used

4 in laboratory and dispensing 11
Study of different Types of Solutions and their calculations

5 according to clinical scenarios 15
6 Introduction of solid dosage forms used to administer the drugs 21
7 Introduction of liquid dosage forms used to administer the drugs 23
Introduction of Semisolids and others dosage forms used to

8 administer the drugs 26
Biostatistics and Dose Calculation
9 Introduction to Biostatistics and its applications 28
10 Study of different formulas used for Dose Calculations in paeds 34
Understanding of loading and maintenance dose and dose

11 adjustment in renal patients 38
Prescription Writing
Study of different parts of prescription and learning of

12 prescription writing 43
General Pharmacology
Practical Manual of Pharmacology v

13 Handling of Laboratory Animals 67
Appropriate and inappropriate drug administration through

14 different routes 69
15 To Study the Effects of Drugs on Rabbit’s Eye 71
16 To Study the Phenomenon of Potentiation on Rabbit’s Eye 75
Autonomic Pharmacology
To Study of the Effects of Cholinergic Drugs (Agonism) on

17 Rabbit’s Intestine 77
To Study of the Effects of Cholinergic Drugs with anticholinergic

18 drugs (antagonism) on Rabbit’s Intestine 80
Cardiovascular System
19 To study the effects of drugs on frog's heart 83
To study the effect of coffee on pulse rate blood pressure on

20 normal human volunteers 85
Blood and Inflammation
Study the anticoagulant activity of different anticoagulants in

21 blood collected from orbital sinus or plexus of rat 88
Evaluation of Analgesic Activity of Diclofenac Sodium against

22 Formalin Induced Pain Model 91
Central Nervous System
Determination of Anti-Depressant Activity of Fluoxetine in Rat

23 94
by Tail Suspension Test
Determination of Anxiolytic Effect of Diazepam in Rat by
24 Light/Dark Exploration Test 96
Determination of Anxiolytic Effect of Diazepam in Rat by

25 Elevation plus Maze Test 98
26 To study the effects of drugs on reflex time in frog 100
Practical Manual of Pharmacology vi

Clinical Pharmacology
27 Identification of drug 103
28 Identification of unknown drug 104
Determine the frequency of analgesic and anti-inflammatory drugs

29 in medical / surgical out patients department 109
Determine the frequency of hypoglycemic drugs in medical/

30 surgical ward/ OPD 110
Determine the frequency of Antihypertensive drugs in medical/

31 surgical ward/ OPD 111
Evaluation of effects produced by physical activity on ECG with

32 and without drug 112
33 To study the effect of drug on lung functions using spirometer 114
Chemotherapy
Determination of antimicrobial activity of different antibiotics

34 using disk diffusion method 118
Practical Manual of Pharmacology vii

Experiment No. 01
Objectives
Understanding and Introduction to Equipment used in Pharmacology laboratory
Kymograph and Organ Bath Assembly
The most commonly used equipment in undergraduate pharmacology laboratory to study the
effects of various drugs on GIT, isolated tissues and heart is kymograph attached with organ bath
assembly.
1. Kymograph
Kymograph is an apparatus consisting of a rotating drum for recording wavelike motions,
variations, or modulations, such as muscular contractions. Kymograph is generally used to study
the effects of drugs on tissue preparations. It is a standalone recording apparatus used alongside
other apparatus such as organ bath. Writing levers are used to trace the recording from muscle
contractions. It mainly consists of following parts
Cylinder Clamp: It is a lock, which is used to fix the drum on the central shaft so that the
drum and the cylinder shaft rotate together.
Drum: A metallic cylinder covered with a paper for recording the activity.
Spindle: A metallic rod at the top of the kymograph that holds the drum and the shaft.
Kymograph Arm: A moveable, adjustable arm on the top of kymograph.
Main Switch: Present on the front of the kymograph, directly connected to the socket of
external switch.
Drum Motor Control Switch: Starts/stops the drum motion with an indicator light.
Speed Regulator: Are used to adjust the speed of the drum. The drum should be moved
manually only in neutral position, indicated by N on the gear.
Start/Pause lever: A small lever connected with the drum movement. If it is at the "start"
position drum moves freely, and if it is at "pause" position, the drum takes one rotation and then
stops.
Practical Manual of Pharmacology 1

Kymograph

2. Organ Bath Assembly
The organ bath assembly is a traditional experimental set-up that is commonly used to
investigate the physiology and pharmacology of in vitro tissue preparations. Perfused tissues can
be maintained for several hours in a temperature controlled organ bath. It has following main
parts.
Organ bath: It is a hollow glass tube that is connected with inlet and outlet tubes and is
fixed in the center of the outer water bath. It may have varying capacity, generally from 10
to 50ml. Physiological solution is added into bath through the inlet tube (coiled around the
tissue bath forming outer jacket) from a reservoir. The coiling tube makes the solution warm
and maintains the temperature inside the organ bath. The organ bath is drained out by
opening the outlet stop cork when washing of the tissue is required and fresh solution is
filled into the organ bath by opening the stop cork of inlet tube from the reservoir.
Outlet Pipe: An outlet drainage attached at the base of the organ bath with pinching stop cork
on it. When opened, it drains out the fluid from the organ bath.
Inlet Pipe: An inlet pipe originates from the reservoir and attached with coiling pipe around
the organ bath. It is used to add the physiological solution to organ bath when required.
Tissue fixing hook: I t is attached with the "L" shaped metal tube, dipped in the organ bath.
One end of the isolated tissue is attached with the help of thread with the hook and the other
end is tied with the frontal writing lever.
Reservoir: It is usually a glass bottle of 5 liters capacity and is kept 2 to 3 feet high above the
level of organ bath. It contains the physiological solution.
Aeration pipe: An "L" shaped metal or glass tube, shorter arm of which is attached to the
oxygen cylinder and longer end of pipe is dipped into the organ bath and releases oxygen in
the perfusion fluid. Tissue fixing hook is also attached at the end of longer arm.
Frontal writing lever: is attached to the sidewall of the water bath or may be mounted to a
stand through an adjustable assembly. Length of the arm of the lever and its position can be
adjusted according to the position of the drum of the kymograph, keeping in mind that the

Organ Bath Assembly

tissue should be attached with the lever at right angles and pen side of the lever should also
be at right angle to surface of the drum.
Ink pen: is attached to the end of the writing lever. Make sure that ink is filled in it.
Thermostat & heater: is provided with in the water bath to maintain the temperature of the
tissue in the organ bath at 35-37°C. The switch of the thermostat is located on the front wall
of the water bath and calibrated control switch is available to adjust the temperature of the
water in the organ bath and hence in the tissue bath.
Procedure
1. Adjust the height of the drum on spindle by losing the clamp and tight the cylinder
clamp.
2. Paste graph paper on the drum smoothly.
3. Connect the instrument to the power supply.
4. Adjust the speed by positioning the gear and the speed-knob appropriately.
5. Switch on the main supply to start the drum rotation.
6. Adjust the bath assembly to the adjustable arm.
7. Mount the given tissue preparation in the bath assembly.
8. Attach the pen assembly with writing lever arm and adjust the position of adjustable
arm with respect to the drum properly.
9. Fill up ink in the stylus
10. Record the tissue activity according to, the requirements of the experiment .
Precaution
Don't pause for a long time because it stops the drum immediately while the motor inside
remains on and could be burnt.

Experiment No. 02
Objectives
Understanding of different systems of measurements and their interconversion used in medical

profession
Metrology
Metrology is defined by the International Bureau of Weights and Measures (BIPM) as "the
science of measurement, embracing both experimental and theoretical determinations at any
level of uncertainty in any field of science and technology".
Importance of Measurement Systems
One of the most essential functions of a Medical professional is the ability to perform accurate
pharmaceutical measurements, calculations and conversions. Without this ability, a medical
professional is not able to apply their knowledge of pharmacology in a practical manner during
their everyday work functions. This is important as one incorrect calculation, conversion or
measurements will affect a dosage, and can potentially harm a patient. Possessing a working
knowledge of the pharmaceutical systems of measurement will benefit a medical professional.
There are three measurement systems in pharmacy calculations, which a medical professional
must learn to carry out the critical functions used in the pharmacy
Metric System
The metric system is a decimal system with all multiples and divisions based on a factor of 10.
This system is also the most commonly used system of measurement for pharmacy calculations,
as it allows for quick and easy conversions between different systems of measurement. As the
metric system is based on multiple of 10 so following prefix are used:
Deca = 10 times
Hecta = 100 times
Kilo = 1000 times
Mega = 1000000 times
Deci = 1/10
Centi = 1/100
Milli = 1/1000

Micro = 10-6
Nano = 10-9
Pico = 10-12
Apothecary System
The apothecary system is one of the three systems of measurement used in pharmacy
calculations, which uses weight and volume as divisions of measurement. This includes
measurements of ounces, gallons, pints and quarts.
1 fluid ounce = 29.573 milliliters (mL)

8 fluid ounces = 1 cup = 16 tablespoons (tbsp)
2 cups = 1 pint = 473 ml
2 pints = 1 quart = 0.946 liter
4 quarts (qt) = 1 gallon = 3.785 liters (L)
1 tablespoon =1/2 fluid ounce = 3 teaspoons (tsp)
Avoirdupois System
The avoirdupois system is similar to the apothecary system, however, the avoirdupois system
exclusively measures weight based on 16-ounce equaling 1 pound (lb). This system of
measurement is the everyday weight measuring system most people recognize
1 pound (lb) =16 ounces (oz) =0.454 kg

1 ounce = 28.35 grams (g)
1 kilogram = 2.2 pounds
1 gram = 0.035 ounce
Household Conversion
1 teaspoon (t) = 5ml

1 tablespoon (T) = 15 ml
Temperature Conversion
To convert Fahrenheit to centigrade: °C = (F – 32) x 5/9.

To convert centigrade to Fahrenheit: F = (°C x 9/5) + 32.

Calculations
1. How many granules of 50 micrograms can be prepared from 1 kg powder
2. Convert 5 lb into grams
3. Convert 4 fluid Ounces into milli liters.

Experiment No. 03
Objectives
Study of different biological Salt Solutions used in vitro studies and their functions and
composition
Biological solution is an artificially prepared solution similar to blood plasma in salt
composition and osmotic pressure. Biological solutions are used in physiological experiments
with isolated organs and in clinical practice. It is important to select a particular solution in
which a tissue survives longest. The functions of a salt solution are:
 To maintain the medium within physiological pH range
 To maintain intracellular and extra cellular osmotic balance
 Modified with a carbohydrate, such as glucose serves as an energy source
Following biological solutions are used commonly in laboratory.
1. Tyrode’s solution
Tyrode solution is a solution that is roughly isotonic with interstitial fluid and is used in
physiological experiments and tissue culture. It resembles lactated Ringer's solution, but
contains magnesium, a sugar (usually glucose) as an energy source and uses bicarbonate and
phosphate as a buffer instead of lactate. It is used to perfuse intestine.
2. Ringer’s solution
Ringer solution is a solution of several salts dissolved in water for the purpose of creating an
isotonic solution relative to the body fluids of an animal. Ringer's solution typically
contains sodium chloride, potassium chloride, calcium chloride and sodium bicarbonate, with the
last used to balance the pH. It is used for amphibian tissues; i.e. frogs.
3. Kreb’s Solution
It has been used experimentally, for instance to study arteries ex vivo and during isolated muscle
testing of mammalian skeletal muscles.
4. Ringer Lock Solution
This solution is used specially for heart tissues.

