BP 181212054815
BP 181212054815
BP 181212054815
ICS
PHARMACOKINET
ICS PRACTICAL
MANUAL
RESHMA FATHIMA.K
ASSISTANT PROFESSOR
GRACE COLLEGE OF
PHARMACY,
PALAKKAD
1
EXP.NO:1
DATE:
INVI
TRO
DISS
OLU
TION
STUD
Y OF
COM
PRES
SED
TABL
ET
AIM
To study
the
invitro
drug
release of
given
compress
ed tablet
of
paracetam
ol.
CROSS THE MEMBRANE SOLUTION
&
REQUIR
REACH THE
EMENTS
BLOOD
Various types’ dissolution test apparatus are official in B.P, I.P& USP.
Paracetamol pure, paracetamol tablets, sodium hydroxide, potassium hydrogen
phosphate, dissolution test apparatus, U.V spectrometer.
TYPE -I Apparatus: Paddle
According to I.P
TYPE- II Apparatus: Basket
THEORY
Dissolution is defined as the amount of drug substance that goes into
TYPE-III Apparatus: Basket
According to U.S.P
solution per unit time of solvent composition.
2
It is one of the quality control tool for solid dosage form .The
bioavailability of a drug from its dosage form is dependent upon the dissolution
TYPE-II Apparatus: Paddle
Choice of apparatus is effect on the physicochemical properties of
dosage form. Compression tablet are standard uncoated tablet made by
direct compression or wet granulation method they may used for local action on
GIT as systemic action. A drug product with rapid dissolution (compressed
tablet) one when greater than or equal to 85% of the labeled amount of drug
dissolved with on 13 minutes using U.S.P apparatus I &II in volume of less
than or equal to 900ml buffer solution.
PROCEDURE
a) Preparation of phosphate buffer of pH5.8
3
REPORT
Invitro dissolution Calculation curve Standard solution
% of drug release of
tablet I 103.5% 99.7%
% of drug release of
tablet II 105.4% 103.8%
% of drug release of
tablet III 108% 105.3%
REFERENCES
Laboratory manual of Biopharmaceutics &pharmacokinetics by V.B
Phase, R.J.Duas page no: 18-21
Indian Pharmacopoeia 1996 vol I
4
EXP.NO:2
DATE:
INVITRO DISSOLUTION STUDY OF MARKETED SUSTAINED
RELEASE TABLET
AIM
To study the invitro drug release of given diclofenac sodium(sustained
release).
REQUIREMENTS
Diclofenac sodium, dissolution medium, apparatus, U.V spectrophotometer
THEORY
SUSTAINED RELEASE TABLET
The main aim behind formation of this dosage form into the medicaments
for a long duration after administration of a single tablet.
A wide spread are of this type of tablet is mainly because of improvement in
the patent complaints as the dose frequency is reduced patient can be an
undisturbed sleep at night. It also beneficial for psychological patients who
forget to take their tablet regularly and the dose related side effect and toxicity
are reduced. Any adjuvants that can water uptake rate swelling and feeling
characteristics of matrixing agents can after the release of rate of API Eg:
electrolyte on hydroxyl propyl methyl cellulose (HPMC) matrix tablets. It’s also
possible to achieve pulsed drug release. Weakly basic drugs exhibits good
solubility at low pH while less soluble drugs shows at high pH, conditions
which can results on incomplete drug release co-sustained release formulation.
The drug release can be modified by providing suitable micro environment pH
in the tablet.
Eg: Acidic polymers, succinic acid etc.
Similarly inclusion of alkaline polymer results in desirable drug release of
acidic drug. Classic approaches are usually based on adaptation of either film
5
coated or multiparticle technologies or those including slow release matrixes.
Which are discussed below
Coating technologies
It combines semi permeable coating and osmotic tablets core to produce
zero
order release. Technology attention s also focused to triggered drug release at
critical time point Eg: To achieve drug release 1-2 hours before patient
awakeness also prolific research activities have yield a technology called ring
cap which is based on tab, preferentially film coated and partially coated with a
series of ring whose respective thickness provides the means of moderating the
rate which the drug is released from final dosage form.
Matrix technology
Classically the matrix products exhibits order (or perhaps sq. root of
times).
Drug released characteristics in order to achieve zero order release
characterization it’s necessary to employed specially designed materials of
strategies that seen to manipulate tablet structures of geometry. Combination of
conventional HPMC matrix technology with upper and lower layer helps to
moderate drug release by increasing in surface area with reduction in drug
concentration within the device .Release of drug can follow various
mechanisms:
Diffusion is rate limiting
Dissolution is rate limiting
Osmotic pressure is rate limiting
Release is controlled by ion exchange
PROCEDURE
Dissolution test for commercial ibuprofen was performed using dissolution test
apparatus [USP 23] . In this paddle type was used to carry out the test.The
set
6
condition was 900ml of purified water at temperature 370c ± 0.50C , 50 rpm. A
tablet was introduced into dissolution test apparatus. Samples were
every 1hr and same volume of fresh dissolution medium was replaced. Samples
were diluted suitably and analyzed by UV spectrophotometer at 254nm.
