Parenteral products III

•Part- III
• Compiled By
• Vaibhav Tripathi Dr. Anil K. Sahu
Dr. Deepak K. Dash

• Quality Control of Parenteral Products
• Mandatory quality of injection
1. Sterile
2. Isotonic
3. Free from foreign particles
• These significant qualities are ensured by
employing a set of specific tests
• Such tests are performed throughout the
production operation.

• There are 3 phases for employing tests:
1. Raw material testing
2. Production material testing (intermediate
products) before filling
3. Finished product testing

1. Raw material testing
Test for purity
Conductivity
testSolid content
test
Stability testing
Pyrogen tests
Rabbit test
LAL test

2. Production material testing
Volume check
Strength
pH
Turbidity
Color
Pyrogen test
Sterility (direct inoculation &
indirect method )

3. Finished product testing
Sterility test
Pyrogen test
Clarity test
Leak test
Assay

1. Raw material testing:
I. Test for purity: purity of API is checked with
the aid of assay as per monograph
II. Conductivity test: Conductivity is measured
by conductometer.
• It measures the conductivity of vehicle used in
sterile preparation.

III. Pyrogen test:
• Fever producing, metabolic by-products of
microbial growth and death.
• Bacterial Pyrogens are called “Endotoxins”.
• Gram negative bacteria produce more potent
endotoxins than gram + bacteria and fungi.

III. Pyrogen test (Rabbit test):
• Test is conducted using rabbit as a test
animal.
• healthy adult rabbits of either sex, each
weighing 1.5 kg are selected
• Do not use any rabbit
• having a temp higher than 39.8 ℃

• The volume of injection is NLT 0.5ml/kg &
NMT 10ml/kg of body weight
• Temperature is recorded by inserting the
thermometer in rectum of rabbit to depth of
about 5 cm

• Inject the solution under examination slowly
into the marginal veins of the ear of each
rabbit over a period not exceeding 4 mins.
• Record the temperature of each animal at half
hourly 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.

No. of Rabbits
Individual Temperature rise
(°c)
Temperature rise
in groups (°c) Test
3 0.6 1.4 Pass
If above not
passes
3+5 =8 rabbits
0.6 3.7 pass

III. Pyrogen test: Limulus amebocyte lysate
(LAL)
• To detect or quantify endotoxins of gram-ve
bacterial origin
• Reagent: amoebocyte lysate enzyme from
horseshoe crab

(LAL)
• The test is based on the gelling properties of
enzyme extracted from the horseshoe crab of
Limulus polyphemus.
• enzymes when come in contact with solution
containing bacterial endotoxin
• Produces turbidity, precipitation or gelation of
the mixture.

(LAL)
• Degree of Gelling related to amount of
Endotoxin present.
• Procedure:
• equal Volume of LAL reagent and test solution
(usually 0.1 ml of each) are mixed in a de-
pyrogenated test-tube
• Incubation at 37°C, 1 hour remove the tube - invert
at (180°) observe the result

(LAL)
• Quantitative estimation is done with the help
of spectrophotometer.
• Advantages
• Fast - 60 minutes vs. 180 minutes
• Greater Sensitivity ,Less Variability
• Much Less False, Positives ,Much Less
Expensive

(LAL)
• Advantages
• Alternative to Animal Model, cheaper,
• Particularly useful for:
• Radiopharmaceuticals and cytotoxic agents
• Blood products
• Water for injection

2. Production material testing (before filling)
I. Volume check: final volume of the container
is checked to ensure the uniformity of final
volume Labelled Size Mobile Liquid (ml) Viscous Liquid
(ml)
0.5 ml 0.1 0.12
1 ml 0.1 0.15
2 ml 0.15 0.25
5 ml 0.3 0.5

2. Production material testing (before filling)
II. pH: it should be maintained as per the
standard parameter.
• Two different types of methods used in
measurement of pH.
A. Dip a piece of pH paper into the sample.
B. pH meter

3. Finished product testing:
I. Sterility test:
• It is a procedure carried out to detect and
• conform absence of any viable form of
microbes in pharmacopoeial preparation or
product.
• PRINCIPLE : If the microorganism are present
in the product can be indicated by a turbidity in
the clear medium.

