Sterile preparations, parenterals, Zones by gazette of India, Aseptic techniques

1. MANUFACTURING OF
STERILE PREPARATION
Prepared by: SHRUTI TYAGI (LECTURER IN PHARMACY)

2. INTRODUCTION
Parenteral preparations are sterile, pyrogen-free liquids (solutions,
emulsions, or suspensions) or solid dosage forms containing one or
more active ingredients, packaged in either single-dose or multi dose
containers. They are intended for administration by injection, infusion,
or implantation into the body.
The dosage form for conveying a drug by means of injection through
the skin or mucous membranes. Parenteral drugs are administered
directly into the veins, muscles or under the skin or more specialized
tissues such as the spinal cord. Circumvented the highly efficient first
line body defense that is skin and mucus membrane. Thus they should
be free from microbial contamination and should have high purity
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3. TYPES
• There are four main forms of parenteral preparations:
• Injections,
• Intravenous infusions (large volume parenterals),
• Powders for injections, and
• Implants.
Certain injections and intravenous infusions may be presented in the form of sterile
concentrated solutions, which must be suitably diluted before use.
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4. FACILITIES REQUIRED FOR PARENTERAL PRODUCTION
Parenteral preparations may contain excipients such as solvents, suspending agents, buffering agents,
substances to make the preparation isotonic with blood, stabilizers, or antimicrobial preservatives.
• The addition of excipients should be kept to a minimum. When excipients are used, they should not
adversely affect the stability, bioavailability, safety, or efficacy of the active ingredient(s), or cause toxicity
or undue local irritation. There must be no incompatibility between any of the components of the dosage
form.
• Water for injections is used as the vehicle for aqueous injections. It should be freshly distilled by the
process described under "Aqua pro Injection", be free from carbon dioxide, and comply with Test for
bacterial endotoxins. Sterilization at this stage may be omitted, provided that the solution or preparation is
immediately sterilized upon finalization. For non-aqueous injections, fixed oils of vegetable origin are used
as vehicles.
• Unless otherwise specified in the individual monograph, sodium chloride or other suitable substance(s),
may be added to an aqueous solution for injection in order to render the preparation isotonic.
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5. 5

6. 6

7. TERMINALLY
STERILIZED/
STERILIZATION BY
FILTERATION
MANUFACTURING OF STERILE PREPARATIONS: KEY REQUIREMENTS
CHANGE
ROOMS
PERSONNEL
FLOW
ASEPTIC
FACILITIES-
ZONES
WALLS AND FLOOR
TREATMENTS
LIGHTNING
FIXATURES
As per Gazette of
India: Black, Grey
and White
As per CGMP:
ZONES 7-1
STERILITYTESTING:
a)Direct Transfer
b) Membrane Filtration
c) Rabbit test
d)LAL test
e) Leaker test
f) Particulate matter testing
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8. MANUFACTURE OF STERILE PREPARATIONS
1. Terminally sterilized: usually involves filling and sealing product
containers under high-quality environmental conditions. Products are
filled and sealed in this type of environment to minimize the microbial
and particulate content of the in-process product and to help ensure that
the subsequent sterilization process is successful. In most cases, the
product, container, and closure have low bio-burden, but they are not
sterile. The product in its final container is then subjected to a
sterilization process such as heat or irradiation.
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9. TERMINAL
STERILIZATION
OF PRODUCTS
STERILIZATION OF THE
PRODUCT IS DONE
USING HEAT OR
RADIATIONS
PRODUCT IS FILLED IN
SUITABLE CONTAINERS
AND CONTAINER IS
SEALED
PRODUCT IS PREPARED
UNDER CONTROLLED
ENVIRONMENT
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10. 2. Sterilization by Filtration:-
1. Previously
sterilized container
are taken
2. Filters having
nominal pore size
0.22 μm or less are
used for filtration
5. No fiber
shedding or
asbestos filters
4. Double filter
layer or second
filtration
3. Remove bacteria
and moulds but
Not viruses &
Mycoplasmas
6. Filter integrity
testing
META FILTERS OR MEMBRANE FILTERS ARE USED
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11. 3. Aseptic Preparation: -
1. The drug product, container,
and closure are first subjected
to sterilization methods
separately
2. Containers are filled and
sealed in an extremely high-
quality environment
5. In area occupied by personal, the air must be exchanged with the frequent intervals. Fresh outside or recycled air
must be first filtered to remove particulate matter and then HEPA filters are used to get CLASS-100 air systems.
3. Before aseptic assembly into a final
product, the individual parts of the
final product are generally subjected
to various sterilization processes
4. Any manual or mechanical manipulation of the sterilized drug, components, containers, or closures
prior to or during aseptic assembly poses the risk of contamination and thus necessitates careful control.
