OUR LADY OF FATIMA UNIVERSITY

COLLEGE OF PHARMACY

POST LAB DISCUSSION

CHROMATOGRAPHY
 Unit Outcomes

At the end of this unit, the students are

expected to:
1.Define Chromatography and its
underlying
principle
2.List the types of chromatography and the
analyte involved in each type.
3.Enumerate the different examples of
derivatizing agent.
CHROMATOGRAPHIC METHOD
• is a process in which a solution of mixture containing inert materials,
drug principles and impurities is separated into its components while moving
through a bed of fixed porous solid having different and reversible affinities
for the substance being separated.
2 Phases of chromatography
- Stationary phase – a fixed bed core of large surface
area
→ ADSORBENT (SILICA GEL G-GYPSUM) (POLAR)
- Mobile phase – liquid, mixture or gas
- a fluid w/c moves through/ or over the surface/
of the stationary phase.
→ ORGANIC SOLVENT (NON-POLAR)
REVERSED PHASE CHROMATOGRAPHY
→ STATIONARY PHASE – NON-POLAR
→ MOBILE PHASE – POLAR
CLASSIFICATION BASED ON TECHNIQUES:
COLUMN CHROMA The simplest type of chromatographic column
(COLUMN, MOBILE consists of a separation flask and a cylindrical
PHASE, glass tube constricted at one end
DEVELOPER) MP- ORG SOLVENT
SP-SILICA GEL
PAPER CELLULOSE OF THE FILTER PAPER IS USED AS
CHROMATOGRAPHY ADSORBENT
MP- ORG SOLVENT (NON-POLAR)
SP- WATER (POLAR)

TLC FOLLOW THE SAME PRINCIPLE AS PC

→ 30 MINS MOST DISTINCT RESULT
→ 20 mg MOST PRECISE
MP- ORGANIC SOLVENT
SP- SILICA GEL G
When the compounds are radioactive, their position
on the column may be determined by the use of a
. GEIGER MUELLER COUNTER

NERNT’S LAW → According to this law, when two

practically immiscible solvents are in contact with
each other and substance which is soluble in each is
added, the substance distributes itself in such a way
that at equilibrium and at a given temperature the
ratio of the concentrations of the two solutions is
constant
Give the use of the ff:
1. H2SO4, Iodine Crystals and UV/Fluorescence →
VISUALIZING AGENT
2. Silica Gel G and Alumina --. ADSORBENT
3.Benzene or chloroform with 10% ethanol → MOBILE
PHASE WHEN THE COMPOUNDS BEING
SEPARATED IS UNKNOWN
 Derivatizing Spot Color Compounds
Reagent Present
Bromocresol Green Yellow Spot Acids

2,4 dinitrophenyl Yellow/Red Spot Aldehydes,

hydrazine Ketones
Derivatizing Agents
Ninhydrin Fluorescent Amino Acids,
Amines
Mercuric Nitrate Yellow Brown Alkaloids
Spots
Bromothymol Blue Light Green Spots Lipids
Aniline phthalate Gray Black Spots Carbohydrates
BASED ON MECHANISM OF PRINCIPLE
PARTITION - L-L CHROMA
CHROMA - DIFFERENCE IN SOLUBILITY
- EX: PAPER CHROMA
ADSORPTION - L-S CHROMA
CHROMA - EX: TLC, COLUMN CHROMA
MOLECULAR - BASED ON MW
EXCLUSION/
GEL
FILTRATION
CHROM

ION- - BASED ON CHARGE

EXCHANGE - BOTH CONTAINS BEADS
GAS CHROMA MP- INERT GAS (CHHeAN)
ANALYSIS OF VOLATILE COMPOUNDS
- RETENTION TIME AND RETENTION VOLUME
Retention time (tR)
➢Is defined as the time required by an average
molecule of component to pass from the injection
point through the column to the detector.
Retention Volume (vR)
➢Is defined as the volume of carrier gas necessary
to carry an average molecule of the component from
the point of injection to the detector.
HPLC (4 PRINCILES) - RETENTION: POSITION OF THE PEAK
HIGH
P – PERFORMANCE, - Solvents must be degassed → TO REMOVE
POWER, PARTITION, BUBBLES before they can be used, or detector will
PRESSURE, PRECISION
adsorb bubbles and erroneous readings will be
LIQUID
CHROMATOGRAPHY obtained

