C.

Freeze-dried or Lyophilized Parenteral Products

Parenteral solutions intended to be freeze-dried must be aqueous because
the drying process involves the removal of water from the product, by sublimation
after it is frozen. This process is only applicable to thermolabile or unstable drugs in
liquid.
Characteristics of Freeze-dried/lyophilized products
1. Of uniform color and texture
2. Contains a supporting matrix of solids, sufficient to maintain the original
volume of the product after drying
3. With sufficient strength to prevent crumbling during storage
4. Rapid reconstitution
D. Parenteral Suspensions
The principal problem in parenteral emulsions is the attainment of
maintenance of uniform size of droplets ranging from 1 to 5 microns (for the internal
phase). Intravenous nutrient emulsions have been made containing cotton seed oil
dextrose and lecithin.
F. Effect of Route of Administration
The intended route of administration has a marked effect on the formulation
of a parenteral product. This factors to be considered are:
1. Volume in which the dose of a drug is given
For intracutaneous route, not more than 0.2 ml is given due to slow
absorption. For subcutaneous route, about 1ml or less. For intraspinal, 10ml or less.
For intravenous large volume parenterals may be given by setting the infusion set
to 250 and above by syringe about 20 ml or less.
2. Isotonicity
It is important for the comfort of the patient, especially for intraspinal
injection where the CSF fluid flow is slow and disturbances of osmotic pressure will
cause headache and vomiting.
3. Stability
This factor is considered by the Product development division and should
evaluate the effect of the components particularly if the product is subjected to
thermal sterilization.
Second Division: Production Area
Production Area includes all the steps in:
1. Accumulation of raw materials and packaging materials
2. Preparation of the product
3. Filling and finishing of the final product

The process, equipment containers closures and other components should be

sterilized after cleaning and prior to use
Environmental Control
Excellence in environmental control could be achieved by:
1. Careful designing of the floor plan such that traffic in and out of the aseptic
areas and sterile areas are minimized.
2. Access by personnel to the aseptic corridors compounding area and filling
rooms is only through air lock area.
3. Personnel are allowed to enter the aseptic area and sterile rooms only after
the following procedures
a. Removal of street clothing
b. Washing of hands and taking a shower
c. Wearing the prescribed aseptic uniform (complete with gown, shoes,
masks/ face hoods, head coverings gloves etc.)
4. Once inside the aseptic or sterile area, they are not permitted to move in and
out the area.
Maintenance of the aseptic or sterile room is done by normal routine of
cleaning of these area at the end of the working day or during the
night. This includes all surfaces of the ceilings walls, floors counters
and equipment. After cleaning and scrubbing surface disinfection is
done.
Laminar flow Enclosures
The development of laminar flow enclosures is a marked improvement in the
environmental control of aseptic and sterile areas. This equipment has been
developed to allow for the draft free flow of clean, filtered air over the work area.
These hoods are commonly found in hospitals and parenteral production
areas for both manufacture and the incorporation of additives into parenteral and
ophthalmic products.
Furthermore, this hood provides a total sweep of the confined area, since the
entire body of air moves with uniform velocity along line originating through the
HEPA filter (High Efficiency Particulate Air) occupying an entire side of the confined
area
Ultraviolet Radiation
This type of radiation control microorganisms in the air by installing
ultraviolate lamps in the cleanup area, aseptic area and preparation room.
However, ultraviolet rays are irritating to the skin and eyes of human beings.
The personnel in the area must be protected from direct exposure by installing
deflectors that directs the UV rays above the head. Further protection is made by
providing clothing to cover the skin and UV absorbing goggles to protect the eyes.
Methods of Environmental Control Tests