Percentage drug release was found out by the following formula:
𝑡𝑒𝑠𝑡𝑎𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐
Percentage drug release = 𝑒 𝑑𝑎𝑟𝑑 × 100
𝑠𝑡 𝑎𝑛
𝑎𝑏 𝑠𝑜𝑟𝑏 𝑎 𝑛 𝑐𝑒
REPORT
The percentage drug release from given ibuprofen 100mg at the end of 10hrs
was found to be
REFERENCES
Indian Pharmacopoeia 1996 vol I
Laboratory manual of Biopharmaceutics &pharmacokinetics by V.B
Phase, R.J.Duas page no: 18-21
7
EXP.NO:3
DATE:
DETERMINATION OF PARTITION COEFFICIENT AND
DISSOCIATION CONSTANT
AIM
To determine Ka, pKa and partition coefficient (pc) value of salicylic acid and
study their relationship
REQUIREMENTS
Separating funnel, 10ml standard flask, u.v spectrophotometer
THEORY
Salicylic acid is a relatiely polar, poorly aqueous soluble material. The salt form
however is quite water soluble by changing PH of the aqueous buffer, you are
able to alter the ratio between the ionized and unionized form of the acid. Since
the unionized form is extracted into the organic phase, the fraction extracted
will vary with PH of the aqueous solution. The definition of ionization constant
(Ka) can be useful.
In aqueous buffer H+
[𝐻+][𝐴−] [𝐻𝐴]
Ka= OR 𝐾𝑎
[𝐻𝐴]
[𝐻+]
This given in an equation with Pc” as function of [H+] with two unknown
parameter, Pc and Ka .This can be converted into a straight line equation by
taking the reciprocal of both sides of the equation.
Thus,
𝑘
1 [𝐻+]
𝑃 𝑐 = 𝑃 𝑐 [𝐻+]
+ 𝐴
𝑃𝐶 [𝐻+]
1
𝑃𝑘𝑐𝐴
= 1+
′ 𝑃𝑐
𝑃𝐶 [𝐻 +]
PROCEDURE
1. Prepare 100ml of buffer at PH 2.5
2. Weigh accurately 200mg of salicylic acid and transfer to 100ml
volumetric flask and adjust volume by buffer of pH 2.5.similarly prepare
0.02% solution of salicylic acid with buffer of pH3 and 4
3. Take 4ml of buffer pH of 2.5 containing salicylic acid stock solution and
add 1ml of ferric nitrate solution. Allow the colour to form and measure
the absorbance at 540nm using spectrophotometer. This is the absorbance
of absorbance I
9
4. Next test 5ml of buffer pH2.5 containing salicylic acid stock solution and
add 5ml of aqueous solvent hexane/ethyl acetate stopper and shake the
test tube for 5 min to complete the extraction.
5. Allow the two phase to settle
6. Remove 4ml of aqueous phase add 1ml ferric nitrate solution(0.55%ferric
nitrate in 0.4m nitric acid)allow the color to form and measure the
absorbance at 540 nm. This is absorbance II
7. Using the two absorbance reading ,calculate the apparent partition
coefficient
𝑡𝑜𝑡 𝑎𝑙 𝑖 𝑛 𝑜𝑟𝑔𝑎 𝑛 𝑖𝑐 𝑝 ℎ𝑎 𝑠𝑒
𝑃𝑐′
𝑡 𝑜𝑡 𝑎 𝑙 𝑖 𝑛 𝑎 𝑞 𝑢 𝑒 𝑜 𝑢 𝑠 𝑝 ℎ𝑎 𝑠𝑒
REPORT
Extraction of salicylic acid in organic layer at pH 2.5 was
The partition coefficient of salicylic acid was found to be
Pka of salicylic acid was found to be
REFERENCE
Dr.S.B. Bhise, Dr.R.J.Dias laboratory manual of Biopharmaceutics
and pharmacokinetics
10
EXP.NO:4
DATE
DETERMINATION OF PARTITION COEFFICIENT AND
DISSOCIATION CONSTANT OF IBUPROFEN
AIM
To determine Ka, pka, and partition coefficient (Pc) of ibuprofen and study their
relation ship
REQUIREMENTS
Separating funnel, 10ml standard flask, u.v spectrophotometer
THEORY
PH- partition hypothesis was put forth by Brodie et al., which states that
drugs
are absorbed from the gastrointestinal track by passive diffusion depending on
the fraction of undissociated drug at the pH of the intestine. Thus, the process of
absorption is governed by:
1. The dissociation constant(pKa) of the drug
2. The lipid solubility of the unionized drug(PC)&
3. The pH at the absorption site
PROCEDURE
1. Prepare 100ml of buffer at pH2.5
2. Weigh accurately 20mg of ibuprofen and transfer to 100ml volumetric
flask and adjust volume by buffer pH 2.5. Similarly prepare 0.02%
solution of ibuprofen with buffer of pH 3&4.
3. Take 4ml of buffer pH of 2.5 containing ibuprofen solution.measure the
absorbance at 540nm using spectrophotometer. This is taken as
absorbance I
4. Next, take 5ml of buffer pH2.5 containing ibuprofen solution and
add
5ml of aqueous solvent hexane/ ethyl acetate. Stopper and shake the test
tube for 5min to complete the extraction. Allow the two phase to settle.
Remove 4ml of aqueous phase and measure the absorbance at 540nm.
This is absorbanceII
11.Using the two absorbance reading ,calculate the apparent partition
coefficient
𝑡 𝑜𝑡 𝑎𝑙 𝑖 𝑛 𝑜𝑟𝑔 𝑎𝑛 𝑖𝑐 𝑝 ℎ𝑎 𝑠 𝑒
𝑃𝑐′
𝑡 𝑜𝑡 𝑎𝑙 𝑖 𝑛 𝑎 𝑞 𝑢 𝑒𝑜 𝑢 𝑠 𝑝 ℎ𝑎 𝑠𝑒
[( 𝐴𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒−𝐼 ) −(𝑎𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒−𝐼𝐼)]
Pc”=
𝑎 𝑏𝑠𝑜𝑟𝑏𝑎 𝑛𝑐𝑒 𝐼
REFERENCE
Dr.S.B. Bhise, Dr.R.J.Dias laboratory manual of Biopharmaceutics
and pharmacokinetics page no1-6
14
EXP.NO:5
DATE:
PROTEI
N
BINDIN
G
STUDY
USING
DYNAM
IC
DIALYSI
S
METHO
D
AIM
Study the
protein
binding of
ibuprofen
by
dynamic
dilution
method.
