PARENTERALS

INTRODUCTION
Parenterals :- are Sterile, Pyrogen free preparations injected through skin or mucous
membrane into internal body compartment.

Parenteral products
A. IV Admixtures consist of one or more sterile drug products added to an
IV fluid. Used for
▶ Drugs intended for continuous infusion
▶ For drugs that may cause irritation or toxicity when given by direct IV
injection.
B. IV fluids
These fluids have multiple uses,
▶ Vehicles in IV admixtures
▶ Provide means for reconstituting sterile powders
▶ Serve as the basis for correcting body fluids and electrolyte disturbances
▶ For administering parenteral nutrition
Dextrose : Generally, a solution of 5% dextrose in water
▶ pH of 5% dextrose ranges from 3.5-6.5. Instability may result if it is combined
with an acid sensitive drug.
▶ In higher conc. (e.g. 10% solution in water), dextrose provides a source of
carbohydrate in parenteral nutrition solutions.
▶ Should used cautiously in patients with diabetes mellitus.
Sodium chloride : usually given as 0.9% solution called as normal saline
solution.
▶ Sterile sodium chloride for injection:
o Used as vehicle in IV admixtures and fluid for electrolyte replacement.
▶ Bacteriostatic sodium chloride for injection:
o It contains an agent that inhibits bacterial growth (e.g. Benzyl alcohol,
Propyl paraben. Methyl papaben), allowing its use in multiple dose
preparation.
Water
▶ Used for reconstitution and for dilution of IV solutions such as dextrose and
sodium chloride.
▶ Water suitable for parenteral preparations include sterile water for injection
and bacteriostatic water for injection.
 Ringer solutions
▶ Used for fluid and electrolyte replacement.
▶ Commonly administered to post surgical patients.
▶ It contains sodium lactate, sodium chloride, potassium chloride, and calcium
chloride.
C. Electrolyte preparation
▶ Ions present in both intracellular and extra cellular fluid.
▶ Surgical and medical patients who can not take food by mouth or who need
nutritional supplementation require the addition of electrolytes in hydrating
solutions or parenteral nutrition solutions.
D. Dialysate
▶ Used in patients with disorder as renal failure, poisoning, and electrolyte
disturbances.
▶ In peritoneal dialysis, a hypertonic dialysis is infused directly into peritoneal
cavity via a surgically implanted catheter. It contains Dextrose and
electrolyte, which removes the harmful substances by osmosis and diffusion.
E. Irrigating solutions
▶ Not intended for infusion into the venous system.
 Topical administration
▶ Used in irrigating wounds, moistening dressings, and cleaning surgical
instruments.
 Infusion of irrigating solutions
▶ Surgeons performing urological procedure often use irrigating solution to
perfuse tissues in order to maintain the integrity of surgical field, remove
blood, and provide a clear field of view.

GMP Requirements for Sterile Products

▶ Specific points relating to minimizing risks of contamination.

– Microbiological
– Particulate matter
– Pyrogen

General Requirements
▶ Production in clean areas
▶ Airlocks for entry
– Personnel entry.
– Material entry
▶ Separate areas for operations

– Component preparation
– Product preparation
– Filling
– Sealing etc…

▶ Level of cleanliness

▶ Filtered air

▶ Air classification: Grade A, B, C and D.

▶ Laminar air flow:

– Air speed (horizontal versus vertical flow)

– Number of air changes
– Air samples
▶ Conformity to standards
▶ Work station and environment

▶ Barrier technology and automated systems

Types of sterile products processing
1 Terminally sterilised
→ prepared, filled and sterilised
2 Sterilised by filtration
3 Aseptic preparation

Manufacture of sterile preparations

1. Terminally sterilised:- 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.

2. Sterilisation by Filtration:-
o Previously sterilized container are taken.
o Filters having nominal pore size 0.22 µm or less are used for filtration
o Remove bacteria and moulds but Not viruses & Mycoplasmas
o Double filter layer or second filtration
o No fibre shedding or asbestos filters
o Filter integrity testing

3. Aseptic Preparation :- In an aseptic process, the drug product, container,

and closure are first subjected to sterilization methods separately, as appropriate, and
then brought together. Because there is no process to sterilize the product in its final
container, it is critical that containers be filled and sealed in an extremely high-quality
environment Before aseptic assembly into a final product, the individual parts of the final
product are generally subjected to various sterilization processes. 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.

Note:- 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 than HEPA filters are used to get CLASS-100 air systems.

