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Quality control and evaluation of parenteral

Three general areas of quality control are:
Raw material / incoming stock control
Manufacturing / in-process control
Finished product control

Incoming stock control:

routine tests on all ingredients
pyrogen tests on WFI
glass tests on glass containers
tests on rubber closures
Microbial load (bioburden) tests etc.

Manufacturing / in-process control:

Conductivity measurements during the distillation of
Confirmation of volume of fill in product containers
Monitoring of time and temperature for thermal
sterilization of the product etc.

Finished product control:

Final assays and other tests like pH, tonicity,
preservative concentration
Sterility test
Clarity test/ Particulate matter
Leakage test
Pyrogen test

1) Final assays and other tests like pH, tonicity,

preservative concentration: These tests are done
as per product requirement and specification/s.
2) Sterility tests:- The methods which are used to
perform sterility tests are
a) Direct transfer method.
b) Membrane filtration method.

A) Direct Transfer method:

it is a traditional sterility test method which involves
a direct inoculation of required volume of a sample in
two test tubes containing a culture medium [fluid
thioglycollate media (FTM), Soya-bean casein digest

This method basically involves filtration of sample
through membrane filters of porosity 0.45 micron and
diameter about 50 mm. The filtration is assisted under
vacuum or pressure difference.
After filtration, the membrane is washed with a sterile
diluting fluid to remove traces of bacteriostatic agents
etc. and aseptically removed.
The membrane is cut into 2 halves One-half of the
membrane is placed in a suitable volume (usually 100
ml) of FTM, and the other membrane half in SCDM.

All test containers should be incubated at temperatures
specified by the pharmacopoeia for at least 14 days,
regardless of whether filtration or direct inoculation test
methodology is used.
*Interpretation: If no visible evidence of microbial
growth in culture medium in test tubes is found, then it
is interpreted that the sample representing the lot is
Appropriate positive and negative controls must be
included in the sterility testing experiments

The number of containers tested per batch and
quantity tested from each container should be in
accordance with the pharmacopoeial specification.

3) Particulate matter testing:- Particulate matter is

of primary concern in the parenteral products given
by I.V. Route. All parenteral products should be free
from insoluble particle.
Biological risk associated with particulate matter:
-Inflammatory response
-Antigenic response
-Occlusion of blood vessels
Further U.S.P. states that GMP Requires that all
containers be visually inspected and that with visible
particle be discarded.

Methods for monitoring particulate matter

Visual method
Coulter counter method
Filtration method
Light blockage method

4) Leaker Test: - The leaker test is intended to detect

incompletely sealed ampoules, so that they may be discarded.
This test is usually performed by producing a negative pressure
within an incompletely sealed ampoule, while the ampoule is
submerged entirely in a deeply colored dye solution.
Most often, approximately 1% methylene blue solution is
employed. After carefully rinsing the dye solution from the
outside, color from the dye will be visible within a leaker.
Vials and bottles are not subjected to such a leaker test, because
the sealing material (rubber stopper) is not rigid.

5. Pyrogen Test: - pyrogens are lipopolysacchrides chemically

and heat stable and are capable of passing through bacteria
retentive filter.

When these pyrogens are introduced into a body they produce a

mark response of fever with body ache and vasoconstriction
within an onset of 1 hour.

The USP evaluates the presence of pyrogens in parenteral

preparations by a qualitative fever response test in rabbits, the
Pyrogen Test, and by the Bacterial Endotoxins Test LAL Test.


Why Rabbit?
Reproducible pyrogenic response
Other species not predictable
Similar threshold pyrogenic response to humans

For the test:

Rabbits must be healthy and mature
New Zealand or Belgian Whites used
Either sex may be used
Must be individually housed between 20 and 23C

Preliminary test (Sham Test)

Intravenous injection of sterile pyrogen-free saline solution
To exclude any animal showing an unusual response to the
trauma (shock) of injection
Any animal showing a temperature variation greater than 0.5
C is not used in the main test
All glassware, syringes and needles must be pyrogen free.

Main test:
Group of 3 rabbits
Preparation and injection of the product:
-Warming the product to 372C
-Dissolving or dilution
-Injection site: ear vein
-The injected volume: about 10 ml per kg of body weight
over 10 min. duration
-Record temperature at 30-min intervals for 3 hours.
-Site of temperature measurement- rectal
Instrument-thermometers/ thermocouples

Interpretation of the results:

The sample may be judged nonpyrogenic if no single rabbit
shows a rise in temperature of 0.5C or greater above its control
If this condition is not met, the test must proceed to a second
stage. In the second stage, five additional rabbits are given a new
preparation of the same test sample as the original three rabbits.
The solution may be judged non-pyrogenic if not more than
three of the eight rabbits show individual temperature rises of
0.5C or more.

High variability in response.
Difficulty in controlling all factors.
Antipyretic drugs such as aspirin, acetaminophen and
morphine mask pyrogenic effect (i.e.,misleading
Some other drugs have their inherent pyrogenic effect.

LAL test:The test was developed in 1960s by Drs. Bang and Levin and
is based on clotting rection of horseshoe crab blood to
Limulus- genera of crab
Amebocytes- crab blood cell from which active component is
Lysate- component is obtained by separating amebocytes from
the plasma and lysing them.


