Simple Notes on Biopharmaceutics and

Pharmacokinetics
(As per PCI Syllabus, for
IV Pharm. D students)
 Index
Page
NO
Unit I: Introduction to Biopharmaceutics
1.1 Journey of a tablet in the body 5
1.2 Definitions 5
1.3 Examples showing the relation between properties of drug, dosage 6
form characteristics, route of administration and bioavailability
1.4 Structure of cell membrane and its significance (importance) in 6
drug absorption
1.5 Mechanisms of drug absorption 6
1.6 Biological Factors affecting absorption of drugs from GIT 9
1.7 Physico - chemical factors of drug affecting drug absorption and 11
bioavailability
1.8 Pharmaceutical factors / Dosage form factors / Formulation 12
factors affecting drug absorption and bioavailability
1.9 First pass effect and its significance 13
1.10 Fick’s first law of diffusion and in vivo sink condition 13
1.11 Rate limiting steps in drug absorption 14
1.12: BCS classification of drugs 15
1.13: Introduction to distribution of drugs 16
1.14: Physiological Barriers for Drug Distribution 16
1.15: Factors Affecting Distribution of Drugs 18
1.16: Volume of distribution Vd 20
1.17: Protein Binding of drugs 20
1.18: Binding of drugs with albumin 21
1.19 Tissue Binding of Drugs 21
1.20: Factors affecting protein binding of drugs 22
1.21: Importance of protein binding of drugs 23
1.22: Introduction on drug metabolism 23
1.23: Phase I reactions 24
1.24: Phase II reactions 26
1.25: Factors influencing metabolism of drugs 27
1.26: Introduction on Excretion of drugs 28

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1.27: Renal excretion of drugs 28
1.28: Clearance 30
1.29: Factors Affecting Renal Excretion of Drugs: 31
1.30: Non Renal Routes of Drug Excretion 32
1.31: Biliary excretion of drugs / Entero hepatic cycling of drugs 32
1.32: Factors Affecting Biliary Excretion of Drugs 33
1.33: Pulmonary excretion of drugs 34
1.34: Salivary Excretion of drugs and Salivary Cycling 34
1.35: Mammary Excretion of drugs 35
1.36: Skin Excretion of drugs 35
1.37: Gastro intestinal excretion of drugs 35
1.38: Genital excretion of drugs 35
Unit 2: Pharmacokinetics
2.1: Basics of kinetics 36
2.2: Introduction to Pharmacokinetics / definitions 36
2.3: Mathematical model / Pharmacokinetic model / Compartment 38
model / Pharmacokinetic study
2.4: Blood level curves 40
Unit 3: One compartment open model
3.1: One compartment open model – Intravenous injection (Bolus) 41
3.2: One compartment open model – Intravenous infusion 43
3.3: One compartment model extra vascular administration 44
3.4: Wagner Nelson method 47
Unit 4: Multi Compartment model
4.1 Two compartment model- IV bolus 48
4.2: Two compartment model- Extra vascular administration 50
4.2: Two compartment model IV Infusion 52
Unit 5: Multiple – Dosage Regimens
5.1: Kinetics of Multiple Dosing / Dosage regimens 54
/Pharmacokinetics of a drug following repeated administration (IV /
extra vascular)
Unit 6: Non Linear Pharmacokinetics
6.1: Introduction 56
6.2: Evidence for non linear pharmacokinetics 56

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6.3: Causes for Non Linear Pharmacokinetics 57
6.4: Non Linear Pharmacokinetics Equation / Michaelis-Menten 58
Equation
Unit 7: Non Compartmental Pharmacokinetics
7.1: Introduction 59
7.2: Determination of Pharmacokinetic Parameters, MRT – IV 59
administration
7.3: Determination of Pharmacokinetic Parameters MRT – Extra 60
Vascular administration
7.4: Advantages of Non Compartmental Analysis 61
Unit 8: Bioavailability and Bioequivalence
8.1: Definitions 62
8.2: Objectives of bio availability studies 62
8.3: Methods for assessing bioavailability 63
8.4: Experimental design in bioavailability / bioequivalent studies / 66
evaluation of bioavailability studies
8.5: In vitro and in vivo correlation methods 67

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 Unit 1: Introduction to Biopharmaceutics
1.1 Journey of a tablet in the body:
1. When a tablet is taken orally, the following steps take place.
2. Disintegration of tablet into
granules.
3. Granules disintegrate to give fine
drug particles.
4. Dissolution of drug particles in
gastro intestinal fluids.
5. The dissolved drug crosses the
gastro intestinal membrane and
reaches the blood. This is called
absorption.
6. The blood carries the drug to all parts of the body. The drug leaves the blood
and enters into tissues. This is called distribution of drugs.
7. The drug that reaches the liver is destroyed by enzymes. This is called
metabolism of drugs.
8. The drug that reaches the kidney is sent into urine. This is called excretion of
drugs. Excretion of drugs also takes place in saliva, sweat, etc.
9. Hence a drug undergoes absorption, distribution, metabolism and excretion
(ADME).
10. In case of intra venous injections, there will be no absorption step, because
the injection is directly given into the vein.
11. In case of intra muscular injections, topical products, etc., there will be an
absorption step.
1.2 Definitions:
1. Absorption: Movement of drug from site of administration into systemic
circulation (blood).
2. Distribution: Movement of drug from blood to all parts of the body is called
distribution.
3. Metabolism: Destruction of drug in the body by liver enzymes is called
metabolism.
4. Excretion: Removal of drugs from the body by kidney and other organs is called
excretion.

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5. Biopharmaceutics: It is a study of relation between properties of drug,
dosage form, route of administration and bioavailability of drug.
6. Bioavailability: The rate and extent of absorption of drug into blood is called
bioavailability.
7. Biopharmaceutics helps us to design dosage forms in a rational manner so that
the drug will be delivered to the site of action.
1.3 Examples showing the relation between properties of drug, dosage form
characteristics, route of administration and bioavailability:
1. If a drug is unstable in acidic fluids of the stomach, it cannot be given orally. We
have to prepare an injection for the drug.
2. Hydrophilic drugs have good solubility in GIT fluids, but cannot cross biological
membranes easily.
3. Lipophilic drugs have low solubility in biological fluids, but can cross biological
membranes easily.
4. Hence a drug should have a balanced hydrophilic and lipophillic nature for good
bioavailability.
5. Drug release is faster from capsules when compared to tablets. Hence onset of
action is quick with capsules.
6. I.V injections act quickly because there is no absorption step.
7. Bioavailability of a drug from different dosage forms is given below.
Injections > Oral solutions > Oral suspensions > Capsules > Tablets

Unit 1 (a): Absorption of drugs from Gastro Intestinal Tract (GIT)

1.4 Structure of cell membrane and its significance (importance) in drug
absorption:
1. The structure of cell membrane is shown below. It has a bi layer of phospholipid
molecules.
2. This phospholipid molecule has a lipophilic tail and hydrophilic head. The tails
are towards each other and form the lipophilic region of the membrane.
3. The heads form the outer and inner boundary of cell membrane.
4. Globular proteins are found on the outer and inner surface of the membrane
and are also interspersed within membrane structure.
5. Hydrophilic drug molecules cannot cross this phospholipid bi layer easily.
6. Lipophilic drug molecules can easily pass through this phospholipid layer.

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7. Aqueous filled pores of 4 to
10 angstroms are present
in the cell membrane.

8. Inorganic ions and small water soluble molecules pass through these pores.
9. The cell membrane is a semi permeable membrane. It allows the transport of
some molecules like glucose, amino acids, vitamins etc., and prevents the entry
of toxins and certain drugs.
1.5 Mechanisms of drug absorption:
1. The different mechanisms of absorption are
a. Passive diffusion e. Ionic or electrochemical diffusion
b. Pore transport f. Ion pair transport
c. Facilitated diffusion g. Endocytosis
d. Active transport
2. Passive diffusion: 90 % of the drugs are absorbed by this mechanism. Difference
in concentration of drug on both sides of the membrane is responsible for
passive diffusion. Example: When we take a tablet orally, the concentration of
drug is more in the GIT and less in blood. So, the drug moves from higher
concentration to lower concentration. This is called passive diffusion. The drug
dissolves in GI fluids, partitions into GI membrane and then goes into blood.
The equation for passive diffusion is given below.
dQ/dt = rate of drug diffusion
D = Diffusion coefficient of drug
A = Area of GIT membrane

Km/w = membrane water partition coefficient of drug

h = thickness of GIT membrane
CGIT – C = Difference in drug concentration between GIT and blood
Rate of drug diffusion (absorption), is directly proportional to D, A, Km/w and
concentration gradient. It is inversely related to thickness of GIT membrane.
3. When concentration of drug in GIT and blood becomes equal, concentration
gradient will become zero and diffusion will stop, but this doesn’t happen in the
body.

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4. As soon as the drug enters the blood, it is carried away and distributed to the
entire body. The concentration of drug will be always very less when compared
to the concentration in GIT. This is called sink condition.
5. Pore transport: Small water soluble molecules like urea, sugar, pass through the
pores in cell membrane to blood. The pressure of GIT fluids is responsible for
pore transport.
6. Carrier mediated transport: The GIT membrane has carriers. These carriers
attach to nutrients (vitamins, amino acids), carry them to other side of
membrane and leave them in blood. Then, they return to the GIT side to carry
another nutrient molecule. If our drug molecule has a structure similar to
nutrients, they will be carried by these carriers. This is called carrier mediated
system.
7. Carrier mediated transport is of two types, facilitated diffusion and active
transport.
8. Facilitated diffusion takes place from high concentration to low concentration; it
involves passive diffusion and carriers.
9. Active transport also involves carriers and takes place from low concentration to
high concentration.
10. Ionic diffusion: The GIT membrane has a positive charge, hence cationic
drugs are repelled. Anionic drugs having a negative charge are attracted by GIT
membrane and are absorbed by passive diffusion.
11. Endocytosis is cell eating. The GIT membrane engulfs the drug and releases
it on other side.
12. The below diagram shows the different mechanisms of absorption.