REQUIR
EMENTS
Ibuprofen, egg albumin, egg membrane, u.v spectrophotometer, magnetic stirrer
etc.
THEORY
The phenomenon of complex formation with proteins is called as protein
binding of a drug. Binding of drugs generally involves weak chemical
bonds
such as hydrogen bonds, ionic bonds or Vander waal’sforces and therefore is as
reversible process. 15
PROCEDURE
Preparation of egg shell membrane
Soak a whole chicken egg in 0.5N HCl solution. The outer calcaraceous shell
gets dissolved then cut off part of egg shell membrane and remove the inner
contents. Wash the obtained membrane thoroughly in distilled water and store
in refrigerator.
Protein binding study of ibuprofen
Tie the egg membrane at one end of open ended glass cylinder and use it
as protein compartment[donor]
Use glass beaker capacity of 25ml as non-protein compartment receptor
and fill it with 20ml distilled water.
16
Place the drug solution to the inner solution tube an immerse into the
beaker. Take care to maintain the level of drug solution coincide with
buffer in outside compartment and fixed with stand.
Keep this whole setup on magnetic stirrer and sir the outer compartment
continuously at an optimal speed. Maintain the temperature 350c ±20c for
the whole experiment.
At predetermined time intervals remove 1ml of the sample from the
beaker an replace it with same volume of 6.8 phosphate buffer.
Determine the concentration of ibuprofen spectrophotometrically at
254nm.
REPORT
REFERENCE
Dr.S.B.Bhise, Dr.R.J. Dias laboratory manual of Biopharmaceutics
and pharmacokinetics
17
EXP.NO:6
DATE:
DETERMINATION OF PROTEIN BINDING USING EQUILIBRIUM
DIALYSIS METHOD
AIM
To determine the protein binding efficiency of ibuprofen.
REQUIREMENTS
Egg membrane, open ended cylinder, beaker, magnetic stirrer, and ibuprofen.
THEORY
In this method human serum albumin is placed in egg membrane tube and the
drug is placed in the outside vessel which is having Ph 6.8 phosphate
buffer
solution.As its dialysis continues the drug molecule permeate through the
membrane and enter into egg membrane tube.As a result drug protein binding is
observed.Albumin being macromolecules can’t pass through the membrane to
outside the vessel.As the time elapses more amount of drug permeate and
available in the tube.If binding occurs the drug concentration in the
bag
containing protein should be greater than its concentration in outside vessel.
PROCEDURE
Take an open ended cylinder and the one end is tied using egg
membrane.
Prepare 5% albumin solution and this solution was introduced [5ml into
egg membrane containing cylinder].
The cylinder is immersed into the beaker containing buffer solution with
Ph6.8. 18
𝑡𝑒𝑠𝑡
Percentage protein binding =
𝑠𝑡 𝑎𝑎𝑛𝑏𝑑𝑠𝑜𝑟𝑏
𝑎𝑟 𝑑 𝑎𝑛 𝑐 𝑒
𝑎 𝑏 𝑠𝑜𝑟𝑏 𝑎𝑛 𝑐 𝑒
REPORT
Percentage of protein binding of ibuprofen at the end of 30min was found to be
REFERENCE
Dr.S.B.Bhise, Dr.R.J. Dias laboratory manual of Biopharmaceutics
and pharmacokinetics
19
EXP.NO:7
DATE:
DESIGN AND EVALUATION OF TRANSDERMAL PATCHES
CONTAINING DICLOFENAC SODIUM
AIM
To prepare and evaluate transdermal patches containing diclofenac sodium and
evaluate their properties.
REQUIREMENTS
Diclofenac sodium, ethyl cellulose, HPMC, polyvinyl alcohol, propylene
glycol, methanol.
THEORY
Transdermal therapeutic systems are defined as self-contained discret dosage
forms which when applied to the intact skin, deliver the drugs through skin for a
controlled rate to systemic circulation.Transdermal drug delivery systems
are
topically administered medicaments in form of patches that deliver drugs for
systemic effects at a predetermined and controlled rate. Transdermal drug
delivery is the noninvasive delivery of medications from surface of skin. Drugs
can be delivered across the tissue adjacent to the site of application
[topical
delivary] or to skin to have an effect after distribution to circulatory system,
inorder to deliver therapeutic agents through human skin for systemic
effects.
The comprehensive morphological biophysical and physiochemical properties
of skin are to be considered. Transdermal drugs provides a leadin edge
over
injectables and oral routes by increasing
20 patient compliance and avoiding first
pass metabolism respectively. Transdermal drug delivery not only provides
controlled, constant administration of drug but also allows continous input
drug with short biological halflife and eliminates pulsed entry into systemic
circulation. It often causes undesirable side effect.
PROCEDURE
Solvent evaporatory method
Polymer of different ratio are accurately weighed and dissolved in 5ml of
methanol
In this solution add 1.5ml of propylene glycol [plastiscer]
After preparation of polymeric solution 10mg of dichlofenac sodium is
dispersed using magnetic stirrer for 2hrs
The preparation is done on PVA backing membrane and dried at 400c for
4hrs
Finally this patches was removed and wrap with aluminium foil and kept
in dessicator
21
REPORT
Transdermal patches was prepared and evaluated
REFERENCES
Laboratory manual of Biopharmaceutics &pharmacokinetics by V.B
Phase, R.J.Duas page no: 18-21
Indian Pharmacopoeia 1996 vol I
22
EXP.NO:8
DATE:
FORMULATION AND EVALUATION OF TRANSDERMAL PATCHES
OF IBUPROFEN
AIM
To formulate and evaluate transdermal patches of ibuprofen.