SPECIFIC REQUIREMENTS FOR MANUFACTURE OF PARENTERAL

PREPARATIONS (SMALL VOLUME INJECTABLES AND LARGE
VOLUME PARENTERALS)
As per schedule –M .

[1] General :- Sterile products, being very critical and sensitive in nature a very
high degree of precautions, prevention are needed for it‟s preparation.Dampness, dirt
and darkness are to be avoided to ensure aseptic conditions in all there shall be strict
compliance in the prescribed standards especially in the matter of supply of water, air,
active materials and in the maintenance of hygienic environment.

[2] Building and Civil Works: –

→The building-built on proper foundation with standardized materials
→Location of services like water, steam, gases etc. shall be such that their servicing or
repair shall not pose any threat to the integrity of the facility. Water lines shall not
pose any threat of leakage to any of the manufacturing area.
→The manufacturing areas-clearly separated into support areas & preparation areas.
→Operations like removal of outer cardboard wrappings of primary packaging materials
shall be done in the de-cartoning areas which are segregated from the washing areas.
→Wooden pallets, fiberboard drugs, cardboard and other particle shedding materials
shall not be taken inside the preparation areas.

General points to be in consideration for ASEPTIC Areas

→ Walls, floors and ceiling-impervious, non-shedding, non-flaking and non-cracking.
→ Flooring-unbroken, provided with a cove both at the junction between the wall and
the floor & wall and ceiling. Epoxy should done in aseptic area, Walls-shall be flat.
→ Light-fittings and air-grills-shall flush with the walls and not hanging from ceiling.
→ Doors & Windows-made of non-shedding material preferably Aluminium or Steel
material. Doors shall open towards the higher-pressure area so that they close
automatically due to air pressure.
→ The furniture-smooth, washable and made of stainless steel or any other appropriate
material
[3] Garments
→The garments-made of non-shedding and tight weave material, single piece with
fastenings at cuffs, neck and at legs to ensure close fit. Trouser legs shall be tucked
inside the cover boots.
→Design-include a hood (head-cover) or a separate hood which can be tucked inside
the over-all. Zips (if any) shall be of plastic material.
→Gloves-made of latex or other suitable plastic materials & long enough to cover wrists
completely and allow the over-all cuff to be tucked in.
→footwear- of suitable plastic or rubber material, daily cleaned with a bactericide.
→Garment changing procedures shall be documented and operators trained in this
respect.

[4] Area planning: Depends on

1). Type of production

 Batch operations: suited to small production volume & minimum financial
investment.
Advantages:
1.Product quality, consistency, and homogeneity are relatively easily controlled.
2. Production documentation is easy.
Disadvantages:
1.Economically undesirable because it is labor intensive and does not exploit the
economies of volume.

 Continuous operations: it is suited to very high volume production requirements. it

requires more space and more complex equipments.
Advantages:
1. Minimizes shortcoming of batch operations; labor, production time, and
environmental exposure of the product.
2. Since the intermediate material handling steps are eliminated, the potential for
product contamination during those steps no longer exists.
Disadvantages:
1. Product quality assurance is difficult.
2. It is very difficult to document the ingredients or process cycle for a product
produced in a continuous process.

2). Container size

→SVPs and LVPs obviously requires different space considerations.
→All the production equipment has container size limitations- large container requires
large equipments and more space.
3). Environment control needs

4). Product characteristics

Liquids are probably the easiest product to handle.
Emulsion may require compounding areas close to filling lines to ease transfer
problems. Pumping systems will be very critical.
Suspension will require a means of maintaining a homogenous mixture prior to
filling.
To minimize the time the suspension resides in piping, reservoir, and pump
system,filling rate should be kept high and the distance from compounding to
filling should be minimized.
5). Space requirements
[QUANTITATIVE LAYOUT OF PARENTERAL MANUFACTURING ]
FUNCTION Area
Square meter Percentage

Production 11,094 45.1

Warehouse 7,606 30.9

Utility 1,716 4.1

Quality control 1,716 7.0

Administration 1,018 4.1

Maintenance 1,014 4.5

Employee services 1,014 4.1

Security 39 0.9

Total 24,607 100.0

6). Personnel Movement

 The movement of personnel should be planned during the design of individual plant
areas.
 Discontinuous and crowded flow patterns can decrease production efficiency,
increase security problems, and increase the problems of maintaining a clean
environment. Personnel flow path from zone to zone must be such that access to
higher level of cleanliness is only through change rooms, gowning rooms, locker
rooms, or other areas as may be required to prepare the personnel for the cleaner
area.
 In a parenteral plant degree of access should be restricted.
 Planning for visitors and nonproduction employees in advance can prevent or
lessen many future problems, particularly in critical area.
 A glassed mezzanine or balcony provides absolute isolation, yet may give
excellent view of process.
[5] Environmental control zone groupings
1st. Zones as per the cGMP:- 1st. Zones as per Gazette of India
 Zone 7:- Filling line
 Zone 6:- Filling area
 Zone 5:- Weighing, mixing & WHITE
transfer area. BLACK
 Zone 4:- Clean area
 Zone3:-General production GRAY
 Zone 2:- Warehouse
 Zone 1:- Exterior