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1.6 Biological Factors affecting absorption of drugs from GIT: The
movement of the drug from the G.I.T fluids into blood is called absorption. The
biological factors affecting drug absorption are discussed below.
1. Area of G.I.T: Stomach has less surface area when compared to small
intestine. Area of small intestine is almost equal to a tennis court. This is
due to presence of villi in the small intestine. Hence absorption of drugs will
be more in small intestine than in stomach.
2. The area of large intestine and rectum is less and absorption of drugs will be
less in this area.
3. The pH of GIT, pKa, and lipophillicity of drug affects absorption of drugs.
This is explained by pH partition theory.
4. Unionized form of drug is lipophillic and absorbed easily in GIT. If drug is
ionized, it is poorly absorbed.
5. The extent of ionization of a drug depends on its pKa and pH at site of
absorption and is given by Henderson- Hasselbalch equation.
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6. pH of the stomach is around 1 to 3, weakly acidic drugs are un ionized here
and absorption will be fast. This is because un ionized drug molecules are
more lipophillic and can easily cross biological membranes.
7. pH of the intestine is around 7.4, weakly basic drugs are un ionized here
and absorption will be fast. This is because un ionized drug molecules are
more lipophillic and can easily cross biological membranes.
8. Very weak acidic and very weak basic drugs are unionized at all pH values
and are absorbed along the entire GIT.
9. Strongly acidic and strongly basic drugs are ionized at all pH values and are
poorly absorbed along entire GIT.
10. A drug should have hydrophilic nature to dissolve in G.I.T fluids and
lipophillic nature to cross GIT. Lipophillic nature can be measured using
octanol water partition coefficient experiments. Octanol represents the GIT
membrane.
11. A drug should have an optimum Ko/w value of 1000 / 1 for dissolution
and absorption in GIT.
12. If a drug is strongly hydrophilic, it will dissolve in GIT fluids but will not
cross GIT easily.
13. If a drug is strongly lipophillic, it will dissolve with difficulty in GIT fluids.
It accumulates in the GIT membrane because of its high lipophillic nature.
14. Food may alter the absorption of drugs. Example: Milk reduces
absorption of iron. Vitamin C enhances the absorption of iron.
15. Gastro intestinal motility: If G.I motility is more, drug absorption may get
reduced. Example: In diarrhea, absorption of drugs will be reduced.

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1.7 Physico - chemical factors of drug affecting drug absorption and
bioavailability:
1. Drug solubility: If a drug has good solubility in water, it will have good
dissolution rate and bio availability will be good. A drug should have a
minimum of 1 % solubility in water to avoid bio availability problems.
2. Particle size of the drug: Fine drug particles have more surface area and
dissolve fast. So bio availability will be good. Example: Tetracycline and
chloramphenicol has been micronized to improve dissolution rate and bio
availability.
3. Polymorphism: When a drug can exist in more than one crystalline form, it
is called polymorphism. Polymorphs have same medicinal action but
different solubility and dissolution rate. Example: Chloramphenicol
palmitate exists as crystalline form A, B and C. Polymorph B has good bio
availability.
4. Amorphism: Drugs can exist in crystalline form and in amorphous form (no
crystalline structure). Amorphous drugs have good bioavailability.
Example: Amorphous novobiocin is 10 times more soluble than crystalline
form.
5. Hydrates: When water is trapped within the crystalline structure of drug, it
is called hydrate. Hydrates may exist in different crystalline forms and are
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 called pseudo polymorphs. This phenomena is called pseudo poly
morphism. Example: Anhydrous ampicillin has more solubility than
ampicillin tri hydrate.
6. Organic solvates have more water solubility. Example: Chloroform solvate of
griseofulvin is more soluble than non solvated form.
7. Salt form of drug: Most of the drugs are weak acids or weak bases and have
poor solubility in water. Their salts have good solubility, dissolution rate and
bioavailability. Example: Sodium salicylate has 400 times greater solubility
than salicylic acid in water. Example: Tetracycline hydrochloride has better
solubility than tetracycline.
1.8 Pharmaceutical factors / Dosage form factors / Formulation factors
affecting drug absorption and bioavailability:
1. Type of dosage form: The bioavailability of a drug depends on the type of
dosage form. Example: Bioavailability is more from elixirs and syrups when
compared to tablets. The bioavailability of drug from various dosage forms is
in the below order.
Solutions > Emulsions > Suspensions > Capsules > Tablets > Coated tablets
> Sustained release products.
2. Manufacturing process: Bioavailability of a drug depends on the
manufacturing method of dosage form. Example: Tablets made by direct
compression are more bioavailable than tablets made by granulation
method.
3. Excipients or additives used in the dosage form: If hydrophilic additives are
used in tablets, bioavailability will be good. If hydro phobic additives are
used, bioavailability will be poor.
4. Binder: If a strong binder is added in a tablet, tablets will have more
hardness, tablets will not disintegrate quickly and bioavailability will be less.
5. Disintegrant: If a weak disintegrant is used in tablets, they will not
disintegrate easily and bioavailability will be less.
6. Lubricants and glidants: They are hydrophobic in nature. If excess
lubricants and glidants are used in tablets, disintegration and dissolution
will be less and bioavailability of drug will be less.

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1.9: First pass effect and its significance:
1. It is also called first pass metabolism or pre systemic metabolism.
2. When a tablet is taken orally, it disintegrates into granules. These granules
are broken down into fine drug particles.
3. The drug particles dissolve in the G.I fluids and are absorbed into blood. The
portal vein carries the drug first to the liver. The liver is rich in enzymes and
converts some amount of the drug into hydrophilic metabolites. This bio
transformation is called first pass effect.
4. Enzymes in the GIT fluids, enzymes in the GIT membrane and liver enzymes
are responsible for first pass effect.
5. Example: Morphine, diazepam, alcohol are metabolized in liver.
1.10: Fick’s first law of diffusion and in vivo sink condition:
1. Fick’s first law of diffusion: Rate of diffusion is directly proportional to the
concentration gradient.
2. Example: When we take a tablet orally, it disintegrates and drug dissolves in
the GIT fluids. Concentration of drug in the GIT is very high and
concentration of drug in blood is zero. The drug moves from higher
concentration (GIT) to lower concentration (blood).
3. Once the drug reaches the blood, it is rapidly distributed to all parts of the
body. So, concentration of drug in blood is always less than the
concentration in GIT. This is called sink condition.
4. The equation for passive diffusion is given below.

dQ/dt = rate of drug diffusion

D = Diffusion coefficient of drug
A = Area of GIT membrane

Km/w = membrane water partition coefficient of drug

h = thickness of GIT membrane
CGIT – C = Difference in concentration between GIT and blood
Rate of drug diffusion (absorption) is directly proportional to D, A, Km/w
and concentration gradient. It is inversely related to thickness of GIT
membrane.

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5. When concentration of drug in GIT and blood becomes equal, concentration
gradient will become zero and diffusion will stop, but this doesn’t happen in
the body.
6. As soon as the drug enters the blood, it is carried away and distributed to
the entire body. The concentration of drug will be always very less when
compared to the concentration in GIT. This is called sink condition.
1.11: Rate limiting steps in drug absorption:
1. The slowest step in a process is called rate limiting step.
2. The speed of the entire process will depend upon the rate limiting step.
3. When a tablet is taken orally, the following steps take place.
4. Disintegration of tablet into granules.
5. Granules disintegrate to give fine drug particles.
6. Dissolution of drug particles in gastro intestinal fluids.
7. The dissolved drug crosses the gastro intestinal membrane and reaches the
blood. This is called absorption.
8. Dissolution is rate limiting step for poorly water soluble drugs.
9. Absorption is rate limiting step for drugs having good solubility in water.

1.12: BCS classification of drugs:

1. BCS means biopharmaceutics classification system of drugs.
2. After dissolution in GIT fluids, drugs have to cross the GI membrane into
blood. This is called permeability.
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3. Basing on solubility and permeability nature of drug, the BCS has four
classes of drugs.
4. Class 1: High Solubility – High Permeability, Ex: Chloroquine phosphate.
5. Class 2: Low Solubility – High Permeability, Ex: Azithromycin
6. Class 3: High Solubility – Low Permeability, Ex: Aspirin
7. Class 4: Low solubility – Low permeability, Ex: Cefixime
8. A drug substance is considered highly soluble, when the highest dose
strength is soluble in 250 ml or less of aqueous media over the pH range of
1-7.5.
9. A drug substance is considered to be highly permeable when 90 % of dose
gets absorbed in humans.
10. Class I drugs can be formulated as oral tablets, capsules, solutions etc.,
because they have good solubility and permeability.
11. Class II drugs have to be modified to improve their solubility and
dissolution rate, then, they can be formulated as tablets and capsules etc.
As class III and IV have poor permeability, they have to be formulated as
injections.
Unit 1 B: Distribution of Drugs
1.13: Introduction to distribution of drugs:
1. Reversible transfer of drugs between compartments is called distribution.
2. Due to absorption, the drug enters into the blood, and now it is distributed
to different parts of the body as shown below.

3. Distribution is a passive process, and takes place from higher concentration

to lower concentration.
4. Initially, the blood concentration is high and tissue concentration is low.
Hence drug moves from blood into tissues until equilibrium is obtained.
5. The drug in the blood also undergoes elimination from the body by kidneys.
As a result drug concentration in blood decreases.

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6. Hence the drug comes back from the tissues (high drug concentration) to
blood (low drug concentration).
7. Hence distribution of drugs between blood and tissues is a reversible
process.
1.14: Physiological Barriers for Drug Distribution:
1. The different barriers for drug distribution are blood brain barrier, blood –
cerebrospinal fluid barrier, blood placenta barrier and blood testis barrier.
2. Blood capillaries carry blood to all tissues. The blood capillary walls are very
thin and are made up of a single layer of endothelial cells. Drugs easily cross
this thin layer and enter into the tissue.
3. The blood capillaries of the brain have a layer of endothelial cells with tight
junctions. They are surrounded by astrocytes as shown in the below figure.
This layer of endothelial cells with astrocytes is called blood brain barrier.

4. The BBB is highly lipophillic in nature and only small, lipophillic drugs can
pass through the BBB by passive diffusion.
5. The blood – brain barrier allows the passage of water, some gases, lipid
soluble molecules by passive diffusion, and selective transport of molecules
such as glucose, vitamins, hormones and amino acids that are necessary to
the brain.
6. Example: Penicillin’s cannot cross BBB.

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7. Blood cerebro spinal fluid barrier: The choroid plexus of the CNS produce
the CSF. The choroid plexus cells have tight junctions and do not allow
transport of drug molecules from blood into CSF.