REVERSED PHASE - MP; POLAR

CHROMA - SP; NON-POLAR
1. If one component of a mixture travelled 9.6 cm from the base
line while the solvent had travelled 12.0 cm, then the Rf value for
that component is
 REFERENCES

à Jenkins, Glenn L, Jenkins Quantitative

Pharmaceutical Chemistry, N.Y, N.Y. : Mc Graw,
Latest Edition, (Chapter in Chromatography)
à Watson, David G., Pharmaceutical Analysis: A
Textbook for Pharmacy Students & Pharmaceutical
Chemist, Churchill Livingstone, Edinburgh, 2012
(Chapter in Chromatography)
POST-LABORATORY
DISSOLUTION TEST
 UNIT OUTCOMES
At the end of this unit, the students are expected to:
• describe the instrument and functions of the essential
parts of dissolution apparatus
• understand the principle involved
• identify useful application of the instruments in
pharmaceutical quality control
 Dissolution Testing <711>

• Is the process by which a solid of only fair

characteristics enters into a solution.
• Considered to be one of the most important quality
control test for solid dosage form.
• Dissolution Rate is the amount of active ingredient
in solid-dosage form dissolved in unit time under
standardized conditions of liquid/solid interface,
temperature and media composition.
 Noyes-Whitney Equation:
dw
------ = KS (Csat – Csoln)
dt
• where:
• K is dissolution constant
• S is surface area of solid
• Csat is concentration of saturated solution
• Csoln is concentration at any given time
 Importance of Dissolution
Testing:
• To meet legal requirements for compendia
drugs.
• As QC tool for drug product manufacturing.
• As dosage formulation optimization tool.
• For stability – bioavailability of drug over
shelf – life.
 PRINCIPLE FUNCTION:
• Optimisation of therapeutic effectiveness during product
development and stability assessment.
• Routine assessment of production quality to ensure
uniformity between production lots.
• Assessment of ‘bioequivalence’, that is to say, production
of the same biological availability from discrete batches of
products from one or different manufacturers.
• Prediction of in-vivo availability, i.e. bioavailability (where
applicable).
• Film Theory by Nernst aims to explain the
mechanism that under the influence of no
reactive or chemical forces, a solid particle
immersed in a liquid, undergoes two
consecutive steps:
• (1) The solution of the solid at the interface,
forming a firm stagnant layer or film, h, around
the particle.
• (2) The diffusion from this layer at the
boundary to the bulk of the fluid.
USP Apparatus 1 and 2
 USP Apparatus 3 and 4

Reciprocating Disk • Flow-through Cell

 USP APPARATUS 7

USP APPARATUS 5

USP APPARATUS 6
 Factors that affect drug
dissolution:
• Intrinsic properties of the API (e.g., solubility,
wettability, particle size, surface area, morphology,
polymorphs),
• Formulation composition and characteristics (e.g.,
excipients, hardness, manufacturing process),
• Dissolution method used for drug assessment (e.g.,
apparatus, medium, test conditions, sampling, and
sample analysis)
Media Selection
• Distilled water (preferred medium)
• Buffered aqueous solution having pH
between 4 – 8, dilute acid may be selected.
• pH of 6.8 or less, add purified pepsin
• pH of 6.8 or greater, add pancreatin
 SINK CONDITIONS
• Is the ability of the dissolution media to dissolve at least 3
times the amount of drug that is in your dosage form.
• In vivo condition, there
is no conc. build up in
the bulk of the solution
and hence no retarding
effect on the dissolution
rate of the drug i.e.
Cs>>Cb and sink
condition maintain.
Controlling variables:
• Common operating speeds:
100 rpm for basket type
50 rpm for paddle type
• Temperature: 37+/- 0.5°C
• Usual volume of medium: 500 to 900
ml
 Acceptance Table
S Number Acceptance Criteria
Tested
S1 6 Each unit is nlt Q + 5%
S2 6 Average of 12 is equal to or greater
than Q, no unit is less than Q - 15%

S3 12 Average of 24 is equal to or greater

than Q, not more than 2 units are
less than Q-15%, no unit is less tha n
Q - 25%
 Computing for Q value or %
dissolved

abs. sx conc. std. vol of medium x vol for dilution

------------- = --------------- DF= -------------------------------------------
vol of sample
abs. std. conc. sx.