REQUIREMENTS
Ethyl acetate, methanol, ethyl cellulose, dibutyl phthalate, ibuprofen.
THEORY
Transdermal patches drug delivery system administered topically in the form of
patches that deliver the drug for a predetermined and controlled rate.
Components of transdermal drug delivery system include drug, polymer,
plastisizer, penetration enhancer, backing membrane.It is very helpful for the
avoidance of first pass metabolism for long duration of action of drugs and
also
suitable for drugs having very short half-life. Narrow therapeutic index, poor
oral bioavailability. The drugs used in transdermal drug delivery system
should
have:
Water solubility greater than 1µg/ml.
Oil solubility greater than 1µg/ml.
Molecular weight less than 1000.
Dose less than 10mg.
PROCEDURE
Weigh separately 400mg ethyl cellulose and 100mg ibuprofen.
Take 100ml beaker and add 16ml ethyl acetate and 4ml methanol in
ratio 8:2.
Keep the beaker on top of magnetic stirrer and add slowly ethyl
cellulose to form complete dispersion.
To this add 100mg drug and again stir to get a uniform solution.
Finally add required quantity of butyl phthalate, pour the solution to
petridish having area 9.4cm2.
Put a funnel in an inverted position inorder to give controlled
evaporation.
After 24hrs scrap the film and do the evaluation study using Franz
diffusion study.
24
REPORT
Transdermal patches of ibuprofen was prepared and submitted.
REFERENCES
Laboratory manual of Biopharmaceutics &pharmacokinetics by V.B
Phase, R.J.Duas page no: 18-21
Indian Pharmacopoeia 1996 vol I
25
EXP.NO:9
DATE:
MODELLING OF DRUG RELEASE FROM DELIVERY SYSTEM
USING KINETICS SOFTWARE
AIM
To determine the release pattern and release mechanisms of given invitro
release data.
REQUIREMENTS
Kinetics software, dissolution data.
THEORY
Invitro release kinetics
The results of invitro release profile obtain from the formulation could be
plotted in models of data treatment as follows:
1. Zero order kinetics model –cumulative percentage drug release Vs. time
2. First order kinetics model-log cumulative percent drug remaining Vs. time.
3. Higuchi’s model-cumulative % drug released Vs. square root of time.
4. Korsemeyer equation or Peppa’s model-cumulative %drug released Vs log
time.
At=A0-K0t
Where,At=drug release at time t.
A0=initial drug concentration K0 =zero
order rate constant.
26
When the data plotted as cumulative percent drug release Vs time ,if the plot is
linear ,then the data obeys zero order kinetics and its slope is equal to zero order
release constant.
27
When the data is plotted according to equation i.e,cumulative drug release Vs
square root of time yields a straight line indicating that drug was released by
diffusion mechanism,then slope is equal to k.
Korsemeyer equation or Peppa’s method
To study mechanism of drug release from controlled release multiparticles of
drug the released data were also fitted to the well known exponential
equation.ie, which is often used to describe the drug release behavior from
polymeric systems.
Mt//Ma =ktn
Mt//Ma=fraction of drug released at time t
k=constant incorporating structural and geometrical characteristics of drug or
polymer system.
n=diffusion exponent related to mechanism of release.
Above equation can be simplified by applying log on both sides,
𝑙𝑜𝑔 𝑀𝑡
𝑀 =log K+ n Log t
𝑎
REPORT
The given formulation was sustained release and obeys first order and zero
order kinetics. The mechanism is Fickian diffusion.
REFERENCE
Indian journal of pharmaceutical sciences
28
EXP.NO:10
DATE:
DETERMINATION OF RELEASE FOR KINETICS FOR THE
DISSOLUTION DATA
AIM
To determine the invitro release kinetics profiled of given data using release
kinetics software.
REQUIREMENTS
Kinetics software,dissolution data.
THEORY
Invitro release kinetics
The results of invitro release profile obtain from the formulation could be
plotted in models of data treatment as follows:
1. Zero order kinetics model –cumulative percentage drug release Vs time
2. First order kinetics model-log cumulative percent drug remaining Vs time.
3. Higuchi’s model-cumulative % drug released Vs square root of time.
4. Korsemeyer equation or Peppa’s model-cumulative %drug released Vs log
time.
At=A0-K0t
Where,At=drug release at time t.
A0=initial drug concentration K0 =zero
order rate constant.
29
When the data plotted as cumulative percent drug release Vs time ,if the plot is
linear ,then the data obeys zero order kinetics and its slope is equal to zero order
release constant.
30
When the data is plotted according to equation i.e,cumulative drug release Vs
square root of time yields a straight line indicating that drug was released by
diffusion mechanism,then slope is equal to k.
Korsemeyer equation or Peppa’s method
To study mechanism of drug release from controlled release multiparticles of
drug the released data were also fitted to the well known exponential
equation.ie, which is often used to describe the drug release behavior from
polymeric systems.
Mt//Ma =ktn
Mt//Ma=fraction of drug released at time t
k=constant incorporating structural and geometrical characteristics of drug or
polymer system.
n=diffusion exponent related to mechanism of release.
Above equation can be simplified by applying log on both sides,
𝑙𝑜𝑔 𝑀𝑡
𝑀 =log K+ n Log t
𝑎
REPORT
The given formulation was found to be controlled release and obeys zero order
kinetics. The mechanism is super case II transport.
REFERENCE
Indian journal of pharmaceutical sciences
31
EXP.NO:11
DATE:
A
B
S
O
R
P
T
I
O
N
S
T
U
D
Y
F
O
R
D
I
C
L
O
F
E
N
A
C
S
O
D
I
U
M
AIM
To carry out
absorption
study for 32
diclofenac
sodium.
solution (100mg/100ml) and the samples were withdrawn from flask at
appropriate intervals and analyzed the drug content by UV method at 268nm.