8. The blood testes barrier is a physical barrier between blood capillaries and
seminiferous tubules of testes. It prevents the passage of drugs to sperm
cells.
9. The placental membrane separates the mother blood capillaries and fetal
blood vessels. Many drug molecules up to a molecular weight of 1000
Daltons cross this membrane and reach the fetus. This barrier is not
effective like BBB.
1.15: Factors Affecting Distribution of Drugs: The various factors
influencing distribution of drugs are
1. Physico – Chemical Properties drug:
a. Drugs with molecular weight less than 500 Daltons easily cross biological
membranes by passive diffusion.
b. If drug molecule is ionized at blood pH, it will have poor tissue permeability.
Example: Penicillin is ionized at blood pH and cannot cross blood brain
barrier.
c. If drug molecule is un-ionized at blood pH, it will have good tissue
permeability. Example: Thiopental is un-ionized at blood pH and can easily
cross blood brain barrier.
d. Lipophillic drugs can easily cross biological membranes. Example:
Thiopental is highly lipophillic and distributes 80 times faster than salicylic
acid into CSF (cerebro spinal fluid).
2. Nature of tissue membrane: Distribution of drugs depends on the nature of
tissue membrane. Example: BBB is highly lipophillic, only highly lipophillic

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 drugs can cross the BBB. Example: Thiopental can cross BBB but
penicillin cannot cross BBB.
3. Organ size / Tissue size and blood perfusion rate:
a. Distribution of a drug to a particular organ depends on its tissue size and
blood perfusion rate.
b. Lungs, kidney, liver, heart and brain have high blood perfusion rate.
c. Muscles and skin have medium perfusion rate.
d. Fat and bone have poor perfusion rate.
e. Greater the perfusion rate, greater will be the distribution.
4. Protein binding of drugs:
a. If the drug is highly bound to albumin in blood, its distribution will be
less. Proteins cannot cross biological membranes and hence protein bound
drug also cannot cross biological membranes.
5. Tissue binding of drugs:
a. If the drug is highly bound to tissue components, accumulation will take
place in that tissue. Example: Tetracycline binds with calcium in bones and
teeth.
b. Example: Thiopental accumulates in adipose tissue.
6. Age:
a. BBB is poorly developed in infants and drugs easily cross them.
b. Fat content is high in infants and elders which influences drug distribution.
c. Proteins are low in infants and elders which influences drug distribution.
7. Pregnancy: In pregnancy, fetus is another compartment into which drugs
can be distributed by crossing placental barrier.
8. Obesity: In obese persons, fat tissue is more and lipophillic drugs
accumulate in fat tissue.
9. Diet: A diet rich in fats will increase fatty acid levels in blood and influence
protein binding of drugs. This will influence drug distribution.
10. Disease states: Altered albumin levels, altered perfusion rates and altered
tissue pH influence distribution of drugs. Example: In meningitis, BBB
becomes more permeable to polar antibiotics like penicillin.
11. Drug interactions: They alter protein binding and distribution of drugs.

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1.16: Volume of distribution Vd:
1. It gives an idea about the distribution of drug in the body.
2. It is defined as hypothetical volume of body fluid in which the drug is
distributed.
3. Vd = Dose / C0
4. The body water is made up of three compartments
a. Blood : 6 liters
b. Extra cellular fluid : 12 liters
c. Intracellular fluid : 24 liters
d. Total : 42 liters
5. If the drug is highly protein bound, it will be present in the blood without
being distributed, so plasma concentration is very high and Vd will be less.
Example: Vd of warfarin is 10 litres.
6. If the drug accumulates in a tissue, plasma concentration will be very less
and Vd will be high. Example: Vd of Chloroquine is 15000 Litres.
1.17: Protein Binding of drugs:
1. When a drug forms a complex with proteins in the blood, it is called protein
binding of drugs.
2. Proteins are very large molecules and cannot cross biological membranes.
Hence protein bound drugs also cannot cross biological membranes.
3. Hence protein bound drugs are not distributed / metabolized / eliminated
from the body.
4. Only the free drug is distributed / metabolized / eliminated from the body.
5. Half-life of protein bound drugs is high.
6. Protein binding is reversible. Weak bonds such as hydrogen bonds,
hydrophobic bonds, Van Der Waals forces are responsible for protein
binding.
7. Example: warfarin, diazepam, digitoxin binds to albumin in blood.
8. Glycoproteins, lipo proteins, globulins and hemoglobin are also involved in
protein binding of drugs.

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1.18: Binding of drugs with albumin:
1. Human serum albumin (HSA) is a very large molecule, having a molecular
weight of 65,000 Daltons.
2. It is present in large quantities in blood.
3. Many drugs (weak acids, weak bases and neutral) bind with albumin.
4. HSA has four different binding sites.
5. Site I – Warfarin binding site – NSAIDS and phenytoin bind at this site.
6. Site II – Diazepam binding site – Benzodiazepines bind at this site.
7. Site III – Digitoxin binding site.
8. Site IV – Tamoxifen binding site.
9. A drug can bind to more than one site. In such cases, the main binding site
is called primary binding site and the other site is called secondary site.
10. If two drugs bind to the same site, there will be competition between the
two drugs for binding.
1.19: Tissue Binding of Drugs:
1. A drug can bind to tissue components also.
2. If binding occurs in tissues, drug gets accumulated in the tissue and acts as
a store house for drugs. Sometimes toxic reactions may be seen.
3. Thiopental accumulates in adipose tissue.
4. Generally tissue binding is reversible, but sometimes it may be irreversible.
5. Example: paracetamol, chloroform bind irreversibly with liver tissue.
6. If a drug undergoes protein binding and tissue binding, there will be
competition between them for drug.
7. The order of binding in extravascular tissues is liver > kidney > lung >
muscle.
8. Examples: Liver – paracetamol, chloroform.
9. Lungs – Anti histamines.
10. Kidneys – heavy metals like lead, mercury, etc.
11. Skin – Chloroquine, phenothiazines.
12. Eye - Chloroquine, phenothiazines.
13. Hair – arsenicals, chloroquine, phenothiazines deposit in hair shafts.
14. Bones – tetracycline, lead.
15. Fats – lipophillic drugs like thiopental and pesticides like DDT.
16. Nucleic acids – DNA interacts with chloroquine and quinacrine.

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1.20: Factors affecting protein binding of drugs:
1. Drug related factors:
a. As concentration of drug increases, protein binding increases until
proteins are saturated.
b. As lipophillic nature of drug increases, extent of protein binding increases.
c. Anionic/acidic drugs bind to albumin.
d. Basic drugs bind to globulins.
e. Neutral and un ionized drugs bind to lipo proteins.
f. A drug may have affinity for a particular tissue. Example: Digoxin has
more affinity for proteins of cardiac muscle.
2. Tissue related factors:
a. Lipoproteins and adipose tissue bind with lipophillic drugs.
b. As concentration of albumin is more in blood, protein binding is more in
blood.
3. Drug interactions:
a. If two drugs bind to the same site, there will be competition between the two
drugs for binding.
b. One drug may displace another drug from albumin binding site.
c. Example: Phenyl butazone displaces warfarin from albumin. This increases
free drug concentration of warfarin and may cause toxic effects.
d. This interaction is called displacement interaction.
e. Warfarin is called displaced drug and phenyl butazone is displacer.
f. Sometimes the drug will alter the protein structure and modify its binding
capacity.
g. Example: Aspirin changes structure of albumin and increases its capacity to
bind with phenyl butazone.
4. Patient related factors:
a. With age, protein content varies and protein binding changes.
b. In new born and elders, protein content is less and protein binding is less.
c. In elders, protein content is less and protein binding is less.
d. Inter subject variability is less and is due to genetic factors.
e. In disease states like CCF, burns, liver diseases, kidney diseases, surgery,
cancer, protein content is less and protein binding is less.

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1.21: Importance of protein binding of drugs:
1. Absorption: Drug gets absorbed from the GIT into blood by a passive
diffusion process. Drug moves from higher concentration to lower
concentration. Due to protein binding in blood, the free drug concentration
will be less and sink conditions are maintained. Hence absorption will be
fast.
2. Distribution: Due to protein binding, only the free drug is available for
distribution and elimination. When free drug has been eliminated from the
body, the protein drug complex dissociates and free drug is available for
distribution and elimination. In this manner, the free drug is released from
protein drug complex for long periods of time. The Complex acts as a store
house for drug. Hence half life of protein bound drugs is high.
3. Elimination: Only free drug can be eliminated from the body by kidneys. As
a result elimination of protein bound drugs is slow.
Example: Tetracycline – 65% protein bound – half life – 8.5 hours.
Example: Doxycycline – 93% protein bound – half life – 15 hours.

Unit 1 C: Metabolism and Excretion of drugs

1.22: Introduction on drug metabolism:
1. The liver enzymes convert the drug into simple water soluble molecules. This
is called drug metabolism or bio-transformation.
2. Drug + enzyme ------- Metabolite
3. Most of the drugs are lipophillic and stay in the body for long periods of
time. So, the body converts them into simple hydrophilic substances so that
they are easily excreted in urine.
4. Generally the metabolites are inactive, but sometimes they have drug action.
5. Example: Phenytoin is converted into p – hydroxyl phenytoin which is
inactive.
6. Example: Aspirin is converted into salicylic acid which is active.
7. Phenacetin is inactive which is converted into paracetamol active drug.
8. Diazepam is a tranquilizer; it is converted into oxazepam (anti convulsant).
9. Metabolism by liver is called hepatic metabolism.
10. Metabolism by other organs is called extra hepatic metabolism.

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11. The enzymes that metabolize drugs in the liver are microsomal and non
microsomal enzymes.
12. There are two pathways for drug metabolism, phase I reactions and
phase II reactions.
1.23: Phase I reactions:
1. Phase I reactions are known as functionalization reactions. In these
reactions, a polar functional group like –OH, - NH2, - SH, -COOH, is
introduced into the drug molecule or unmasked if already present in the
drug molecule.
2. Microsomal and non-microsomal enzymes are responsible for phase I
reactions.
3. Phase I reactions include oxidation, reduction or hydrolysis.
4. Oxidative reactions are catalyzed by mixed function oxidase enzymes.
5. Reduction reactions: Aliphatic aldehydes, aromatic aldehydes, acids are
reduced by enzymes called aldo – keto reductases.
6. Ester drugs are hydrolyzed by esterase enzymes.
7. Amide drugs are hydrolyzed by amidase enzymes.
8. Examples of phase I reactions are given below.