Abs sx ave wt of sx
% Dissolved (Q) = ---------- x Conc Std x DF x -----------------
Abs std wt of ind sx
X100
Label Claim
 Sample Problem: Pyrazinamide
500 mg Tablet
In the dissolution data for Pyrazinamide Tablet, 500 mg. The
test used 3 paddle type apparatus set at the following parameters:
Medium - water, 900 mL
Speed - 50 rpm
Time - 45 minutes
Dilution - withdraw 10 ml of sample and dilute to 500-
VF ml
Wavelength – measure absorbance reading at 264 nm using
visible spectrophotometer.
PZA 500 mg Dissolution Profile

• Absorbance Conc wt of tablet

• Std 0.557 0.01mg/ml
• Sx 1 0.538 745.9 mg
• Sx 2 0.547 747.0 mg
• Sx 3 0.555 746.5 mg
• Dissolution Apparatus Demo:
https://www.youtube.com/watch?v=KPTx9yRxM6
w
• Dissolution Principle;
https://www.youtube.com/watch?v=FpkU123LVTc
 OUR LADY OF FATIMA UNIVERSITY
College of Pharmacy

Drug Stability
Pharmaceutical Analysis with Quality Control 2
(PHAN212)
 Unit Outcomes

◉ Explain principles of
stability studies, its vital
role in the manufacture of
drugs
◉ Differentiate the types of
overages
◉ Apply the principles of
stability studies in
computing for shelf-life
and assigning of
expiration dates
2
 Outline

◉ Principles and
Requirements of a Stability
Study
◉ Physical and Chemical
Stability
◉ Overages
◉ Calculations, Evaluations,
Application

3
 Stability
◉ It can be defined as the ability of a particular
formulation in a specific container or closure
system to remain with in its physical, chemical,
microbiological, therapeutic and toxicological
specifications.

The period of stability of a

preparation is the time from
the date of manufacture of
the formulation until its
chemical or biological act.
and is NLT 90% of the
labelled potency
5
 Shelf Life
◉ It indicates the period when the
formulation is expected to remain “fit
for use” under ordinary conditions of
handling and storage in the
environment such as warehouse,
home, hospital and pharmacy shelf
Duration of time during
which a drug preparation will
remain stable (possessing
NLT 90% of the labeled
potency)
 Expiration Date

◉ It is the direct application

and interpretation of
knowledge gained from
stability testing

◉ It limits the period during

which a preparation may
be expected to have its
labeled potency, provided
the product has been
stored as directed on the The date placed on the container
labeling label, after which it must not be
used
● Date of manufactured + shelf life
 Factors Affecting Stability of a
Pharmaceutical Product

◉ Stability of Active Ingredients

◉ Potential Interactions between active and inactive
ingredients
◉ Manufacturing process
◉ Container closure system
◉ Environmental conditions
◉ Storage
◉ Handling
◉ Length of time between handling manufacture
and usage
 Product Stability Evaluations

◉ Physical Stability is important to

formulators for three primary reasons:
○ Appearance

○ Uniformity

○ Availability
 Product Stability Evaluations

◉ Chemical Stability – causes chemical

deterioration incompatibilities which
may be:
○ Physical

○ Chemical

○ Therapeutic
 Important Parameters

◉ Drug Products
○ Loss of activity or potency of the active
ingredient
○ Amount of degradation product
◉ Cosmetics
○ Retention of the physical qualities of
freshly manufactured product
○ Instability that is gauged by its loss of
elegance
12
 Types of Stability Studies

