1. Pharmaceutical Quality
Management
7th Semester Syllabus
Submitted to:
Dr. Sajid Bashir

2. Introduction to Quality
Management and Quality
Control
Presented by:
M. Talha Afzal

3. Quality:-
"the totality of features and characteristics of a product or service
that bears its ability to satisfy stated or implied needs.’’
Quality Management:-
“The act of overseeing all activities and
tasks needed to maintain a desired level of excellence. This includes the
determination of a quality policy, creating and implementing quality
planning and assurance, and quality control and quality improvement.”

4. Quality Management System:-
A quality management system (QMS) is a formalized system that
documents processes, procedures, and responsibilities for achieving quality
policies and objectives. A QMS helps coordinate and direct an organization’s
activities to meet customer and regulatory requirements and improve its
effectiveness and efficiency on a continuous basis.
Quality management systems serve many purposes, including:
 Improving processes
 Reducing waste
 Lowering costs
 Facilitating and identifying training opportunities
 Engaging staff
 Setting organization-wide direction

5. Quality Assurance:-
Quality assurance is the totality of the arrangements made with the
object of ensuring that pharmaceutical products are of the quality required
for their intended use. It incorporates GMP and other factors, including
those outside the scope of this guide such as product design and
development.
Quality Control:-
Quality control (QC) is a procedure or set of procedures intended to ensure
that a manufactured product or performed service adheres to a defined set of
quality criteria or meets the requirements of the client or customer. QC is
similar to, but not identical with quality assurance.

6. Difference b/w Quality Assurance and Quality Control
 The difference is that QA is process oriented and QC is product
oriented.
 Quality Assurance makes sure you are doing the right things, the right
way.
 Quality Control makes sure the results of what you've done are what
you expected.
 Quality Assurance is focuses on defect prevention, while quality
control is focuses on defect identification.

7.  Quality assurance(QA) in pharmaceutical industry is used to ensure that
product or service will satisfy requirement for “Quality” and covers all
matters related to factors influencing quality of a product.
 Quality control(QC) is concerned with sampling, specifications,
testings' , organization, documentation and release procedures which
ensures that necessary and relevant tests are actually carried out and
that materials are not released for sale, supply until their quality has
been judged to be satisfactory.

8. Comparison chart
Features Quality Assurance Quality Control
Definition QA is a set of activities for ensuring
quality in the processes by which
products are developed.
QC is a set of activities for ensuring
quality in products. The activities
focus on identifying defects in the
actual products produced.
Focus on to prevent defects with a focus on
the process used to make the
product. It is a proactive quality
process.
to identify (and correct) defects in the
finished product. Quality control,
therefore, is a reactive process.
Goal is to improve development and test
processes so that defects do not
arise when the product is being
developed.
is to identify defects after a product is
developed and before it's released.

9. Features Quality Assurance Quality Control
Responsibility Everyone on the team involved in
developing the product is
responsible for quality assurance.
Quality control is usually
the responsibility of a
specific team that tests the
product for defects.
As a tool QA is a managerial tool QC is a corrective tool
Orientation QA is process oriented QC is product oriented
Example Verification is an example of QA Validation/Software Testing
is an example of QC

10. Quality Control Test of Tablets
And Capsules

11. TABLETS
• Tablets may be characterized or described by number of specifications.
• These include: shape, diameter, thickness, weight variation, hardness,
friability, disintegration time and dissolution characteristics.
General Appearance:
Size, shape, and thickness
Organoleptic properties:
Color and odor of the tablets.

12. OFFICIAL AND UNOFFICIAL TESTS:
Non-Official Tests:
Hardness, Friability
Official Tests:
Weight variation, Disintegration,
Dissolution, Content uniformity

13. GENERAL APPEARANCE
 Diameter and Shape:
• The diameter and shape depend on the die and punches selected for the
compression of the tablet.
• Generally, tablets are discoid in shape, although they may be oval,
oblong, round, cylindrical, or triangular.
• Their upper and lower surfaces may be flat, round, concave or convex
to various degrees.
• The top or lower surface may be embossed or engraved with a symbol
or letters which serve as an additional mean of identifying the source
of the tablets.

14.  Tablet Thickness:
• The thickness of the tablet is the only dimensional variable related to the
compression process. Tablet thickness is consistent batch to batch if the
tablet granulation or powder blend is adequately consistent in particle size
and size distribution.
 The thickness of individual tablets may be measured with a micrometer.
 Thickness should be controlled within ± 5% variation of a standard value.
 Thickness must be controlled for consumer acceptance of the product, and
to facilitate packaging.