1. Air Sampling Techniques

a. Collection of particulate matter from air by drawing air sample through a
clean sterile membrane filter. A new membrane filter should be used at each
location of the area.
The filters are examined microscopically for particulate matters as lint and
dust. The filters may also be placed on the culture media and incubated for the
detection of microorganisms.
b. Collection of air sample into a measured volume of nutrient broth in an
impinger. Microorganisms in the broth may be collected by filtration on the
membrane filter and incubated.
c. Drawing a measured volume of air through a salt sampler which causes the
air to impinge on the surface of a slowly rotating agar plate. This sampler is portable
and can be disinfected and hand-carried whenever needed.
2. Exposure of nutrient agar plates to the settling of micro-organisms from the air.
If pathogenic microorganisms are particularly of interest, blood agar plates
may be needed. The exposure period may vary as deemed needful for given
circumstances, but even a period of one hour may not collect one microorganism
under conditions of use in a well-controlled aseptic area.
3. Total Sterility Test the best indication of the efficiency of the aseptic filling
process
Thi is done by filling and sealing sterile fluid thioglycollate or trypticase soy
broth in sterile containers under the same conditions used for an aseptic film of a
product.
The entire lot is incubated and examined for the appearance of growth of
microorganisms. Such growth is indicative of contamination from the environement
and equipment. It also maybe used to measure the efficiency of a particular
operator.
4. Use of Instrumental scanners
There are instruments correctly being utilized to obtain particle counts from a
measured volume of air as a means of indicating the level of particle contamination
in the environment.
These instruments operate on the principle of the measurement of light
scattered from particles passed through the optical system.
Filtration of Parenteral Solutions
Solutions must be filtered for the following primary objectives:
1. Clarification is also termed as polishing and a highly polished solution would
require the removal of particulate matter down to at least three microns in size.

2. Sterilization by Filtration is the removal of approximately 0.2 to 0.45 microns,

the size of viable microorganisms and spores; sad are used for pharmaceutical
solutions that are heat labile.
The five (5) filters used for sterilization with their comparative characteristics are:
Principal
Components

Filter Type

Constituents
Liberated
Alkaline
substances, heavy
metals and fibers

Effects on
Solutions
Absorb large
charged ions
Raises pH.

1. Asbestos pad
(disposable)

Asbestos fibers or
wood cellulose

2. Cellulose ester
membrane

Cellulose acetate
and/or cellulose
nitrate

None

None

3. Diatonaceous
earth candles

Diatoms of S1O2,
Fe2O2 and others
(about 80%)

Alkaline
substance, heavy
metals fractured
atoms

Absorb lage
charged ions.
Raises pH

Borosilicate glass

Glass beads

Negligible

Clay, silicon
dioxide

Clay particles

Negligible

4. Sintered glass
candles/discs
5. Unglazed
porcelain/
seals candles

Cellulose ester membrane is officially recognized in the USP and BFAD since it
provides the best flow rate although problems of surface clogging is also present,
but remedied by a porous pre-liter.
Membrane filters retain maicroorganisms on the surface of the filter and not
distorted by pressure, it requires no pre-treatment and may be autoclayed or gassterilized.
Filling and Packaging of Parenterals
In filling of parenteral products into individual containers the following points
are considered:
1. Filling of Parenteral Solutions
a. In filling solutions, a means is provided for repetitively foreing a
measured volume of liquid through the orifice of the delivery tube,
which is introduced into the container.
The size of the delivery tube will vary from that of about 20 gauge
hypodermic needle to a tube at 0.5 in or more in diameter. The size of
a delivery tube that enters the opening of the container is determined
by.
i. Physical characteristics of the liquid (such as viscosity and
density)

ii. Speed of delivery desired

iii. Inside diameter of the neck of the container
The tube must freely enter the neck of the container and deliver the
liquid deep enough to permit air to escape the tube should have the
maximum possible diameter to reduce delivery force which causes
splashing.
b. In filling ampules, withdrawal of the delivery tube should not allow the
hanging drop from the tip to touch the neck of the ampule, by using a
filling machine with retraction device.
c. For large volume parenterals like infusion, solutions filling may be
made by gravity a pressure a pump filling machine or vacuum filling
machine. Gravity filling is slow pressure pump filling is the best method
since, it is faster and can be rendered automatic.
A vacuum is produces in the bottle when a nozzle gasket makes a scale
against the lip of the container. The liquid is drawn by the vacuum from
the reservoir through the delivery tube and the vacuum is released at
the adjusted capacity is reached.
2. Filling of Sterile Solids such as antibiotics are more difficult to subdivide
evenly into containers than liquids because the rate of flow is slow and
irregular.
One type of machine used for filling free-flowing solids into ampules and vial
employs an auger in the stem of the funnel at the bottom of the hopper. By the
controlling the size of the auger and its rotation a regulated volume of granular
material can be delivered from the funnel stem into the ampule or vial.
Sealing of Ampule, Vials and Cartridges
Parenteral containers should be sealed in the aseptic immediately to prevent
the contents from contamination. Tamperproof or filterproof seals are used.
A. Types of ampule sealing methods
A.1 tip sealing method
Ampules are sealed by melting enough glass ath the tip of the neck of
the ampule to form a bead and close the opening. Such seals are made
rapidly in a high temperature gas oxygen flame.
To produce a uniform bead the ampule neck must be heated evenly
on all sides. This is accomplished by means of stationary burners on opposite
sides of stationary ampules.