◉ Short Term / Accelerated Stability Studies

○ This involves the use of exaggerated
conditions of temperature, light, moisture,
pH and humidity to test the stability of drug
formulations
■ 3 months acceptable data at 37-40°C/75%
RH for 2 year expiry date
○ The purpose is to determine kinetic
parameters, if possible and/or to predict the
tentative expiration
FDA 2014dating
bulletin: period
One year Controlled RT
samples and stored for three months using
accelerated stab testing = 1 year + 2 years
 Significance of Accelerated Stability Studies

◉ To intensify the degradation loss with time

◉ To enable researchers to predict the shelf
life of a product within a short period of time
◉ To determine the most stable formulation for
a particular therapeutic actives (in pre-
formulation studies)
 Types of Stability Studies

◉ Long Term/Real Time Stability Studies

○ Conducted under the usual/normal
conditions of the environment, transport and
storage expected during product distribution
■ 25°C / 60% RH ± 5%. Duration of at least
2yrs
○ It makes use of different “climatic zones” also
called Global Assessment of Stability of
Exposure
■ Zone 1 = Temperate
■ Zone 2 = Subtropical
■ Zone 3 = Hot and Dry
■ Zone 4 = Hot and Humid
 CLIMATIC ZONE

Climatic Definition Long term Conditions

Zone

I Temperate 21°C / 45% RH

II Subtropical and 25°C / 60% RH
Mediterranean

II Hot and Dry 30°C / 35% RH

IVA Hot and Humid 30°C / 65% RH
IVB Hot and very humid 30°C / 75% RH

16
Sample of countries of various
zones:

◉ Zone I : Britain, North Europe, Russia,

Canada
◉ Zone II : U.S.A, Japan , South Europe
◉ Zone III : Iran, Iraq, Sudan
◉ Zone IV :Brazil, Ghana, Indonesia,
Philippines
17
 Types of Stability Studies

◉ Stress Tests (done on API)

○ Involves the use of elevated
temperatures in 10 degrees increments
higher than those in ASS.
○ This test is performed until the total
physical and chemical degradation of the
product is reached
○ This stability testing has a duration of 6-12
months.
 Summary of Stability Conditions

Source: Rushing, W. (2017) ICH Stability Requirements: Overcoming the Challenges. EAG Laboratories Inc.

19
Stability Testing Frequency

Ex. March 2021–June/21-

Sept/21- Dec/21-Mar/22
Sept/22-Mar/23-Mar/24
 Storage Temperatures

Cold NMT 8°C (46°F)

Refrigerator 2°-8°C (36-46°F)
Freezer -20°C to -10°C (-4° to 14°F)
Cool 8-15°C (46-59°F)
Room Temperature Temperature prevailing in the
area (Ambient)
Controlled Temperature 15-30°C (59-86°F)
Warm 30-40°C
Excessive Heat Above 40°C (104°F)
 Overage

◉ Defined as “the voluntary introduction of a

specific excess during the manufacture of
pharmaceutical forms of medicaments that are
unstable by nature and difficult to stabilize, in
order to maintain during their period of use an
active content within the limits compatible
with therapeutic requirement”
 Types of Overage
◉ Manufacturing Overage
○ Overage added to a preparation to
compensate loss during
manufacturing of the preparation
○ Decrease stability profile during
manufacturing process
◉ Stability Overage
○ Is the excess added to a
preparation to extend its shelf life
 Overages are Justifiable when:

◉ The labile active ingredient cannot

possibly be standardized
◉ The overage allows an even equilibrium of
the content of the active ingredient within
the acceptable limits
◉ The overage would not present a
possibility of a therapeutic overdosage if
the preparation were used during the
early part of the product’s shelf life
◉ The clinical studies show that overage is
safe therapeutically
 Rules on Overages for Vitamins

◉ A loss of NMT 10% of the labeled potency

is considered normal at the end of the
term of validity of the product
◉ Different galenical dosage form is
considered separately with a distinction
made between simple and complex
preparations including a separate study of
preparations containing higher doses
◉ The added overage is limited to NMT 30%
of the labeled potency of the particular
ingredient
 Allowable Overage