15. ORGANOLEPTIC PROPERTIES:
• Colour:
Many pharmaceutical tablets use color as a vital means of rapid identification
and consumer acceptance.
The color of a product must be uniform within a single tablet
Non-uniformity of coloring not only lacks esthetic appeal but could be
associated with poor quality of the product.
• Odour:
The presence of odor in a batch of tablets could indicate stability problems,
such as the odor of acetic acid in degrading aspirin tablets.
However, the presence of an odor could be characteristic for the drug
(vitamins), added ingredients (flavoring agents) or the dosage form (film-coated
tablets).

16. NON-OFFICIAL TESTS
• HARDNESS
Tablet hardness can be defined as the force required to break a tablet in
a diametric compression test. OR It is the load required to crush the
tablet when placed on its edge.
• Why do we measure hardness?
To determine the need for pressure adjustments on the tableting machine.
Hardness can affect the disintegration. So if the tablet is too hard, it may
not disintegrate in the required period of time. And if the tablet is too
soft, it will not withstand the handling during subsequent processing such
as coating and packaging.

17. • Factors Affecting the Hardness:
Compression of the tablet and compressive force.
Amount of binder.
Method of granulation in preparing the tablet
• Hardness Test Apparatus
Monsanto tester, Pfizer tester, strong cobb hardness tester are
manually used & Heberlien schleuniger, Eweka, Casburt hardness
tester are motor driven testers.

18. • PROCEDURE:
MONSANTO TESTER: The apparatus consists of 2 jaws facing
each other, one of which move towards the other. Measurements is
carried out on 10 tablets, taking care to remove all the fragments of
the broken tablets before each determination & then take the average
hardness.
• LIMITS:
Normal tablet hardness ranges from 4 – 8 Kg (1 Kg = 10 Newton)

19. • FRIABILITY:
It is expressed as a percentage loss in weight OR It is the tendency of
tablets to powder, chip, or fragment and this can affect the elegance
appearance, consumer acceptance of the tablet.
The measurement of friability is made by Roche friabilator. A number
of tablets are weighed and placed in the tumbling apparatus where they
are exposed to rolling and operated shocks resulting from free falls
within the apparatus. After a given number of rotations, the tablets are
weighed and the loss in weight indicates the ability of the tablets to
withstand this type of wear.

20. • Method:
Select 20 tablets randomly, dedust and weigh (WO).
Place the tablets in the Roche friabilator drum, switch on the apparatus
adjusting the timer at 4 min and the speed at 25 rpm.
At the end of this operation, remove the tablets from the friabilator,
dedust and reweigh (W). (Any tablet that breaks up should be rejected
before reweighing).
friability is measured by using formula as:
• Limit:
If the value of friability (% loss) is less than or equal to 1%, the batch is
accepted.
%loss = 100x
W
WW
O
O 

21. OFFICIAL TESTS
• DISINTEGRATION:
It is the time required for the tablet to break into particles,
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.

22. • The Disintegration Apparatus:
This apparatus consists of a basket rack containing 6-open-ended glass
tubes held in a vertical position. A number 10-mesh stainless steel wire
screen is attached to the bottom.
One tablet is placed in each tube and the basket rack is positioned in a 1-
L beaker of water, simulated gastric fluid (pH 1.2), or simulated intestinal
fluid (pH 7.4), at 37 oC. A standard motor driven device is used to move
the basket assembly containing the tablets up and down through a
distance of 5 to 6 cm at a frequency of 23 to 32 cycles per minute,
perforated discs may also be used in the test. These are placed on the top
of the tablets. These discs are useful for tablets that float.
To be in compliance with standards, the tablets must disintegrate, and all
of the particles must pass through the 10-mesh screen in the time
specified.

23. •Limits:
 Uncoated tablets should disintegrate within 30 minutes
 Enteric coated tablets are to show no evidence of
disintegration after 1 hour in simulated gastric fluid. These
tablets are then tested in simulated intestinal fluid and are
to disintegrate in 2 hours plus the time specified in the
monograph.
If one tablet fails to disintegrate, the disintegration test
is repeated on 12 additional tablets. Not less than 16 out oh
the total 18 tablets tested disintegrate completely within 30
minutes.

24. • Uniformity of Active Ingredients:
It is measured to ensure a constant dose of drug between individual
drugs
Traditionally, dose variation between tablet is tested in two separate
tests namely
A) Weight variation
B) Content uniformity

25. • WEIGHT VARIATION:
Uniformity of weight is an in-process test parameter
which ensures consistency of dosage units during
compression.
The weight of the tablets being made should be
routinely measured to help ensure that the tablet contains
the proper amount of drug.

