PRINCE SATTAM BIN ABDULAZIZ UNIVERSITY COLLEGE OF PHARMACY DEPARTMENT OF PHARMACEUTICS LABORATORY MANUAL PHARMACEUTICS IV PHT-414 LEVEL VII EXPERIMENTAL COORDINATOR - Dr. MUQTADER MOHAMMED Email:- [email protected]Academic Year 2016-17 Semester II PSAU-COP, Al-Kharj- KSA
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COLLEGE OF PHARMACY DEPARTMENT OF ... OF PHARMACY DEPARTMENT OF PHARMACEUTICS LABORATORY MANUAL PHARMACEUTICS IV PHT-414 LEVEL VII EXPERIMENTAL COORDINATOR - Dr. MUQTADER MOHAMMED
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calculate and determine the partition coefficient.
3 Disintegration , study of disintegration time for
different type of tablet formulations.
3.2 Disintegration of uncoated tablets.
3.3 Disintegration of coated tablets.
3.4 Disintegration of enteric coated tablets.
4 Drug Dissolution time profile study.
4.1 Drug dissolution of Paracetamol/Acetaminophen
tablet marketed formulation.
4.2 Drug dissolution of ciprofloxacin tablet
marketed formulation
5 Enhancement of solubility by preparation of
solid dispersion techniques.
5.1 Preparation of solid dispersion by physical
mixture method.
5.2 Preparation of solid dispersion by solvent
evaporation techniques.
6 Influence of vehicle on the solubility of
Ciprofloxacin HCl.
7 Demonstration of in-vitro diffusion study of
Tran dermal drug delivery systems.
2
CERTIFICATE OF LAB PERFORMANCE
SL# ACTIVITY NAME A B C
1.
2.
3.
4.
5.
Sl.
no
Content &
Learning outcomes
EVALUATIONS Attendance Performance Worksheet GLP
P A E G S C I A B C
1 PH Partition Theory, , you will be able to
calculate and determine the partition coefficient.
3 Disintegration , study of disintegration time for
different type of tablet formulations.
3.2 Disintegration of uncoated tablets.
3.3 Disintegration of coated tablets.
3.4 Disintegration of enteric coated tablets.
4 Drug dissolution time profile study.
4.1 Drug dissolution of Paracetamol/Acetaminophen
tablet marketed formulation.
4.2 Drug dissolution of ciprofloxacin tablet
marketed formulation
5 Enhancement of solubility by preparation of
solid dispersion techniques.
5.1 Preparation of solid dispersion by physical
mixture method.
5.2 Preparation of solid dispersion by solvent
evaporation techniques.
6 Influence of vehicle on the solubility of
Ciprofloxacin HCl.
7 Demonstration of in-vitro diffusion study of
Tran dermal drug delivery systems.
Marks Awarded:
3
Introduction & grading about the Course Outline
Pharmaceutics IV: PHT-414
Grading
Final Exam 15
Viva/synopsis/other activities 5
Record, GLP, ACTIVITIES 5
Total 25
4
Partition Coefficient
Practical-1: Date
Aim:
Determination of the Partition Coefficient of Citric Acid (drug) between Water (aqueous phase) and 2-Methylpropan-1-ol (isobutyl alcohol) (non aqueous phase).
Introduction
If a solute is added to two immiscible solvents, A and B. in contact with each other, the solute distributes itself between the two and equilibrium is set up between the solute molecules in solvent A and the solute molecules in solvent B. The ratio of the concentration of the solute in the two solvents is
where K is known as the partition coefficient or distribution coefficient.
Chemicals
2-Methylpropan-1-ol (density = 0.805 gdm-3), 0.2 M Citric acid (drug already dissolved in the aqueous phase – water), 0.1 M NaOH, phenolphthalein indicator
Apparatus
100 ml separating funnel, titration apparatus, 5 ml pipette
Procedure
1. Record the room temperature.
2. Using suitable apparatus pour 25 ml of the 0.2 M Citric acid solution and 25 ml of 2-methylpropan-1-ol into a 100 cm3 separating funnel. Stopper the funnel and shake vigorously for 6 minutes. (Release pressure in the funnel by occasionally opening the tap.)