Composition of physiological salt solutions (Quantities for 10 liters)
Tyrode Ringer-
Chemicals Frogs Ringer Kreb's solution
solution Lock
NaCl 65 g 69 g 80 g 90 g
KCI 1.4 g 3.5 g 2g 4.2 g
MgCl2 - - 1g 2g
Mg SO4.7 H2O - 1.4 g - -
Na.H2P04.2H2O 0.65 g - 0.65 g -
KH2PO2 - 1.6 g - -
Glucose 20 g 20 g 10 g 10 g
NaHCO3 4g 21 g 10 g 5g
CaCl2 (1 M) 10.8 ml 25.2 ml 18 ml (2 g) 10.8 ml
Aerating Gas Air O2+ 5% CO2 O2 or air O2 + 5% CO2

Experiment No. 04
Objective
Understanding and calculations of Serial and Stock Dilutions used in laboratory and dispensing
Serial Dilutions
A serial or series of dilution is the stepwise dilution of a concentrated substance into less
concentrated solution. It is very difficult to measure accurately minute quantities; i.e.
micrograms. Usually the dilution factor at each step is constant. Serial dilutions are used to
accurately create highly diluted solutions as well as solutions for experiments resulting
in concentration curves with a logarithmic scale. A tenfold dilution for each step is called
a logarithmic dilution or log-dilution.
Dilution Factor: The dilution factor or the dilution is the initial volume divided by the final
volume. For example if 0.2 ml stock solution is added to 9.8 ml diluent, the dilution factor will
be: Initial volume= 0.2, final volume= 0.2 + 9.8= 10ml, so factor will be 0.2/10= 1/50. To
calculate the final concentration, concentration stock solution is multiplied with dilution factor of
that particular dilution.
Final Concentration = Stock sol. Conc. X dilution factor
Stock Solution Dilutions
Stock solution is a concentrated solution prepared, subsequently used in different dilutions. It is
a common practice to prepare stock solutions in large amount for laboratory procedures. It is
more convenient and less time consuming than weighing and dissolving a compound every
time when required for dispensing a mixture. The chances of error are less when very small
amount of compounds are required. The only disadvantage is short shelf life and chances of
mixing.
There are many ways to dilute or calculate the unknown concentration but following equation
used more frequently.
C1V1=C2V2
Where
C1 = concentration of stock solution
V1 = volume of stock solution
C2= concentration of required solution
V2= volume of required solution

Calculations
1- A stock solution with a concentration of 10.00 ppm was diluted to make three other
solutions. For each dilution, 10.00 mL of the previous solution was diluted in a 100.0 mL
volumetric flask. What were the concentrations of the 3 diluted solutions?
2- What is the dilution factor if you add 0.5 mL of a stock solution to 5.5 mL of diluent?
What will be the concentration of 4th dilution if the stock solution concentration was
10g/ml?

3. A vial of a drug contains 10 mg drug /ml. What volume of this stock solution is required
to prepare one liter solution containing 2µg/ml?
4. A teaspoonful (5 ml) of syrup has Ibuprofen 7.5 mg, Pseudoephedrine 9.0 mg and
flavored syrup 5.0 ml. calculate the amount of each ingredient in 120 ml syrup.

5. How many grams of paracetamol would be required for 60 ml paediatric suspension? The
composition of the suspension is 100 mg in each dropper full (2.5 ml).
6. A 154 lb patient is advised ceftriaxone 20 mg/Kg /day. How many millilitres of the
infusion should be given daily from a dosage-form of 2 gram in each 1000 ml.

Experiment No.05
Objectives
Study of different Types of Solutions and their calculations according to clinical scenarios
The quantity of a solute in any solution can be given as
1. Percentage.
2. Fractional
3. Moles / Millimoles
4. Equivalents/milli-equivalents.
5. Osmole/milli-osmoles
1. Percentage Solutions
One part of any substance mixed in 100 parts of another substance makes one percent
concentration. Percent has no unit. It is a type of ratio expressed as %. It may be weight by
volume (W/V), weight by weight (W/W) or volume by volume (V/V).
2. Fractional solutions
These are the solutions in which the relative quantities of solute and solvent are expressed as
ratio e.g. 1:100, 1:1000 etc. It means 1 gm of a drug is dissolved in 100 or 1000 ml of distilled
water respectively.
3. Moles/ Millimoles
Molar Solutions
In case of fractional or percentage solutions, one cannot determine the exact amount of active
drug present in a solution, e.g. 1 % atropine sulfate solution and 1 % atropine hydrochloride
solution will not contain the same amount of atropine base, because of the difference in the
molecular weights of the two salts. The molar system has been devised, to overcome this
problem.
Mole
A mole is defined as the molecular weight of a drug expressed in grams. One mole of a solution
means one gram molecular weight of a drug dissolved in one liter. It has been established that
gram molecular weight of any substance contains 6.02 x 1023 particles. For example: 180 g of
glucose molecule, 23 g of sodium ion, 35.5 g of chloride ion, 17 g of hydroxyl ion, all are equal
to 1 mole or 6.02 x l023 molecules or particles. It is called Avogadro’s number. Mole is indicated
by M.

1 Millimole (mM) = 1/1000 M
Or l Millimole is = 0.001 M (i.e 1 formula weight in milligram)
1 Micromole (uM) = 0.001 mM (i.e 1 formula weight in microgram)
4. Equivalents/milli-equivalents.
Equivalent (eq) solutions
An equivalent is the weight of a substance which combines with or replaces one gram atomic
weight of Hydrogen ion. It can be calculated by dividing the molecular weight of a radical by its
valency. Gram equivalent weight of a solute dissolved in one litre of solvent gives one
Equivalent (Eq) solution. One milliequivalent (mEq) is 1/1000 of an equivalent.
Interconversion of mole into equivalent.
The valency of a radical determines the number of electrons released or accepted by it in a
reaction medium. An equivalent of radical corresponds to molecular weight divided by its
valency. Therefore, one mole of a monovalent ion, half mole of a divalent ion and one third mole
of a trivalent ion are all equal to an equivalent solution of these respective ions.
For example Eq weight of K+ (which has a valency = 1)
Eq wt = 39/1 = 39 gm
1 mEq wt = 39/1000 = 0.039 = 39 mg
5. Osmole/milli-osmoles
Osmolar Solutions
Osmole is gram molecular weight divided by number of particles or ions into which a substance
dissociates in solution.
Osmolarity is one of the colligative properties of a solution which is based on the concentration
of solute particles. One Mole in case of non-electrolytes is equal to 1 Osmole. In case of
electrolytes it is equal to Mole X number of particles into which it dissociates in solution.
Milliosmole is 1/1000 of an Osmole.

Calculations
1. Calculate the amount of KCl required to prepare 1000 ml Tyrode solution. The composition
of the solution is 0.55 %.
2. A patient of 110 lb is having an acute attack of ventricular fibrillation after cardiac

resuscitation. How much volume of lignocaine you will administer to this patient if the dose
is 1.4 mg/kg and dosage form contains 2 % lignocaine?

3. Calculate the concentration of pilocarpine in 50-ml organ bath if we dissolve 2 ml of
1:6000 stock solution.
4. A patient in gynecology unit has developed an acute attack of anaphylaxis after Imferon
injection. How much of injection epinephrine you will give if the dose is 0.5 mg and dosage
form is I: 1000.

5. Calculate the amount of CaCl2 required to make 150 ml of 8.5 mMo1 solution (Molecular
weight of CaCl2 = 111)
6. Calculate the mMol concentration of ORS.

BP Formula
Glucose = 40 g
NaC1 =7g
NaHCO3 =5g
KCl =3g
(MW glucose 180, Na 23, K 39, HCO3 61, Cl 35.5)

7. A patient is taking injection ticarcillin disodium one gram daily. How many mEq of Na“ is
being injected? (M.W of ticarcillin disodium 428.4).
8. How many grams of glucose are required to prepare 500 ml solution having 300 milli
Osmole per liter concentration. What is the percentage of this solution?
[MW = 180]

Experiment No. 06
Objectives
Introduction of solid dosage forms used to administer the drugs

Dosage forms are the means by which drug molecules are delivered to sites of action within the
body. Dosage forms are also known as the drug delivery systems.
Solid preparations
Tablets
These are compact products containing medicaments in compressed or molded form. They are
usually circular in shape with either flat or convex surfaces. They are prepared by compressing a
granulated preparation of medicaments by means of punches in suitable dies. Examples: Aspirin
Tablets, Co-trimoxazole Tablets, Phenobarbitone Tablets.
Capsules
They consist of a medicament enclosed in a shell. They are convenient for medicaments which
have unpleasant taste. Capsules may be cylindrical with hemispherical ends, spherical or ovoid
in shape. Shell is composed mainly of gelatin. There are two types of capsules on the basis of
shells; hard gelatin and soft gelatin capsules. Hard gelatin capsules enclosed powders; i.e.
Amoxicillin and Cefixime, and granules; i.e. Omeprazole. Soft gelatin capsules have oils and
liquids; i.e. vitamin A & D and E Capsules.
Granules
These are preparations of medicinal substances usually in the form of small irregular particles
ranging from 2 to 4 millimeters in diameter. Example: Bephenium Granules.
Powders
These are usually mixtures of two or more medicaments, in finely divided form intended for
internal use. The minimum weight of powder for internal use is 120mg (2 grains), if the quantity
of the drug given in the prescription is very small, then the bulk should be made up with inert
substance such as lactose. Examples: dusting powders, tooth powders.
Application: Zinc oxide and boric acid dusting powder in starch base has antiseptic properties.
They are used for application on wounds and abscesses. Tooth powder (e.g Dentonic) is used for
its antiseptic and tooth cleaning properties.

Lozenges
These consist of medicaments incorporated in a flavored basis. They are intended to dissolve or
disintegrate slowly in the mouth. Example Pulmonol Lozenges, Strepsil throat lozenges.
Suppositories
These are solid bodies suitably shaped for rectal administration and usually containing
medicaments.
Application: glycerin suppositories are used to relieve constipation.