26. • Procedure:
Weigh 20 tablet selected at random, each one individually X1, X2,
X3… X20
Determine the average weight. X= (X1+X2 +X3+…+ X20 )/20
Not more than 2 of the individual weights deviate from the average
weight (x) by more than the % deviation given below & none
deviates by more than twice that %.
• Limits:
Weight of tablet 80 mg or less, then % deviation = ±10%
Weight of tablet >80 - <250 mg, then % deviation = ±7.5
 Weight of tablet 250 mg or more, then % deviation = ±5%

27. • CONTENT UNIFORMITY:
Uniformity of Content is a pharmaceutical analysis parameter for the
quality control of tablets. Multiple tablets are selected at random and a
suitable analytical method is applied to assay the individual content of the
active ingredient in each tablet.
• Method:
Randomly select 30 tablets. 10 of these assayed individually.
The Tablet pass the test if 9 of the 10 tablets must contain not less than 85%
and not more than 115% of the labeled drug content and the 10th tablet may
not contain less than 75% and more than 125% of the labeled content.
If these conditions are not met, remaining 20 tablet assayed individually and
none may fall out side of the 85 to 115% range.

28. • DISSOLUTION TEST:
The release of drug from the tablet into solution per unit time under
standardize condition is called dissolution test.
Media used in dissolution testing may be purified water, simulated gastric
fluid, simulated intestinal fluid or others.
•Dissolution Apparatus
1. Rotating basket method 2. Paddle method
3. Reciprocating cylinder method 4. Flow through cell method
5. Paddle over disk method 6. Cylinder method
7. Reciprocating disk method

29. • PROCEDURE:
APPARATUS-1 (BASKET TYPE):
A single tablet is placed in a small wire mesh basket attached to the
bottom of the shaft connected to a variable speed motor.
The basket is immersed in a dissolution medium (as specified in
monograph) contained in a 1000 ml flask.
The flask is cylindrical with a hemispherical bottom. The flask is
maintained at 37±0.5 0 C by a constant temperature bath. The motor is
adjusted to turn at the specified speed and sample of the fluid are
withdrawn at intervals to determine the amount of drug in solutions.

30. APPARATUS-2 (PADDLE TYPE):
• It is same as apparatus-1, except the basket is replaced by a paddle.
• The tablet is allowed to sink to the bottom of the flask before stirring.
• For dissolution test U.S.P. specifies the dissolution test medium and
volume, type of apparatus to be used, rpm of the shaft, time limit of
the test and assay procedure.
• The test tolerance is expressed as a % of the labeled amount of drug
dissolved in the time limit

31. QUALITY CONTROL TEST
for Capsules
Presented by:
M. Talha Afzal

32. CAPSULES
• QUALITY CONTROL TEST
for Capsules includes:
Weight variation
Content uniformity
Disintegration
Dissolution

33. • CONTENT UNIFORMITY TEST:
This test is performed when specified by individual monographs.
In this case, 30 capsules are selected, 10 of which are assayed by the
specified procedure.
The requirements are met if 9 of the 10 are within the specified
potency range of 85 to 115 %, and the tenth is not outside 75 to 125 %.
If more than 1, but less than 3, of the first 10 capsules fall out side the
85 to 115 % limits, the remaining 20 are assayed The requirements are
met if all 30 capsules are with in 75 to 125 % of the specified potency
range, and not less than 27 of the 30 are within the 85 to 115% range.

34. • DISINTEGRATION:
The disintegration of capsules is different from those of tablets because
the determination of end point is difficult owing to the adhesive nature of
shell. The shell pieces after disintegration may agglomerate forming large
mass of gelatin taking more time to dissolve and may adhere to the mesh
thus, blocking the holes.
 According to USP, place one dosage unit in each of the tubes of the basket
with water or any other specified medium (depends on individual
monograph) maintained at 37 + 2C.
Observe the capsules for a time limit (specified in individual monograph),
at the end of prescribed time, all of the capsules must have been
disintegrated excluding the fragments from the capsule shell.
• The capsules pass the test if:
No residue remains on the screen of the apparatus,
If a residue remains, it consists of fragments of shell.

35. • WEIGHT VARIATION:
For hard capsules: Accurately weigh 10 capsules. By suitable means the
contents of each capsule should be removed. The weights of emptied shells
should be recorded individually. The difference of both the weights will
yield the net weight of the contents. Then calculate acceptance value.
For soft capsules: pre weigh 10 capsules. Cut the capsules by suitable means
(either scissors or any open blade) remove the contents by washing with a
suitable solvent and let the solvent evaporate by placing them at room
temperature for about 30 mins. Weigh the individual shells. Calculate the
acceptance value
• Limit:
Not more than two of the individual weights deviate from the average
weight by more than the percentage deviation shown in the the table, and
none deviates by more than twice that percentage.