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12. GMP REQUIREMENTS FOR STERILE PRODUCTS
MINIMISING THE RISK
OF MICROBIOLOGICAL
CONTAMINATION AND
PARTICULATE MATTER
OR PYROGEN
PRODUCTION IN CLEAN AREAS,
AIRLOCKS FOR ENTRY (PERSONAL
AS WELL AS MATERIAL), SEPARATE
AREAS FOR OPERATIONS SUCH AS
PRODUCT PREPARATION, FILLING
AND SEALING ETC.
LEVEL OF
CLEANLINESS SUCH
AS FILTERED AIR, AIR
CLASSIFICATION-
GRADE A, B, C AND D
LAMINAR AIR FLOW: AIR
SPEED (HORIZONTAL
VERSUS VERTICAL
FLOW), NUMBER OF AIR
CHANGES AND AIR
SAMPLES
CONFORMITY TO STANDARDS,
WORK STATION AND
ENVIRONMENT AND BARRIER
TECHNOLOGY AND AUTOMATED
SYSTEMS
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13. ZONES AS PER THE C GMP
• ZONE 7:- FILLING LINE
• ZONE 6:- FILLING AREA
• ZONE 5:- WEIGHING, MIXING & TRANSFER AREA.
• ZONE 4:- CLEAN AREA
• ZONE 3:-GENERAL PRODUCTION
• ZONE 2:- WAREHOUSE
• ZONE 1:- EXTERIOR
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14. Zone-7:-filling line:-
• The walls of the filling area are the last physical barrier to the ingress
of contamination, but within the filling area a technique of
contamination control known as laminar flow may be considered as
the barrier to contamination.
Zone-6:-filling area:-
• Zone 6 is a distinct zone of the controlled environment area for an
aseptic filling process but may not be distinct zone for non-aseptic
filling processes.
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15. Zone-5:-weighing, mixing, and transfer area:-
• Zone5 encompasses those activities of “weighing, mixing, filling or transfer
operations” addressed by c GMP section 212.81 which are not handled as zone 6 but
which require a controlled environment.
Zone-4 :-clean area:-
• Activities in these may include washing and preparations of equipment or
accumulation and sampling of filled product.
Zone-3:-general production and administration area:-
• The third zone of environmental control is formed by the periphery of the general
production area. Openings into the area are usually well sealed and large enough for
only essential material-handling equipment and personnel.
Zone-2:-warehouse:-
• Basic warehousing functions include receiving, shipping, and in-process storage.
Receiving areas include unpacking, sampling and incoming quarantine.
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16. Zone-1:-plant exterior:-
• The environmental with in which a plant located is first environmental
control zone. It is a base point from which to work in determining the
requirements for the various control barriers. Management actions to control
zone 1 might include the maintenance of sterile areas around the facility
where weeds, insects and rodents are controlled or eliminated.
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17. ZONES AS PER GAZZETE OF INDIA
• White zone:-Final step (filling of parenteral)
• Grey zone:-weighing, Dissolution & filtration.
• Black zone:-Storage, Worst area from
contamination view point
• a) Have a per-cubic-particle count of not more
than 100 in a size range of 0.5 micron and
larger through the entire work area upstream of
the work piece.
• b) Be supplied at the point of use as specified
in section 212.77.
• The layout of the plant must be carefully
developed in coordination with the needs of
the HVAC (Heating Ventilation and Air
Conditioning) system
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18. 2. WALL & FLOOR TREATMENT :
The design of filling areas or more generally, controlled environment
areas involves attention to many seemingly minor details. The basic
clean-ability requirement includes smooth, cleanable walls, floors,
ceilings, fixtures, and partition exposed columns, wall studs, bracing,
pipes, and so on are unacceptable. The need for clean-ability also
eliminates the open floor system commonly used in the microelectronics
industry for laminar airflow rooms. The goal of the designer, when
creating the details for the architectural finishes and joining methods, is
to eliminate all edges or surfaces with in the room where dirt may
accumulate.
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19. 3. LIGHTNING FIXTURES :
Lighting fixtures should be reduced flush with the ceiling. Since most
lighting fixtures are not tightly sealed, the diffuser should be sealed
integrally with the ceiling, and the lamps changed from outside the
room. Either recessed or surface mounted fixtures can be used. Special
“wash-down” fixtures are well sealed, but protrude obtrusively into the
room and have clips and sealing lips which are difficult to sanitize.
Areas having a full HEPA ceiling obviously cannot accommodate
recessed lighting fixtures. In these areas, fixtures are of a special
“teardrop” shape which minimizes disruption to the laminar airflow
pattern.
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20. 4. CHANGE ROOMS:
Personnel access to all controlled areas should be through change
rooms. Change rooms concepts and layouts vary from single closet size
rooms to expensive multi-room complexes.
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21. 5. PERSONNEL FLOW:- The movement of personnel should be
planned during the design of individual plant areas. Each individual
production area may have a smooth and efficient personnel flow pattern,
a discontinuous or crowded pattern may develop when several
individual production area plants are combined. The separation of
people and products is greatly facilitated by the use of the third
dimension. Security concerns about personnel flow may include
minimizing access to controlled substances and minimizing the
personnel traffic in or near work areas where controlled substances are
handled.