Experiment No. 07
Objective
Introduction of liquid dosage forms used to administer the drugs

Syrups
These are concentrated aqueous solutions of sucrose or other sugars to which medicaments or
flavoring agents may be added. Example: Codeine Phosphate Syrup
Application: Brufen syrup is used for its analgesic antipyretic activities in fever and pain.
Emulsions
lt is a mixture of two immiscible liquids in which one liquid is broken down into small globules
and dispersed uniformly in the other liquid with the help of an emulsifying agent, The liquid
which is broken down into small globules is called the internal phase and the other liquid as the
continuous or the external phase. For example in case of castor oil emulsion, castor oil is the
disperse phase and water is the continuous phase. The aqueous phase may contain water soluble
drugs, preservatives, coloring and flavoring agents. Purified water should be used for making
emulsion because Mg or Ca ions present in water can have adverse effect on the stability of
emulsion. The oil phase of emulsion consists of fixed or volatile oils and drugs that exist in the
form of oils e.g. oil soluble vitamins. Oil used in the preparation of an emulsion should be kept
free from bacteria.
Types of emulsions:
1. Oil in water (O/W): In this type of emulsion the oil is in the disperse phase and water in
the continuous phase e.g. Castor oil emulsion. This t is used internally because oil is more
readily absorbed in a finely divided state and the preparation is more palatable when water forms
the continuous phase. Oil in water emulsions are usually milky white in color.
2. Water in Oil (W/O): In this type of emulsion, water is the disperse phase and oil is in the
continuous phase e.g. cold creams. These are used externally.
Application: Griseofulvin emulsion is used for its antifungal activity.
Suspensions
These are mixtures of insoluble or sparingly soluble drugs in water or other vehicles, in which
particles of insoluble drugs are kept in suspended state by means of suitable suspending agents,
Example: Bismuth Chalk Suspension.
Suspensions can be administered through following routes:

1. Oral suspensions e.g. Bismuth Carbonate Suspension.
2. Intramuscular suspension e.g Procaine Penicillin.
3. Ophthalmic suspensions (for eyes) e.g. Cortisone acetate.
Solutions
Solutions are liquid preparations containing one or more-soluble ingredients usually dissolved in
water. A solution is a perfectly homogenous mixture of two substances, solute and solvent
dissolved in the solvent to make the solution. They are for internal or external use for instillation
into body cavities. Example: Chloroxylenol solution, Weak Iodine Solution.
Application: Povidone iodine (Pyodine) solution is applied on wounds due to its antiseptic
properties.
Elixirs
These are pleasantly flavored and sweetened liquid preparations of potent or nauseous
medicaments. These contain a high proportion (5%-40%) of alcohol or glycerin or propylene
glycol.
Example: Cough elixirs containing acefyline and diphenhydramine.
Spirits
Spirits are alcoholic or hydro-alcoholic solutions of volatile substances that contain 50% to 90%
alcohol. This high alcoholic content maintains the water-insoluble volatile oils in solution. If
water is added to a spirit, the oils separate. Some spirits are medicinal (e.g., aromatic ammonia
spirit). Many spirits (e.g., compound orange spirit, compound cardamom spirit) are used as
flavoring agents. Spirits should be stored in tight containers to reduce loss by evaporation.
Infusions
These are dilute solutions containing the readily soluble constituents of crude drugs. They are
prepared by diluting one volume of a concentrated infusion to ten volumes with water.
Example: Saline Infusion
Injections
These are sterile solutions or suspensions intended for parenteral administration. They are used
to administer drugs which may be therapeutically inactive or not tolerated when given by mouth,
to produce a rapid, localized, or prolonged action, or for diagnostic purpose.
Examples: Hydrocortisone Sodium phosphate injection, Paraldehyde injection, Streptomycin
injection

Mouth washes
These are aqueous solutions (in concentrated form) of substances with deodorant, antiseptic,
local analgesic, or astringent properties.
Application: Compound thymol Glycerin Mouth wash, Listerine mouth wash (antibacterial)
used due to its disinfectant properties.
Nasal drops
These are liquid preparations for instillation into the nostrils by means of a dropper. They may be
aqueous or oily solutions and usually contain substances with antiseptic, local analgesic, or
vasoconstrictor properties.
Application: Ephedrine nasal drops are used for its nasal decongestant effects.
Eye drops
These are fluid preparations containing drugs dissolved or suspended in water, glycerin, diluted
alcohol, propylene glycol or other suitable solvent, and are intended for instillation into the eye.
Application: Hydrocortisone eye drops are used to reduce swelling and redness of eye.
Lotions
Lotions are “Liquid preparations intended for application to the skin. These are applied on or
rubbed on the skin with a brush. They are usually aqueous solutions or suspensions which cool
diffusely inflamed unbroken skin. They cool by evaporation and should be reapplied frequently.
The addition of alcohol in a lotion hastens its drying and accounts for its cooling effect whereas
the inclusion of glycerin keeps the skin moist for a considerable time.
Application: Salicylic acid lotion used for its keratolytic anti-acne and anti- dandruff activity.
Enemas
These are aqueous or oily solutions or suspensions ‘for rectal administration. They are given for
their anthelmintic, anti-inflammatory, nutritive, purgative or sedative effect or for X-ray
examination of the lower bowel. Example: Prednisolone enema is used for the rectal diseases and
ulcerative colitis.

Experiment No. 08
Objective
Introduction of Semisolids and others dosage forms used to administer the drugs
1. Semisolids
Creams
These are viscous emulsions of semi-solid consistency, which may be of oil in water (washable)
or of water in Oil (oily) type. These are prepared with a suitable emulsifying agent and are
intended for external use.
Example: Fusidic Acid, Permethrin, Sulphasalazine creams used in skin allergy and burns. Cold
creams and vanishing creams are used for their moisturizing and whitening effects respectively.
Ointments
Ointments are semisolid preparations intended for external application to the skin or mucous
membranes. Ointments may be medicated or not. Unmedicated ointments are used for the
physical effects they provide as protectants, emollients, or lubricants. Medicated ointments are
used for their therapeutic applications. For example sulfur ointment for the treatment of scabies.
Applications: protection of skin from dry cold air, sunlight, dust and bacteria.
Liniments
These are liquid or semi-liquid preparations intended for external application. These may contain
substances possessing analgesic, rubefacient, soothing or stimulating properties.
Example: Turpentine Liniments.
Pastes
These are semi-solid preparations for external application. They generally contain a larger
proportion of solid material (25%) than ointments and therefore are stiffer. They consist of
medicaments mixed with soft or liquid paraffin or with a non-greasy base made with glycerol
mucilage or soap. They are used as antiseptic, protective or soothing dressings which are spread
on lint before being applied.
Example: Zinc, Coal Tar paste and tooth pastes.

2. Inhalation dosage forms
Sprays
These are preparations of medicaments in aqueous, alcoholic, or glycerol containing media to be
applied to the nose by means of an automizer. Example: Normal Saline and Pseudoephedrine
spray.
Aerosols
They consist of solutions or suspension of a medicament in a mixture of inert propellants which
is held under pressure in an aerosol dispenser, which consists of a suitable container fitted with a
special metering valve.
Application: Salbutamol aerosol inhalation is used in asthma due to its bronchodilator properties

Experiment No. 09
Objectives
Introduction to Biostatistics and its applications

Statistics
By definition, it is the science of compiling, classifying & tabulating numerical data &
expressing results in mathematical or graphical form.
Biostatistics:
It is medical statistics which deals with development and application of the most appropriate
methods for the:
 Collection of data.
 Presentation of the collected data.
 Analysis and interpretation of the results.
Purpose & Uses:
 To be familiar with the problems arising in the planning of projects & making it presentable
with elimination of errors.
 To enable a scientist to place his results in more precise & scientifically acceptable manner.
Observation:
It is an event, which is seen to occur. The event seen is observation. Usually the observation is
measurement of expression of both the event and its measurement.
 We take five students of a class and record their weight
 These are five observations
 The number of observation is taken as “n”
Data:
A set of information is called data. It is classified as qualitative and quantitative data.
Qualitative data: Arise when individuals may fall into separate classes, such as diagnosis or sex.
For example:
 Male pt. or female pt.etc
 Cancer, Hepatitis etc
 Intensity of pain etc
Quantitative data: are numerical values, arising from counts or measurements.
 4 Patients got fever 100, 102, 100, 103

Methods of presentation of data
 Numerical presentation
 Graphical presentation
 Mathematical presentation
Numerical presentation
 Tabular presentation (Simple-Complex)
 Simple frequency distribution table (S.F.D.T)
Graphical presentation
 Line graphs
 Statistical maps
 Pi Chart
Mathematical presentation
 Measures of location
Variable:
In statistics, we use the turn variable to mean a quality or quantity which varies from one
member of a sample or population to another.
 Systolic blood pressure is a variable, which varies both from person to person
and from measurement to measurement within the same.
Arithmetic Mean:
It implies arithmetic average or arithmetic mean which is obtained by summing up all the
observations and diving by the total number of observations.
E.g. ESR of 7 patients is: 7,5,4,6,4,5,9
̅ = ∑𝑿 ⁄ 𝒏
𝑿
𝑋̅ = 7 + 5 + 4 + 6 + 4 + 5 + 9 ⁄ 7 = 40/7 = 5.71
Median: The middle item of the arranged data is called Median. . In this when all observations
are arranged in either ascending or descending order, the middle observation is called Median;
i.e. mid value of series.
For odd number of values: Arrange them in ascending or descending order, the middle value is
median.
68, 69, 79, 80, 84, 93, 96
Median = 80

For even number of values: Arrange them in ascending or descending order, add middle two
values and divide them by two.
E.g. 2, 2, 4, 3, 2, 1, 7, 6
Median = 2+3/ 2 = 2.5
Mode:
The value which occurs with the greatest frequency; i.e. the most common value.
E.g. In the given number of values,
85,75,51,79,71,95,75,77,75,90,71,75,79,95,75,77,54,75,81,75
Mode = 75
Range of dispersion: It is defined as difference between the highest and lowest figures in a
given samples.
Range = Max. Value- Minimum Value
Deviation:
It is difference of individual observation from their arithmetic mean
̅
D = X- 𝑿
Standard Deviation (SD):
It is defined as positive square root of AM (Arithmetic Mean) of the square deviation from the
mean of distribution.
̅ )²/ 𝐧 − 𝟏
S.D = √∑(𝐗 − 𝑿
Standard error SE:

S.E = S.D / √𝒏
T-test: A t-test is a type of inferential statistic which is used to determine if there is a significant
difference between the means of two groups which may be related in certain features.
̅1 - 𝑿
t=𝑿 ̅ 2 /√𝐒𝐄𝟏 + 𝐒𝐄𝟐
Degree of freedom:
DOF = n1+ n2 – 2
P-Value: It is calculated via frequency distribution table

A small p-value (P ≤ 0.05) indicates strong evidence against the null hypothesis, so you reject
the null hypothesis. A large p-value (P>0.05) indicates weak evidence against the null
hypothesis, so you fail to reject the null hypothesis
Example:
Take the healthy human volunteers and divide them equally into two groups. Measure the blood
pressure of the volunteers and arrange the data. Calculate the Mod, Median and apply the student
t test.

