Environmental Monitoring

1. Particulate Matter
Particle Counter
2. Differential Pressure
Positive pressure differential of 10-15 Pascal should
be maintained between adjacent rooms of different
classification (with door closed).
Most critical area should have the highest pressure
3. Air Changes and Air flow patterns
Air flow over critical areas should be uni-directional

4. Temperature and Relative Humidity

Ambient Temperature and RH.

5. Microbiological Testing of Air

Two types of Microbiological Air testing Methods
A. Sedimentation method
B. Impaction method

Microbiological Testing of Air

1. Sedimentation method

The principle involved is Gravitational fallout in a

given time on a given area.

Irregularities in counts due to wild air currents,

physical movement of personnel etc.
2. Impaction Method
Different types of air samplers use this principle
The particles in air Impact onto the medium
plates by
the application of Electrical or
mechanical forces.
Slit Samplers
Sieve Samplers
Centrifugal Samplers

Microbiological Testing of Water

1.
2.
3.
4.

Membrane filtration technique

Standard Plate Count technique
Most Probable Number Test
Biochemical Oxygen Demand (BOD)
Test

1. Membrane Filter Technique

100 ml water sample is passed through
membrane filter. Filter pad is then
transferred to a bacteriological growth
medium
Bacteria trapped in the filter grow on the
medium and form colonies when incubated.
By counting the colonies, an estimate can
be made of the number of bacteria in the
original 100 ml sample.

Diagram for Membrane filtration

technique

2. Standard Plate Count Technique

Samples of water are diluted in jars containing
99-ml sterile water
Samples are placed in Petri dishes with
nutrient agar medium.
After incubation, the colony count is taken and
multiplied by the dilution factor to obtain
the total number of bacteria per ml of sample.

3. Most Probable Number (MPN) Test

Test tubes of lactose broth are inoculated with

water samples measuring 10 ml, 1 ml, and 0.1
ml.
During incubation, coliform organisms produce
gas
Depending upon which tubes from which
water samples display gas, an MPN table is
consulted and a statistical range of the
number of coliform bacteria is determined.
The MPN test is easy to perform but is not
exact as standard plate count method.

4. The Biochemical Oxygen Demand

(BOD)Test
The BOD is the amount of oxygen required by the
microorganisms during their growth.
The BOD test is begin by noting the oxygen
concentration in a sample of water before
incubation
The water is then incubated in an air-tight,
stoppered bottle for a period of about five days at
a temperature between 5 and 20C
The oxygen concentration in the water is then
noted again and the difference in the dissolved
oxygen is the BOD

Remember that
A higher BOD indicates presence of
a higher amount of organic matter

Evaluation of aseptic operations

Media Fill Test (MFT):
Most useful method for evaluating SAL of an aseptic
filling operation is Media Fill Test (MFT).
MFT is the exact simulation of an aseptic process except
that a microbiologic growth medium is substituted for
the active ingredients in a sterile product.
MFT is used to identify the potential weakness in an
aseptic processing operation that might contribute to
microbiologic contamination of the drug product.
Minimum of 3 MFTs should be performed for initial
validation of Aseptic Processing Operations.

Media and Incubation Conditions for MFT :

Ideal Medium for MFT
Soyabean Casein Digest Medium (SCD) also
known as Triptic Soya Broth (TSB).
Commercially available TSB is Tripticase
Fluid Thioglycollate Medium (FTM)
Incubation Conditions
14 days at ambient temperature (20 -25 C)

PARENTERAL FORMULATION
VEHICLES
A. Aqueous vehicles
i. Water for injection USP
ii. Water for injection free from CO2 (Barbiturates &
Sulfonamides)

iii. Sterile water for injection USP

iv. Bacteriostatic water for injection USP
v. Sodium chloride injection USP
vi. Bacteriostatic Sodium chloride injection USP
vii. Ringers injection USP
viii. Lactated Ringers injection USP

---

Non aqueous vehicles

i. Fixed oils
ii. Ethyl alcohol, propylene glycol, PEG
ADDED SUBSTANCES
i. Solubilising agents e.g. Surfactants
(polysorbate, tweens)
ii. Stabilizers e.g. Antioxidants (thiourea,
ascorbic acid, sodium bisulphite, sodium
metabisulphite)
iii. Buffering agents e.g. Acetates, citrates and
phosphates

--added substances-iv. Anti bacterial agents (e.g. Phenol or cresol,

chlorocresol, phenyl mercuric nitrate, bezalkonium
chloride)
v. Tonicity contributors (e.g. Sodium chloride,
Dextrose, Boric acid)
vi. Chelating agents e.g. EDTA
(Ethylenediaminetetraacetic acid)
vii. Suspending agents (e.g. Methyl cellulose, CMC,
gelatin, acacia)
viii. Emulsifying agents (e.g. Lecithin)
ix. Wetting agent (e.g. tween 80 , sorbitan trioleate )

CONTAINERS AND CLOSURES

1. Glass: Glass containers Type I, are best for
aqueous preparations (vials & ampoules).
Siliconization inside surface is done to prevent
interaction.
2. Plastics polypropylene containers can
withstand autoclaving.
3. Rubber closures permit needle into multipledose vial.
It is held by aluminum band.

PROCESSING OF PARENTERALS

STERILIZATION METHODS
Evaluation of Sterilization Method
Sterile
products
possess
several
unique
properties, such as freedom from microorganism,
pyrogens, particulates and high standards of
purity and quality. This ultimate goal in the
manufacture of sterile products can be attained
by evaluation of sterilization procedure. The
Sterilization processes are likely to be subjected
to the most detailed and complex validation
procedures. Evaluation of processing includes
equipments, process, personnel, material etc.

The principle involve in the evaluation of sterilization

process are:

i. To build sterility into product.

ii. Perform a maximum level of probability.
iii. Establish specification and performance characteristic.
iv. To provide greater assurance of support of the result.
v. Specific methodology, process and equipment.
vi. Final product testing using validated analytical method
&
vii. Verification, calibration and maintenance of
equipments used in the processes.