The basic procedure is the combination of 0.1 ml of test

sample with 0.1 ml LAL Reagent.
After incubation for 1 hr at 370C, the mixture is analyzed for
the presence of Gel clot.
The LAL test is positive, indicating the presence of endotoxin,
if the gel clot maintains its integrity after slow inversion of the
test tube containing the mixture.
This method has several advantages of Rabbit test like Greater
sensitivity and
reliability specificity, less variation, wider application, less
expensive and simplicity.

Gel/ clot reaction

Preparations for IV Fluids:

LVPs which are administered by IV route are commonly

called as IV fluids.
Purpose: Body fluids and electrolyte replenisher
Volume supplied: 100 to 1000 ml
According to their basic use, LVPS can be classified into:
1-Basic nutrition.
2-Restoration of electrolyte imbalance.
3-Bodys fluid replacement.
4-Blood and blood products.
5-Drug carriers.
6-Parenteral nutrition.
7-Special (miscellaneous) use.

Examples of LVPs:
Dextroseinjection: Available in 2 , 5 , 10 , 25 & 50 % w/v solution.
Used for: Fluids replenisher, Electrolyte replenisher
Contains 0.11 to 0.9 % Sodium chloride and 2.5 to 5.0 % Dextrose
Used for : Fluids replenisher, Electrolyte replenisher, Nutrient
Contains 0.9 % conc., Also known as normal saline solution
Used as : Isotonic vehicle , Fluids replenisher, Electrolyte replenisher
Contains 1.75 to 1.95 % w/v of sodium lactate
Used as : Fluids replenisher, Electrolyte replenisher

Contains 5, 10 , 15, 20 % of mannitol
Used as : Diagnostic aid, Renal function determination , as a diuretic
Contains 5, 10 , 15, 20 % of mannitol & 0.45 % of Sodium chloride
Used as :As a diuretic
Is a sterile solution of sodium chloride, potassium chloride and calcium
chloride in water for injection (WFI).
It is isotonic with physiological fluids
Used as vehicle for other drugs or alone as electrolyte replenisher and plasma
volume expander.
Lactated Ringer's solution is abbreviated as "LR" or "RL". It is also known as
Ringer's lactate solution

Total Parenteral Nutrition (TPN)

This is a complete form of nutrition, containing
protein, sugar, fat and added vitamins and minerals as
needed for each individual.
Total Parenteral Nutrition (TPN) may be defined as
provision of nutrition for metabolic requirements and
growth through the parenteral route.

Components of TPN solutions:

(1) Protein as crystalline amino acids.
(2) Fats as lipids.
(3) Carbohydrate as glucose.
(4) ElectrolytesSodium, potassium, chloride,
calcium and magnesium.
(5) Metals/Trace elementsZinc, copper,
manganese, chromium, selenium.
(6) Vitamins A, C, D, E, K, thiamine, riboflavin,
niacin, pantothenic acid, pyridoxine, biotin,
choline and folic acid.

TPN might be necessary if:

A patient is severely undernourished, and needs to have
surgery, radiotherapy or chemotherapy;
A patient suffers from chronic diarrhea and vomiting;
A baby's gut is too immature;
A patient's (their "gastrointestinal tract") is paralysed, for
example after major surgery.

The solution is administered through superior venacava which is

accessed by the subclavian vein near the heart.
The preferred method of delivering TPN is with a medical
infusion pump.
A sterile bag of nutrient solution, between 500 ml and 4 L is
provided. The pump infuses a small amount (0.1 to 10 mL/hr)
continuously in order to keep the vein open.

They are topical solutions used to flush (clean)
open wounds or body cavities.
Although they often labeled using the same terms
for injections, they are never given parenterally.
Container size is usually larger than 1 liter.

Dialysis is the process in which substances are
separated from one another due to their
differences in diffusibility (distribution) through
the membrane.
The fluids used in dialysis are known as
dialysis fluids.

General uses
Renal failure: to remove waste products and maintain
Transplantation of kidney:
Poisoning cases
Dialysis is a life saving procedure is done either by
haemodialysis or intraperitoneal dialysis.

Haemodialysis is used to remove toxins from

In haemodialysis, the blood from artery is
passed through artificial dialysis membrane,
bathed in dialysis fluid.
The dialysis membrane is permeable to urea,
electrolytes & dextrose but not to plasma
proteins & lipids. So excess of urea is passed
out from blood through dialysis fluid.
After dialysis blood is returned back to the body
circulation through vein.

A kidney unit may require more than 1200 litres of solution /

week. So, haemodialysis fluid is prepared in concentrated
form then it is diluted with deionised water or dist. water
before use.
Composition of Concentrated Haemodialysis Fluid BPC.
Dilute 1 liter of conc. solution with 39 liters of water to
make 40 litres.
Storage: store in warm place as it is liable to convert into
crystals on storage.


Intraperitonial dialysis is used to remove the toxic substances

excreted by the kidney.

Definition: When two or more sterile products are added to
an IV fluid for their administration, the resulting
combination is known as IV admixture.
In hospitals, prepared by nurses by combining or mixing
drugs to the transfusion fluids.
The drugs may be admixed in one syringe or are
incorporated into bottles of LV transfusion fluids.
Must be prepared under ASEPTIC conditions.