Observations and Calculations
Systolic blood pressure Deviation from mean

Obs.no d2
(mmHg) d
Group 1
Mean value= 𝑋̅1 ∑d2 =
Group 2
Mean value= 𝑋̅2 ∑d2 =


Experiment No. 10
Objectives
Study of different formulas used for Dose Calculations in paeds

Dose: The amount of a particular medication to be administered.
e.g. A patient with chest pains needs to be given 4 mg of Morphine Sulfate for Pains.
4 mg is the Desired Dose
Strength/Potency
Strength or potency of a drug is the amount or quantity of the active pharmaceutical ingredient
(API) in milligram or gram at which any dosage form available.
For example:
500 mg of paracetamol
75 mg of aspirin
Pediatric Dose calculation:
Young's Rule: It is pediatric formula based on Age. This one is valid for a patients under the
age of 12.
Age in years
Child Dose = Adult Dose ×
Age in years + 12
Clark’s Rule: It is not used clinically, but it is a popular dosage calculation formula for
pediatric nursing instructors
Weight (lb) × 𝐴𝑑𝑢𝑙𝑡 𝑑𝑜𝑠𝑒

Child Dose =
150
Fried's Rule: It is another method used to calculate the correct dose of medication for the
pediatric patient when given only the adult dose.
Age (months) × 𝐴𝑑𝑢𝑙𝑡 𝑑𝑜𝑠𝑒

Child Dose =
150

Dose calculation on basis of Body Surface Area
Dose calculation on basis of BSA is required in paeds and oncology. BSA can be calculated by
following formula
𝑨 = √𝑾. 𝑯/𝟑𝟔𝟎𝟎
Where
A is BSA in m2
W is weight in kg
H is height in centimeters
3600 is a factor
Patient ′ s BSA m²
Patient Dose = × Adult Dose
1.73 m²

Calculations
1. Normal dose of paracetamol of 60 year old is 500 mg. calculate the dose of paracetamol
for a 2 month infant
2. A 70 kg patient is receiving 500 mg ciprofloxacin b.i.d. How much volume of syrup

containing ciprofloxacin 100 mg/5 ml will be required for child of 22 lb.

Calculations
3. If the adult dose of a drug is 25 mg, what would be the dose for a child weighing 40 lb.
and measuring 32 in. in height
4. The physician prescribed Benadryl 150mg/m²/ day for an 8-year old child who weighs 75
pounds and is 4 feet 2 inches tall. The normal adult dose is 25 mg q. i. d. How many mg
of Benadryl will be administered four times a day for the child?

Experiment No 11
Objective
Understanding of loading and maintenance dose and dose adjustment in renal patients
Loading dose
When the time to reach steady state is appreciable, as it is for drugs with long half-lives, it may
be desirable to administer a loading dose that promptly raises the concentration of drug in plasma
to the target concentration (Tc). If a loading dose is to achieve the target concentration, then from
equation:
Loading dose = volume of distribution (Vd) ×Tc
Maintenance dose
In most clinical situations, drugs are administered in such a way as to maintain a steady state (ss)
of drug in the body, ie, just enough drug is given in each dose to replace the drug eliminated
since the preceding dose. Thus, calculation of the appropriate maintenance dose is a primary
goal. Clearance is the most important pharmacokinetic term to be considered in defining a
rational steady-state drug dosage regimen. At steady state, the dosing rate (“rate in”) must equal
the rate of elimination (“rate out”). Substitution of the target concentration (Tc) for concentration
(C) in equation given below
If intermittent doses are given, the maintenance dose is calculated from given formula
Dosing rate (ss) = CL×Tc
CL = K×Vd
Whereas K= elimination rate constant
K= 0.693
t(1/2)
if the desired target concentration is known, the clearance in that patient will determine the
dosing rate. If the drug is given by a route that has a bioavailability (F) less than 100%, then the
dosing rate (ss) must be modified. For oral dosing, dosing rate is:
Dosing rate (oral) = Dosing rate (ss)

F (oral)
If intermittent doses are given, the maintenance dose is calculated from formula given below.
Maintenance dose = Dosing rate ×Dosing interval

Dose adjustment
Dose adjustment for certain drugs is required in patients with reduced renal function to avoid
toxicity as many drugs are eliminated by the kidneys. Renal disease or reduced cardiac output
often reduces the clearance of drugs that depend on renal elimination. Alteration of clearance by
liver disease is less common but may also occur. Impairment of hepatic clearance occurs (for
high extraction drugs) when liver blood flow is reduced, as in heart failure, and in severe
cirrhosis and other forms of liver failure. Because it is important in the elimination of drugs,
assessing renal function is important in estimating dosage in patients. The most important renal
variable in drug elimination is glomerular filtration rate (GFR), and creatinine clearance (CLcr)
is a convenient approximation of GFR. Renal function is altered by many diseases and is often
decreased in older patients. CLcr can be measured directly, but this requires careful measurement
of both serum creatinine concentration and a timed total urine creatinine. A common method that
requires only the serum (or plasma) creatinine measurement (Scr) is the use of Cockcroft Gault
equation.
For males:
CLcr (mL/min) = (140 - Age) × body weight (kg)

72× Scr
For females:
CLcr (mL/min) = (140 - Age) × body weight (kg) ×0.85

72× Scr
The dosage in a patient with renal impairment may be corrected by multiplying the average
dosage for a normal person times the ratio of the patient’s altered creatinine clearance (CLcr) to
normal creatinine clearance (approximately 100 mL/min, or 6 L/h in a young adult).
Corrected dosage= Average dosage× Patient s CLcr

100 mL/min
This simplified approach ignores non-renal routes of clearance that may be significant. If a drug
is cleared partly by the kidney and partly by other routes, formula given below should be applied
to the part of the dose that is eliminated by the kidney. For example, if a drug is 50% cleared by
the kidney and 50% by the liver and the normal dosage is 200 mg/d, the hepatic and renal
elimination rates are each 100 mg/d. Therefore, the corrected dosage in a patient with a
creatinine clearance of 20 mL/min will be:
20ml/min
Dosage = 100𝑚𝑔 (𝑙𝑖𝑣𝑒𝑟) + 100mg/d × 𝑘𝑖𝑑𝑛𝑒𝑦
100𝑚𝑙/𝑚𝑖𝑛
Dosage= 100 mg/d + 20 mg/d = 120 mg

Calculations
1. A patient is administered with IV theophylline injection and target plasma concentration
is 20 mg/L to relieve acute bronchial asthma in a patient. Mean clearance for 70 kg
patient is 2.8 L/h/70 kg. After the relieve of asthma attack, the clinician might want to
maintain this plasma level using oral theophylline, which might be given every 12 hours
using an extended-release formulation to approximate a continuous intravenous infusion.
Calculate the maintenance and loading dose.
2. Thom, a 70 kg patient has been prescribed an anti-inflammatory drug with the following
information printed on its monograph: Vd = 50 L, Half-life (t ½ ) = 3 h, Target
concentration = 3.5 mg/L. What is dosing interval? Calculate the maintenance dose of
this drug.

3. Neena, a 20 kg child, has strep throat and was prescribed an antibiotic drug every 12 hr to
treat the infection. The drug monograph gives you the following information: Vd = 0.5
L/kg, Half-Life (t 1/2) = 2 h, Dosage interval = 3 h, Target concentration = 150 mg/L
Bioavailability = 70%. Calculate the loading and maintenance dose of antibiotic.
4. A patient weighs 65 kg. He is taking an NSAID with the following properties: Vd = 1.5
L/kg, Half-life (t 1/2) = 5 h, Target concentration = 4.5 mg/L. Calculate the loading dose
of this drug.

5. Mr Jones (age=40 yrs; wt=76 kg) has zero kidney function and is undergoing
hemodialysis while awaiting a kidney transplant. He takes metformin for type 2 diabetes
mellitus and was previously stabilized (while his kidney function was adequate) at a
dosage of 500 mg twice daily, given orally. Metformin excretion through urine is 90%.
The plasma concentration at this dosage with normal kidney function was found to be 1.4
mg/L. He has been on dialysis for 10 days and metformin toxicity is suspected. A blood
sample now shows a metformin concentration of 4.2 mg/L. Calculate the corrected
dosage for the patient.

Experiment No. 12
Objectives
Study of different parts of prescription and learning of prescription writing

“A prescription is a written order from the physician to the pharmacist so that a drug or a
combination of drugs from pharmacy is dispensed to patient. It contains directions to the
pharmacist and also for the patient regarding its use”.
Types of prescription
Prescriptions are of two types based on availability of prescribed drugs
i) Pre-compounded prescription
ii) Extemporaneous (compounded) prescription
Pre-compounded prescription
A pre-compounded prescription order is one that calls for a drug or mixture of drugs supplied by
the pharmaceutical company by its official or proprietary name and pharmacist dispense the
same in the form available without making any pharmaceutical alteration.
Extemporaneous prescription
An extemporaneous or compounded prescription is the one in which physician selects the drugs,
doses and form of the preparation that he/she desires and the pharmacist prepares the medication
accordingly.
Format: Traditionally a prescription is written in a definite order that facilitates its
interpretation. Prescription begins with name, age, address, and diagnosis of the patient on the
left hand side and date on the right side.
Parts of prescription
Main body of prescription consists of following parts
1. Superscription
2. Inscription
3. Subscription
4. Signatura
5. Physician’s signature

1. Superscription
This is the sign Rx which is instruction to the pharmacist. It is derived from Latin word recipe,
meaning “take thou”.
2. Inscription
It is the main part of the prescription that contains name of drug, dosage form and strength.
3. Subscription
It contains direction for the pharmacist including size of each dose, amount to be dispensed and
form of drug.
4. Signatura
It contains direction for the patient, usually precede by the symbol Sig: the place where the
physician indicates instructions. These may include
(a) The method of administration and application
(b) The dose if the preparation is for internal dose
(c) The time of administration or application
(d) The diluents (e.g. water) if relevant, or means of application (e.g. brush)
(e) The part of body where the preparation is to be applied, in case of external use
5. Physician’s signatures
These are written on the right hand side at the end of prescription. It includes;
i). Physician signature.
ii). Professional qualification.
iii). License Number
iv). Address
General considerations
1) It should be in National language.
2) It should be legible.
3) Abbreviations should be avoided.
4) Preferably the doses should be written in metric system.
5) Medicines should be written in serial number.
6) Every prescription should be signed.
7) Form & stregtn of every medicine should be written.
8) Full name rather than formula of the medicine should be written.