Unstable Antibiotics NMT 15%

Dry Dosage Forms NMT 15%
Liquid Dosage Forms NMT 20%
Ointment
Suppositories
Aerosols } NMT 25%
Creams
Foams
 Predicting Shelf Life

◉ The technique of estimating the shelf life

of a formulation from its accumulated
stability data has evolved from examining
the data and making an educated guess
through plotting the time, temperature
points on appropriate graph paper and
crudely extrapolating a regression line to
the application of vigorous physical
chemical laws, statistical concepts and
computers to obtain meaningful reliable
estimates
 Methods of Predicting Shelf Life

◉ Free Hand Method

◉ Least Square Method
○ Linear regression, use of retained
samples
y = a + bx
where: y = 90 (constant)
 Steps in Least Square Method
x y x2 xy
1. Arrange the data in columns (mos) potency

2. Compute for the sum of the four values

3. Solve for a (Potency)
4. Solve for b
5. Assign y=90 (from the definition of shelf life: ∑= ∑= ∑= ∑=
the time from the date of manufacture until
its chemical or biological activity is NLT 90%
of its labeled potency)
6. Solve for x (shelf life in months)
7. Assign the expiration date using the
computed shelf life

Expiration Date = Date of Manufacture + Shelf Life

 FORMULA

y = a + bx
where: y = 90 (constant)

a = y - bx
30
Sample Problem 1:
A drug product (Label claim-120mg) was
manufactured by December 2012. The stability studies
results are listed below:
Potency should always be in %
Months (x) Initial 3 6 9
(0)
Assay (y) 108.0 103.0 105.0 103.0
Result (%)
x (months) y (potency) x2 xy
0 108 0 0
3 103 9 309
6 105 36 630
9 103 81 927

∑= 18 31
∑= 419 ∑= 126 ∑= 1866
a. Compute for a
b. Compute for b
c. Compute for x
d. What is the expiration date of the
product?

32
 ∑y∑x2 - ∑x∑xy
a = ------------------------------
N (∑x2) – (∑x)2

(419 x 126) - (18 x 1866)

a = ------------------------------------
(4 x 126) - (18) 2

(52,794) - (33588)
a = ------------------------------------
(504) - (324)

19,206
a = ----------------------
180
a=106.7
33
 N∑xy - ∑x∑y
b = ------------------------
N (∑x2) – (∑x)2
(4 x 1866) – (18 x 419)
b = ------------------------------
(4 x 126) – (18) 2
7464 - 7542
b = ------------------------
504 - 324

-78
b = ----------------
180
b= -0.43
34
 Compute for X

106.7 – 90
x = ----------------
0.43

16.7
x = ----------------
0.43

x = 38.84 3 years and 3 months

= 39 months March 2016
35
 Responsibility of Pharmacist

1. Observe FEFO and expiration dates

2. Observe proper storage conditions
3. Observe the products for evidence of instability
4. Properly treat and label repackaged, diluted or
mixed products
5. Dispense using the proper container with
proper closure
6. Inform and educate patients regarding the
proper storage and use of products including
proper disposition on outdated prescriptions
36
 References

◉ Rushing, W. (2017) ICH Stability Requirements: Overcoming

the Challenges. EAG Laboratories Inc.
◉ https://www.who.int/medicines/areas/qu
ality_safety/regulation_legislation/icdra/
WI-2_2Dec.pdf?ua=1
◉ https://www.fda.gov/media/71524/downl
oad
◉ https://www.fda.gov.ph/wp-
content/uploads/2021/03/ASEAN-
Guidelines-on-Stability-of-Drug-Product-
Version-6.0-Draft-5-May-2013.doc

37
38
Post-Lab
Spectrophotometry

Quality Control II- Laboratory

 UNIT OUTCOMES

At the end of this unit, the students are expected

to:
1. Understand the concept and principles
of Spectrophotometry.
2. Identify the laws governing Spectrophotometry.
Spectrophotome ry
 SPECTROMETRY