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9. Oxidation Phase I reactions:
S.NO Reaction Reaction Pathway Example
1 Aromatic Phenylbutazone to
hydroxylation oxy phenyl butazone
2 Aliphatic Pentobarbital
hydroxylation
3 Oxidative Amphetamine
deamination
4 N–dealkylation Morphine

5 O-dealkylation Phenacetin

6 Alcoholic Ethanol
Oxidation
7 Aldehyde Acetaldehyde
oxidation

10. Reduction Phase I reactions:

S.NO Reaction Reaction Pathway Example
1 Azo Sulfasalazine
reduction
2 Nitro Chloramphenicol
reduction
3 Aldehyde Chloralhydrate
reduction
11. Hydrolytic Phase I reactions:
S.NO Reaction Reaction Pathway Example
1 De esterfication Procaine

2 De amidation Lidocaine

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1.24: Phase II reactions:
1. The drug /metabolite obtained in phase I reaction is subjected to phase II
reaction. In these reactions, a small polar group like glucuronic acid,
sulfate, glycine etc., are attached to the phase I product.
2. The phase II product is called conjugate and hence these reactions are
called conjugation reactions.
3. Phase II products are more water soluble than phase I products and are
easily eliminated in urine.
4. Enzymes of phase II reactions are called transferases, because they transfer
a polar group to the phase I metabolite.
5. Glucuronidation is a common phase II reaction. Glucuronic acid is available
in the liver due to glucose metabolism.
6. Uridine di phosphate glucuronic acid (UDPGA) conjugates with a number of
drugs / metabolites under the influence of enzyme glucuronyl transferase.
7. Drugs or their phase I metabolites containing hydroxyl / carboxyl groups
form ester/ether glucuronides which are highly polar and easily eliminated
in urine.
8. Glycine and glutamine conjugation reactions occur with drugs or their
phase I metabolites to form amides by trans acylase enzymes.
9. Gluthathione conjugation reactions occur by gluthathione transferase
enzymes.
10. Sulfate conjugation reactions occur by sulfo transferase / sulfo kinase
enzymes.
11. Methylation conjugation reactions occur by methyl transferase enzymes.
The methylated products are less polar than the original drug or phase I
metabolite.
12. Acetylation conjugation reactions occur by acetyl transferase enzymes.
The acetylated products are less polar than the original drug or phase I
metabolite.

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 S.NO Reaction Examples
1 Glucuronidation Salicylates, oxazepam, sulfamethoxazole
2 Glycine conjugation Salicylic acid, fenfluramine
3 Glutathione conjugation Paracetamol
4 Sulfate conjugation Steroids
5 Methylation Oxprenolol
6 Acetylation Procainamide

1.25: Factors influencing metabolism of drugs:

1. Physico chemical properties of the drug: Size of the molecule, pKa, acidity /
basic nature, lipophillicity influence the interaction of drug with enzyme. As
a result metabolism is affected.
2. Drug interactions: In multiple drug therapy, one drug increases or decreases
the metabolism of other drugs.
e. When a drug increases the metabolism of other drugs it is called enzyme
induction. Example: Barbiturates increase the metabolism of anti
coagulants. Induction occurs by increasing the synthesis of enzymes or by
decreasing the degradation of enzymes responsible for drug metabolism.
f. When a drug decreases the metabolism of other drugs, it is called enzyme
inhibition. Example: Para amino salicylic acid decreases the metabolism of
phenytoin. Inhibition occurs by decreasing the synthesis of enzymes or by
increasing the degradation of enzymes responsible for drug metabolism.
3. Biological factors:
a. Qualitative differences are differences in the actual metabolic pathway. This
is due to genetic deficiency of a particular enzyme. Example: In man and
rabbit amphetamine is metabolized by oxidative deamination, where as in
rats aromatic oxidation is the main route.
b. Quantitative differences are differences in the extent of metabolism. This is
due to differences in quantity of enzyme present from person to person and
species to species. The metabolic pathway is same.
4. Sex: Drug metabolism variation in male and females is due to difference in
sex hormones. Example: women metabolize benzodiazepines slowly than
men.
5. Age: Enzyme content varies with age and metabolism varies.

26 | P a g e
a. In neonates (upto 2 months age), enzyme systems are not fully developed
and many drugs are metabolized slowly. Example: Caffeine has half life of 4
days in neonates, it is 4 hours in adults.
b. As age increases, metabolizing capacity increases.
c. In very elderly persons, the liver size is reduced and enzyme activity is
decreased. Hence drug metabolism is reduced.
a. Diet: Protein rich diet increases enzymes in body and increases metabolism
of drugs. Deficiency of vitamins and minerals decrease activity of enzymes.
6. Pregnancy: High levels of steroid hormones in last stages of pregnancy
reduce the activity of enzymes.
7. Disease states:
a. Any disease condition with liver will reduce activity of enzymes.
b. In congestive cardiac failure, blood flow to liver decreases and enzyme
activity is reduced.
c. If albumin levels in blood changes, protein binding changes, free drug
concentration changes and metabolism changes.
8. Circadian rhythm: Variation in enzyme activity with light cycle is called
circadian rhythm. It has been found that enzyme activity is more in early
morning (6 to 9 a.m) and minimum in afternoon (2 to 5 p.m.) Example:
Plasma levels of theophylline and diazepam are less in night than in day
time.
9. Route of administration: In oral route, first pass effect is seen. In I.V and
sublingual route first pass effect is not seen.
1.26: Introduction on Excretion of drugs:
1. It is a process by which the drugs and its metabolites are sent out of the
body.
2. Excretion of drugs is mainly done by kidneys and is called renal excretion.
Excretion by other organs like lungs, salivary glands, biliary system, sweat
glands, skin and intestine is called non renal excretion.
1.27: Renal excretion of drugs:
1. Excretion of drugs is mainly done by kidneys and is called renal excretion.
2. Nephron is the basic functional unit of kidney. Each kidney has around one
million (10 lakhs) nephrons.

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3. Each nephron has glomerular capsule, proximal tubule, loop of Henle, distal
tubule and collecting tubule.
4. The steps involved in urinary excretion of drugs are glomerular filtration,
active tubular secretion and tubular reabsorption.
5. Glomerular filtration:
a. Every minute, around 1.2 liters of blood goes to the kidney and gets
filtered. The filtrate volume is 130 ml/min. This is called glomerular filtration
rate (GFR).
b. Filtration takes place through the semi permeable walls of glomerular
capsule. RBC, WBC, platelets, albumin and globulin are not filtered.
c. The filtrate contains water, minerals, salts, glucose, hormones, drugs and
their metabolites.
d. The filtrate volume is 130 ml/min that is 180 liters per day.
e. Even though filtrate volume is 180 liters per day, only 1.5 liters urine is
excreted per day. Remaining water is reabsorbed from the renal tubules.
f. Creatinine is excreted by glomerular filtration only and is used to estimate
GFR. If GFR is 130 ml/minute, it indicates that the person kidney is
functioning properly.

6. Active tubular secretion:

a. As blood does not stay for sufficient time in glomerulus, the drugs are not
completely removed by filtration.
b. Drugs that have escaped in the above filtration step undergo active
secretion in proximal renal tubule by carriers.
c. Carrier systems are available for weakly acidic and basic drugs.

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7. Tubular re absorption:
a. Tubular re absorption takes place in the entire renal tubule.
b. Drugs and other nutrients like glucose, electrolytes, vitamins, amino
acids that are required for the body undergo tubular re absorption by
active or passive process.
c. As most of the water filtered in glomerulus is reabsorbed in renal tubule,
the drug concentration in renal tubule is very high than that in blood,
hence passive diffusion of drugs from renal tubule to blood takes place.
1.28: Clearance:
1. Clearance is defined as the hypothetical volume of body fluids from which
the drug is removed completely.
2. Total body clearance = Renal clearance + hepatic clearance + lung clearance
+ biliary clearance.
3. Clearence = Total elimination rate /Cp
4. Renal clearance is defined as the hypothetical volume of body fluids from
which the drug is removed completely by kidneys.
5. ClR = Urinary excretion rate of drug / Cp
6. ClR = [Rate of drug filtration + rate of drug secretion –rate of drug re
absorption] / Cp
7. The below table gives the relationship between renal clearance values and
mechanism of clearance.
S.NO Renal Mechanism Example
Clearence,
ml/min
1 0 Drug filtered and reabsorbed Glucose
completely
2 Less than 130 Drug filtered and partially reabsorbed Lipophillic
drugs
3 Greater than Drug filtered and secreted actively Polar, ionic
130 drugs
4 = 130 Drug is filtered only Creatinine,
inulin

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1.29: Factors Affecting Renal Excretion of Drugs: The various factors
influencing renal excretion of drugs are
1. Physico – Chemical Properties drug:
a. Small drug molecules with molecular weight less than 500 daltons are easily
filtered in glomerulus.
b. Hydrophillic drugs are easily excreted in urine. Unionized and lipophillic
drugs undergo re absorption in renal tubules. They stay for long time in the
body.
2. Plasma concentration of drug: If Cp is more, excretion rate of drug will be
more. More drug will be filtered in kidneys.
3. Distribution and binding characteristics of the drug: If a drug is highly
protein bound, it cannot be filtered in kidney and renal clearance will be
less. Example: Tetracycline is 93 % bound to plasma proteins; hence it has
low renal clearance.
4. Tissue binding of drugs:
a. If the drug is highly bound to tissue components, accumulation will take
place in that tissue. Example: Tetracycline binds with calcium in bones
and teeth.
b. Example: Thiopental accumulates in adipose tissue.
5. Old Age: In old age, GFR decreases and renal tubular secretion is altered,
hence renal clearance is decreased.
6. New Born: In new born, renal function is 30 to 40 % less, hence renal
clearance is less.
7. Renal clearance is 10 % less in females than in males.
8. Drug interactions: Any drug interaction which alters protein binding, renal
blood flow, active secretion, urine pH, will alter renal clearance of drugs.
a. Diuretics increase urine flow rate and increase renal clearance of drugs.
b. Furosemide reduces protein binding of gentamycin in blood, so more
gentamycin is excreted in urine.
c. Acidification of urine with ascorbic acid or ammonium chloride increases
excretion of basic drugs.
d. Alkalinisation of urine with citrates, tartrates, bicarbonates, increases
excretion of acidic drugs.

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9. Disease states: In kidney failure, renal elimination of drugs decreases.
Hence GFR should be determined using creatinine clearance. Dose should
be adjusted depending on creatinine clearance. If it is half of normal value of
130 ml/minute, reduce the dose by half.
1.30: Non Renal Routes of Drug Excretion: The other routes of excretion of
drugs are biliary excretion, pulmonary excretion, salivary excretion, mammary
excretion, skin or dermal excretion, GI excretion, genital excretion.
1.31: Biliary excretion of drugs / Entero hepatic cycling of drugs:
1. Bile juice is produced by liver and is stored in gall bladder.
2. It is secreted into the duodenum for digestion of fats. The bile flow rate is
0.5 to 1 ml per minute. 90 % of the bile acids are reabsorbed from the
intestine and is transported back to the liver for re secretion.
3. Drugs and their metabolites enter from the plasma to bile juice through
hepatocytes (liver cells). This bile juice carries the drug and secretes in
duodenum. Thus the drug comes out along with fecal matter. Some of the
drug is again reabsorbed from intestine into blood and goes back to the
liver. Again it is excreted by bile juice. This cycling of drugs is called entero
hepatic cycling of drugs.
4. Entero hepatic cycling is important for conservation of important substances
such as vitamin B12, vitamin D3, folic acid, steroid hormones, and bile
salts.