9) Instruction to patient for intake of medicine should be complete.
10) Instruction regarding diet & medicine should be complete.
11) Instruction regarding diet & medicine should be written at the end.
Abbreviation used in Prescription Writing
Following are the most commonly used abbreviations while writing the prescription
Abbreviations Latin Meaning

R, Recipe You take
b.d. Bis-die Twice a day
B.i.d Bis in die Twice a day
T.i.d. Ter in die Thrice a day
t.d.s. ter die soinendum Thrice a day
q.i.d. quarter in die 4 times a day
o.d. Once daily Once daily
s.o.S si-opus-sit If necessary
P.r.n. Pro-re-wata When necessary
Stat Statin Immediately
a.c. Ante cibos Before meal
p.c. Post cibos After meal
H.S. Hora somni At bed time
hor.dec Hora decabitus At bed time
q.d. Qnaqua die Every day
q.h. Qnaqua hora Every hour
Omn. hor Omni hora Every hour
Omn. bih Omin bi horio Every 2 hour
Omn 4 hor Omin Quarta hora Every 4 hour

C cum with
S sine without
Q.S. Quantum stis sufficient quantity
q.lib Quantum libitum Quantity as much as one like
q.n.s. Quantum not status Quantity not sufficient
a.a. Ana of-each
Ad Ad to, upto
Sig Sigma Label
p.o. per os By mouth
noct nocte At night
nebul nebula A spray
Placebo Placebo To satisfy
Garg Gargarisma Gargle
Agit Agita Shake

Prescriber’s Name:
License No:
Timing:
Contact No:
Patient’s Information
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
Presenting Complains
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

License No:
Timing:
Contact No:
Name _________________ Age: ________________
Gender________________ Weight______________
Address _________________________ Date: _______________
History ℞
Diagnosis
Refill Information
Sig.----------------------

Experiment No. 13
Objectives
Handling of Laboratory Animals

Material Required
Rabbits, Rats, Gloves

The most commonly used animal in undergraduate pharmacology laboratory are rabbits and rats.
It is very important to learn the animal handling properly to carry out the practical and reproduce
the results.
Rabbit Handling
 Animals should be handled o n l y in the animal house. Always wear lab coat before
handling the animals. Do not panic the animal and grasp the animal gently
without harming.
 Rabbits can be grasped by gripping the scruff of the neck firmly with one hand and their
body with the other hand or they may be held in a wooden animal box for the purpose
of injections. If the animal is frightened, put a cloth around the legs to prevent
scratching (Fig. 12a).
 The area should be rough, in case of slippery area, the rabbit may get irritated due to
slipping of feet. Hold the animal firmly and gently and never let it go if the animal get
frightened or irritated.
 If the rabbit is in a box which restrains movements without undue pressure, then only
one person is needed for giving injection or instillation of eye drops. Sometimes if the
animals are much frightened and aggressive, it is advisable to wear the woolen gloves.
Rat Handling
 Frequent handling of rats in a firm, yet gentle, manner will accustom them to manipulation.
Rats may be removed from their cage or primary enclosure by grasping the tail 1–2 cm
from the base (Fig. 12b). One may restrain the rat by any of the following means;
 Grasp the loose skin over the back and neck with the non-dominant hand (Fig. 12c).
 Firmly grasp the rat around the thorax with one thumb or index finger under the mandible
to prevent being bitten (Fig. 12d). Grasping with the non-dominant hand will give the
restrainer use of the dominant hand for animal manipulations or to support the animal’s

hind-quarters. With single hand restraint, the tail may be held between the fourth and fifth
finger
 The rat may be palmed over its back with the index and middle finger placed between its
mandible to minimize the chance of being bitten
12a
12b 12c
12c 12d
12a
12a

Experiment No. 14
Objectives
Appropriate and inappropriate drug administration through different routes.

Theory
Route of administration affects the onset and duration of action of drugs. So it is important to
select the appropriate routes of administration.
The selection of routes of drug administration is governing by various factors:
 Physical and chemical properties of the drug. The physical properties are solid, liquid and
gas. The chemical properties are solubility, stability, pH, irritancy etc.
 Site of desired action: the action may be localized and approachable or generalized and
not approachable.
 Rate of extent of absorption of the drug from different routes.
 Effect of digestive juices and the first pass metabolism of drugs.
 Condition of the patient.
Ketamine and Xylazine are used in combination (50 mg/kg: 5mg/kg) for anesthesia
intraperitoneally.
Materials
Ketamine, Xylazine, NG tubes, gloves, Disposable syringe (1 ml)
Subject
Rats
Procedure
 Arrange all the material required to perform the practical
 Prepare the cocktail of ketamine and Xylazine by mixing the 10:1 ratio
 Divide the rats into two groups
 Weight the rat and administer the cocktail 0.2ml/100 g of rat intraperitoneally
 Administer the same dose to rat of other group orally with NG tube
 Observe the onset and duration of action of the drugs in both groups
Observations
Observe the animals of both groups for the onset of action of the drug by loss of reflexes. To
check the reflexes, pinch or press the paw or tail of the rate forcefully after every 2 minutes upto
10 minutes and observe any movement of the rat. Note down the time of loss of reflex.
Continuously observe the animal and note the times when reflexes are restored

Observations
Route of Weight of Dose of Time of Onset Reflex Duration

administration rat drug admn. time restoration time of action
Results:

Experiment No 15
Objectives
To Study the Effects of Drugs on Rabbit’s Eye

Principle:
The parameters like corneal reflex, light reflex, and pupillary diameter are studied before and
after adding drug.
 Corneal reflex is studied by touching the cornea with a cotton probe. The rabbit immediately
closes the eye lid.
 The light reflex is studied by directing the light rays into the rabbit eye and pupillary
diameter is reduced.
 The diameter of the pupillary size is measured by a scale
Apparatus:
Scissors, Scale, Dropper, Torch, and Cotton plug.
Subject:
Rabbit
Drugs:
Pilocarpine, Atropine, Phenylephrine, Proparacaine
Procedure:
1. Hold the animal from trunk
2. Trim the eye lashes of both eyes with scissors
3. Consider one eye as control and other as experimental
4. Test and record the size of pupil, color of conjunctiva, corneal reflex and light reflex of both
eyes of the rabbit
5. Then tilt the head of the animal to opposite side and block naso-lacrimal duct with thumb
6. Put a few drops of drug in experimental eye
7. Allow the drug to be absorbed fully in eyes
8. Then record the observations after 0, 5, 10, 15, 30 and 45 min and tabulate the results

Precautions:
 Care should be taken not to injure the eyes
 Before cutting, moisten the eye lashes to avoid eye lashes from getting into eye of animal
 Mark one eye as standard/control for comparison with experimental
 Block the naso-lacrimal duct for 1-2 min to avoid drainage of drug into nose
 Head of the animal should be tilted backwards allowing the drug to remain in the eye for few
min for its adequate absorption and action
 The observation should be taken in dim light and entry of light rays directly into the eyes
should be avoided

Observations & Calculations
1. Pilocarpine HCL:
Time Size of Pupil Light Reflex Corneal Conjunctival Dryness of
(min) (mm) Reflex Color Eye
C E C E C E C E C E
0
5
10
15
30
45
2. Atropine Sulphate:
C E C E C E C E C E
0
5
10
15
30
45

3. Phenylephrine HCL:
C E C E C E C E C E
0
5
10
15
30
45
4. Proparacaine HCL:
C E C E C E C E C E
0
5
10
15
30
45
Result:

Experiment No 16
Objective
To Study the Phenomenon of Potentiation on Rabbit’s Eye

Principle:
The parameter pupillary diameter is studied before and after adding drug.
 The diameter of the pupillary size is measured by a scale.
Apparatus:
Scissors, Scale, Dropper
Subject:
Rabbit
Drugs:
Tropicamide 1%, phenylephrine 2.5%
Procedure:
 Take three rabbits in each group and mark them as A, B and C
 Hold the animal from trunk. Trim the eye lashes of both eyes with scissors.
 Measure and record the size of pupil of both eyes of the rabbit before addition of drug
 To animal A, mark one eye as control and other as experimental. Then tilt the head of the
animal to opposite side and block the naso-lacrimal duct with thumb. Put a few drops of
tropicamide 1% in experimental eye. Allow the drug to be absorbed fully in the eyes. Then
record the observations of both eyes after 0, 5, 10, 15, 30 and 45 min and tabulate the results
 To animal B, add few drops of phenylephrine 2.5% to experimental eye and record the
observations
 To animal C, add few drops of tropicamide and phenylephrine with interval of 5 minutes in
both eyes and record the observations.
Precautions:
 Care should be taken not to injure the eyes
 Before cutting, moist the eye lashes to avoid eye lashes from getting into eye of animal
 Block the naso-lacrimal duct for 1-2 min to avoid drainage of drug into nose
 Head of the animal should be tilted backwards allowing the drug to remain in the eye for few
min for its adequate absorption and action
 The observation should be taken in dim light

Time
Animal A Animal B Animal C
(min)
Tropicamide Tropicamide
Control Tropicaide Control Phenylephrine + +
Phenylephrine Phenylephrine
0
5
10
15
30
45
Results:

Experiment No. 17
Objective
To Study of the Effects of Cholinergic Drugs (Agonism) on Rabbit’s Intestine

Apparatus:
Dissection box, kymograph, smoke drum, isolated organ bath, petri-dish, thermometer, syringe,
beakers, stitching needle, thread, stop watch, water bath, funnel.
Subject:
Rabbit
Physiological salt solution:
Tyrode solution
Drugs
Acetylcholine, Pilocarpine
Procedure:
1. Set kymograph and organ bath on table, following the washing of organ bath. Organ bath
consist of two jackets outer and inner. Fill outer jacket with water and turn it ON. Maintain
the temperature at 38-40°C. Then fill the inner jacket with Tyrode solution and mark the
level. Maintain this level and temperature throughout the experiment. Aerate the Tyrode
solution with oxygen continuously.
2. Dissect the rabbit and dissect out the rabbit intestine, cut a piece of rabbits intestine 2cm in
length and keep it in petri dish containing Tyrode maintained at temperature of 37 0C. Clean
and remove the fecal matter from this tissue. Then pass a thread through one end of the tissue
and tie a knot. Similarly pass another thread from other end of this tissue and tie it.
3. Now suspend this tissue in inner jacket of organ bath with the help of tissue suspending hook
by tying one end of thread (loop) with hook and other with the lever then adjust the lever so
that it moves smoothly against the kymograph drum. Wash the tissue by replacing the Tyrode
solution and allow it to take rest for at least 2-3 min. before the start of experiment. Take
muscle contractions without adding any drug until become regular and uniform.
4. Add few drops of acetylcholine into inner jacket of the organ bath and record the muscle
contractions for few seconds.