3. Separate approximately 15 ml of EACH layer and collect them in TWO
clean beakers. (Discard the fraction near the junction of the two layers.)
4. Pipette 5 ml of the aqueous layer into a titration flask and titrate it with 0.1 M sodium hydroxide solution using 2 drops of phenolphthalein indicator.
5. Using another pipette, deliver 5 ml of the non-aqueous layer into a
titration flask and titrate it with 0.1 M sodium hydroxide solution using 2 drops of phenolphthalein indicator.
5
Observation :
Trial Volume of 0.1 M NaOH
titre for aqueous layer/
cm3
Volume of 0.1 M
NaOH titre for non-
aqueous layer/ cm3
Calculation:
M1V1=M2V2
M1(UNKNOWN MOLARITY/ CONC), V1( VOLUME IN CONICAL FLASK)
M2( MOL OF BUREETE SOLUTION ), V2(VOL CONSUMED IN TITRATION)
FOR NON-AQUOUS LAYER
M1= M2V2
V1
FOR AQUOUS LAYER
M1= M2V2
V1
7. For each experiment, calculate the ratio of the concentration of citric acid in the non-aqueous layer in relation to the aqueous layer.
If calculated Partition Coefficient: K = O/W is ≤ 10 relatively hydrophilic, K = O/W
is ≥ 104
Hydrophobic in nature.
Concentration of solute
(unknown)
M1
In aqueous layer
In non-aqueous layer
6
Report : The partition coefficient for the given drug sample was found to be ________ . Inference : As per the result the compound was found to be hydrophilic / hydrophobic in nature.
Student ID / Reg No
Comment on your results.
********************************
7
Exp.No: Date:
Effect of pH on the Partition Coefficient of acidic and basic drugs
Aim
Determination of the effect of pH on Partition Coefficient of Citric Acid (drug)
between (aqueous phase) and 2-Methylpropan-1-ol (isobutyl alcohol) (non
aqueous phase).
Introduction (pH - Partition Theory)
For a drug to cross a membrane barrier it must normally be soluble in the
lipid material of the membrane to get into membrane it has to be soluble in the
aqueous phase as well to get out of the membrane. Many drugs have polar and non-
polar characteristics or are weak acids or bases. For drugs which are weak acids or
bases the pKa of the drug, the pH of the GI tract fluid and the pH of the blood
stream will control the solubility of the drug and thereby the rate of absorption
through the membranes lining the GI tract.
Brodie et al. stated that when a drug is ionized it will not be able to get through the
lipid membrane, but only when it is non-ionized and therefore has higher lipid
dissolved in the aqueous phase – Buffer solutions with different pH ), 0.1 M NaOH,
phenolphthalein indicator
Apparatus
100 ml separating funnel, titration apparatus, 5 ml pipette
8
Procedure
1. Record the room temperature.
2. Using suitable apparatus pour 25 ml of the 0.2 M Citric acid solution and
25 ml of 2-methylpropan-1-ol into a 100 cm3 separating funnel.
Stopper the funnel and shake vigorously for 6 minutes. (Release pressure in
the funnel by occasionally opening the tap.)
3. Separate approximately 15 ml of EACH layer and collect them in TWO
clean beakers. (Discard the fraction near the junction of the two layers.)
4. Pipette 5 ml of the aqueous layer into a titration flask and titrate it with 0.1 M
sodium hydroxide solution using 2 drops of phenolphthalein indicator.
5. Using another pipette, deliver 5 ml of the non-aqueous layer into a
titration flask and titrate it with 0.1 M sodium hydroxide solution using 2
drops of phenolphthalein indicator.