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1.32: Factors Affecting Biliary Excretion of Drugs: The various factors
influencing biliary excretion of drugs are
1. Physico – Chemical Properties drug:
a. Drug molecules with molecular weight greater than 500 Daltons are
excreted in bile.
b. Drug molecules with molecular weight less than 300 Daltons are excreted in
urine.
c. Drug molecules with molecular weight between 300 to 500 Daltons are
excreted in bile and urine.
2. Drug metabolism: Polar drugs are readily excreted in bile. Drugs are
metabolized in the liver and polar metabolites are produced. They are readily
excreted in the liver.
3. Miscellaneous factors: Age, sex, species differences, protein binding,
disease states and drug interactions influence biliary excretion of drugs.
a. High molecular weight drugs show good biliary excretion in rats, dogs and
hen. Biliary excretion is poor in rabbits, guinea pigs and monkeys.
b. In cholestasis, bile flow rate is reduced and biliary excretion of drugs is
reduced.
c. Orally administered drugs after absorption go to liver and are excreted more
in bile when compared to parenteral products.
d. Diet also influences biliary excretion of drugs. Protein and fat rich diet
increases bile flow.
Biliary Clearance: The ability of the liver to excrete the drug in the bile is
expressed as biliary clearance. Biliary clearance = Biliary excretion rate / Cp
1.33: Pulmonary excretion of drugs:
1. Gaseous and volatile substances such as general anesthetics are absorbed
through the lungs by simple diffusion. Similarly excretion of drugs through
the lungs is also possible. Pulmonary blood flow, rate of respiration,
solubility of volatile substance etc. influence pulmonary excretion of drugs.
2. Example: Alcohol is excreted via lungs slowly. Nitrous oxide has less
solubility in blood and is excreted rapidly.

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1.34: Salivary Excretion of drugs and Salivary Cycling:
1. Excretion of drugs into saliva is a passive process and can be predicted by
pH partition theory. Un ionized, lipophillic drugs are readily excreted in
saliva. This depends on pKa of drug and pH of saliva and blood.
2. The s/p ratios have been found to be less than one for weak acids and
greater than one for weak bases. Basic drugs are excreted more in saliva as
compared to acidic drugs.
3. As s/p ratios are fairly constant for a drug, it can be used to determine
blood concentration from saliva concentration.
4. Example: Caffeine, theophyliine, phenytoin are excreted in saliva.
5. Some drugs are actively secreted, example: penicillin and phenytoin.
6. Bitter after taste in the mouth of a patient is due to salivary excretion of
drugs. Some drugs inhibit salivary secretion and are responsible for dryness
of mouth.
7. Drugs excreted in saliva can undergo cycling as shown below.

1.35: Mammary Excretion:

1. Drugs excrete into mother’s milk also. Excretion of drugs in milk is a
passive process and is dependent on pH partition behavior, molecular
weight, lipid solubility, degree of ionization.
2. The pH of milk is around 7.0 and free, unionized, lipid soluble drugs diffuse
into mammary cells by passive diffusion.
3. The extent of drug excretion in milk is given by milk / plasma drug
concentration ratio.
4. As milk is acidic when compared to plasma, weakly basic drugs concentrate
more in milk and have M/P ratio more than one.

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5. The amount of drug excreted in milk is generally less than 1% of the dose
and will not produce toxic effects in infants.
6. Some examples of drugs producing toxic effects in infants are,
a. Chloramphenicol : bone marrow suppression
b. Diazepam: sedation
c. Tetracycline: permanent staining of teeth
1.36: Skin Excretion:
1. Drugs are excreted through the skin via sweat and follows pH partition
hypothesis. This is the reason for skin reactions and dermatitis.
2. Compounds such as benzoic acid, salicylic acid, alcohol and anti pyrine,
heavy metals like lead, mercury and arsenic are excreted in sweat.
1.37: Gastro intestinal excretion of drugs: Excretion of drugs into the GIT
occurs after parenteral administration, by passive diffusion process. This is
reverse process of GI absorption.
Orally administered drugs can also be reabsorbed and excreted in the GIT.
Drugs excreted in the GIT are reabsorbed into the systemic circulation and
undergo recycling.
1.38: Genital excretion: Reproductive tract and genital secretions may
contain the excreted drugs. Some drugs have been detected in semen.
Drugs can also be secreted in lachrymal fluid.

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 Unit 2: Introduction to Pharmacokinetics
2.1: Basics of kinetics:
1. Kinetics deals with speed of a process. A process may be zero order or first
order.
2. If the speed of a process does not depend on any concentration term, it is
called zero order process. Example: Active absorption of riboflavin in GIT.
3. The equation for a zero order process is Ct = C0 – K0 t.
4. The time taken for half of the process to be completed is called half-life.
C0 = Initial concentration of drug at zero time.
Ct = Concentration of drug remaining at time ‘t’.
K0 = Zero order rate constant.
A graph of Ct versus t will give a straight line with a
negative slope. Slope is equal to - K0. The Y
intercept is equal to C0.
5. If the speed of a process depends on one concentration term, it is called first
order process. Example: ADME of drugs.
6. The equation for a first order process is
log Ct = log C0 – [ K1 t/2.303 ].
7. The time taken for half of the process to be
completed is called half-life, t1/2 = 0.693/ K1
8. First order kinetics graph: A plot of log Ct versus t
will give a straight line with a – ve slope equal to
– K1 / 2.303. The Y intercept is equal to log C0.
2.2: Introduction to Pharmacokinetics and its importance in Bio
availability and clinical practice / Concepts, definition and explanation of
terminologies used:
1. Pharmacokinetics deals with kinetics of ADME of drugs.
2. Once a drug is administered, it gets absorbed, distributed, metabolized
and eliminated. The speed of these ADME processes should be studied to fix
dose and dosage regimen for a patient.
3. The various important pharmacokinetic parameters of a drug are as follows.

35 | P a g e
a. Absorption rate constant
b. Elimination rate constant
c. Elimination or biological half life
d. Volume of distribution
e. Clearance
f. Area under the curve
4. If we know, how fast the drug is getting absorbed, distributed, metabolized
and eliminated; we can fix the dosage regimen (dose and dosing time table)
for a drug.
5. Example: Paracetamol biological half-life is 4 hours, so we take a
paracetamol tablet every 4 or 6 hours.
6. It is difficult to measure the concentration of drug at the site of action.
Example: The site of action for digoxin is heart muscle.
7. The concentration of drug in the site of action is directly proportional to the
concentration of drug in blood. Hence we can take blood samples to know
how fast the drug is getting absorbed, distributed, metabolized and
eliminated.
8. Example: We give a tablet to a person, and collect his blood samples, every
one hour, until the drug is completely eliminated from the body. Then,
graphs are constructed to know the kinetics of ADME.
9. The different pharmacokinetic parameters are calculated from graphs, which
will be useful for fixing dosage regimen.
10. Clinical pharmacokinetics: Application of pharmacokinetic principles for
fixing dose and dosage regimen in individual patient.
11. Clinical pharmacokinetics is used to fix dose and dosage regimen in
infants, elderly persons, patients suffering with hepatic disease, renal
disease etc.
12. Example: If a person creatinine clearance is half of the normal value, it
indicates that the kidneys are functioning by 50 %, so the dose will be
reduced by half.
13. Pharmacodynamics deals with concentration of drug in blood / receptor
and its pharmacological response.
14. Pharmacokinetics is a study of what the body does to the drug.
15. Pharmacodynamics is a study of what the drug does to the body.

36 | P a g e
2.3: Mathematical model / Pharmacokinetic model / Compartment model
/ Pharmacokinetic study:
1. The drug movement in the body is a complex process. To estimate the
pharmacokinetic parameters of a drug, we require compartment models.
There are mammillary compartment model and catenary model.
2. Mammillary compartment model: In this model, the body is divided into one
or more compartments.
3. In one compartment model, the entire body is considered as a single
compartment.
4. In two compartment model, the body is divided into two compartments.
Brain, liver, blood, heart, kidney, lungs have high blood supply and is
considered as first compartment. The rest of the body is second
compartment.
5. In case of three compartment model, the body is divided into three
compartments, basing on blood supply.
6. Brain, liver, blood, heart, kidney, lungs – first compartment
7. Muscle tissue – second compartment.
8. Adipose tissue and bone tissue – third compartment.
9. The drug enters into the first compartment and distributes to other
compartments in a reversible manner.
10. Drug is eliminated from first compartment.
11. By dividing the body into compartments, calculation of pharmacokinetic
parameters become easy.
12. In mammillary model, we have one or more peripheral compartments
connected to a central compartment. It is also called satellite model.
13. Caternary model: In this model, the compartments are joined to one
other like the compartments of a train. It is shown below.
14. Physiological modeling: In this model, the body is divided into a series
of organs or tissue spaces. The model describes the uptake and elimination
of drug from these organs. This model is used in animals. Example: A drug
is given to rats, after specified time, it is killed, organs are separated, and
amount of drug distributed in each tissue is found out.

37 | P a g e
2.4: Blood level curves:
1. A blood level curve is constructed by taking time on x axis and blood /
plasma drug concentration on y axis.
2. An IV injection or oral tablet is given to a human volunteer and his blood
samples are analyzed for drug concentration. Then the graph is constructed
by taking time on x axis and drug concentration on Y axis. The following
curves will be obtained.

38 | P a g e
3. Cp is concentration of drug in blood / plasma. After an IV injection, the
drug gets distributed to different parts of the body. Elimination of drug takes
place in the kidney and hence blood concentration of drug decreases. This
can be seen in the below graph.
4. In case of oral administration, the blood level curve is shown in the below
graph.
5. The tablet breaks into pieces and the drug dissolves in the GIT fluids. It gets
absorbed into blood.
6. Once the drug is absorbed into blood, elimination of drug also starts.
Initially absorption speed is greater than elimination speed, so the Cp
increases as shown in the graph.
7. After some time, absorption speed is equal to elimination speed and the
graph is flat at the top.
8. Later elimination dominates absorption and the graph shows a decrease in
Cp concentration.
9. The drug will show its action as long as the drug concentration is between
MEC (minimum effective concentration) and MSC (maximum safe
concentration).