I. Sterility test:
• Sterility test is destructive test thus it is
impossible to test every item for sterility.
• That’s why, a fixed percentage of the final
container are selected
• According to Indian Pharmacopoeia

I. Sterility test:
Number of items in the batch Minimum number of items to be tested
for each medium
Not more than 100 containers 10 per cent or 4 containers
More than 100 but not more than 500 10 containers
More than 500 containers . 2 per cent or 20 containers (10
containers for large-volume
parenterals) whichever is the less

I. Sterility test:
Media to be used in the
sterility test
Fluid Thioglycollate Medium
(FTM)
Soyabean-casein Digest
Medium
(SCDM)
Methods
Method - A
Membrane filtration method
Method- B
Direct inoculation method

I. Sterility test:
• Method – A (Membrane Filtration Test)
• 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 Vacuum
• after filtration completion the membrane is cut into 2
halves

I. Sterility test:
• Method – A (Membrane Filtration Test)
• one halve is placed in a test tube containing FTM
medium
• Second halve is dipped into another test tube
containing SCDM medium
• Incubate the media for not less than 14 days.
• All steps of this procedure are performed aseptically
in a Laminar Flow Hood

• Membrane is washed
Thrice with 100 ml sterile water
Prior to test.
• Fatty formulations can be
Diluted with isopropyl
myristate, if needed
Membrane filter
0.45μ porosity
Filter the test
solution
After filtration
remove the filter
Cut the filter
into two halves

First halves
(For Bacteria)
Transfer in 100 ml
culture media
(FTM)
Incubate at 30-35
℃ for not less
than 14 days
Observe the
growth in the
media
Second halves
(For Fungi)
Transfer in 100
ml culture media
(SCDM)
Incubate at 20-
25 ℃ for not less
than 14 days
Observe the
growth in the
medium

I. Sterility test:
• Method – B (Direct Inoculation Test)
• It involves a direct inoculation of required volume of
a sample in two tests tube
• containing a culture medium that is FTM, SCDM.
• Volume of the preparation under examination is not
more than 10% of the volume of the medium.
• Incubate the inoculated media for not less than 14
days.

I. Sterility test:
• INTERPRETATION OF RESULTS
• The following observations are possible:
1. No evidence of growth, the test is passed for
sterility.
2. There is evidence of growth(preserved)
• Re-testing is performed following the initial
procedure for not less than 4 days
• No evidence of growth the test is passed for sterility.
• There is evidence of growth the test is failed for
sterility

• CONTENT UNIFORMITY TEST
• 30 sterile units are selected from each batch.
• The weight of 10 individual sterile units is noted
• 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 form gross weight.

• The dose uniformity is met if the amount of active
ingredient is within the range of 85-115.0% of label
claim.
• 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 NMT 7.8%.

• Clarity Test
• Performed to ensure that the parenteral products are
free from foreign particles.
• Visual Method
• Coulter Counter Method
• Filtration Method

• Clarity Test
Particle Size (μm) Max. no. of particles per
ml
10 50
25 05
50 Nil

• Leak Test
• Leaker test for ampoules is intended to detect
incompletely sealed ampoules so that they can be
discarded in order to maintain sterile condition of the
medicines.
• Tip seals are more likely to be incompletely closed
than pull seals.
• Open capillaries or cracks at the point of seal result
in LEAKERS.

• Leak Test
• The leaker test is performed by immersing the
ampoules in a dye solution, such as 1% Methylene
blue
• and applying at least 25 inches of vacuum for a
minimum of 15 mins.
• Containers are removed & dye spotted area is
investigated.

• Quality control should be a fundamental segment of
parenteral products manufacturing.
• All tests are essential and have its own importance in
parenteral production.
• All of these tests ensure that product meet its quality
which has been judged to satisfactory also.
• Each test is unique and provides detailed
assessment of quality control for parenteral products.

1. Remington, The Science and Practice of Pharmacy,21st edition,Vol-I,p:
812-813,831-833.
2. Kenneth E.Avis,Herbert A.Lieberman and Leon Lachman,“
Pharmaceutical Dosage Forms: Parenteral Medication", Vol.III, 2nd
edition, p :40-41,58-62.
3. Sandeep Nema, John D. Ludwig (eds.)" Pharmaceutical Dosage Forms:
Parenteral Medication", Third Edition, Vol-2,p :122-128, 153-157.

Parenteral products III

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