FORMULA
𝑡𝑒𝑠𝑡 𝑎𝑏𝑠𝑜𝑟𝑏 𝑎𝑛 𝑐 𝑒×𝑠𝑡 𝑎𝑛 𝑑 𝑎𝑟𝑑 𝑐 𝑜𝑛𝑐 𝑒 𝑛𝑡 𝑟𝑎 𝑡𝑖 𝑜𝑛
Amount of drug absorbed =
𝑠𝑡 𝑎𝑛 𝑑 𝑎 𝑟 𝑑 𝑎𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒×𝑡𝑒𝑠𝑡 𝑐 𝑜𝑛 𝑐𝑒 𝑛 𝑡 𝑟𝑎𝑡 𝑖𝑜𝑛
REPORT
The absorption study of diclofenac sodium by everted sac technique was
performed.
The percentage drug absorbed was found to be
REFERENCES
S.P.Vyas, R.K.Khar, “Targeted and Controlled drug delivery”, novel carrier
system, CBS publishers and distributors, New Delhi, page no: 122
33
EXP.NO:12
DATE:
DETERMINATION OF DIFFUSION RATE AND PERMEABILITY CO-
EFFICIENT OF DICLOFENAC SODIUM USING DIFFUSION CELL
AIM
To determine the diffusion rate and permeability coefficient of diclofenac
sodium using cellophane membrane or semipermeable membrane.
REQUIREMENTS
Diclofenac sodium, coloring reagent, cellophane membrane, open diffusion
cell, burette stand, beaker, glass rod etc.
THEORY
Stratum corneum is the principle barrier for cutaneous penetration allowing
slow absorption for majority of drugs. In any case, the permeability of stratum
corneum is increased by using appropriate vehicle. The best vehicle for
topical
use has been described as the one which contribute to reversible decrease in
the stratum corneum resistance and allows diffusion of molecules into the
vehicle
itself.
The most common invitro design is one where a membrane separates
two
compartments-one compartment contains the drug in a vehicle, possibly a
single aqueous or buffer solution and the other compartment contains a receptor
(receives). Solution that provides sink condition after sufficient time, steady
state permeation across the membrane is constant under this condition following
equation can be used.
𝑑𝑚 𝐷𝐶𝑜
= -(1) 34
𝑑𝑡 ℎ
Where m = cumulative mass of the drug that passes through per unit area of the
membrane in time t
Co = concentration of the drug in the first layer of membrane
h = membrane thickness
In practical terms it is very difficult to measure Co, concentration of drug in
first layer of the membrane, removal of the outer layer for assay is problematic
and contamination from the applied donor solution is almost inevitable.
However concentration of the drug in the vehicle bathing the stain membrane
and is usually known or can be determined relatively easily. Since Co and Cv are
simply related.
35
INVITRO DIFFUSION STUDY USING OPEN DIFFUSION CELL
Cellophane membrane was prepared and it is tied to one end of open tube.
50ml of receptor solution in a beaker was taken.
5ml of sample was taken in the open tube and immersed in the receptor
compartment.
Periodically at 0, 15, 30, 45, 60 min 1ml of sample was taken and replaced with
fresh medium.
Measured the absorbance at 268nm by UV spectrophotometrically.
The graph was plotted and flux of permeability coefficient was determined.
REPORT
The flux was found to be
The permeability co-efficient of diclofenac sodium was found to be
REFERENCE
Laboratory manual of Biopharmaceutics &pharmacokinetics by V.B Phase
,R.J.Duas page no:18-21
36
EXP.NO:13
DATE:
DETERMINATION OF PERMEABILITY COEFFICIENT AND
DIFFUSION RATE OF PARACETAMOL USING DIFFUSION CELL
AIM
To report the permeability coefficient and diffusion rate of paracetamol using
cellophane.
REQUIREMENTS
Paracetamol, cellophane membrane, open diffusion cell, burette stand,
beaker, glass rod etc.
THEORY
Stratum corneum is the principle barrier for cutaneous penetration allowing
slow absorption for majority of drugs. In any case, the permeability of stratum
corneum is increased by using appropriate vehicle. The best vehicle for
topical
use has been described as the one which contribute to reversible decrease in
the stratum corneum resistance and allows diffusion of molecules into the
vehicle
itself.
The most common invitro design is one where a membrane separates
two
compartments-one compartment contains the drug in a vehicle, possibly a
single aqueous or buffer solution and the other compartment contains a receptor
(receives). Solution that provides sink condition after sufficient time, steady
state permeation across the membrane is constant under this condition following
equation can be used.
𝑑𝑚 𝐷𝐶𝑜
= -(1) 37
𝑑𝑡 ℎ
Where m = cumulative mass of the drug that passes through per unit area of the
membrane in time t
Co = concentration of the drug in the first layer of membrane
h = membrane thickness
In practical terms it is very difficult to measure Co, concentration of drug in
first layer of the membrane, removal of the outer layer for assay is problematic
and contamination from the applied donor solution is almost inevitable.
However concentration of the drug in the vehicle bathing the stain membrane
and is usually known or can be determined relatively easily. Since Co and Cv are
simply related.
38
INVITRO DIFFUSION STUDY USING OPEN DIFFUSION CELL
Cellophane membrane was prepared and it is tied to one end of open tube.
50ml of receptor solution in a beaker was taken.
5ml of sample was taken in the open tube and immersed in the receptor
compartment.
Periodically at 0, 15, 30, 45, 60 min 1ml of sample was taken and replaced with
fresh medium.