36. • DISSOLUTION TEST:
The dissolution test is carried out using the dissolution apparatus
The capsule is placed in a basket , and the basket is immersed in the
dissolution medium and caused to rotate at a specified speed
The dissolution medium is held in a covered 1000ml glass vessel and
maintained at 37c +-0.5 by means of a constant temperature suitable water
bath .
The stirrer speed and type of dissolution medium are specified in the
individual monograph

37. Quality Control tests for
Syrups And Elixirs

38. Syrup:-
Syrup:-
“Syrups are concentrated aqueous preparations of a sugar or sugar substance
with or without flavoring agents and medicinal substances.”
Components of Syrup:
(1) Sugar or its substitute (2) Medicinal agent
(3) Flavourent (4) Colourent etc.
Elixirs:-
“A clear, sweetened, hydro-alcoholic liquid intended for oral
use; elixirs contain flavoring substances and are used either as vehicles or for
the therapeutic effect of the active medicinal agents.”

39. Viscosity Measurement
Viscosity is a property of liquids that is directly related to the resistance to
flow.
viscosity measurement is very important quality control test in case of
syrups and elixirs.
viscosity and consistency directly relates with stability of solutions.
viscosity chance of stability.

41. Method 1 (U tube viscometer)
Apparatus:-
The apparatus consists of a glass U tube viscometer made of clear
borosilicate glass and constructed in accordance with the dimensions given
in official books. The monograph states the size of viscometer to be used.

42. Method
• Fill the viscometer with the liquid being examined
through tube L to slightly above the mark G, using a long
pipette to minimize wetting the tube above the mark.
• Place the tube vertically in a water bath and when it has
attained the specified temperature, adjust the volume of the
liquid so that the bottom of the meniscus settles at the mark
G.
• Adjust the liquid to a point about 5 mm above the mark E.
• After releasing pressure or suction, measure the time
taken for the bottom of the meniscus to fall from the top
edge of mark E to the top edge of mark F.

43. Method II (Capillary viscometer method)
(Ph. Eur. method 2.2.9)
• The determination of viscosity using a suitable capillary viscometer is
carried out at a temperature of 20 ± 0.1 °C, unless otherwise prescribed.
• The time required for the level of the liquid to drop from one mark to the
other is measured with a stop-watch to the nearest one-fifth of a second.
• The result is valid only if two consecutive readings do not differ by more
than 1 per cent.
• The average of not fewer than three readings gives the flow time of the
liquid to be examined.

44. Calculate the dynamic viscosity ƞ in mili-pascal seconds
using the formula:
ƞ=kρt
k= constant of the viscometer
ρ= density of the liquid to be examined expressed in milligrams per
cubic millimeter.
t= flow time, in seconds, of the liquid to be examined.
The constant k is determined using a suitable viscometer calibration
liquid.

45. Method III (Rotating viscometer method)
(Ph. Eur. method 2.2.10)
The principle of the method is to measure the force acting on a rotor (torque)
when it rotates at a constant angular velocity (rotational speed) in a liquid.
• Rotating viscometers are used for measuring the viscosity of Newtonian
(shear-independent viscosity) or non-Newtonian liquids (shear dependent
viscosity or apparent viscosity).
• Rotating viscometers can be divided in 2 groups, namely absolute and
relative viscometers.
• In absolute viscometers the flow in the measuring geometry is well defined.
The measurements result in absolute viscosity values, which can be compared
with any other absolute values.

46. In relative viscometers the flow in the measuring geometry is not
defined. The measurements result in relative viscosity values, which cannot
be compared with absolute values or other relative values if not determined
by the same relative viscometer method.
Different measuring systems are available for given viscosity ranges as
well as several rotational speeds.

47. Weight/mL
• Important for tinctures and extracts and defined as:
“The weight per milliliter of a liquid is the weight expressed in grams of 1
milliliter of a liquid when weighed in air at the specific temperature”.
• Almost equal to density of the liquid. Expressed in g/ml
Procedure: For this,
– Weigh a clean and dry pycnometer accurately
– Fill it with liquid, remove any access and weigh
– Determine the weight of liquid by subtraction
• Determine wt./ml by dividing the weight of liquid that filled the pycnometer
by the capacity of pycnometer

49. SUPPOSITORIES
“Suppositories are solid dosage forms intended for
insertion into body orifices where they melt, soften, or
dissolve and exert localized or systemic effects".
It comes under semi solid preperation because it is
prepared by melting all ingredient.

