EVALUATION OF PARENTERALS

FINISHED PRODUCT QUALITY CONTROL

TEST

There are mainly five quality control test for parenterals

are performed.
 Content Uniformity Test
 Leaker test
 Pyrogen test
 Sterility test
 Particulate test
CONTENT UNIFORMITY TEST
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 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.
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 NMT 7.8%.
LEAKER 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.
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 vaccum for a
minimum of 15 mins.

Detection of leaker is prominent when ampoules are

immersed in a bath of dye during autoclaving as this
has advantage of acomplishing both leaker detection
and sterilization in one operation.
Another means of testing for leakers is a high frequency
spark test system which detect presence of pinholes in
ampoules.
Bottles and vials are not subjected to such a vaccum test
because of the flexibility of the rubber closure.
PYROGEN TEST
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 mins.
Test animals: Use healthy, adult rabbits of either sex,
preferably of the same variety.
Recording of temperature:
Clinical thermometer, thermistor.
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°
Record the temperature of the animals,beginning at least
90 mins 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
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.
PROCEDURE
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.
INTERPRETATION OF RESULT

No. of Rabbits Individual Temperature rise Test

Temperature rise in groups (°c)
(°c)

3 Rabbits 0.6 1.4 Passes

If above not 0.6 3.7 Passes

passes
3+5 =8 rabbits
BACTERIAL ENDOTOXIN TEST
BET measures the concentration of bacterial endotoxin
that may be present in the sample using a lysate derived
from the amoebocytes of the horsehoe crab, Limulus
polyphemus.

The addition of a solution containing endotoxins to a

solution of a lysate produces turbidity,precipitation or
gelation of the mixture.
METHODS:

Method A-Gel clot limit test method.

Method B-Semi quantitative gel clot method.

Method C-Kinetic Turbidimetric method.

Method D-Kinetic Chromogenic Method

Method E- End point chromogenic method.

Gel clot limit test method

 Preparation of test solution-

Preparation of test solution by dissolving or diluting
active substances

Adjust the pH of test solution(about 6 to 8)

pH adjust with use of acid ,base ,buffer.

Prepare a sample solution at any dilution at or below
Maximum Valid Dilution.

Use water bacterial endotoxin as –ve control and 2

positive control.

One of the +ve control consist of the control standard

endotoxin at a conc. Of 2λ.

Other consist of test solution spiked with control

standard endotoxin to give a conc. Of 2λ (ppc)
Interpretation Of Result
The product under examination complies with the
bacterial endotoxin test if the +ve control is +ve and –
ve control as well as test solution are –ve.

The test is not valid if the +ve control is –ve or if the

–ve control is +ve.
KINETIC TURBIDIMETRIC METHOD-
A photometric assay measuring the increase in
turbidimetry caused by the reaction of the endotoxin
with the lysate.

KINETIC CHROMOGENIC METHOD-

A photometric assay measuring a colour developed
by the chromophore released from a chromogenic
substrate by the reaction of the endotoxin with the
lysate.
STERILITY TEST
Sterility is defines as freedom from the presence of
viable microorganism.
Sterility test is define as microbiological test applied
to sterile product to show are the product
manufactured and processed under specification
guided by cGMP.
Sterility test is destructive test thus,it is impossible to
test every item for sterility.
Media to be
used in the Methods:
sterility test

Method A-
Fluid
Membrane
Thioglycollate
filtration
Medium
method

Method B-
Soyabean-casein Direct
Digest Medium inoculation
method
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 Vacuum, after filtration
completion the membrane is cut into 2 halves and one
halve is placed in two test tubes containing FTM,
SCDM medium.
Incubate the media for not less than 14 days.
DIRECT INOCULATION METHOD

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.
Minimum quantity to be used for each medium
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 20 ml

greater than 100 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
Minimum number of items to be tested

Number of items in the batch Minimum number of items to be

tested for
each medium
Parenteral preparations
Not more than 100 containers 10 per cent or 4 containers whichever
is the greater
More than 100 but not more than 500. 10 containers
2 per cent or 20 containers (10
More than 500 containers . containers for large-volume
parenterals) whichever is the less
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.
PARTICULATE TEST
Particulate matter refers to the extraneous, mobile,
undissolved particles, other than gas bubbles,
unintentionally present in the solutions.

2 methods are used:

Method A-Light Obscuration Particle Count Test
Method B-Microscopic particle count test
LIGHT OBSCURATION 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.
Limits :

Sample Particle size in μm Maximum no. of

paricles.

LVP ≥ 100 ml Average in the units

tested
10 25 per ml
25 3 per ml

SVP – 100 ml and less 10 6000 per container

than 100 ml 25 600 per container
MICROSCOPIC PARTICLE COUNT TEST
Wet the inside of the filter holder fitted with the
membrane filter with several millilitre 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
Limits :

Sample Particle size in μm Maximum no. of

paricles.

LVP ≥ 100 ml Average in the units

tested
10 12 per ml
25 2 per ml
SVP – 100 ml and less 10 3000 per container
than 100 ml 25 300 per container
CONCLUSION

Quality control  should  be a fundamental segment 

of parenteral  products  manufacturing. 
 All  of  the  5 basic  tests  which  are performed 
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.