Measured the absorbance at 257nm by UV spectrophotometrically.
The graph was plotted and flux of permeability coefficient was determined.
REPORT
The flux was found to be
The permeability co-efficient of paracetamol was found to be
REFERENCE
Laboratory manual of Biopharmaceutics &pharmacokinetics by V.B Phase
,R.J.Duas page no:18-21
39
EXP.NO:14
DATE:
DETER
MINAT
ION OF
PHAR
MACO
KINET
IC
PARA
METE
RS-1
AIM
To determine the following pharmacokinetic parameter of drug (Cmax, tmax, AUC
(0-α), Ke, t1/2, which is administered orally (500mg)
THEORY
Pharmacokinetics is the study of the time course of absorption, distribution,
biotransformation, and excretion of drugs.
Pharmacokinetic Parameters
The predictive capability of a pharmacokinetic model lies in the proper
selection and development of mathematical functions called parameters
that
govern a pharmacokinetic process. In practice pharmacokinetic parameters
are determined experimentally from a set of drug concentrations collected over
various times known as data. Parameters are also called as variables. Variables
are of two types.
Independent variables which are not affected by any other parameter, for
example time.
Dependent variables which change as the independent variables change, for
example, plasma drug concentration
40note regarding application of
Certain points, which are important to
parameters in pharmacokinetic studies, include-
For the pharmacokinetic parameters to be valid, the number of data points
should always exceed the number of parameters in the pharmacokinetic
model.
DATA:
TIME (hrs.) Plasma drug concentration(µg)
0 0
0.5 5.4
1 10
2 17.2
4 25.8
8 29.8
12 26.6
18 19.4
24 13.3
36 5.9
48 2.6
REPORT
Cmax =
tmax =
AUC
(0-α) =
Ke =
of Biopharmaceutics and Pharmacokinetics by D.M.
t1/2 =
Brahmankar, Sunil.B.Jaiswal
REF
ER Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
EN
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
CES
T 41
e
x
EXP.NO:15
DATE:
DETER
MINAT
ION OF
PHAR
MACO
KINETI
C
PARAM
ETERS-
II
AIM
To determine the following pharmacokinetic parameters
Peak plasma concentration Cmax
Time of peak concentration tmax
Elimination rate constant Ke
Biological half-life (t1/2)
Area under curve AUC (0-α)
Area under first moment
curve AUMC (0-α)
THEORY
Pharmacokinetics is the study of the time course of absorption, distribution,
biotransformation, and excretion of drugs.
Pharmacokinetic Parameters
The predictive capability of a pharmacokinetic model lies in the proper
selection and development of mathematical functions called parameters
that
govern a pharmacokinetic process. In practice pharmacokinetic parameters
are determined experimentally from a set of drug concentrations collected
over 42
various times known as data. Parameters are also called as variables.
Variables are of two types.
The number of parameters needed to describe the pharmacokinetic model
depends upon the complexity of the pharmacokinetic process and on the
route of drug administration.
More the number of parameters more are the difficulties in accurate
estimation of these parameters.
For the pharmacokinetic parameters to be valid, the number of data points
should always exceed the number of parameters in the pharmacokinetic
model.
DATA:
TIME (hrs.) Plasma drug concentration(µg)
1 3.3
2 6.8
3 8.8
4 9.1
5 9.9
6 14.3
7 11.3
8 1.9
9 0.9
REPORT
Cmax =
tmax =
AUC (0-α) =
AUMC (0-α) =
MRT
Ke =
t1/2 = 43
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
44
EXP.NO:16
DATE:
DETERMINATION OF PHARMACOKINETIC PARAMETERS
INTRAVENOUS BOLUS –ONE COMPARTMENT OPEN MODEL
AIM
To determine the pharmacokinetic parameters (Ke, t (1/2) , Vd, AUC,
clearance)
THEORY
One compartment open pharmacokinetic model depicts the body as a single
kinetically homogenous unit that has no barrier to the movement of drug.
Also,
the final distribution equilibrium between the drug in plasma and other body
fluids is attained instantaneously and maintained at all time. This model
applies
only to those drugs that distributes rapidly.
The plasma drug concentration is representative of all body tissues
concentration. The term open indicates the input and output are unidirectional.
Intravenous bolus administration
When a drug that distributes rapidly in the body is given in the form of rapid
IV
bolus or slug, it takes about 1-3 minutes for complete circulation, therefore the
absorption rate is not considered in calculation.
Following intravenous bolus administration of a drug, by considering
following assumptions the fundamental pharmacokinetic parameters of a drug
can be obtained.
One compartment model, first order process and passive diffusion are
operative
No metabolism takes place(elimination is 100% via renal excretion)
The drug is being monitored in blood (plasma /serum) and urine.
45
Important Pharmacokinetic parameters
1. Elimination rate constant Ke
2. Elimination half-life (t1/2)
3. Apparent Volume of Distribution (Vd)
4. Clearance (Clt)
5. Total area under curve(AUC)
6. Calculation of K from urinary excretion data
DATA
The plasma drug concentration time profile, after IV administration of an
antibiotic (250mg) is given below. Plot the data and describe the model.
Time(hr.) Plasma drug concentration(µg/ml)
1 8
2 6.3
3 4.9
4 4
5 3.2
6 2.5
7 1.9
REPORT
AUC (0-α) graphically =
AUC (0-α) theoretically =
Volume of distribution
(Vd) =
Elimination rate
constant =
46
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
47
EXP.NO:17
DATE:
DETERMINATION OF PHARMACOKINETIC PARAMETERS IV
BOLUS ONE COMPARTMENT OPEN MODEL- II
AIM
To determine the following pharmacokinetic parameters (Ke, t1/2, Vd, ClT,
AUC(0-α)
THEORY
One compartment open pharmacokinetic model depicts the body as a
single
kinetically homogenous unit that has no barrier to the movement of drug. Also,
the final distribution equilibrium between the drug in plasma and other body
fluids is attained instantaneously and maintained at all time. This model applies
only to those drugs that distributes rapidly.