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4 3
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4 2
3
NO YES
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22. STERILITY TESTING OF PARENTRALS
The Basic quality control tests which are performed on sterile parenteral
products include:-
1) Sterility Tests.
2) Pyrogen Tests.
3) Leaker Tests.
4) Particulate matter testing.
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23. 1) Sterility tests:- Sterility is the most important and Absolutely Essential characteristics of Parenteral products.
Sterility means complete absence of all viable Micro-organism. It is an absolute term. The methods which are used to
perform sterility tests are
• a) Direct transfer method.
• B) membrane filtration method.
• A) Direct Transfer method:– it is an traditional sterility test method which involves a direct inoculation of required
volume of a sample in two tests tube containing a culture medium that is FTM, SCDM.
• This method is simple in theory but difficult in practice when the demand for repetition in opening container, sampling
Transferring, and mixing increases causes potential error in operator technique
• B) Membrane Filtration method:– It is official in U.S.P. 1970. This method basically involves filtration of Sample
through membrane filters of porosity 0.22 micron and Diameter 47mm. 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.
• *Interpretation: – If no visible evidence of microbial growth in culture medium in test tube then it is interpreted that the
sample representing lot is without intrinsic contamination
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24. 2) Pyrogen Test: – Pyrogens are products of metabolism in microorganisms Gm-ve
bacteria produces most potent pyrogens. These 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. Basically there are test performed to
detect the presence of pyrogens in sterile parenteral products they are C) Rabbit Test D)
LAL Test.
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25. • C) Rabbit test:– This test basically involves the injection Sample solution which is to be tested
into a Rabbits Which are use as test animals through ear vein. The Temperature sensing probe
(Clinical Thermometer, Thermistor or similar probe) into a rectum cavity of Rabbit at the depth of
7.5 cm, the test solution must be warmed at 37 degrees prior to injection. Then Rectal temperature
is recorded at 1,2,3 hr subsequent to injection. This test is performed in separate area designed
solely for this purpose under environmental conditions similar to animal house should be free from
disturbances that likely to excite them. Initially this test is performed on 3 Rabbits but if required
results are not obtained this test is repeated on 5 additional Rabbits with same sample solution
administer to initial 3 rabbits. Prior to 1hr of injecting sample solutions the control temperatures of
rabbits are determined. Use only those rabbits whose control temperature is no vary by more than 1
degree Celsius.
• *Interpretation:- The solution is judged to be non pyrogenic if no single rabbit show rise in
temperature of 0.5 degree Celsius but if this condition is not met then the test if repeated on 5
additional rabbits with same preparation administer
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26. D) LAL test:– It is an recently developed in vitro test method for pyrogen utilizing
gelling property of lysates of amebocytes of limulus Polyphemus which is found
only at specific locations along the east coast of North America and along
southeast Asia. It is derived from horse shoe crab; the basic procedure is the
combination of 0.1 ml of test sample with LAL Reagent after incubation for 1 hr at 37
degree Celsius the mixture is analyzed for the presence of Gel clot. The LAL Test is
positive indicating that the presence of endotoxin. Its applications are mainly to
Pharmaceutics, Biological, devices, disease states, food, and validation of heat cycles.
This method has several advantages of Rabbit test they are Greater sensitivity and
reliability specificity, less variation, wider application, less expensive and simplicity.
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27. 3) Leaker Test: – The leaker test is intended to detect incompletely sealed
ampoules, so that they may be discarded. Tip sealed ampoules are more prone to
leak than pull sealed. In addition to that crack my present around seal or at the base
of ampoule as a result of improper handling leakers are usually detected by
producing negative pressure within the incompletely sealed ampoule usually into a
vacuum chamber while those ampoules are submerged into a colored dye solution
of 0.5 to 1% methylene blue. Vials and bottles are not subjected to such leaker test
because rubber closure is not rigid however bottles are often sealed while vacuum is
pulled so that bottle remains evacuated during its shelf life.
The presence of vacuum is detected by striking at the base of bottle sharply with the
heel of hand to produce typical water hammer sound. Another test is to apply
a spark tester probe outside to the bottle moving form liquid layer into air space a
blue spark discharge occur is air space is evacuated.
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28. 4) Particulate matter testing:–
Particulate matter is primary concern in the parenteral products
given by I.V. Route, all parenteral products should be free from
insoluble particle. Further U.S.P. states that GMP Requires that all
containers be visually inspected and that with visible particle be
discarded. The visual inspection is done by holding the ampule by its
neck against highly illuminated screens. White screens for the detection
of black particle and black screens for the detection of white particles to
detect heavy particles it may be necessary to invert container but care
must be exercised to avoid air bubble. The instrumental methods are
based on principles of light scattering, light absorption, electrical
resistance as in coulter counter. A method which utilizes a video image
projection could detect a moving particle without destruction of product
unit.
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29. THANK YOU
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