6. Repeat steps (2) to (5) with another separating funnel using either one of
the following as aqueous solutions
a. Acetate buffer pH 4.5
b. Phosphate buffer Ph 7.2 Observation
Trial Buffer (pH) Volume of 0.1 M NaOH
titre for aqueous layer/
cm3
Volume of 0.1 M
NaOH titre for non-
aqueous layer/ cm3
9
Calculation:
M1V1=M2V2
M1(UNKNOWN MOLARITY/ CONC), V1( VOLUME IN CONICAL FLASK)
M2( MOL OF BUREETE SOLUTION ), V2(VOL CONSUMED IN
TITRATION)
FOR NON-AQUOUS LAYER
M1= M2V2
V1
FOR AQUOUS LAYER
M1= M2V2
V1
7. For each experiment, calculate the ratio of the concentration of citric acid in
the non-aqueous layer in relation to the aqueous layer.
If calculated Partition Coefficient: K = O/W is ≤ 10 relatively hydrophilic, K = O/W is ≥ 10
4 Hydrophobic in nature.
Increase in P
H increases or decreases the Partition Coefficient
Report: The partition coefficient for the given drug sample was found to be ________ . Inference : As per the result the compound was found to be hydrophilic / hydrophobic in nature and the effect of P
H was found to
be_____________________________________________
Concentration of solute
(unknown)
M1
In aqueous layer
In non-aqueous layer
PH =
10
Student ID / Reg No
Comment on your results.
Report:
###############
11
Disintegration & Dissolution
Both tests are a measure of Wetting – How quickly the tablet is exposed to the media
Disintegration – the speed and nature of particle release
Dissolution- the speed and nature of API dissolved
12
DISINTEGRATION TEST
Introduction:
The disintegration test is a measure of the time required under a given set of conditions for a
group of tablets to disintegrate into particles which will pass through a 10 mesh screen.
Generally, the test is useful as a quality assurance tool for conventional dosage forms.
Apparatus
The apparatus consists of a basket-rack assembly, a 1000 mL, low-form beaker, 138-160 mm
in height and having an inside diameter of 97-115 mm for the immersion fluid, a thermostatic
arrangement for heating the fluid between 35 °C and 39 °C, and a device for raising and
lowering the basket in the immersion fluid at a constant frequency rate between 29 and 32
cycles per minute, through a distance of not less than 53 mm and not more than 57 mm. The
volume of the fluid in the vessel is such that at the highest point of the upward stroke the wire
mesh remains at least 15 mm below the surface of the fluid and descends to not less than 25
mm from the bottom of the vessel on the downward stroke. At no time should the top of the
basket-rack assembly become submerged. The time required for the upward stroke is equal to
the time required for the downward stroke and the change in stroke direction is a smooth
transition, rather than an abrupt reversal of motion. The basket-rack assembly moves vertically
along its axis. There is no appreciable horizontal motion or movement of the axis from the
vertical.
Basket-rack assembly
The basket-rack assembly consists of six open-ended transparent tubes, each 75.0-80.0 mm
long and having an internal diameter of 20.70-23.00 mm and a wall 1.0-2.8 mm thick; the
tubes are held in a vertical position by two plates, each 88-92 mm in diameter and 5.00-8.50
mm in thickness, with six holes, each 22-26 mm in diameter, equidistant from the centre of the
plate and equally spaced from one another. Attached to the lower surface of the lower plate is
a woven stainless steel wire mesh, which has a plain square weave with 1.8-2.2 mm apertures
and with a wire diameter of 0.570-0.660 mm. The parts of the apparatus are assembled and
rigidly held by means of three bolts passing through the two plates. A suitable means is
provided to suspend the basket-rack assembly from the raising and lowering device using a
point on its axis.
The design of the basket-rack assembly may be varied somewhat provided the specifications
for the glass tubes and the screen mesh size are maintained. The basket-rack assembly
conforms to the dimensions shown in Figure 1.