Blood level curve after IV Blood level curve after oral tablet
administration administration
MEC = Minimum effective 1 – Absorption phase
concentration 2 – Post absorption phase
MSC = Maximum safe concentration 3 – Elimination phase

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 Unit 3: One compartment open model
3.1: One compartment open model – Intravenous injection (Bolus):
1. In this model, the entire body is considered as a single compartment.

2. A single dose is given intravenously. It is assumed that the drug is

distributed in the body in two to three minutes. Hence there is no
absorption step.
3. Blood samples are taken at different time intervals and they are analyzed for
drug concentration and graphs are drawn. The elimination rate constant is
found from the slope of the graph (zero order or first order graph).
4.

Zero Order: Y intercept = C0, First Order: Y intercept = log C0,

Slope = – KE Slope = – KE / 2.303
C0 = Plasma concentration at zero Cp = Plasma concentration at time t.
time KE = Elimination rate constant

5. Biological Half Life, t 1/2: The time taken for half of the drug to be
eliminated from the body is called biological half life. In zero order
elimination, t ½ = C0 / 2 KE , in first order elimination, t ½ = 0.693/ KE
6. If a drug eliminates slowly from the body, t½will be more. Example:
Paracetamol half life is 4 hours and diazepam half life is 20 hours.
7. Volume of Distribution, Vd: It gives an idea about the distribution of drug
in the body. It is defined as hypothetical volume of body fluid in which the
drug is distributed. Vd = Dose / C0
8. If the drug is present in the blood without being distributed, plasma
concentration is very high and Vd will be less.
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9. If the drug accumulates in a tissue, plasma concentration will be very less
and Vd will be high. Vd of warfarin is 10 liters, Vd of Chloroquine is
15000 Litres.
10. Clearance, CL: It gives an idea about the elimination of drug in the body.
It is defined as theoretical volume of body fluid from which the drug is
eliminated completely. CL = Elimination Rate/ Cp
11. CL = KE VD
12. If the clearance is more, it indicates that the drug is getting eliminated
from the body quickly. Example: Clearance of aspirin is 39 L/hr,
paracetamol is 20 L/hr.
13. In kidney failure, clearance of drug will be less and dose of drug should
be reduced. If clearance reduces by half due to kidney failure in a patient,
decrease dose of drug by half.
14. Area under the Curve, AUC: The area under the plasma concentration
versus time curve is directly proportional to the administered dose. Greater
the dose, greater will be the AUC. If dose is doubled AUC will also double.
15. AUC can be found by trapezoidal rule or by the following formulas.
16. AUC = C0 / KE = Dose / KE VD = Dose / CL
17. AUC is used to compare bio
availability of the same drug
administered by IV and other
routes.

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3.2: One compartment open model – Intravenous infusion:
1. In this model, the entire body is considered as a single compartment.

2. Drug is given intravenously continuously for long periods of time.

3. Intravenous infusion is an example of zero-order absorption and first-order
elimination.
4. Blood samples are taken at different time intervals and they are analyzed for
drug concentration.
The plasma drug concentration at
any time after the start of an
intravenous infusion is given by the
following equation:
5. R is the zero-order rate of infusion (milligrams per min).
6. The various pharmacokinetic
parameters in this model are
calculated by drawing graphs
between log Cp and time t.

7. If the intravenous infusion is discontinued, the plasma drug concentration

decreases by a first order process. The elimination rate constant, KE, can be
obtained from the slope of the curve.
8. As the drug is infused, the plasma drug concentration increases to a
plateau, or steady-state concentration (CSS). Under steady-state
conditions, the rate of infusion equals the rate of drug elimination from the
body.
9. The plasma concentration at steady
state (Css) is given by the following
equation:
10. The rate of drug infusion (R) may be calculated from a rearrangement of
the above equation if the desired Css, the VD, and the KE are known.

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11. A loading dose (DL) is given as an initial intravenous bolus dose to
produce the Css as rapidly as possible. The intravenous infusion is started at
the same time as the DL.
12. The time to reach Css depends on the elimination half-life of the drug.
Reaching Css without a DL takes around 6 half lives. Thus, for a drug with
an elimination t½ of 8 hrs, it will take 48 hours hrs, to reach Css if no
loading dose is given.
13. DL is calculated by the following equation:

The IV infusion keeps the plasma drug concentration between the minimum
toxic concentration (MTC) and the minimum effective concentration (MEC).
3.3 One compartment model extra vascular administration:
1. In this model, the entire body is considered as a single compartment. The
product is taken orally and there is an absorption step.

2. Blood samples are taken at different time intervals and they are analyzed for
drug concentration.
3. Once the drug is absorbed into blood, elimination of drug also starts. Initially
absorption speed is greater than elimination speed, so the Cp increases as
shown in the graph.
4. After some time, absorption speed is equal to elimination speed and the
graph is flat at the top.
5. Later elimination dominates absorption and the graph shows a decrease in Cp
concentration.

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 6. The various pharmacokinetic
parameters in this model are
calculated by drawing graphs.
1 – Absorption phase
2 – Post absorption phase
3 – Elimination phase

MEC = Minimum effective concentration

MSC = Maximum safe concentration
6. Plasma Concentration Cp: The plasma concentration of drug at different
time intervals can be calculated using the below equation.
Cp = Plasma concentration of drug
F = Fraction of drug absorbed

D0 = Dose taken orally K E = Elimination rate constant

K A = Absorption rate constant Vd = Volume of distribution

7. Tmax: The time taken for the peak plasma concentration to occur is called tmax.
The tmaxcan be obtained from the below equation.
A shorter tmax indicates faster rate of
absorption.

8. Elimination rate constant: By drawing log Cp versus time we get the below
graph. The terminal portion (line 2) is the elimination phase. Slope of the
terminal portion is equal to– KE / 2.303.
10. Biological half life: The time taken for half of the drug to be eliminated
from the body is called biological half life. It is given by the equation
t½ = 0.693/KE.
11. Absorption rate constant: The absorption kinetics of a drug into blood is
given by the absorption rate constant. It can be determined by method of
residuals or feathering or peeling or stripping or curve fitting method.
12. Draw log Cp versus time, we will get the below graph.

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13. Line 1 represents the absorption phase.
14. Line 2 represents the elimination phase. Extend this line to cut y axis
(line 3). This line is called back extrapolated (extended) line.
15. Calculate the difference between points on extrapolated line 3 and
corresponding points on line 1, we get residuals.
16. Plot the values of residuals versus time. We will get line 4. Slope of this
line is equal to – KA / 2.303.
17. Absorption Half Life, t 1/2: The time taken for half of the drug to be
absorbed into blood is called absorption half life. t ½ = 0.693/ KA
18. Area Under the Curve: The rate and extent of absorption is called
bio availability. It is measured using the area under the curve. The greater
the AUC greater the bio availability. The AUC is measured by trapezoid rule.
The curve is divided into a number of trapezoids and area of each trapezoid is
calculated by the below formula.
19. Area of trapezoid = W (a+b) /2
20. Calculate the tail area using the formula, Area of tail = Clast / KE

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 Fraction of drug absorbed F :
It is calculated using the below
formula. If F is more, it indicates that
bio availability of the tablet is more.

21. The volume of distribution and clearance are given by the below formula.

3.4 Wagner Nelson method:

1. The absorption rate constant can also be determined by Wagner Nelson
method.
2. Absorption of drugs may be zero order or first order.
3. This method requires that elimination of drugs follow first order kinetics.
4. In this method, the cumulative amount of drug absorbed into the blood at
different time intervals is calculated.
5. Now the amount of drug remaining to be absorbed (DRA) is calculated.
6. If a plot of DRA versus time gives a straight line, it indicates that the drug is
absorbed according to zero order kinetics.
7. If a plot of Log DRA versus time gives a straight line, it indicates that the
drug is absorbed according to first order kinetics.
8. From the slope of the graphs KA can be calculated.

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Slope = - KA Slope = - KA / 2.303

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 Unit 4: Multi Compartment model
4.1 Two compartment model- IV bolus:
1. In this model, the body is divided into two compartments.
2. Brain, liver, blood, heart, kidney, lungs have high blood supply and is
considered as first compartment. The rest of the body is second
compartment.

3. A single dose is given intravenously.

4. The drug enters into the first compartment, distributes quickly to all the
tissues of the first compartment, because the blood supply is very high.
Simultaneously elimination also starts from the first compartment.
5. The drug distributes slowly from first compartment to second compartment
in a reversible manner.
6. Blood samples are taken at different time intervals. They are analyzed for
drug concentration.
7. A plot of log Cp versus time is shown below.

8. Line 1: The Cp falls sharply because the drug is getting distributed from
first compartment to second compartment and getting eliminated at the
same time.
9. At time x in above graph, distribution is complete.
10. Line 2: Distribution is complete, this line represents only elimination
phase.
11. Line 3: This line shows the increase in concentration of drug in tissue
compartment. As drug comes from first compartment to second

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 compartment concentration of drug increases.
12. Line 4: As concentration of drug decreases in central compartment drug
goes back from second compartment to first compartment. Hence its
concentration decreases with time.
13. It is assumed that distribution and elimination follows first order
kinetics.
14. The various pharmacokinetic parameters in this model are calculated by
drawing graphs.
15. The equation for the below graph is Cp = A e – t +Be – t

Cp = Plasma concentration at time t A,B = Hybrid coefficients

= hybrid first order rate constant = hybrid first order rate constant for
for distribution process elimination process

16. The slope of line 2 in the graph is equal to – / 2.303.

17. By extrapolating line to cut y axis, we get y intercept equal to B.
18. Method of residuals can be used to determine A and . Calculate the
difference between points on extrapolated line 3 and corresponding points
on line 1, we get residuals.
19. Plot the residuals versus time, and we get line 4.
20. The y intercept of line 4 is equal to A and slope is equal to – / 2.303.
21. Once A, B, , and are known, other pharmacokinetic parameters can
be calculated.
22. Volume of central compartment = Vc = Dose/ A+B

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23. , 24. CLt = Dose / AUC
4.2: Two compartment model- Extra vascular administration:
1. In this model, the body is divided into two compartments.
2. Brain, liver, blood, heart, kidney, lungs have high blood supply and is
considered as first compartment or central compartment. The rest of the
body is second compartment.