50. 1.APPEARANCE
 It includes
Odour
 Colour
Surface condition
Shape.

51. WEIGHT UNIFORMITY
 Weigh 20 suppositories individually. w1, w2, w3….w20
 Weigh all the suppositories together
 Calculate the average weight = W/20.
 Limit: Not more than 2 suppositories differ from the average
weight by more than 5%, and no suppository differs from the
average weight by more than 10%.

52. MELTING RANGE TEST
 1-The suppository is completely immersed in the constant
temperature water bath, and the time for the entire
suppository to melt or disperse in the surrounding water is
measured.

53. DISINTEGRATION TEST…
 The suppository is considered disintegrated when:
A- It is completely dissolved or
B- Dispersed into its component part.
C- Become soft “change in shape” with formation of
• core which is not resistant to pressure with glass rod.

54. Method:
 Place a suppository on the lower perforated disc of metal device
and then insert the device into the cylinder and attach this to the
sleeve.
 Repeat the same operation with further two suppositories, metal
device and sleeve. Place each piece of apparatus in a vessel
containing at least five liters of water at 37C and fitted with a
slow stirrer and by means of holding the top of apparatus 90
mm below the surface of water. After every 10 minutes invert
each apparatus without removing it from water.

55. Disintegration is complete when molded suppositories are:
 Completely dissolved.
 Dispersed into its components.
 Have become soft.
 Disintegration occurs in not more than 30 minutes
for fat based suppositories and for water-soluble
suppositories disintegration occurs in not more than
60 minutes.

56. LIQUEFACATION TIME OR
SOFTENING TIME TEST
 In this test a U tube is partially immersed in a constant temperature
bath and is maintained at a temperature between 35 to 37°C. There is a
constriction in the tube in which the suppository is kept and above the
suppository, a glass rod is kept. The time taken for the glass rod to go
through the suppository and reach the constriction is known as the
liquefaction time or softening time
 Another apparatus is there for finding “softening time” which mimics
in vivo conditions. It uses a cellophane tube, and the temperature is
maintained by water circulation. Time taken for the suppository to melt
is noted.

57. DISSOLUTION TEST
 By using different types of apparatus such as wire mesh
basket, or dialysis tubing is used to test for in vitro release
from suppositories

58.  Two types of apparatus are avaliable for testing the
dissolution rate. They are,
 SUPPOSITORY DIALYSIS CELL - lipophilic
suppositories are tested using suppository dialysis cell.
 STATIONARY BASKET – rotating paddle apparatus
( USP dissolution test apparatus). Hydrophilic suppositories
are tested using stationary basket.

59. STABILITYTESTING
 Cocoa butter suppositories on storage, “bloom”; i.e., they form a white powdery
deposit on the surface. This can be avoided by storing the suppositories at
uniform cool temperatures and by wrapping them in foils.
 Fat based suppository harden on storage.
 There is an upward shift in melting range due to slow crystallization to the
more stable polymeric forms of the base.
 The softening time test and differential scanning calorimetry can be used as
stability indicating test methods.
 if we store the suppositories at an elevated temperature, just below its melting
range, immediately after manufacture, the aging process is speeded.

60. QUALITY CONTROL TESTS FOR
PARENTRALS
Presented by:
M. Talha Afzal

61. Contents
• Uniformity of content
• Test for volume of liquid
• Test for pyrogen
• Test for sterility
• Clarity of solution
• Test for bacterial endotoxin
• Leakage test

62. PARENTERALS
Term derived from Greek words “Para” outside & “Enteron” intestine.
• Parenterals are sterile solution/suspension of drug in aqueous or oily vehicle.
• Parenteral drugs are administered directly into the veins, muscles or under the
skin, or more specialized tissues such as the spinal cord.
• Term parenteral used for any drug/fluid whose delivery does not utilize the
alimentary canal for entering into the body tissues.
• Parenteral preparations are supplied in glass , plastic container and
prefilled syringes with closures are made up of plastic or elastomer.

63. Categories of parenteral preparations:
 Injections or infusions
 Concentrates for injections or infusions
 Powders for injection or infusions
 Gels for injections
 Implants
Unique Characteristics of Parenterals:
Sterile Particulate-free Pyrogen free
Stable for intended use pH – not vary significantly
Osmotic pressure similar to blood

64. QUALITY CONTROL TESTING

65. General areas of Quality
control
Three General Areas are:
• 1. Incoming Stock:
Routine work testing
• 2. Manufacturing:
Include numerable tests, reading and observations through out the
manufacturing process
• 3. Finished Products:
Sterility test, Pyrogen test, Clarity test, Leaker test.