Discs
The use of discs is permitted only where specified or allowed. Each tube is provided with a
cylindrical disc 9.35-9.65 mm thick and 20.55-20.85 mm in diameter. The disc is made of a
suitable, transparent plastic material having a specific gravity of 1.18-1.20. Five parallel 1.9-
2.1 mm holes extend between the ends of the cylinder. One of the holes is centered on the
cylindrical axis. The other holes are centered 5.8-6.2 mm from the axis on imaginary lines
perpendicular to the axis and parallel to each other. Four identical trapezoidal-shaped planes
are cut into the wall of the cylinder, nearly perpendicular to the ends of the cylinder. The
trapezoidal shape is symmetrical; its parallel sides coincide with the ends of the cylinder and
are parallel to an imaginary line connecting the centres of two adjacent holes 6 mm from the
cylindrical axis. The parallel side of the trapezoid on the bottom of the cylinder has a length of
1.5-1.7 mm and its bottom edges lie at a depth of 1.50-1.80 mm from the cylinder’s
circumference. The parallel side of the trapezoid on the top of the cylinder has a length of 9.2-
13
9.6 mm and its centre lies at a depth of 2.5-2.7 mm from the cylinder’s circumference. All
surfaces of the disc are smooth. If the use of discs is specified, add a disc to each tube and
operate the apparatus as directed under procedure. The discs conform to the dimensions found
in Figure 1.
The use of automatic detection employing modified discs is permitted where the use of discs is
specified or allowed. Such discs must comply with the requirements of density and dimension
given in this chapter.
Procedure
Place one dosage unit in each of the six tubes of the basket and if specified add a disc. Operate
the apparatus using water as the immersion fluid unless another liquid is specified and
maintain its temperature at 35-39 °C. At the end of the specified time, lift the basket from the
fluid and observe the dosage units: all of the dosage units have disintegrated completely. If
one or two dosage units fail to disintegrate, repeat the test on 12 additional dosage units. The
requirements of the test are met if not less than 16 of the 18 dosage units tested are
disintegrated.
14
Figure 1. Diagram for disintegration apparatus
(dimensions are expressed in millimeters)
15
FORMULATION INDIAN PHARMACOPOEIA 1996 (IP) BRITISH PHARMACOPOEIA 2009 (BP)
UNITED STATES
PHARMACOPOEIA 2009
(USP)
Uncoated ed tablets
Uses disintegration apparatus with 250ml or 1 L
of specified medium & for specified time as
per monograph. If 1-2 fail, repeat for 12
different units. 16 out of 18 indicate test is
affirmative.
Uses disintegration apparatus Uses liquid medium
water R or specified medium for 15 min. If 1-
2 fail, repeat for 12 different units. 16 out of
18 indicate test is affirmative.
Uses disintegration apparatus with specified
liquid medium & for specified time as
per monograph. If 1-2 fail, repeat for
12 different units. 16 out of 18
indicate test is affirmative.
Chewable tablets -NA- Disintegration test not applicable.
Same as uncoated tablets. Uses
disintegration apparatus with specified
liquid medium & for 4 hours or as per
monograph. If 1-2 fail, repeat for 12
different units. 16 out of 18 indicate
test is affirmative.
Non-film coated tablets -NA-
Uses disintegration apparatus with water R or
specified liquid medium & for 60 min., if
there is no cracking of coat; continue by
replacing the medium with 0.1 M HCl for
specified time as per monograph. If 1-2 fail,
repeat for 12 different units. 16 out of 18
indicate test is affirmative.
Same as uncoated tablets. Uses
disintegration apparatus with specified
liquid medium & for specified time as
per monograph. If 1-2 fail, repeat for
12 different units. 16 out of 18
indicate test is affirmative.