▪ is a method of analysis which deals

with the measurement of spectra.
 SPECTROPHOTOMETRY
•is a branch of spectrometry which embraces
the measurement of the absorption by chemical
species of radiant energy of definite and
narrow wavelength approximating
monochromatic radiation.
ELECTROMAGNETIC SPECTRUM
• complete system of energy
propagated in a wave form
• Energy of this nature:
✓ Radiant energy
ELECTROMAGNETIC SPECTRUM
PHOTOMETER
Electromagnetic wave parameters:

Wavelength (λ): Wavelength is the

distance between the consecutive peaks
or crests
 The ranges of the wavelength
UV 220 – 380 nm
Visible 380 – 780 nm
Near IR 780 – 3000 nm
Medium IR 3.0 – 15 um
Far IR 15 – 300 um
Frequency (n): Frequency is the
number of waves passing through any
point per second.
PRINCIPLES
❖Every substance absorbs or transmits
certain wavelengths of radiant energy but
not other wavelengths

❖Intensity of the transmitted radiant

energy is a function of the concentration
 CHROMOPHORE
• Are functional groups which absorb
radiant energy in the uv or vis regions
❑ EXAMPLES:
✓ Ethylene, Methylene, Organic acids,
Ketones, Aldehydes
 COLORIMETRY
• Branch of spectrophotometry in
which absorption takes place in the
vis region
• Uses a FILTER instead of a prism
• USEFUL IN DETECTINGTHE
PRESENCE OF GLUCOSE INTHE
URINE
Tungsten lamp Conde:nsing tens CUV\!'tte
I
II V'

*
I;
/
...... ....
... ...

A /I;. A,,
•·
I Ii
Slit FHter Photocell
 TRANSMITTANCE, ABSORBANCE
AND THE BEER LAMBERT’S LAW
▪ TRANSMITTANCE- the ratio of the
amount of light transmitted to the
amount of light that initially fell on
the surface.
 TRANSMITTANCE, ABSORBANCE
AND THE BEER LAMBERT’S LAW
▪ ABSORBANCE – the negative
logarithm of transmittance.
– Absorbance and transmittance bear
inverse relationship
 Beer’s Law states that the power of a
transmitted radiant beam decreases
exponentially as the concentration of the
solution containing the absorbing chemical
species increases arithmetically.

 Lambert’s / Bouguer’ s Law states that the

power of a transmitted radiant beam
decreases exponentially as the thickness of
the solution containing the absorbing
chemical species increases arithmetically.
Beer – Lambert or Beer –
Bouguer’ s Law is a combination
of the above law and relates the
power of the incident and the transmitted
radiant beam to the thickness and
concentration of the solution containing
the absorbing chemical species.
LIGHT SOURCE

UV spectro
▪ Hydrogen gas lamp
▪ Mercury lamp

Visible spectrometry
▪ tungsten lamp
Principle:
▪ Visible and ultraviolet spectroscopy is a study of electronic
spectra of organic molecules which are found in the
wavelength region of 100nm-400nm (UV region) and 400nm-
750nm (Visible region)
UV-VIS Spectrophototmeter
 UV-VIS Spectrophototmeter:
Instrumentation
▪ Light source
– Tungsten filament lamps and Hydrogen-Deuterium
lamps are most widely used and suitable light source as
they cover the whole UV region.
▪ Monochromator
– generally is composed of prisms and slits.
▪ Sample and reference cells
– These cells are made of either silica or quartz. Glass can’t
be used for the cells as it also absorbs light in the UV
region.
 UV-VIS Spectrophototmeter:
Instrumentation
▪ Detector
– Generally two photocells serve the purpose of detector
in UV spectroscopy.
▪ Amplifier
– Generally current generated in the photocells is of very
low intensity, the main purpose of amplifier is to amplify
the signals many times to get a clear and recordable
signals.
▪ Recording devices
– Computer stores all the data generated and produces
the spectrum of the desired compound.
 UV-VIS Spectrophototmeter:
Applications
1. Detection of Impurities
2. Structure elucidation of organic compounds
3. Quantitative determination of compounds that
absorb UV radiation.
4. Detection of presence or absence of functional
group in the compound.
5. In the study of kinetics of reactions
6. Drug and raw materials assay
7. Measurements of molecular weights.
Sample Problem:

Rifampicin 500mg Capsule

Weight of Reference Standard:
=55.9mg dilute to 100ml take 5ml dilute to 100ml
Weight of Sample
=60.02mg dilute to 100ml take 5ml dilute to 100ml
Total Weight of 10 Capsules = 6030mg
Weight of Empty Gelatin Capsule =101mg
Peak Area of Standard = 90651 nm2
Peak Area of Sample = 84414 nm2
Specification= NLT 90% but NMT 110%
28
1. DETERMINE THE CONCENTRATION OF ACTIVE
CONSTITUENT IN THE FINAL DILUTION

Concentration of Sample (Actual)

Peak Area of Sample
= ------------------------------- X Conc. of Standard
Peak Area of Standard
Answer:
84414 nm2
= ------------------------------- X 27.95 mcg/ml
90651 nm2

= 26.03mcg/ml 29
2. DETERMINE THE CONCENTRATION OF THE SAMPLE
TAKEN IN THE FINAL DILUTION.

Concentration of Sample (Theoretical)

Answer:
Concentration of Sample in the Final Dilution
60.02mg 5ml 1000ug
= ---------- x ------ x -------
100ml 100ml 1mg
= 30.01mcg/ml

30
3. DETERMINE THE QUANTITY OF RIFAMPICIN
PRESENT PER CAPSULE.

Amount (mg/dosage form)

Conc. of Sample (Actual) X Total Weight
# of units
=
Conc. of Sample (Theoretical)
Answer:
26.03mcg/ml x 502mg
=
30.01mcg/ml
= 435.42mg

31
4. DETERMINE THE PERCENT LABELLED
AMOUNT.

% Potency (% Assay)
Amount present per dosage form
= X 100
Label Claim
Answer:
435.42mg
= ------------- x 100 = 87.084%
500.00mg
32
5. DETERMINE THE PERCENT PURITY OF THE
SAMPLE.
% Purity
Conc. of Sample (Actual)
= x 100
Conc. of Sample (Theoretical)
Answer:
26.03mcg/ml
= ------------------ x 100 = 86.74%
30.01mcg/ml
33
 Abs Sx
SUMMARY:
------------- x Conc Std
Abs Std
% Purity of sample = ------------------------------ x 100
Conc of Sx (Theoretical)

mg /tab = Pure sample x Ave wt.

Abs sx
------------- x Conc Std x Ave wt
Abs Std

Conc of Sx (Theoretical)
% Assay = x 100
Label Claim
 REFERENCES

→ Jenkins, Glenn L, Jenkins Quantitative

Pharmaceutical Chemistry, N.Y, N.Y. : Mc
Graw, Latest Edition, (Chapter in
Chromatography)
→ Watson, David G., Pharmaceutical Analysis: A
Textbook for Pharmacy Students &
Pharmaceutical Chemist, Churchill Livingstone,
Edinburgh, 2012 (Chapter in Chromatography)
 Post – Lab
Quality Control II with
Instrumentation

DISINTEGRATION TESTING
USP <701>
Unit Outcomes:
At the end of this unit, the
students are expected to:
❑ Define the disintegration
test and its underlying
principle.
❑ Enumerate the different
examples of disintegrants.
❑ Identify the parts of the
apparatus.
• For a drug to be readily available to the body , it must
be in solution.
Most tablets, the first important step toward solution is
break down of the tablet into smaller particles or granules,
a process called disintegration.
 Disintegration test is provided to
determine whether tablets or capsules
disintegrate within the prescribed time
when placed in a liquid medium under the
experimental conditions presented below.

 Complete disintegration is defined as

that state in which any residue of the unit,
except fragments of insoluble coating or
capsule shell, remaining on the screen of
the test apparatus or adhering to the
lower surface of the disk, if used, is a soft
mass having no palpably firm core
(USP).
 Disintegrants – substances or
agents added to compressed tablets
to cause them to break apart or
disintegrate when placed in an
aqueous medium.