3. A single dose is given orally and there is an absorption step.

4. It is assumed that absorption, distribution and elimination of drug follow
first order kinetics in the body.
5. Elimination is from central compartment only.
6. The drug is absorbed into the first compartment, distributes quickly to all
the tissues of the first compartment, because the blood supply is very high.
Simultaneously elimination also starts from the first compartment.
7. The drug distributes slowly from first compartment to second compartment
in a reversible manner.
8. Blood samples are taken at different time intervals. They are analyzed for
drug concentration.
9. In this model the plasma concentration is given by the below tri exponential
equation.
10. Cp = N e – Ka t +Le – t +Me – t

Cp = Plasma concentration at time t L, M, N = Hybrid coefficients

= hybrid first order rate constant = hybrid first order rate constant for
for distribution process elimination process
Ka = absorption rate constant

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11. A plot of log Cp versus time is shown below. The three exponents can be
found out by step wise application of method of residuals.

12. Line 1: The Cp rises because the drug is getting absorbed.

Simultaneously drug is distributed and eliminated, but since absorption
dominates, line 1 shows an increase in Cp with time.
13. Line 2 represents the elimination phase of the drug and the slope of line
2 in the above graph is equal to – / 2.303.
14. Back extrapolate line 2, we will get line 3. It cuts the y axis and y
intercept is equal to log M.
15. Find the difference between points on line 1 and line 3 and plot these
residuals, we will get curve 4. It is first residual curve. The slope of the
terminal phase line 5 is – / 2.303.
16. Back extrapolate line 5 to cut y axis, the y intercept is equal to log L.
17. The back extrapolated line is called line 6. It is back extrapolated
distribution curve.
18. Using curve line 6 and 4 calculate residuals and plot a second residual
line. The slope of this residual line 7 is = - Ka/2.303.
19. Hence curve 1 has been resolved into three lines 3, 6 and 7, to find out
the pharmacokinetic parameters.

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4.3: Two compartment model IV Infusion:
1. In this model, the body is divided into two compartments.
2. Brain, liver, blood, heart, kidney, lungs have high blood supply and is
considered as first compartment or central compartment. The rest of the
body is second compartment.

3. A single dose is given by constant rate IV infusion and there is no

absorption step.
4. It is assumed that distribution and elimination of drugs follow first order
kinetics in the body.
5. Elimination is from central compartment only.
6. The drug distributes quickly to all the tissues of the first compartment, and
slowly to the tissues of the second compartment.
7. Simultaneously elimination also starts from the first compartment.
8. Blood samples are taken at different time intervals. They are analyzed for
drug concentration.
9. Initially the blood concentration will be zero at zero time. As infusion
continues, drug concentration in blood rises. After some time, steady state is
achieved. Under steady-state conditions, the rate of infusion equals the rate
of drug elimination from the body.
10. At steady state, there is equilibrium between drug in first and second
compartment.
The plasma drug concentration at
steady state after the start of an
intravenous infusion is given by the
following equation:
14. R is the zero-order rate of infusion given in units as milligrams per
hour or milligrams per minute. Vc is volume of the central compartment
and Ke is the elimination rate constant of drug.

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15. The various pharmacokinetic
parameters in this model are
calculated by drawing graphs
between log Cp and time t.

16. If the intravenous infusion is discontinued, the plasma drug

concentration decreases by a first order process. The elimination rate
constant, KE, can be obtained from the slope of the declining plasma drug
concentration versus time curve.
17. A loading dose (DL) is given as an initial intravenous bolus dose to
produce the Css as rapidly as possible. The intravenous infusion is started at
the same time as the DL.
18. The time to reach Css depends on the elimination half-life of the drug.
Reaching Css without a DL takes around 6 half lives. Thus, for a drug with
an elimination t½ of 8 hrs, it will take 48 hours hrs, to reach Css if no
loading dose is given.
19. DL is calculated by the following equation:

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 Unit 5: Multiple – Dosage Regimens
5.1: Kinetics of Multiple Dosing / Dosage regimens /Pharmacokinetics of
a drug following repeated administration (IV / extra vascular):
1. Dosage regimen is the manner in which a drug is taken.
2. Example: Paracetamol tablets are taken four times a day.
3. In chronic diseases, the same dose is administered repeatedly at fixed
dosing intervals. Dose and dosing interval should be designed basing on the
pharmacokinetic parameters of the drug and disease state of the patient.
4. Example: In diabetes, B.P, tablets have to be taken daily at a definite time in
the day.
5. The Cp versus time graph on repeated administration is given below.

6. Generally, the next dose is administered before the previous dose is

completely eliminated.
7. The dosing interval is equal to the half life of the drug. For example half life
of paracetamol is four hours. So every four hours, a paracetamol tablet is to
be taken.
8. On repeated administration drug accumulates in the body. As concentration
of drug increases in the blood, elimination speed also increases according to
first order kinetics.
9. When the rate of administration is equal to rate of elimination, steady state
reaches in the body. Generally steady state is reached after five to six half
lives.
10. At steady state, the amount of drug eliminated during one dosing interval
is equal to the dose administered.
11. At steady state, Cp values will fluctuate between Cpss min and Cpss max.
12. In repeated IV administration, steady state Cpss min and Cpss max can be
calculated using the below equations.
D = dose and τ = dosing interval.

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13. In repeated EV administration, steady state Cp min and Cp max can be
calculated using the below equations.

15. In case of emergency a loading dose is given followed by maintenance dose.

IV administration: Loading dose = Cp desired x Vd
EV administration: Loading dose = Cp desired x Vd / F
16. The average steady state plasma concentration is directly proportional to
dosing rate FD/ τ.
17. By increasing F, D and decreasing τ we can increase CpssAv . It should be
between MEC (minimum effective concentration) and MSC (maximum safe
concentration).

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 Unit 6: Non Linear Pharmacokinetics or Dose Dependent
Pharmacokinetics or Mixed Order Pharmacokinetics or Capacity Limited
Pharmacokinetics or Saturation Pharmacokinetics
6.1: Introduction:
1. If the rate (speed) of ADME of a drug depends on drug concentration, the
ADME is said to follow first order or linear pharmacokinetics.
2. If the rate of ADME of a drug does not depend on drug concentration, the
ADME is said to follow zero order pharmacokinetics.
3. If the rate of ADME of a drug follows first order kinetics at low drug
concentration and zero order at high drug concentration, the ADME is said
to follow mixed order pharmacokinetics.
4. Example: Riboflavin is absorbed by carriers in GIT. At low doses, absorption
rate follows first order kinetics. At high doses, carriers become saturated
and absorption rate is constant. Hence riboflavin absorption is following
mixed order kinetics.
6.2: Evidence for non linear pharmacokinetics:
1. In linear pharmacokinetics, the various pharmacokinetic parameters like F,
Ka, Ke, t1/2, Cl, do not change with change in dose of drug. In non linear
pharmacokinetics these pharmacokinetic parameters change with the dose
given to the patient.
2. AUC is not proportional to the dose of drug. In linear kinetics, if dose is
doubled, AUC will be doubled. If dose is increased by three times AUC will
increase by three times. In case of non linear kinetics the AUC will not
change in a direct proportional manner.
3. Steady state plasma concentration Css is not proportional to administered
dose. In linear kinetics, if dose is doubled, Css will be doubled. If dose is
increased by three times, Css will increase by three times. In case of non
linear kinetics, the Css will not change in a direct proportional manner.
Example: when the dose of phenytoin is increased from 300 mg/day to 450
mg/day, the Css increased by 10 times.

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6.3: Causes for Non Linear Pharmacokinetics: Non linearity can be observed
in absorption, distribution, metabolism and elimination of drugs.
1. Non linear drug absorption: Riboflavin is absorbed by carriers in GIT. At
low doses, absorption rate follows first order kinetics. At high doses carriers
become saturated and absorption rate is constant. Hence riboflavin
absorption is following mixed order kinetics.
2. Non linear drug metabolism: Some drugs are metabolized in liver by
enzymes. At low doses, metabolic rate follows first order kinetics. At high
doses, enzymes become saturated and metabolic rate is constant. Hence
liver metabolism is following mixed order kinetics. Example: Phenytoin,
alcohol shows mixed order metabolism.
3. Non linear drug distribution: Some drugs bind to proteins in blood. At low
doses, protein binding follows first order kinetics. The free drug available for
distribution increases linearly with dose. At high doses, proteins are
saturated with drug and no more protein binding occurs. As a result, free
drug concentration increases greatly and high amount of free drug is
available for distribution. This may even cause toxic effects. Example:
salicylate, phenylbutazone, saturate albumin in blood.
4. Non linear drug elimination: Drugs that undergo tubular secretion and re
absorption by carriers in kidney show non linear pharmacokinetics. At low
doses, tubular secretion and re absorption follows first order kinetics. At
high doses, carriers become saturated and tubular secretion and re
absorption rate is constant. Hence elimination is following mixed order
kinetics. Example: Penicillin G is eliminated by tubular secretion and its
elimination follows non linear kinetics.

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6.4: Non Linear Pharmacokinetics Equation / Michaelis-Menten Equation:
Michaelis - Menten Equation is used to calculate pharmacokinetic parameters
in mixed order kinetics.

Vmax = Maximum speed possible with the

process. C = Concentration of drug,
Km = Michaelis constant which depends on
interaction between drug and carrier or
enzyme.

Three situations can be considered, depending on values of Km and C.

a. When Km = C, then Speed = Vmax / 2.
b. When Km is very large than C, then, Speed = Vmax C/2. The process is
following first order kinetics.
c. When Km is very small than C, then, speed = Vmax. The process speed is
constant, i.e. it is following zero order kinetics.
The other pharmacokinetic parameters can be calculated using the below
equations.

CLT = Total Clearence, D = dose,

t1/2 = Biological half life,
F= bio availability, τ = dosing interval

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 Unit 7: Non Compartmental Pharmacokinetics
7.1: Introduction:
1. It is also called model independent method.
2. Some drugs follow two compartment model on IV administration and one
compartment model on oral administration. In such cases this non
compartmental model will be useful.
3. Calculation of pharmacokinetic parameters is difficult in compartment models
and computers are required.
4. No assumption of compartment model is required. It is assumed that drug and
its metabolites follow first order kinetics process in the body.
5. It is based on statistical moment theory. After an IV administration the drug
molecules are randomly distributed in the body. Some molecules are eliminated
quickly while some drug molecules are eliminated slowly. The difference in the
residence time for each molecule depends on chance according to statistical
moment theory. Each molecule has a certain probability to be eliminated at a
certain time ‘t’.
6. From this theory, the mean residence time of the drug in the body can be found
out.
7.2: Determination of Pharmacokinetic Parameters, MRT – IV administration:
1. Give an IV dose to a person and collect his blood samples at different time
intervals.
2. Analyze the blood samples and construct a Cp versus time graph. This curve is
called zero moment curve.
3. Now construct a Cp t versus time curve. This is called first moment curve.
4. Calculate the area under the zero moment curve (AUC) and first moment curve
(AUMC) by trapezoidal rule.
5. The average time that the drug resides (stays) in the body is given by mean
residence time (MRT).
6. MRT = Total residence time for all drug molecules / Number of drug molecules
7. MRT = AUMC / AUC
8. MRT represents the time taken for 63.2 % of drug to be eliminated from the
body. MRT is similar to biological half life in compartment models.
9. If MRT is more, it indicates that the drug is eliminated slowly from the body.
10. If MRT is less it indicates that the drug is eliminated quickly from the body.