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

Environmental control :
Sources Control
People  Total body covering in critical area and partial covering in non
critical area.
 Adequate personal flow and restricted access to aseptic and critical
environment.
 Minimum movement of personal.
 Adequate operation procedure for personal.
Barrier  Adequate sterilization procedure
 Protective laminar flow equipment
 Barrier and separation between high risk and low risk operation.
 Adequate operation procedure to assure proper handling,
cleaning, and sterilization of machinery and equipment
Material  Adequate material control and selection
 Adequate sterilization and filtration procedure
Air  Adequate air filtration system
 Adequate monitoring of air cleanliness level.
 Adequate air system validation procedure.
AIR HANDLING SYSTEM (AHU)

CRITICAL AREAS GRADES

Aseptic feeling area B

Sterilized component unloading area C

Change room D
The filter Configuration in the AHU shall be suitably designed to achieve the Grade of
air as given in Table1.

TABLE I AIRBORNE PARTICULATE CLASSIFICATION FOR MANUFACTURE OF

STERILE PRODUCTS.

Grade Maximum number of permitted particles per cubic metre equal to or

above.
AT REST IN OPERATION

0.5µm 5µm 0.5µm 5µm

A 3520 29 3500 29
B (a) 35,200 293 3,52,000 2,930
C (a) 3,52,000 2,930 35,20,000 29,300
D (a) 35,20,000 29,300 Not (c) Not defined (c)
defined

Notes :

(a) In order to reach the B, C and D air grades, the number of air changes shall be
related to the size of the room and the equipment and personnel present in the room.
The air system shall be provided with the appropriate filters such as HEPA for Grade A,
B and C. the maximum permitted number of particles in the “at rest” condition shall
approximately be as under:

Grade A corresponds with Class 100 or M 3.5 or ISO Class 5; Grade B with Class 1000
or M 4.5 ISO Class 6; Grade C with Class 10,000 or M 5.5 or ISO Class 7; Grade D with
Class 100,000 or M 6.5 or ISO Class 8.

(b) The requirement and limit for the area shall depend on the nature of the operation
carried out.

(c) Type of operations to be carried out in the various grades are given in Table II and
Table III as under.

TABLE II TYPES OF OPERATIONS TO BE CARRIED OUT IN THE VARIOUS

GRADES FOR ASEPTIC PREPARATIONS

Grade Types of operations for aseptic preparations

A Aseptic preparation and filling
B Background room conditions for activities requiring Grade A
C Preparation of solution to be filtered
D Handling of components after washing
TABLE III

Types of operations to be carried out in the various Grades for terminally

sterilized products.

Grade Types of operations for terminally sterilized products.

A Filling of products, which are usually at risk
C Placement of filling and sealing machines, preparation of solutions when
usually
at risk. Filling of product when unusually at risk.
D Moulding, blowing (pre-forming) operations of plastic containers,
preparations of
solutions and components for subsequent filling

The recommended frequencies of periodic monitoring shall be as follows(As per

Schedule - M) :-

Particulate monitoring in air 6 Monthly

Air change rates 6 Monthly
HEPA filter integrity testing (smoke testing) Yearly
Air Pressure differentials Daily
Temperature & Humidity Daily
Microbiological monitoring by settle plates and/or swabs in
Daily
aseptic areas
AIR CLASSIFICATION AS PER CDER – Centre For Drug Evaluation & Research:
Clean Area ISO >0.5 µm Microbiological Microbiological
Classification Designation particles/m3 Active Air Settling Plates
(0.5 um Action Action Levelsc,d
particles/ft3) Levelsc(cfu/m3 ) (diam. 90mm;
cfu/4 hours)
100 5 3,520 1e 1e
1000 6 35,200 7 3
10,000 7 352,000 10 5
100,000 8 3,520,000 100 50