Examples: starch, cellulose

derivatives, CMC, avicel, alginates.
 DISINTEGRATION APPARATUS

CONTROL
PANEL

TEMPERATURE ARM
PROBE

VESSEL
BASKET
RACK
ASSEMBLY
WATER
BATH
DISINTEGRATION APPARATUS
Apparatus:
 Basket rack assembly
 Beaker
 Thermostat
 Device or lever which lowers and
raises the basket (Arm)
Basket rack assembly
includes:
• Six open ended
transparent tubes, each
7.75 +/- 0.25 cm long with
diameter of approx. 21.5
mm and wall thickness of
approx. 2 mm.
• Two plastic plates, with
the lower plate having a
woven stainless steel wire
cloth, plain square weave.
Basket rack assembly
includes:
• Disks – cylindrical in shape,
9.5 +/- 0.15 mm thick and
20.7 +/- 0.15mm in diameter.
PROCEDURE:

1. Place 1 dosage unit in each of the 6 tubes of the basket

2. Place the disk on top of each unit
3. Operate the apparatus using the specified medium kept at
37+/-2°C
4. At the end of the time limit specified, inspect the units if all
units have disintegrated completely
 USP SPECIFICATIONS:

Uncoated Hard
Plain Enteric Buccal Sublingual
tablets gelatin
uncoated coated tablets
-37±2º tablet -apply the and soft
tablet *apply the test for gelatin
-distilled *apply the - Immerse test for uncoated -apply test
water test for the basket uncoated tablets
in water tablets for
- 30 minutes uncoated -time uncoated
tablet for 5 mins.
- 4 hours specified tablets
-simulated by USP
gastric - Attached
fluid TS 10-mesh
removable
- 1 hour wire cloth
Acceptance Criteria

TEST SAMPLES CRITERIA

NMT 1 fails to
disintegrate
Initial Test 6 samples
within the
prescribed time.
+ 12 samples
16/18 should
Second Test disintegrate
Total of 18
completely
samples
Factors affecting disintegration rate:
 physical and chemical properties of
granules
 hardness and porosity of the tablet
 disintegration agent used
 medium used
Solvents for testing:
 distilled water
 gastric juice
 phosphate buffer
 simulated gastric juice
 simulated intestinal juice
 DISINTEGRATION AS A QUALITY
CONTROL TEST
According to decision Tree #7 in the International Council for
Harmonization (ICH) Tripartite Guideline Q6A, the use of disintegration
testing instead of dissolution is allowed when the following criteria are
met:

 Immediate-release dosage form (i.e. no modified

release);
 The drug product contains a drug that is highly
soluble throughout the physiological range
(dose/solubility volume < 250 mL from pH 1.2 to
6.8);
 DISINTEGRATION AS A QUALITY
CONTROL TEST
 Rapidly dissolving products (dissolution > 80% in 15
minutes at pH 1.2, 4.0, and 6.8); and
 Establishment of a relationship between
disintegration and dissolution or when disintegration
is shown to be more discriminating than dissolution.

NOTE: USP Chapter <2>, Oral drug products – Product quality

test, states “only when disintegration has been correlated with
dissolution of a dosage form can a disintegration test be used as a
product performance test”.
Key Points:
1. What is the disintegration test?
2. Give at least 2 disintegrants?
3. What are the acceptance
criteria in initial and second
testing?
4. What is the medium used for
Enteric coated tablets?
Pop-up quiz :
CONTROL
PANEL

TEMPERATURE ARM
PROBE

VESSEL
BASKET
RACK
ASSEMBLY
WATER
BATH
REFERENCES:
 à Jenkins, Glenn L, Jenkins
Quantitative Pharmaceutical
Chemistry, N.Y, N.Y. : Mc Graw,
Latest Edition, (Chapter in
Chromatography)
 à Watson, David G., Pharmaceutical
Analysis: A Textbook for Pharmacy
Students & Pharmaceutical Chemist,
Churchill Livingstone, Edinburgh, 2012
(Chapter in Chromatography)