However, excessive heating results in the expansion of gases within

the ampule against a soft bead seal causing a bubble to form and eventually
burst.
A.2 Pull sealing method
Ampules are sealed by heating the neck of the ampule, below the tip
leaving enough of the tip for grasping with forceps or other mechanical
device. The ampule is rotated in the flame of a single burner.
When the glass has softened the tip is grasped firmly and pulled
quickly away from the body of the ampule, which continues to rotate.
Pull sealing is slower, but the seals are much surer than tip sealing.
Ampules containing powders and ampules with a wider orifice must be
sealed by pull sealing. An improperly of insufficiently sealed ampule is called
a leaker.

B. Sealing of Vials and Bottles

Vials and bottle are sealed by closing the opening with a rubber closure or
stopper. The closure must fit the mouth of the container snugly enough so that its
elasticity will permit adjustment to slight irregularities in the lip and neck of the
container.
When rubber closures are to be inserted mechanically, the surface of the
closure is often halogenated or siliconized to give it less friction. Thus, it is
possible to convey the closures through a chute to the place where it is positioned
over a vial and then inserted by a plunger or other pressurized device.
Mechanical stoppering has been developed to meet the need for high speed
production
Rubber closures are held in place by means of aluminum caps. The aluminum
cap cover the rubber closure and is crimped in place under the lip of the vial or
bottle.
Such confirmation is necessary to assure the integrity of the contents as to
sterility and other aspects of product quality.
Sterilization of Parenteral Products
The term sterilization as applied to pharmaceutical preparations mean the
complete destruction of all living organisms and their spores or their complete
removal from the preparation.
Sterilization of parenterals may be accomplished by either physical or
chemical processes.

I. Thermal Sterilization Methods

a. Dry Heat Sterilization
This is applicable for substance unaffected at a temperature of 148 to 260
degrees in the oven, at an exposure time of 45 minutes. This method kills spores as
well as vegetative forms of microorganisms. This method is ideal for sterilizing
glassware, metal wares and anhydrous oils.
The principle of sterilization involved is the oxidation of microorganisms by
heat.
b. Moist Heat Sterilization
This is more effective than dry heat method. It destroys Spores and
vegetative forms of bacteria at 121 degrees Celsius for 20 minutes at 15 PSI in an
autoclave. This method is ideal for sterilizing rubber stoppers, glassware, bottles
(ampules/vials), uniform and cellulose membrane filters.
The principle of sterilization is the coagulation of the cell protein of the
microorganism.
c. Fractional Sterilization Method
c.1 Tyndallization makes use of moist heat at 100 degrees Celsius using free
flowing steam. It is normally performed by 2 to 3 exposured, alternated with
intervals at room temperature or incubator temperature.
c.2 Inspissation a traditional method of sterilization at 60 degrees Celsius in an
oven alternated with intervals at room temperature or incubation for 2 to 3 days.
Fractional methods of sterilization are effective for vegetative forms of
microorganisms but not for spores. Bacteriostatic agents may be used to improve
this method.
II. Non-thermal Sterilization Methods
a. Ultraviolet Irradiation
This is used to aid reduction of air borne contamination produced by mercury
vapor lamps. This method has poor penetration capability. Its effectiveness
depends on
1. Length of time exposure
2. Intensity of radiation
3. Susceptibility of the microorganism
b. Ionization Radiation
This radiation method makes use of high energy emitted from radioactive
isotopes such as cobalt CO (gamma rays) or by cathode or beta rays
(mechanical acceleration of electrons to high velocity and energy)