Film coated tablets -NA-
Uses disintegration apparatus with water R or
specified liquid medium & for 30 min., if
there is no cracking of coat; continue by
replacing the medium with 0.1 M HCl for
specified time as per monograph. If 1-2 fail,
repeat for 12 different units. 16 out of 18
indicate test is affirmative.
Same as uncoated tablets. Uses
disintegration apparatus with specified
liquid medium & for specified time as
per monograph. If 1-2 fail, repeat for
12 different units. 16 out of 18
indicate test is affirmative.
Gastro resistant tablets
(Enteric Coated)
Uses disintegration apparatus with 250ml or 1 L
of specified medium & for specified time as per
monograph. If 1-2 fail, repeat for 12 different
units. 16 out of 18 indicate test is affirmative.
Uses disintegration apparatus with 0.1 M HCl at
37+-2OC for 2-3 hrs (nlt 1 hour), if there is no
cracking or breaking of coat, continue with
phosphate buffer, pH 6.8 at same temperature for 1
hour or as per monograph. If 1-2 fail, repeat for 12
Street Waltham, Massachusetts, U.S Selection of Wavelength: Scan standard solution in UV spectrophotometer between 200 nm to 400 nm on spectrum mode, using diluents as a blank. Paracetamol shows λ max at 243 or ________ λ max _.
24
###############
Figure : 01 standard graph
25
Exp.No: Date:
DISSOLUTION TEST FOR PARACETAMOL TABLETS, WITH
DIFFERENT FORMULATIONS (U.S.P. 1995)
Conditions:
o Apparatus : II (paddle) o Medium : 900 ml of phosphate buffer pH 5 o Temp. : 37 ± 0.5
oC
o Speed : 50 rpm o Time : 45 min.
Procedure:
1. Place one tablet in the vessel, and then start the apparatus at the above
conditions.
2. At specified time intervals (5, 10, 15, 20, 25, 30, and 45 till 60 min) withdraw 1 ml
sample through a Millipore filtration unit (polyethylene tube with a cotton),
and place the sample in a test tube. Replace the withdrawn sample with 1 ml
fresh phosphate buffer kept at 37 ± 0.5 o
C.
3. Dilute 1 ml of the collected sample with fresh phosphate buffer (in a volumetric flask) ,
mix well.
4. Read the absorbance for the diluted samples at 249 nm against a blank of
phosphate buffer.
5. Calculate the concentration of paracetamol released and express this conc. As a
percentage of the labeled amount.
6. Plot the dissolution curve of paracetamol (% released vs.time).
USP stated that:
The tablet should release not less than 80% of its content within 30 minutes and not less
than 90% of its content within 45 minutes. (For immediate release tablets)
26
Observation of the dissolution for _________________________________ tablets:
Time
(min)
Absorbance
(nm)
Conc.
mcg/ml
Amount of drug dissolve
mg/ml
Cumulative amount of
drug dissolved in
900 ml
Cumulative % drug
dissolved
0
5
10
15
20
25
30
35
40
45
50
55
60
27
Plot the Time versus % release on graph paper and comment on the on the drug released rate?
Report : the % drug release for one hour was found to be __________________________.
28
Student ID / Reg No
Comment on your results.
Report:
###############
29
Exp.No: Date:
DISSOLUTION TEST FOR CIPROFLOXACIN Hcl TABLETS)
Conditions:
o Apparatus : I (basket) o Medium : 900 ml of distilled water o Temp. : 37 ± 0.5
oC
o Speed : 50 rpm o Time : 60 min.
Procedure:
Place one tablet in the basket, immerse in the vessel, and then start the
apparatus at the above conditions.
At specified time intervals (5, 10, 15, 20, 25, 30 and 45,50,55,60 min)
withdraw 1 ml sample from the dissolution medium, through a Millipore
filtration unit (polyethylene tube with a cotton), and place the sample in a test
tube.
Replace the withdrawn sample with 1 ml distilled water kept at 37 ±0.5o
C.