66. In process Quality control tests
 Conductivity measurement
Volume filled
 Temperature for heat sterilized product
Environmental control tests
Visual inspection

67. TESTS FOR PARENTERALS
Finished product Quality control tests:
There are mainly five Quality control test for the parenterals are
performed.
1) LEAKER TEST
2) CLARITY TEST
3) PYROGEN TEST
4) STERILITY TEST
5) CONTENT UNIFORMITY TEST

68. UNIFORMITY OF CONTENT
• 30 sterile units are selected from each batch.
• The weight of 10 individual sterile units is noted and the content is
removed from them and empty individual sterile unit is weighed accurately
again.
• Then net weight is calculated by subtracting empty sterile unit weight
from gross weight.
• The dose uniformity is met if the amount of active ingredient is within the
range of 85-115% of label claim.

69. • Relative standard deviation is equal to or less than 6.0%.
• If one unit is outside the range of 85-115.0%, and none of the sterile unit is
outside the range of 75-125.0% or if the relative standard deviation of the
resultant is greater than 6.0% ,or if both condition prevail, an additional 20
sterile unit should be tested.
• The sterile units meet the requirements if not more than one unit is out side
the range of 85-115%, no unit is outside the range of 75-125.0% and the
calculated relative standard deviation is 7.8%.

70. TEST FOR VOLUME OF LIQUID
Test applies to liquid supplied in single dose , only part of the content is
used.
Empty the contents of one container & determine the volume of contents.
For Emulsions & suspensions, shake the container before the
determination
The volume should not be less than the amount stated on the label.

71. Parenteral
preparations
Minimum number of
items tested
Not more than 100
containers
10% or 4 container
More than 100 but not
more than 500 containers
10 containers
More than 500 containers 2% or 20 containers
whichever is less
For large volume
parenterals
2% or 20 containers
whichever is less

72. 1) LEAKER TEST
 Leakage occurs when a discontinuity exists in the wall of a package
that can allow the passage of gas under the action of a pressure or
concentration differential existing across the wall.
 Presence of capillary pores or tiny cracks can cause microbes or other
dangerous contaminants to enter the ampoules or package or may lead
to the leakage of contents to outside. This may cause contamination of
the sterile contents and also spoilage of appearance of the package.
Changes in temperature during storage can cause expansion and
contraction of the ampoule or package and thereby causing interchange
of its contents if an opening exists.

73. Leaker test is employed to detect incompletely sealed ampoule so
that they may be discarded.
• To test the package integrity.
• Package integrity reflects its ability to keep the product in and to keep
potential contamination out.
Leaker tests are 4 types:
a) visual inspection
b) bubble test
c) dye test
d) vacuum ionization

74. a) Visual inspection
• Visual inspection is the easiest leaker test method
to perform.
• The method is used for the evaluation of large volume parenterals.
• To increase the sensitivity of the method the visual inspection of the
sample container may be coupled with the application of vacuum to
make leakage more readily observable.
• This method is simple and inexpensive.
• Disadvantage: less sensitive
• Sensitivity is increased by applying pressure/vacuum.

75. b)Bubble test
 The test package is submerged in liquids.
 A differential pressure is applied on the container.
 The container is observed for bubbles.
 Sometimes, surfactant added liquid is used for immersion of test package.
 Any leakage is evident after the application of differential pressure as the
generation of foaming in immersion liquid.
 The method is simple and inexpensive.
 The location of the leaks can be observed in this method.

76.  Generation of a differential positive pressure of 3 psi inside
the vial and observation of any leakage using magnifying glass
within a maximum test time of 15 minutes.
 However, it is relatively insensitive and the findings are
operator dependent and are qualitative.
 The optimized conditions can be achieved using a surfactant
immersion fluid along with the dark background and High
intensity lighting.

77. C)Dye test
• The test container is immersed in a dye bath.
• Vacuum and pressure is applied for sometime.
• The container is removed from the dye bath and washed.
• The container is then inspected for the presence of dye either visually
or by means of UV spectroscopy.
• The dye used is usually 0.5% to 1% methylene blue.
• The dye test can be optimized by use of a surfactant and or a low
viscosity fluid.

78. • The dye test is widely accepted in industry and is approved in drug
use.
• The test is inexpensive and is requires no special equipment
required for visual dye detection.
• However, the test is qualitative, destructive and slow.
• The test is used for ampoules.