5. Rinse the tissue with temperature maintained (37 0C) Tyrode solution until contractions
reaches to baseline.
6. Then adds few drops of pilocarpine and record the contraction for few minutes and rinse the
tissue.
7. Add few drops of acetylcholine, at the height of contraction, add the pilocarpine and record
the contractions.
Precautions:
 Wash the inner jacket at least 2 to 3 times with Tyrode solution
 Temperature of outer and inner jacket should be consistently maintained throughout the
experiment at 37 0C
 Clean the tissue properly to avoid the damage of enteric plexus
 Tissue should be suspended vertically in such way to avoid direct contact with inner
walls & suspended loop
 Keep the tissue all the time in Tyrode solution
 Maintain the level of Tyrode soln. before & after washing
 Dosing interval for rabbit intestine is 3 mins
 Different syringes should be used for different drugs
 No air bubble should present during measuring the dose of drug

Magnitude of Magnitude of
Drug contraction contraction after Difference Remarks
before drug drug
Acetylcholine
Pilocarpine
Acetylcholine +
Pilocarpine
Results

Experiment No. 18
Objectives
To Study of the Effects of Cholinergic Drugs with Anticholinergic Drugs (antagonism) on

Rabbit’s Intestine
Apparatus:
Dissection box, kymograph, smoke drum, isolated organ bath, petri-dish, thermometer, syringe,
beakers, stitching needle, thread, stop watch, water bath, funnel.
Subject:
Rabbit
Physiological salt solution:
Tyrode solution
Drugs
Acetylcholine, Atropine
Procedure:
1. Set kymograph and organ bath on table, following the washing of organ bath. Organ bath
consists of two jackets; outer and inner. Fill outer jacket with water and turn it ON. Maintain
the temperature at 38-40°C. Then fill the inner jacket with Tyrode solution and mark the
level. Maintain this level and temperature throughout the experiment. Aerate the Tyrode
solution with oxygen continuously.
2. Dissect the rabbit and dissect out the rabbit intestine, cut a piece of rabbits intestine 2cm in
length and keep it in petri dish containing Tyrode maintained at temperature of 37 0C. Clean
and remove the fecal matter from this tissue. Then pass a thread through one end of tissue
and give a knot. Similarly pass another thread from other end of this tissue and tie it.
3. Now suspend this tissue in inner jacket of organ bath with the help of tissue suspending hook
by tying one end of thread (loop) with hook and other with the lever then adjust the liver so
that it moves smoothly against the kymograph drum. Wash the tissue by replacing the Tyrode
solution and allow it to take rest for at least 2-3 min. before the start of experiment. Take
muscle contractions without adding any drug until it becomes regular and uniform.
4. Add few drops of acetylcholine into inner jacket of the organ bath and record the muscle
contraction for few seconds.

5. Rinse the tissue with temperature maintained (37 0C) Tyrode solution until contractions
reaches to baseline.
6. Add few drops of acetylcholine, at the height of contraction, add the atropine and record the
contractions.
Precautions:
 Wash the inner jacket at least 2 to 3 times with Tyrode solution
 Temperature of outer and inner jacket should be consistently maintained throughout the
experiment at 37 0C
 Clean the tissue properly to avoid the damage of enteric plexus
 Tissue should be suspended vertically in such way to avoid direct contact with inner
walls & suspended loop
 Keep the tissue all the time in Tyrode solution
 Maintain the level of Tyrode soln. before & after washing
 Dosing interval for rabbit intestine is 3 mins
 Different syringes should be used for different drugs
 No air bubble should present during measuring the dose of drug

Magnitude of Magnitude of
Drug contraction contraction after Difference Remarks
before drug drug
Acetylcholine
Atropine
Acetylcholine +
Atropine
Results:

Experiment No. 19
Objectives
To study the Effects of Drugs on Frog's Heart

Theory
Efficiency of the heart as a mechanical pump depends on auto-rhythmicity, conductivity and

contractility, which are inherent properties of the myocardium but are also under the
influence of autonomic nervous system. The parasympathetic supply is inhibitory in its
action, whereas the sympathetic supply causes stimulation.
Drugs
Adrenaline 1: 20,000
Propranolol 1 mg/ml
Acetylcholine 1 : 100,000
Atropine I: 10,000
Digoxin 0.25 mg/ml
Normal Saline (0.6 % NaCl for frog)
Apparatus
Surgical kit, frog board, stands with mounted spring lever, kymograph and revolving drum.
Procedure
Stun and pith the frog and fix it on the frog board with the help of pins. Expose the heart by
cutting the sternum (saving the ventral vein). Fix the hook of the lever in the apex of the
heart. Lever moves with normal cardiac contractions which are recorded on the revolving
drum of the kymograph. Take the normal tracing per minute. Pour a few drops of the
given drug on the exposed heart and record the effects. Compare the effects produced by
various drugs with the preceding normal and write your inference.
Precautions
1. Take a fresh normal tracing before using every drug and compare the effects of
subsequently used drug with it.
2. After using a drug wash the heart thoroughly with normal saline and give rest for five
to ten minutes before applying the next drug.

Observations
Amplitude of Contraction Rate of Contraction

Drugs Used Inference
(mm) (per min)
Results:

Experiment No. 20
Objectives
To study the Effect of Coffee on Pulse rate and Blood Pressure on Normal Human volunteers
Theory
Caffeine is an alkaloid present in coffee, chocolate, tea and cola-drinks. It is a cardiac and CNS
stimulant and also produces diuresis. 30-60 mg caffeine is present in 150 ml coffee and 360 cola
drink. In larger doses, caffeine causes insomnia, palpitations.
Procedure
 Select volunteers randomly without any discrimination of age, sex
 Measure the blood pressure and pulse rate
 Administer a cup of coffee containing a teaspoon of coffee and 2 teaspoons of sugar
mixed in 250 ml of water
 Count the Pulse and measure the blood pressure before taking the drink i.e. zero min
and then at regular intervals of 30 min for 2 hours
 Calculate the standard deviation and standard error for each data
 Compare the 0 minute data of the two groups by applying Student's t test and find out
the statistical significance

Systolic blood pressure Deviation from mean

Obs.no d2
(mmHg) D
Group 1
Mean ∑d2 =
value= 𝑋̅1
Group 2
Mean ∑d2 =
value= 𝑋̅2


Experiment No. 21
Objectives
Study the anticoagulant activity of different anticoagulants in blood collected from Orbital
Plexus or Sinus of rat
Materials:
Blood sampling test tubes, Capillary tube, gauze sponge or swab, Heparin, Sodium citrate,
Ketamine, xylazine
Theory
Blood collection is a common practice in clinical setups as well as in research laboratories. In
research laboratories, blood is collected from animals for clinical trials of drugs and in vitro
blood testing. The blood can be collected from animals by different method; i.e. Orbital Sinus,
Heart Puncture and from veins.
Whenever your skin becomes broken, blood vessels are damaged, blood is released and the
sticky platelets contained in the blood form clots to stop blood flow. As soon as blood from a
wound is exposed to the air, the platelets disintegrate and react with fibrinogen to create fibrin, a
mass of tiny threads. This triggers a whole series of sequential reactions that rely on adequate
levels of calcium and vitamin K to work and this process is known as the ‘clotting cascade’. The
fibrin hardens very quickly to form a scab, sealing the wound. The wound heals and the clot
dissolves. Unless prevented from doing so, blood collected into glass bottles or blood bags
undergoes the same process and will clot.
Procedure:
 Mark the blood sampling test tubes as A (without anticoagulant), B (containing
coagulant, 4% sodium citrate 0.5ml) and C (10-30 USP unit/ml heparin)
 Manually restrain and anesthetize (ketamine 50mg/kg and xylazine 5mg/kg) the animal
 Introduce the end of the micro hematocrit tube/ Capillary tube at the canthus of the orbit
when animal become fully anesthetized
 Lateral canthus: Pick up the rat and restrain it in one hand. Insert the tube into the lateral
canthus. The tube should be at about a 30 degree angle to the side of the head and swirl it
slowly
 Medial canthus: Place the rat on a table or cage lid in lateral recumbancy. The body of the
animal is restrained against the table with the palm of the hand. The thumb and
forefingers of the same hand restrain the hand and gently open the eyelids to expose the
eye. Insert the tube into medial canthus and hold it at a 30 degree angle to the nose
 Insert micro hematocrit tube into the orbital sinus/plexus by quickly rotating the tube.
The eye is not damaged because the tube passes under the eye ball. Blood flow may be
increased by changing the angle of the tube slightly
 After the required amount of blood is obtained, the tube is withdrawn and bleeding
usually ceases by the eye (orbit) pressure alone. If necessary, hemorrhage can be
controlled by direct pressure using cotton over the eyelids

 Observe the animal for recovery from anesthesia
 Observe the sample tubes for clotted and un-clotted blood.
 Place the test tubes in centrifuge machine and centrifuge the blood at 3000 rpm for 15
minutes
 Observe either the serum separated in both tubes

Observations and Calculation
Results:

Experiment No 22
Objectives
Evaluation of Analgesic Activity of Diclofenac Sodium against Formalin Induced Pain Model
Analgesia:
An analgesic, or painkiller, is any member of the group of drugs used to achieve analgesia (relief
from pain without loss of consciousness).
Formalin:
The formalin model is widely used for evaluating the effects of analgesic compounds in
laboratory animals. Injection of formalin into the hind paw induces a biphasic pain response; the
first phase is thought to result from direct activation of primary afferent sensory neurons,
whereas the second phase has been proposed to reflect the combined effects of afferent input and
central sensitization in the dorsal horn. Formalin excites sensory neurons by directly activating
TRPA1, a cation channel that plays an important role in inflammatory pain. Activation of
channel, which is permeable to Na+, Ca2+ and other cations, causing depolarization and initiation
of action potentials.
Diclofenac Sodium:
Diclofenac is a nonsteroidal anti-inflammatory drug (NSAID). It works by reducing
inflammatory mediators in the body that cause pain and inflammation. Diclofenac has analgesic,
anti-inflammatory, and antipyretic properties and also used to treat signs and symptoms
of osteoarthritis or rheumatoid arthritis. The mechanism of action is not completely understood
but involves inhibition of cyclooxygenase (COX-1 and COX-2). Because diclofenac is an
inhibitor of prostaglandin synthesis, its mode of action may be due to a inhibition of
prostaglandins synthesis in peripheral tissues.
Subject:
Albino Rats (150-250g of either sex)
Procedure:
 Animals are randomly divided into 2-groups each consisting of 2-animals

 The first group serves as control group receives only vehicle (Distilled water 5 ml/kg i.p.)
 The second group of animal receives standard treatment Diclofenac (25 mg/kg i.p.)