The plasma drug concentration is representative of all body tissues
concentration. The term open indicates the input and output are unidirectional.
Intravenous bolus administration
When a drug that distributes rapidly in the body is given in the form of rapid
IV bolus or slug, it takes about 1-3 minutes for complete circulation,
therefore the
absorption rate is not considered in calculation.
Following intravenous bolus administration of a drug, by considering
following
assumptions the fundamental pharmacokinetic parameters of a drug can be
obtained.
One compartment model, first order process and passive diffusion are
operative
No metabolism takes place(elimination is 100% via renal excretion)
The drug is being monitored in blood (plasma /serum) and urine.
48
Important Pharmacokinetic parameters
7. Elimination rate constant Ke
8. Elimination half-life (t1/2)
9. Apparent Volume of Distribution (Vd)
10.Clearance (Clt)
11.Total area under curve(AUC)
12.Calculation of K from urinary excretion data
DATA
The following data shows the plasma concentration profile obtained from I.V
bolus administration of amoxicillin injection 250mg.
Time(hr.) Plasma drug concentration(µg/ml)
0 30
5 22
10 11.2
20 9
30 4.8
40 2.8
50 1.5
60 0.86
REPORT
1. Elimination rate constant Ke
2. Elimination half-life (t1/2)
3. Apparent Volume of Distribution (Vd)
4. Clearance (Clt)
5. Total area under curve(AUC)
49
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
50
EXP.NO:18
DATE:
DETERMINATION OF VARIOUS PHARMACOKINETIC
PARAMETERS AFTER IV INFUSION
AIM
To calculate various pharmacokinetic parameters from the given blood data of
IV infusion (one compartment model)
THEORY
When a single intravenous bolus dose of a drug is given, the desired
therapeutic concentrations is achieved immediately. However this mode of
administration is unsuitable when it is necessary to maintain plasma or tissue
concentrations for prolonged duration. Here the aim is to reach the therapeutic
range and then maintaining drug concentration within the therapeutic range for a
longer duration. It is common practice, in the hospital setting to infuse a drug at
a constant rate (constant rate input or zero order input). This method permits
precise and readily controlled drug administration.
DATA
Estimate the volume of distribution, elimination rate constant, half-life,
clearance from the data in the following table obtained on infusing a drug at
the rate of 50mg/hr for 7.5h
51
9 4.6
12 2
15 0.9
52
EXP.N0:19
DATE
DETERMINATION OF ORDER OF PROCESS,RATE CONSTANT AND
BIOLOGICAL HALF LIFE
AIM
To determine the order, rate constant and half life
DATA
53
EXP.NO:20
DATE
DETERMINATION OF ORDER OF PROCESS, RATE CONSTANT,
AND HALF LIFE
AIM
To determine the order, rate constant and half life
DATA
54
EXP.NO:21
DATE
DETERMINATION OF PHARMACOKINETIC PARAMETERS-ONE
COMPARTMENT OPEN MODEL EXTRAVASCULAR
ADMINISTRATION
AI
M
To
deter
mine
the
phar
maco
kineti
c
para
meter
s
THE
ORY
Administration of drug dose by an extravascular route involves passage of the
drug by absorption through a biological membrane. The plasma profile
obtained
following extravascular administration of a drug is different from plasma profile
of same drug obtained after the drug administered as a rapid intravenous
Time(hr.) Plasma drug concentration(µg/ml)
bolus
1 26.501
injection because the entire dose of administered drug is not absorbed all at
2 36.091
once.
DATA 3 37.512
4 36.055
The data given below is obtained from after administration of 500mg
antibiotic5orally. Find out the pharmacokinetic
32.924parameters Ke, Ka, C t
max, max
absorbed.
55
8 22.784
16 7.571
18 5.734
20 4.343
REPORT
REFERENCES
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
56
EXP.NO:22
DATE
DETERMINATION OF ABSORPTION RATE CONSTANT BY
METHOD OF RESIDUALS
AIM
To determine absorption half-life and absorption rate constant (Ka) for given
data by using method of residuals
THEORY
Absorption rate constant can be calculated by method of residuals. The
technique is also known as feathering or peeling and stripping. It is
commonly
used in pharmacokinetics to resolve a multiexponential curve into its individual
components for drugs that follows one compartment kinetics and
are
administered extravascularly.
Time(hr.) Plasma drug concentration(µg/ml)
DATA 0.5 5.36
1 9.35
2 17.18
4 25.78
8 29.78
12 26.63
18 19.40
24 13.26
36 5.88
48 2.56
70 0.49
57
REPORT
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
58
EXP.NO:23
DATE
DETERMINATION OF ABSORPTION RATE CONSTANT BY
WAGNER NELSON METHOD
AIM
To calculate absorption rate constant of using Wagner Nelson method
THEORY
After a single oral dose of a drug, at any time the amount of drug
absorbed into
the systemic circulation Aa, is the sum of the amount of drug in the body A and
the amount of drug eliminated from the body Ae. Thus
Aa =A+Ae (1)
The amount of drug in the body is A = VC while the amount of drug eliminated
at
Aeany time t can be calculated as follows
= KVAUC (2)
(0-t)
59
The method requires collection of blood samples after a single oral dose at
regular intervals of time till the entire amount of drug is eliminated from the
body. K is obtained from semilog plot of C Vs t.