79. D)Vacuum ionization test
 Vacuum ionization test is useful for testing leakage in the vials or
bottled sealed under vacuum.
 This test is used for testing of the lyophilized products.
 High voltage, high frequency field is applied to vials which to cause
residual gas, if present to glow.
 Glow intensity is the function of headspace vacuum level.
 The blue glow is the indicative of vacuum while the purple glow
indicative of no vacuum.

80.  The sensitivity of the method is not documented.
 This test is rapid and is non destructive test.
 However, the proteins present in the test sample may be decomposed.
 This method is used for the lyophilized vials of biopharmaceuticals

81. 2) CLARITY TEST
Definition:
Clarity is a relative term, its mean a clear solution having a high
polish conveys to the observer that the product is of exceptional quality
and purity.
Clarity test is carried out to check the particulate matter in the sample.
It is practically impossible that every unit of lot is perfectly free from
visible particulate matter ,that is, from particles that are 30 to 40
micrometer and large in size.

82. PARTICULATE MATTER TEST
• Particulate matter refers to the extraneous, mobile, undissolved
particles, other than gas bubbles, unintentionally present in the
solutions.
2 methods are used:
(i)Light obstruction Particle Count Test
(ii) Microscopic particle count test

83. LIGHT OBSTRACTION PARTICLE COUNT
TEST
• Use a suitable apparatus based on the principle of light blockage which
allows an automatic determination of the size of particles and the number of
particles according to size.

84. Limits:
Sample Particle size in μm Maximum no. of
particles.
LVP ≥ 100 ml
10
25
Average in the units
tested
25 per ml
3 per ml
SVP – 100 ml and
less than 100 ml
10
25
6000 per container
600 per container

85. MICROSCOPIC PARTICLE COUNT
TEST
• Wet the inside of the filter holder fitted with the membrane filter with
several milliliter of particle-free water .
• Transfer the total volume of a solution pool or of a single unit to the
filtration funnel, and apply vacuum.
• Place the filter in a Petri dish and allow the filter to air-dry.
• After the filter has been dried, place the Petri dish on the stage of the
microscope, scan the entire membrane filter under the reflected light from
the illuminating device, and count the number of particles.

86. Limits :
Sample Particle size in μm Maximum no. of
particles.
LVP ≥ 100 ml 10
25
Average in the units
tested
12 per ml
2 per ml
SVP – 100 ml and
less than 100 ml
10
25
3000 per container
300 per container

87. 3. PYROGENS TEST
PYROGENS:
• Pyrogens are fever producing substances.
• Pyro means ‘pyrexia’, Gen means ‘producing’.
• Pyrogens are the by-products of microorganisms mainly of bacteria,
molds and viruses.
• During the processing these pyrogens may come from water, active
constituent or the excipient or from the equipments.
• Chemically these pyrogens are lipid substances associated with carrier
usually polysaccharides or may be proteins.

88. Types of Pyrogen test
For Detection and quantification of Pyrogens:
Basically there are 2 tests performed to detect the presence of pyrogens
in sterile parenteral products.
they are:
1) In Vivo pyrogen test (Rabbit Test)
2) In Vitro pyrogen test (Limulus Ameabocyte Lysate Test)

89. 1) In Vivo Pyrogen test (Rabbit Test)
PRINCIPLE:
This test consists of measuring the rise in body temperature evoked in
rabbits by the injection of a sterile solution of the substance being
examined.
Procedure:
The test involves measurement of the rise in body temperature
of rabbits following the IV injection of a sterile solution into ear vein
of rabbit.
• Dose not exceeding 10 ml per kg injected intravenously within a
period of not more than 10 min.

90. • Test animals: Use healthy, adult rabbits of either sex,
preferably of the same variety.
• Recording of temperature: Clinical thermometer
PRELIMINARY TEST (SHAM TEST)
If animals are used for the first time in a pyrogen test or have not been used
during the 2 previous weeks condition them 1 to 3 days before testing the
substance by injecting IV 10ml per kg pyrogen free saline solution warmed to
about 38.5°C.
• Record the temperature of the animals beginning at least 90 min before
injection and continuing for 3 hours after injection.
• Any animal showing a temperature variation of 0.6° or more must not be
used in main test.

91. MAIN TEST
• Carry out the test using a group of 3 rabbits.
Preparation of the sample: Dissolve the substance in or dilute with pyrogen
free saline solution . Warm the liquid to approximately 38.5° before injection.

92. PROCEDURE
• Inject the solution under examination slowly into the marginal veins of the
ear of each rabbit over a period not exceeding 4 min.
• Record the temperature of each animal at halfhourly intervals for 3 hours
after injection.
• The difference between the initial temperature and the maximum temperature
which is the highest temperature recorded for a rabbit is taken to be its
response.

