QUALITY CONTROL TESTS OF PARENTERALS

1. QUALITY
CONTROL
TESTING OF
PARENTRALS
PRESENTEDBY GROUP D

2. GROUP
MEMBERS
• Swaira Shakeel
• Khadija Smee
• Fatima Ihsan
• Atifa Shafqat
• Zumar Muhammad Imran
• Aqeela
• Muhammad Bin Naseem
• Ali Ahmed

3. CLARITY TEST

4. 1. CLARITY TEST
• It is used to determine the presence of particulate matter within the parenteral.
The particulate matter includes dust, lint, carbon, rubber fragments; glass particles and fibers
which may result in the formation of pathological granulomas on injection into the body.
• The clarity test in its subjective sense is meant to spot any particulate matter in the ampule or
otherwise, with the naked eye.
• This evaluation is affected by the visual acuity of the observer, the degree of critical thought
preparatory to the inspection, the meaningful perception of what is seen, and the physical
alertness.
• The possibility of contamination exists at every step of the process.

5. METHODS OF CLARITY TESTING
For checking clarity of parenterals, there are three methods;
1. Visual method
2. Automated techniques

6. i. VISUAL METHOD
Particles 50 microns or larger can be detected by visual inspection under suitable conditions of
visibility.
PROCEDURE:
• In this method each injectable is inspected visually against black and white background.
• White background aids in detection of dark color particles and lighter or reflective particles
will appear against dark background.
• A magnifying lens at magnification power of 2.5X set at eye level will facilitate the inspection.
ACCEPTANCE CRITERIA:
• There should be no particulate matter in the parenterals.

7. ii. AUTOMATED TECHNIQUES/ LIMIT TESTS
a. Electron particle counter (EPC)
It is based on these principles
• Change in electrical resistance
• Light blockage principle
• Light scattering principle
Change in electrical resistance involve coulter-counter method, that is used to measure the particle
size against the electrical resistance.
When particles are passed through orifice along with current, electrical resistance is created and
deflection is appeared and so particulate matter is detected in this way.
 Particle detection limit by this method is 0.1-1000 micrometers.

8. PYROGEN TEST

9. 2. PYROGEN TEST
WHAT IS A PYROGEN?
• Term pyrogen is reffered to as gram negative bacterial endotoxins that cause fever when
administered through IV/inhalation route.
• Microbial substance that act as pyrogen includes bacteria, plasmodia, fungi, streptococcal
species.
• According to USP the endotoxin amount that is needed to induce pyrogenicity in human and
rabbit is 1mg/kg as per body weight.
• Naturally occurring endotoxins are protein, carbohydrate and lipids in nature that are present in
the outer membrane of gram negative bacteria.

10. METHODS FOR PYROGEN TESTING
For pyrogen testing these two methods are used
Rabbit test (in vivo)
Limulus amebocyte lysate LAL test (in vitro)

11. i. RABBIT TEST
PRINCIPLE
• It involves measurement in the rise of body temperature after administration of test solution in
sterile form through IV route. Rise in temperature indicates the presence of endotoxins
pyrogen in the test solution.
EQUIPMENT REQUIRED
• Test animal (rabbit), Glassware, 20 30 G syringe, thermometer, rabbit holding box, cotton plug
PRECAUTION
• Thermometer should be thoroughly washed with water for injection and heated in hot air oven
at 25 degrees for not less than 30 minutes

12. TEST PROCEDURE
Usually conducted on group of three rabbits
a. Sample preparation
• Warm the solution at 37 2. While in case of lyophillized products test product is dissolved in normal
saline solution (0.9% of NaCl)
b. Determination of initial temperature of rabbit before injection
• Insert the sterlized thermometer in the rectum of rabbit at depth of 5cm and readings will be noted before
injection. Two such readings are taken to take the mean temperature.
c. Temperature recording
• USP states that always use an accurate temperature measuring device with an accuracy of 0.1 degrees eg
40.6 degree

13. TEST PROCEDURE
d. Determination of response from rabbit
• Test solution is inserted into marginal ear vein at a volume of 0.5-1 ml/kg. Record the temperature after every 30 minutes for 3
hours. Difference between initial and final temperature is taken as response.
• Try to avoid taking rabbit having body temperature beyond 38.1-39.8 degree
e. Interpretation of results
• If there is no change in the temperature of single rabbit of 0.5 degrees or more then the product/test solution meets the
requirements that it is pyrogen free.
• If single rabbit shows the rise in temperature up to 0.5 degrees then repeat test in extra 5 rabbits.
• If not more than 3 out of 8 rabbits show individual rise in temperature and sum of all 8 values doesn't exceed 3.3 degrees then
product is declared pyrogen free.

14. LIMITATION OF PYROGEN RABBIT TEST
• There is temperature variability in living animals like no two rabbits offer same body
temperature or respond identically to pyrogen test.
• Rabbits are sensitive and vulnerable to environment

15. ii. LIMULUS AMEBOCYTE LYSATE
(LAL) TEST