DATA
Bioavailability of phenyl propanolamine hydrochloride was studied in 24 adult
male subjects. The following data represents the mean blood phenyl
propanolamine hydrochloride concentration(ng/ml) after the oral administration
of a single 25mg dose of phenylpropanolamine hydrochloride solution.
Time(hr.) Plasma drug concentration(µg/ml)
0.25 51.33
0.5 74.05
0.75 82.91
1 85.11
1.5 81.76
2 75.51
3 62.92
4 52.32
6 36.08
8 24.88
12 11.83
18 3.88
24 1.27
REPORT
60
REFERENCE
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
61
EXP.NO:24
DATE
DETERMINATION OF RATE OF ELIMINATION CONSTANT AND
HALF-LIFE BY URINARY EXCRETION METHOD
AIM
To determine the elimination rate constant and half-life from urinary excretion
method
DATA
The following data were taken from a graph plotted after IV administration of
100mg of drug.
REPORT
The rate of elimination rate constant from urinary excretion data by urinary
excretion method was found to be
62
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
63
EXP.NO:25
DATE
DETERMINATION OF RATE OF ELIMINATION CONSTANT AND
HALF-LIFE BY SIGMA –MINUS METHOD METHOD
AIM
To determine the elimination rate constant and half-life from urinary excretion
data by using sigma-minus method following IV administration of 100mg
of
drug
DATA
The following data were taken from a graph plotted after IV administration of
100mg of drug.
Sample Time of urine Volume of urine Conc.of
collection collected unchanged drug
0 0
1 0-2 140 250
2 2-4 150 100
3 4-6 90 80
4 6-8 200 20
5 8-12 310 10
6 12-24 600 4
REPORT
The rate of elimination rate constant from urinary excretion data by sigma-
minus method method was found to be
64
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
65
EXP.NO:26
DATE:
DATA ANALYSIS-TWO COMPARTMENT IV BOLUS
ADMINISTRATION
AIM
To determine the following pharmacokinetic parameters
DATA
The plasma concentration Vs time data following administration of
250mg of
IV bolus of dose is given plot the data and find out the pharmacokinetic
parameter (K21,K10,K12,t1/2,Vc,Vd,Vt,AUCtheoretically)
66
REPORT
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
67
EXP.NO:27
DATE:
DET
ERM
INAT
ION
OF
PER
CEN
TAG
E
BIOA
VAIL
ABIL
ITY
AIM
To determine the percentage bioavailability for the following data of oral IV
drug administration
THEORY
Bioavailability means the rate and extent to which the active ingredient or
active moiety is absorbed from a drug product and becomes available at the
site
of action. For drug products that are not intended to be absorbed into the
bloodstream, bioavailability may be assessed by measurements intended
to
reflect the rate and extent to which the active ingredient or active moiety
becomes available at the site of action.
Relative Availability
Relative (apparent) availability is the availability of the drug from a drug
product as compared to a recognized standard. The fraction of dose
systemically
available from an oral drug product is difficult to ascertain. The availability of
68
drug in the formulation is compared to the availability of drug in a
standard
Absolute Availability
PROCEDURE
Cut and weigh method
Take 2 similar types of graph and then plot the graph between time Vs plasma
drug concentration. Extrapolate it to zero and cut the graph and weigh them
and
calculatebioavailability
Absolute absolute bioavailability
𝑒 𝑖 𝑔 ℎ𝑡 𝑜 𝑓using
= 𝑤𝑤𝑒 𝑜 𝑟 𝑎 𝑙 formula
x100
𝑖𝑔 ℎ𝑡 𝑜𝑓
𝐼𝑉
69
Area method
In this method, the squares under the curve are counted. Squares having area
more than half in graph is counted and calculate bioavailability by
Absolute bioavailability = 𝐴 𝑟 𝑒 𝑎 𝑜 𝑓 𝑜𝑟𝑎𝑙
x100 [1 square = 1cm2]
𝐴𝑟𝑒𝑎 𝑜𝑓 𝐼𝑉
Trapezoidal method
This is most accurate method to obtain bioavailability. It
involves breaking up
of plasma drug concentration Vs time profile in several trapezoids. Calculate
the areas of individual trapezoids and the add up these areas to arrive at
cumulative AUC
Area under curve trapezoid = ½(C1+C2) (t2-t1) or 1/2b (h1+h2)
Absolute bioavailability = 𝐴 𝑟 𝑒 𝑎 𝑜 𝑓 𝑜𝑟𝑎𝑙
x100
𝐴𝑟𝑒𝑎 𝑜𝑓 𝐼𝑉
DATA
Sl.no Time (hrs.) Cp IV (µg/ml) Cp oral (µg/ml)
1 0.5 6.82 0.97
2 1 4.47 3.46
3 1.5 3.14 4.02
4 2 2.21 3.32
5 3 1.15 1.62
6 4 0.55 0.85
7 5 0.28 0.39
8 6 0.14 0.19
9 7 0.07 0.09
REPORT
The absolute bioavailability of given drug by trapezoidal method was found to
be
The absolute bioavailability of given drug by area method was found to be
70
REFERENCES
Textbook of Biopharmaceutics and Pharmacokinetics by
D.M. Brahmankar, Sunil.B.Jaiswal
Textbook of Biopharmaceutics and Pharmacokinetics by Dr.Javed Ali,
Dr. Alka Ahuja, Dr. Sanjula Baboota, Dr.Roop.K.Khar
Laboratory Manual of Biopharmaceutics and
Pharmacokinetics by Dr.S.B.Bhise, Dr.R.J. Dias, Dr. S.C Dhawale, Shri.
K. K. Mali
71
CALIBRATION CURVES
72
PREPARATION OF CALIBRATION CURVE FOR PARACETAMOL
73
PREPARATION OF CALIBRATION CURVE FOR DICLOFENAC
POTASSIUM
74