Gamma rays are more reliable because there is no mechanical breakdown,

but it has a disadvantage of rare source and cannot be shut off immediately.
Accelerated electrons provide higher and more uniform dose outfit and can
destroy organisms by stopping its reproduction.
c. Filtration by Membrane Filtration as previously discussed
II. Chemical Sterilization Methods
a. Gas Sterilization
A method of sterilization using:
1. Formaldehyde and sulfur dioxide gas has a problem of leaving residual
deposits which is difficult to remove.
2. Ethylene oxide and basic propiolactone limited to dry powders and plastic
materials.
This is done by autoclaving the substanceat 55 Celsius at 27 PSI the ethylene
oxide gas is introduced, and exposure is timed at 60 minutes.
b. Surface Disinfection
A method of sterilization of work surfaces with the use of chemical
disinfectants for environmental control. Phenolic and quarternary ammonium
compounds are commonly used.
III. Quality Control of Parenteral Products
1. Sterility Tests
Alll lots of parenterals on their final containers must be tested for sterility. The
USP prescribes the requirements for this test for official injections.
1.1 One official method recommended by the BFAD is the Membrane Filtration
technique. The parenteral product is filtered, retaining organisms if any, on the
filter, washed and transferred in a culture media.
1.2 Another official method is Test Tube Inculation Method wherein turbidity is the
measure of the growth of the microorganisms.
2. Pyrogen Tests
There are two (2) wyas of carrying out this test, namely:
2.1 Qualitative fever Response Test in Rabbits rabbits are used as test animals,
since they show physiologic response similar to man.
2.2 Limulus Test an invitro test based on the golling or color development of a
pyrogenic preparation in the presence of lysate on the amebocytes of the horseshoe
crab (limulus polyphemus)
This is a simple, more rapid and of greater sensitivity than the rabbit test
although it detects only gram-negative bacteria.

3. Clarity Test
The USP does not provide specifications for a clarity test. It contains only the
statement that the CGMP requires that each final container of an injection should be
subjected individually to a visual inspection.
The objective of Clarity inspection is to prevent the distribution and use of
parenterals which contain particulate matter, which may be physiologically or
actually harmful to the recipient. Solutions to be introduced intravenously require
the most critical evaluation.
4. Leaker Test
Ampules that have been sealed by fusion must be subjected to a test to
determine whether or not a passageway remains to the outside. If such passageway
remains, all or part of the contents of the ampule may leak to the outside and spoil
the package.
Vials or bottles are not subjected to this test because the sealing material
(rubber stoppers) are not rigid. Therefore, results from such a test will be
meaningless.
5. Safety Test
Safety testing in animals are required for most biological products, since it is
entirely possible for a parenteral drug to pass the routine sterility test, pyrogen test
and chemical analyses, and still cause unfavorable reactions when injected.
A safety test in animals is essential to provide additional assurance that the product
does not have unexpected toxic properties.
IV. Packaging, Labeling and Storage
It is essential that the packaging of parenterals should provide ample protection
against physical damage from shipping handling and storage; and should protect
light sensitive materials from ultraviolet radiation.
Considerations in Packaging Parenterals
1. Parenterals intended for intraspinal, intracisternal or preidural administration
are packaged only in single-dose containers.
2. Unless an individual monogram specifies otherwise, no multiple dose
container shall contain a volume of injection more than sufficient to permit
the withdrawal and administration of 30 ml.
3. Injections packaged for use as irrigation solutions or dialysis solutions may be
packaged with a fill volume in excess of 1 liter.
4. Injections intended for veterinary use are exempt from the packaging and
storage requirements concerning the limitation of single-dose containers and
to the volume of multiple-dose containers.
Considerations in Labeling of Parenterals

1. The container label is so arrange that a sufficient area of container remains

uncovered for its full length or circumference, to permit inspection of the
contents
2. Preparations labeled for use as dialysis, hemofiltration or irrigation solutions
must meet the requirements for injections other than those relating to
volume must also bear on the label statements that they are not intended for
intravenous infection.