 After 30min of treatment, all the group animals are subjected to pain stimulus by
administering 0.05ml of 2.5% formalin subcutaneously into the right hind paw of rats
 The time spent on licking the injected paw by each rat is observed as soon (early phase 0-5
min, post-injection) as the formalin is injected and later (late phase 15-40 min, post
injection). The percentage inhibition is calculated by the given formula:
Percentage Inhibition = N- Nt / N x 100
N = Time spent on licking/ biting of Vehicle group

Nt = Time spent on licking/ biting of Treatment group/ standard

Observation and Calculation:
Sr. Weight Dose Treatment Licking Response %age

No. Inhibition
0-5 15-40 1st 2nd
(min) (min) Phase Phase
Results:

Experiment No 23
Objectives
Determination of Anti-Depressant Activity of Fluoxetine in Rat by Tail Suspension Test

Theory
The most common symptoms of depression are restlessness, sleeplessness, loneliness and give
up behavior. Antidepressant drugs increase the serotonin level in body which can jump-start the
mood and boost the activities of the subject. These drugs increase the mobility activities. For
evaluation of anti-depressant activity in rats, experimental animals are divided into 2-groups
each consisting of 2-animals.
Groups:
Normal Control: Normal saline (10ml/kg, i.p)
Experimental Group: Fluoxetine (10mg/kg, i.p)
Tail Suspension Test

Tail suspension apparatus consists of wooden chamber (70cm height x 47cm width x 25cm
depth)
1. A rod is fitted between side walls of chamber, at a height of 60cm from ground or 10cm
from top of the apparatus
2. Animals are hung with the rod by placing adhesive tape one inch from tip of tail
3. Animals are given a pretest session of 15 minutes each, 24 hours before final test session
4. Animals are treated with respective group treatments immediately after pretest session, 6
hours before final test session and 30 minutes before final test session
5. 30 minutes after final dose or 24 hours after pretest session, each animal was individually
hung with rod for final test session of 5 minutes each
6. Duration of immobility is noted for each animal for 5 minutes. Rat is considered immobile
when it passively hung with rod with no efforts to escape

Duration of Immobility
Group Duration of Mobility (s)
(s)
Control
(N/S 10 ml/kg)
Fluoxetine
(10 mg/kg)
Result:

Experiment No 24
Objectives
Determination of Anxiolytic Effect of Diazepam in Rat by Light/Dark Exploration Test

Theory
Diazepam relieve the symptoms of anxiety such as restlessness, doom, panic and tiredness.
Diazepam increase the duration of rodents stay in light as compare to stay in darkness. For
evaluation of anxiolytic activity in mice, experimental animals are divided into 2-groups, each
consisting of 1-animal.
Groups:
Experimental Group: Diazepam (1mg/kg, i.p)
Light/Dark Exploration Test:
Procedure:
1. The apparatus consists of two adjoining boxes (25 × 25 × 25 cm3) with a wall between them.
2. One box is covered at the top and is painted black (dark room), the other box is open from
top and is painted white (light room)
3. The light box is illuminated with 40 watt lamp placed at 25cm height
4. The center wall has a hole through which mouse can freely pass into both boxes
5. Apparatus is placed in a room having dim illumination
6. Animals are treated with respective group treatment and after 30 minutes, they are placed on
Light/ Dark exploration apparatus at the hole, facing dark box
7. The time (in seconds) spent by each rat in light and dark box will be recorded for 5 minutes
Light/ Dark Exploration Apparatus

Group Time Spent in Light (s) Time Spent in Dark (s)
Control (N/S 10 ml/kg)
Diazepam (1 mg/kg)
Results:

Experiment No 25
Objectives
Determination of Anxiolytic Effect of Diazepam in Rat by Elevation plus Maze Test

Theory
Diazepam increase the duration of rodents stay in open area as compare to stay in closed area.
For evaluation of anxiolytic activity in mice, experimental animals are divided into 2-groups
consisting of 1-animal each.
Experimental Group: Diazepam (1mg/kg, i.p)
Elevated Plus Maze Test:
Procedure:
EPM test is most widely used and validated test to measure anxiety in rodents
1. Apparatus consists of four arms of which two are open and two close respectively
2. Open arms (45 × 10 cm2) are crossed with closed arms (45 × 10 × 30 cm3) at a center point
(10 × 10 cm2)
3. EPM is elevated to a height of 70 cm from the ground and placed in a room having dim
illumination
4. Animals are treated with respective group treatment and after 30 minutes, they are placed on
EPM apparatus at the center, facing one of the closed arms
5. The time (in seconds) spent by each rat on open and closed arms is recorded for 5-miuntes
Elevated Plus Maize Apparatus

Observations and Calculations:
Time Spent in Open Arms Time Spent in Closed

Group
(s) Arms (s)
Control (N/S 10 ml/kg)
Diazepam (1 mg/kg)
Result:

Experiment No. 26
Objectives
To Study the Effects of Drugs on Reflex time in Frog

Theory
Reflex time may be defined as the time that elapses between application of the stimulus
and appearance of the reflex response. The elements involved in such reflex activity are,
excitation of sensory receptors, conduction over afferent nerve fibers and finally, after
specific CNS activity, excitation of the motor nerves supplying the effector m uscle. Reflex
time may be increased by the drugs, which either depress the CNS or block the
neurotransmission of impulses. It is decreased by the drugs that stimulate the CNS
Materials
Frog, Stand, Beakers, Syringe, Dissection Kit, Thread with hook etc.
Drugs
Diazepam 0.5%
Caffeine Citrate Lignocaine 1.0%
Mucilage of Acacia 4.0%
HCl 20.0 %
1 :1000 (Used as stimulus)
Procedure
Take a frog and decapitate it, hang with the stand. Note the normal reflex time of both legs
by dipping them in HCI solution separately. Take the mean of three readings for each leg.
The effects of drugs are studied in the following manner
a. Immerse one leg in 20% mucilage of Acacia for 5 minutes. Note the reflex time and
compare it with the normal reflex time of that [articular leg. Wash the leg with tap water.
Apply 4% lignocaine solution to one of the legs for 5 minute. Note any change in reflex
time
b. Inject 0.5 ml of caffeine citrate into the ventral sac of the frog and note the change
in reflex time, 15 minutes after injection of the drug. Allow sufficient time for recovery
of the animal from the effects of caffeine. Then inject 1 .0 ml of diazepam solution into
the ventral sac of the frog. Note any change in reflex time 1 5 minutes after
administration of the drug

Preca utions
 Dip the same length of leg in HCl and drug solution
 Take a fresh normal reading before using every drug and compare the effect of
subsequently used drug with the preceding normal reading
 After studying effects of a drug, allow sufficient time for recovery from the effects
of drugs used before using the next drug
 After d ipping the leg in HCl solution, wash it thoroughly with tap water
immediately after noting the reflex time

REFLEX TIME (Sec)
DRUG USED RIGHT LEG LEFT LEG INFERENCE
1 2 3 mean 1 2 3 mean
Normal
Mucilage of Acacia
Normal
Lignocaine
Normal
Caffeine Citrate
Normal
Diazepam

Experiment No. 27
Objectives
Identification of Drug
Procedure
1. Visit a pharmacy
2. Collect empty packets and blisters of drugs
3. Identify the generic name, formula/ composition of these preparations
4. Enumerate the classification / sub classification of each group of collected packs of
drugs
5. List uses of each substance / ingredient collected
Substance /
Name of the Classification/ Uses of each substance &
ingredient /formula
drug grouping ingredients
name
Brand Generic Drug 1
name name
a.
b.
c.
d.
Drug 2
a.
b.
c.
d.
Drug 3
a.
b.
c.
d.

Experiment No. 28
Objectives
Identification of Unknown Drug

Procedure
1. Analyze the graph of unknown drug labeled as “A, B, C and D” of frog heart.
2. Enumerate the change in inotropic and chronotropic effect after drug administration.
3. Calculate the heart rate per minute before and after drug administration and change in
heart rate after drug administration.
4. Calculate the percentage change in force of concentration.
5. Name the possible drug “A, B, C and D”.

Calculations

Calculations

Experiment No. 29
Objectives
Determine the Frequency of Analgesic and Anti-inflammatory Drugs in Medical / Surgical / OPD
Procedure
1. Visit on OPD or ward in hospital

2. Take permission from relevant authority
3. Take consent of the patient
4. Select patient taking analgesics and anti-inflammatory drugs
5. Inquire and record following information
6. Analyze the information collected
Generic name Frequency, Relieve in pain &

Patient Name, Age & Drug brand
/Formula / Doses duration inflammation on 1 Unwanted effects
Presenting Complain name
Composition of treatment – 10 scale

Experiment No. 30
Objectives
Determine the Frequency of Hypoglycemic Drugs in Medical / Surgical / OPD

Procedure



Experiment No. 31
Objectives
Determine the Frequency of Antihypertensive Drugs in Medical / Surgical / OPD

Procedure



Experiment No. 32
Objective:
Evaluation of Effects Produced by Physical Activity on ECG with and without Drug
Theory:
Propranolol is beta antagonist used to prevent angina (chest pain), migraine headaches, and to
improve survival after a heart attack. This practical will illicit the effect of Propranolol in normal
subjects.
Material Required:
ECG machine, Treadmill, Inderal (Propranolol) 10mg tablets,
Procedure:
Day 1: The volunteer subjects are selected after obtaining consent, ECG will be recorded after 10
minutes of exercise on treadmill.
Day 2: Tab Propranolol 10 mg is ingested by the subject is allowed to rest for 1 and a half hours
and then after 10 minutes of exercise ECG will be recorded.

Observations and Calculation
Subjects S-T segment T segment Heart rate Unwanted Effects
DAY 1 DAY 2 DAY 1 DAY 2 DAY 1 DAY 2 DAY 1 DAY 2
Results

Experiment No. 33
Objective
To study the Effect of Drug on Lung Functions using Spirometer

 Confirm the presence of airway obstruction
 Help assess response to therapy
 Confirm an FEV1/FVC ratio < 0.7 after bronchodilator
 Provide an index of disease severity
 Aid in predicting prognosis and long-term survival
Theory
Spirometry (meaning the measuring of breath) is the most common of the pulmonary function
tests (PFTs). It measures lung function, specifically the amount (volume) and/or speed (flow) of
air that can be inhaled and exhaled. Spirometry is helpful in assessing breathing patterns that
identify conditions such as asthma, pulmonary fibrosis, cystic fibrosis, and COPD. The
bronchodilator test is a method for measuring the changes in lung capacity after inhaling a short-
acting β2-agonist that dilates the airway.
Parameters
Forced vital capacity (FVC)
Forced vital capacity (FVC) is the volume of air that can forcibly be blown out after full
inspiration, measured in liters. FVC is the most basic maneuver in spirometry tests. The forced
vital capacity depends on the elasticity of the lungs and chest which, in turn, depends on age,
gender, chest size and physical condition. In females the value is 25% lower than males. Athletes
have exceptionally high values, especially swimmers due to their well-developed respiratory
muscles. Vital capacity is small if the elasticity of the lung decreases or if volume capacity
decreases.
Forced expiratory volume in 1 second (FEV1)

FEV1 is the volume of air that can forcibly be blown out in first 1 second, after full
inspiration. Average values for FEV1 in healthy people depend mainly on sex and age,
according to the diagram at left. Values of between 80% and 120% of the average value are
considered normal.