59 | P a g e
 Zero Moment Curve First Moment Curve

300 1000

250 800
200
600

Cp . t
Cp

150
400
100

50 200

0 0
0 5 10 15 20 25 30 0 5 10 15 20 25 30
Time Time

7.3: Determination of Pharmacokinetic Parameters MRT – Extra Vascular

administration:
1. In IV administration, the MRT of a drug in a given patient is generally constant.
2. In oral administration the drug spends additional time at the site of
administration. Mean absorption time MAT is the average time a drug molecule
spends at absorption site.
3. Give an oral dose to a person and collect his blood samples at different time
intervals. Analyze the blood samples and construct zero moment and first
moment curves.
4. MRToral = AUMC / AUC
5. MRToral = MRTiv + MAT
6. MAT can be found out if we know MRToral and MRTiv.
7. MAT of different brands of same drug can be compared for absorption studies.
Example: MRT of paracetamol IV injection is 5 hours, MRToral of crocin is 6 hours
and MRToral of calpol is 7 hours. So MAT of crocin is 1 hour and calpol is 2 hours.
It indicates that crocin is getting absorbed faster than calpol.
Zero Moment Curve First Moment Curve

140
900
120
800
100 700
80 600
Cp

Cp . t

60 500
400
40
300
20 200
0 100
0 5 10 15 20 25 30 0
0 5 10 15 20 25 30
Time
Time

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7.4: Advantages of Non Compartmental Analysis:
1. Calculation of pharmacokinetic parameters is difficult in compartment models
and computers are required. This model is simple for calculating
pharmacokinetic parameters.
2. No assumption of compartment model is required.
3. A detailed description of distribution and metabolism kinetics is not required.
Disadvantages:
1. It does not treat non - linear cases satisfactorily.

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 Unit 8: Bioavailability and Bioequivalence
8.1: Definitions:
1. Bioavailability: The rate and extent (amount) of drug reaching the blood
(or site of action) from the dosage form is called bioavailability. Example:
Bioavailability of an IV injection is 100 %. Bioavailability of a tablet is 80 %.
2. Absolute bioavailability: If the bioavailability of a product is compared with
an IV injection, it is called absolute bioavailability. Example: bioavailability
of diclofenac tablets is compared with diclofenac injection.
3. Relative bioavailability: If the bioavailability of a product is compared with
an oral solution, it is called relative bioavailability. Example: bioavailability
of paracetamol tablets is compared with paracetamol elixir.
4. Bioequivalent products: If the rate and extent of drug absorption is nearly
same for the two products, they are called bioequivalent products. Example:
Crocin and calpol tablets have 500 mg paracetamol and are bioequivalent.
5. Pharmaceutical equivalents are drug products that contain the same drug
and are of the same dosage form. They are identical in strength,
concentration, and route of administration. Excipients may vary. Example:
Calpol and Crocin tablets have 500 mg paracetamol.
6. Pharmaceutical alternatives are drug products that contain the same drug
but are different salts, esters, or complexes. Example: tetracycline
hydrochloride and tetracycline phosphate.
7. Generic product: It is an alternative to innovator product. It will show the
same therapeutic performance as the innovator product. Example: Bayer
company is innovator of ciprofloxacin.
8.2: Objectives of bio availability studies:
1. Development of a suitable dosage form for the newly discovered drug.
Example: If a drug is having poor absorption in GIT, it is formulated as an
injection.
2. To study the effect of excipients on bioavailability of drugs.
3. To study the effect of food and patient related factors on bioavailability of
drugs.
4. Quality control of drug products during manufacturing.
5. Comparison of bioavailability of generic product with innovator product.

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8.3: Methods for assessing bioavailability:
1. Pharmacokinetic methods using blood level data or urinary excretion data of
drugs.
2. Pharmacodynamic methods based on pharmacological or therapeutic
response.
3. Pharmacokinetic method using blood level data:
a. A human subject is given an IV dose and blood samples are collected at
different time intervals. Now a graph is constructed by taking time on x axis
and plasma concentration of drug on y axis.
b. After suitable wash out period, the human subject is given a tablet orally
and his blood samples are collected at different time intervals. Now a graph
is constructed by taking time on x axis and plasma concentration of drug on
y axis.
c. The bioavailability of the product is found out by calculating Cmax, Tmax,
and AUC.
d. The Cmax should be within the therapeutic range.
e. The Tmax will be less if the drug is absorbed rapidly.
f. The AUC is a measure of extent of absorption. If AUC is more bioavailability
will be more.
g. Bioavailability = F = AUC of oral product x Oral dose/AUC of IV product x
IV dose.

Blood level curve for IV injection Blood level curve for oral tablet

63 | P a g e
4. Pharmacokinetic method using urine data:
a. The principle in this method is the urinary excretion rate of drug is directly
proportional to plasma concentration of drug.
b. This method can be used only if at least 20 % of drug is excreted in urine.
c. The human subject is asked to fast over night. Water loading is done by
taking 400 ml of water after fasting. This will help in collecting sufficient
urine samples.
d. After one hour from water loading, the urine bladder is emptied completely,
and this urine sample is used as blank.
e. Now the IV product is administered. 200 ml water must be taken orally
every one hour for four hours.
f. Urine samples are collected at different time intervals. Each time, the total
volume of urine should be measured accurately.
g. Urine samples should be collected for at least seven biological half lives.
h. The urine samples are analyzed for drug content. Then, the total amount of
drug excreted in urine at each time interval is calculated.
i. Rate of urinary excretion of drug is calculated for each time interval.
j. Now a graph is plotted by taking time on X axis and rate of urinary excretion
of drug on Y axis.
k. After suitable wash out period, the human subject is given a tablet orally
and his urine samples are collected at different time intervals. Rate of
urinary excretion of drug versus time graph is constructed.
l. The bioavailability of the product is found out by calculating maximum
urinary excretion rate of drug, Tmax, and AUC.
m. The maximum urinary excretion rate corresponds to Cmax in blood level
data.
n. The time for maximum urinary excretion rate corresponds to Tmax of blood
level data.
o. The AUC of the graph corresponds to AUC of blood level data.
p. The AUC is a measure of extent of absorption. If AUC is more bioavailability
will be more.
q. Bioavailability = F= AUC of oral product x Oral dose/AUC of IV product x IV
dose.

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 IV administration Oral administration

5. Acute pharmacological response method:

a. In this method, the product is given by IV / oral route and the
pharmacological response is measured.
b. Example: ECG readings change, B.P change, pupil diameter change etc are
measured at different time intervals.
c. Then curves are constructed by taking time on x axis and the
pharmacological effect on y axis.
d. Pharmacological response should be measured for at least three biological
half lives.
e. By comparing the AUC for IV product and oral product, bioavailability can
be calculated.
f. The disadvantage with this method is, the pharmacological response is
variable and may not correlate with bioavailability of drug.
6. In Vitro drug dissolution rate and bioavailability:
a. Dissolution rate testing is a simple method to have an idea about
in vivo bioavailability.
b. The dissolution test mimics the in vivo conditions.
c. The dissolution test is carried on the two brands of products and their
graphs are compared. Time is taken on x axis and % of drug dissolved is
taken on y axis. Then, the t50, t90 and AUC are calculated and compared
for the two brands.
d. The difference factor f1 and similarity factor f2 are calculated.
e. If f1 is zero and f2 is 100 both the products will have same bioavailability.
f. If difference factor is greater than 15 and similarity factor is less than 50,
the two products will have different bioavailability.

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8.4: Experimental design in bioavailability / bioequivalent studies /
evaluation of bioavailability studies:
1. Bioavailability / bioequivalent studies are carried out with generic product
and innovator product.
2. These studies are carried out in healthy, young, adult, male volunteers.
3. Latin square cross over design is used for these studies.
4. A two way cross over design has 12 volunteers. They are divided into two
groups. First group receives generic product and the second group receives
innovator product. Blood samples or urine samples are collected and
analyzed for drug.
5. After wash out periods, first group receives innovator and second group
receives generic product. Blood samples or urine samples are collected and
analyzed for drug.
6. Now graphs are constructed by taking time on x axis and Cp on y axis. From
the AUC values, the bioavailability of the product is calculated.
7. The wash out period is generally one week, so that the drug gets completely
eliminated from the body.
8. Statistics is applied to the results to know whether the generic and
innovator product are bioequivalent or not.
9. Randomized block design: In this design, the volunteers are divided into
homogenous blocks.
10. Example: We want to compare the bioavailability of six brands of
paracetamol tablet. We will take 60 volunteers and divide them into 10
blocks. Each block has six homogenous volunteers.
11. Now, each volunteer in a block will receive one brand randomly.
12. This is done for all the 10 blocks.
13. Blood samples are taken and analyzed for drug and graphs are
constructed.
14. Statistical analysis of the AUC, Cmax, tmax is done to know whether all
the six brands are bioequivalent or not.
15. Cmax and tmax gives an idea of rate of absorption and AUC is a measure
of extent of absorption.

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8.5: In vitro and in vivo correlation methods:
1. There are various methods to correlate the in vitro dissolution data and in
vivo data.
2. If there is a good correlation between in vitro and in vivo data, we can
predict in vivo performance from in vitro dissolution data.
3. Correlation is done between rate of dissolution, extent of dissolution and
pharmacokinetic parameters like Cmax, tmax, AUC.
4. The physico chemical properties of dosage form influence the in vitro
dissolution and in vivo performance.
5. In vitro testing criteria must be designed to correlate with in vivo data.
6. There are three levels of correlation, level A, level B and level C.
7. Level A IVIVC is a point to point correlation between in vitro dissolution rate
and an vivo input rate. In vitro data can be used to predict in vivo profile
(Cmax, Tmax, AUC and elimination half life.)
8. In level B correlation, the mean dissolution time is correlated with mean
residence time of the drug. This correlation cannot be used to predict the
plasma concentration profile. Level B correlation is not acceptable for
biowaivers.
9. A level C correlation is between a single in vitro dissolution parameter and a

single in vivo dissolution parameter. Example: Correlation between amount

of drug dissolved at a certain time point and Cmax.

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