Dilute as per the requirement.
Read the absorbance for the diluted samples at 272 nm against a blank
distilled water.
Plot the dissolution curve (% released vs. time).
USP stated that: the tablet should release not less than 70% of its content within
30 minutes and not less than 90% of its content within 45 minutes
30
STANDARD GRAPH OF CIPROFLOXACIN HCL
Abs (nm) Concentration (mcg/ml)
PLOT
31
Observations of the dissolution of ciprofloxacin Hcl tablets: ( CIPRO BAY 500 MG )
Time
(min)
Absorbance
(nm)
Conc.
mcg/ml
Amount of drug dissolve
mg/ml
Cumulative amount of
drug dissolved in
900 ml
Cumulative % drug
dissolved
0
5
10
15
20
25
30
35
40
45
50
55
60
32
Plot the Time versus % release on graph paper and comment on the on the drug released rate?
Report : the % drug release for one hour was found to be __________________________.
33
Pag
e33
Student ID / Reg No
Comment on your results.
Report:
###############
34
Pag
e34
Enhancement in solubility of poor water-soluble drugs by solid dispersion
Solid dispersion (SD) has been widely used to improve the dissolution rate,
solubility, and oral absorption of poor water-soluble drugs. SD refers to the group of solid
products consisting of at least two different components, generally a hydrophilic matrix and
a hydrophobic drug; the matrix can be either crystalline or amorphous. Solid dispersion was
first introduced to overcome the low bioavailability of lipophilic drugs by forming eutectic
mixture of drugs with water soluble carriers. Approximate 40% of new chemical entities
(NCE) being synthesized by combinatorial screening programs possessing superior
pharmacological activities are poorly soluble, which is a great obstacle in formulation
development.
35
Pag
e35
Exp.No: Date:
Preparation of Solid dispersion (SD) by physical mixture
Preparation by physical mixture
The physical mixture is prepared by using hydrophilic carrier in proportions viz., (--------).
Mixing of drug and carrier in a glass mortar. Solid mass is pulverized and passed through
sieve No. 120 to get uniform sized particles.
Observation
Procedure factor Name Grams
Total amount
Ratio
Polymer
Drug
Report : Solid dispersion (SD) of _____________ drug with ____________prepared by
physical mixture and submitted/ preserved for evaluation .
36
Pag
e36
Exp.No: Date:
Preparation of Solid dispersion (SD) by Solvent evaporation techniques
Solvent evaporation method
solid dispersions are prepared by solvent evaporation method using carrier PVP K 30 in
proportions viz., (-------). The drug and carrier are dissolved in methanol in a china dish and
the mixture is heated until the solvent evaporated and clear film of drug and carrier is
obtained. The resultant solid dispersion is scraped out with a spatula. Solid dispersions are
pulverized in a mortar and pestle and passed through a 125 µm sieve before packing in an
airtight container.
Observation
Procedure factor Name Grams
Total amount
Ratio
Polymer
Drug
Report : Solid dispersion (SD) of _____________ drug with ____________prepared by
physical mixture and submitted/ preserved for evaluation .
37
Pag
e37
Student ID / Reg No
Comment on your results.
Report:
38
Pag
e38
Exp.No: Date:
INFLUENCE OF VEHICLE ON THE SOLUBILITY OF CIPROFLOXACIN HCL
The study on solubility yields information about the structure and intermolecular forces of
drugs.
The term ‘solubility’ is defined as maximum amount of solute that can be dissolved in a
given amount of solvent. It can also be defined quantitatively as well as qualitatively.
Quantitatively it is defined as the concentration of the solute in a saturated solution at a
certain temperature. In qualitative terms, solubility may be defined as the spontaneous
interaction of two or more substances to form a homogenous molecular dispersion. A
saturated solution is one in which the solute is in equilibrium with the solvent. The solubility
of a drug is represented through various concentration expression such as parts, percentage,