Tidal volume (TV)
Tidal volume is the amount of air inhaled or exhaled normally at rest.
Apparatus: Spirometer
Subject: Humans
Drug: Ventolin inhaler
Procedure
The subject is asked to take the deepest breath they can, and then exhale into the sensor as hard
as possible, for as long as possible, preferably at least 6 seconds. It is sometimes directly
followed by a rapid inhalation (inspiration), in particular when assessing possible upper airway
obstruction. Sometimes, the test will be preceded by a period of quiet breathing in and out from
the sensor (tidal volume), or the rapid breath in (forced inspiratory part) will come before the
forced exhalation.
During the test, soft nose clips may be used to prevent air escaping through the nose. Filter
mouthpieces may be used to prevent the spread of microorganisms.
After measurement of FVC and FEV1 in subjects, bronchodilator test is performed to check the
effect of short acting β2 -bronchodilator on lung functions.
In the (short-acting β2-bronchodilator) bronchodilator test, the subject fully exhales slowly, and
sprays an albuterol metered dose of 100 µg (1 puff) while biting a valved chamber. The subject
then slowly and deeply inhales until reaching TLC over 3–5 seconds, holds the breath for 5–10
seconds, and exhales. This procedure is repeated four times (total 400 µg of albuterol), at
intervals of 30 seconds. However, if there is any concern of affecting the pulse rate of the subject
or causing occurrence of hand tremors, the dose may be decreased to 200 µg. After inhalation of
the last medication, the spirometry test is conducted again between 10–20 minutes
FVC Predicted value equation

Male: (Height (m)*5.76)- (age*0.026)-4.34
Female: (height (m)*4.43)-(age*0.026)-2.89
FEV1 predicted value equation

Male: (height (m)*4.30)-(age*0.029)-2.49

Female: (height (m)*3.95)-(age*0.025)-2.60
Evaluation criteria
Percentage of predicted FEV1 Value Result

80% or greater Normal
70%-79% Mildly abnormal
60%-69% Moderately abnormal
50%-59% Moderate to severely abnormal
35%-49% Severely abnormal
Less than 35% Very severely abnormal
FVC Result
Is greater than or equal to the lower limit of normal Normal
Is less than the lower limit of normal Abnormal

Observations and calculations
Forced vital capacity (FVC) = tidal volume (Vt)+inspiratory reserved volume (IRV) +
expiratory reserved volume (ERV)
Treatment Tidal volume Inspiratory Expiratory Forced vital

(Vt) reserved volume reserved volume capacity
(L) (IRV) (L) (ERV) (L) (FVC)
(L)
Without drug
(control)
Drug
Forced expiratory volume (FEV1)
Treatment FEV1 (L)

Without drug (control)
Drug
FEV1 % of predicted=FEV1 (measured)/FEV1 (predicted)*100
RESULTS:

Experiment No. 34
Objective
Determination of antimicrobial activity of different antibiotics using disk diffusion method
Theory
Antimicrobials are agents that kill bacteria (bactericidal) or retard their growth (bacteriostatic).
They are widely used in medicine to treat microbial infections. The most common pathogens
detected with a sputum culture are bacteria such as Streptococcus pneumoniae, Haemophilus
influenza, Staphylococcus aureus and Klebsiella species.
Materials and chemicals
Bunsen burner, sputum sample, antimicrobial agents (A, B, C and D), nutrient agar plates,
incubator, airflow hood, filter paper discs sterilized by incubator, sterile forceps, adhesive tapes
and disinfectant solution.
Procedure
 A common, official and standard method used for antimicrobial activity is agar disk-
diffusion method.
 Take a dry and sterile petri dish. Divide the base into four sections by drawing a cross with
the marker pen. Label the sections as A, B, C and D
 Pour the cool, but still molten, MHA (Mueller Hinton Agar) medium in a conical flask or
bottle into the Petri dish in an aseptic environment
 Agar medium is allowed to freeze properly
 Inoculate the medium with sputum sample and spread it smoothly and evenly with the help
of sterile L shaped (hockey stick) glass spreader
 Cut the filter paper into disk shape of size of 6 mm diameter
 Prepare the solution of known concentration of all antibiotic drugs, by grinding the tablet,
if not in solution form and mark them as A,B,C and D
 Work very close to lit Bunsen burner. Flame the forceps and use them to pick up a filter
paper disc and dip the disc into antibiotic A.
 Allow it to dry for 5 minutes on an open, sterile Petri dish, next to a lit Bunsen burner.
 Follow the same procedure for other antibiotics B, C and D.
 Use the agar plate that has already been prepared and seeded with bacteria.
 Flame the forceps and then use them to pick up antibiotic disc A. Raise the lid of the Petri
dish at an angle and place the discs onto the agar in the center of respective sections

 Label the agar plate with your name and date. Tape the lid securely and give pre diffusion
time (30-60 m) and incubate the plates for 16-18 hours at 35±2 °C
 Observe the plates after incubation period without opening them
 Measure the size of the clear zone of inhibition around each antimicrobial disc and
compare with each other
Precautions
 Wear a coat to protect your clothes
 Keep your desk free of unnecessary materials at all times and at the end of the period
leave it free of all materials and equipment
 Since many of the microorganisms with which you will be working are potentially
pathogenic, it is imperative to develop aseptic techniques in handling and transferring
them
 Avoid any hand - to - mouth operations such as smoking, eating, drinking
 Report immediately all accidents such as cuts, burns or spilled cultures to your instructor;
take every precaution to avoid such accidents
 Some of the chemicals employed in the laboratory can be hazardous if not handled
properly, we have selected the experiments to minimize the use of such substances

Observation and calculations
Sample Antimicrobial agent Diameter of zone of inhibition (mm)
Results

NOTES

NOTES

NOTES

NOTES

NOTES

NOTES

NOTES

References
ANSEL, H. C. & PRINCE, S. J. 2004. Pharmaceutical calculations: the pharmacist's handbook,

Lippincott Williams & Wilkins.
ALLEN, L. & ANSEL, H. C. 2013. Ansel's pharmaceutical dosage forms and drug delivery
systems, Lippincott Williams & Wilkins.
ASHOK KUMAR, B. S., LAKSHMAN, K., VELMURUGAN, C., SRIDHAR, S. M. &

GOPISETTY, S. 2014. Antidepressant activity of methanolic extract of amaranthus spinosus.
Basic and clinical neuroscience, 5, 11-17.
ANSEL, H. C. 2012. Pharmaceutical calculations, Lippincott Williams & Wilkins.
JAJU, B.P. Pharmacology Practical Exercise Book, Jaypee Brothers
JOHNSON, C. W., TIMMONS, D. L. & HALL, P. E. 2003. Essential laboratory mathematics:

concepts and applications for the chemical and clinical laboratory technician, Cengage
Learning.
KLEMENT, P., LIAO, P., HIRSH, J., JOHNSTON, M. & WEITZ, J. I. 1998. Hirudin causes
more bleeding than heparin in a rabbit ear bleeding model. The Journal of Laboratory and
Clinical Medicine, 132, 181-185.
Najmi, H.M. and Khan, M.A. Handbook of Applied Pharmacology, M.N Publications.
PARK, J.-H., LEE, Y.-C. & LEE, S.-Y. 2009. The comparison of mydriatic effect between two
drugs of different mechanism. Korean journal of ophthalmology: KJO, 23, 40-42.
PATEL, M., ANTALA, B., BARUA, C. & LAHKAR, M. 2013. Anxiolytic activity of aqueous
extract of Garcinia indica in mice.
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Roll No……………….Class……...............…………………… has carried out the necessary

practical work as per course of studies for the year……………………………………………

Signature of Facilitator…………………… Date…………………

CMH INSTITUTE OF MEDICAL SCEINCES BAHAWALPUR

Practical Manual of Pharmacology i

Practical Manual of Pharmacology ii

1st Edition 2018

2nd Edition 2019

Practical Manual of Pharmacology iii

Practical Manual of Pharmacology iv

Understanding and Introduction to Equipment used in

Understanding of different systems of measurements and their

Study of different biological Salt Solutions used in vitro studies

Understanding and calculations of Serial and Stock Dilutions used

Study of different Types of Solutions and their calculations

6 Introduction of solid dosage forms used to administer the drugs 21

7 Introduction of liquid dosage forms used to administer the drugs 23

Introduction of Semisolids and others dosage forms used to

Biostatistics and Dose Calculation

9 Introduction to Biostatistics and its applications 28

10 Study of different formulas used for Dose Calculations in paeds 34

Understanding of loading and maintenance dose and dose

Study of different parts of prescription and learning of

Practical Manual of Pharmacology v

Appropriate and inappropriate drug administration through

15 To Study the Effects of Drugs on Rabbit’s Eye 71

16 To Study the Phenomenon of Potentiation on Rabbit’s Eye 75

To Study of the Effects of Cholinergic Drugs (Agonism) on

To Study of the Effects of Cholinergic Drugs with anticholinergic

19 To study the effects of drugs on frog's heart 83

To study the effect of coffee on pulse rate blood pressure on

Blood and Inflammation

Study the anticoagulant activity of different anticoagulants in

Evaluation of Analgesic Activity of Diclofenac Sodium against

Central Nervous System

Determination of Anti-Depressant Activity of Fluoxetine in Rat

Determination of Anxiolytic Effect of Diazepam in Rat by

26 To study the effects of drugs on reflex time in frog 100

Practical Manual of Pharmacology vi

27 Identification of drug 103

28 Identification of unknown drug 104

Determine the frequency of analgesic and anti-inflammatory drugs

Determine the frequency of hypoglycemic drugs in medical/

Determine the frequency of Antihypertensive drugs in medical/

Evaluation of effects produced by physical activity on ECG with

33 To study the effect of drug on lung functions using spirometer 114

Determination of antimicrobial activity of different antibiotics

Practical Manual of Pharmacology vii

Practical Manual of Pharmacology 1

Practical Manual of Pharmacology 2

Practical Manual of Pharmacology 3

Practical Manual of Pharmacology 4

Practical Manual of Pharmacology 5

Understanding of different systems of measurements and their interconversion used in medical

Importance of Measurement Systems

Hecta = 100 times

Kilo = 1000 times

Mega = 1000000 times

Practical Manual of Pharmacology 6

1 fluid ounce = 29.573 milliliters (mL)