93. INTERPRETATION OF RESULT
NUMBER
OF
RABBITS
MATERIAL PASSED IF
SUM OF RESPONSE
DOES NOT EXCEED.
MATERIAL FAILED IF
SUM OF RESPONSE
EXCEEDS.
3 RABBITS 1.15ºC 2.65ºC
6 RABBITS 2.80ºC 4.30ºC
9 RABBITS 4.45ºC 5.95ºC
12 RABBITS 6.10ºC 7.60ºC

94. In Vitro Pyrogen test
Limulus Ameabocyte Lysate(LAL Test)
Measures the concentration of bacterial endotoxin.
Test is using lysate derived from hemolymph cells or amoebocytes of
horse shoe crab.
Endotoxin limit calculated by
K/M
K- maximum no.of endotoxin which receive the patient without suffering
toxic reaction.
M- maximum dose administered to a patient/kg/hr

95. Mechanism of LAL Test

96. LAL Reagent
Bleeding adult crabs blood into an anticlotting solution
Washing and centrifuging to collect the amaebocytes.
Lysing in 3% NaCl.
Lysate is washed and lyophilized for Storage.
Procedure:
Test:
Equal volume of LAL reagent and test solution (usually 0.1 ml of each) are
mixed in a depyrogenated test-tube
Incubation at 37ºC, for 1 hour
Remove the tube – invert in one smooth motion (180º)
Observe the result

97. Different Techniques:
Three different techniques:
The gel-clot technique – gel formation
The turbidimetric technique – the development of turbidity after cleavage
of an endogenous substrate
The chromogenic technique – the development of color after cleavage of a
synthetic peptide – chromogen complex

98. Gel Clot Technique
A solid gel is formed in the presence of endotoxins
This technique requires positive and negative controls
Positive controls – a known concentration of endotoxin added to the lysate
solution.
Negative controls – water, free from endotoxins, added to the lysate solution.
Turbidimetric Technique
The test is based on the measurement of opacity change due to the
formation of insoluble coagulin.
• Opacity is directly proportional to the endotoxin concentration
• This technique is used for water systems and simple pharmaceutical
products.

99. Chromogenic Technique
• This is based on the measurement of color change which is caused by
the release of the chromogenic chemical
• p-nitroanilide
• The quantity of the p-nitroanilide produced is directly proportional to
the endotoxin concentration

100. TEST FOR STERILITY
Sterility is defines as freedom from the presence of viable
Microorganism.
METHOD A:
Membrane
filtration
METHOD B:
Direct
inoculation

101. Media to be used in the sterility
test
Two types of media are used.
Fluid
Thioglycolate
Medium
Soyabean casein
digest
Medium

102. MEMBRANE FILTRATION METHOD:-
• A membrane has a nominal pore size not greater than 0.45μ and
diameter of approximately 50mm.
• This method basically involves filtration of Sample through membrane
filters.
• The filtration is assisted under strict aseptic condition, after filtration
complete the membrane is cut into 2 halves and one halve is placed in
suitable volume of ( 100 ml usually) FTM, SCDM medium.
• Incubate the media for not less than 14 days.

103. DIRECT INOCULATION METHOD:-
The DI method is the more traditional sterility test method. Basically,
the DI method involves three steps:
1. Aseptically opening each sample container from a recently sterilized
batch of product.
2. Using a sterile syringe and needle to withdraw the required volume of
sample for both media from the container.
3. Injecting one-half of the required volume sample into a test tube
containing the required volume of FTM and the other half volume of
sample into a second test tube containing the required volume of SCD.

104. MINIMUM QUANTITY TO BE USED FOR EACH
MEDIUM
Minimum quantity used is..
Quantity per container Minimum quantity to be used for
each medium
Liquids
1. less than 1 ml The whole contents of each
container
2. 1-40 ml Half the contents of each
container but not less than 1 ml
3.Greater than 40 ml and not
greater than 100 ml
20 ml
4. Greater than 100 ml 10 per cent of the contents of the
container but not less than 20 ml
Antibiotic liquids 1 ml

105. INTERPRETATION OF RESULTS
• If the material being tested renders the medium turbid so that the presence or
absence of microbial growth cannot be easily determined by visual
inspection,14 days after the beginning of incubation , transfer portion (< 1 ml)
of the medium to fresh vessels of the same medium and then incubate original
and transfer vessel for not less than 4
days.
• If No evidence of microbial growth is found- complies with test for sterility.
• If evidence of microbial growth is found- does not complies with test for
sterility.