1. Air Classification as per Schedule M

Grade Maximum permitted number of particles/m3 equal or above
at rest in operation
0.5µm 5.0µm 0.5µm 5.0µm
A 3,520 29 3,500 29
B 35,200 293 3,52,000 2,930
C 3,52,000 2,930 35,20,000 29,300
D 35,20,000 29,300 Not defined not defined
 2. Air Classifications by USFDA guideline on Sterile Drug Products
Clean Area <0.5 µm <0.5 µm Microbiological Limit
Classification Particles/ft3 Particles/mt3 cfu/ft3 cfu/m3
100 100 3,500 <1 <3
1000 1000 35,000 <2 <7
10000 10000 350,000 <3 <18
100000 100000 3,500,000 <25 <88

3. Air Classifications as per WHO 2002

Grade Maximum Number Permitted / M3
Particles Microorganisms
0.5µm 5.0µm
A (LAF) 3,500 0 <1
B 3,500 0 5
C 3,50,000 2,000 100
D 3,500,000 20,000 500

4. Air Classifications as per ISO

Grade ISO Class Particle/cum Class(SI)
A 5 100 3.5 M 3.5(filling)
B 6 1000 35 M 4.5
C 7 10000 350 M 5.5
(preparation)
D 8 100000 3500 M 6.5

3) DESIGN CONCEPTS

1. CHANGE ROOM
 Entrance to a change room is normally through vestibules whose doors are
electrically interlocked so that both can not open simultaneously, thus maintaining
the necessary air pressure differential to prevent the entry of airborne contamination.
 Upon entry in the change room, wash sinks are provided for scrubbing hands and
forearms. Automatic or foot operated controls for water and soap eliminate hand
contact with contaminated surfaces.
 Next, hands are dried by hot air blowers.
 Commercial hand driers may create undesired airflow patterns. Special filtered
driers are available to minimize the creation of particulate contamination.
 After the hands are dry, garments are taken from dispensers and donned while
moving across a dressing bench.
 As a final gowning step, aseptic gloves are put on and sanitized.

2. FILLING AREA:

It is the most critical area in parenteral plant, where the product & sterilized
components are exposed to room environment. Therefore these areas are specially
constructed, filtered, and maintained to prevent environmental contamination.
 CLEAN ROOM An area with defined environmental control of particulate and
microbial contamination, constructed and used in
such a way as to reduce the introduction, generation and retention
of contaminants within the area
QUALITIES OF CLEAN ROOM
 The room should undergo 15-20 air changes per hour.
 HEPA filters are to clean the air entering the room.
 HEPA filters remove all airborne particles of size 0.3 or larger with an efficiency
of 99.97%.
 Maintaining higher air pressure(+ve pressure) within the critical area to minimize
infiltration of airborne contaminants from outside.
 Care should be taken to ensure that air flows do not distribute particles from a
particle-generating person, operation or equipment to a zone of higher product
risk. A warning system should be provided to indicate failure in the air supply.
 Adjacent rooms of different grades should have a pressure differential of
10 - 15 Pascals.
 Counters in the clean room should be made of stainless steel or other non-
porous, easily cleaned material.
 Walls and floors should be free from cracks or crevices and have rounded
corners. If the walls or floors are to be painted, epoxy paint is used.
 The air flow should move with uniform velocity along parallel lines. The velocity of
the air flow is 90 ± 20 ft/m3.
 Providing temp. & humidity controls appropriate to the product being manufactured.
LAY OUT OF CLEAN ROOM

STANDARDS FOR CLEAN ROOM

Federal Standard 209
MEASURED PARTICLE SIZE (MICROMETERS)
CLASS
0.1 0.2 0.3 0.5 5.0
1 35 7.5 3 1 NA
10 350 75 30 10 NA
100 NA 750 300 100 NA
1,000 NA NA NA 1,000 7
10,000 NA NA NA 10,000 70
100,000 NA NA NA 100,000 700

Selected ISO 209 airborne particulate cleanliness classes for

cleanrooms and clean zones.
numbers Maximum concentration limits (particles/m^3 of air) for particles
(N) equal to and larger than the considered sizes shown below
0.1m m 0.2m m 0.3m m 0.5m m 1m m 5.0m m
ISO 1 10 2
ISO 2 100 24 10 4
ISO 3 1 000 237 102 35 8
ISO 4 10 000 2 370 1 020 352 83
ISO 5 100 000 23 700 10 200 3 520 832 29
ISO 6 1 000 000 237 000 102 000 35 200 8 320 293
ISO 7 352 000 83 200 2 930
ISO 8 3 520 000 832 000 29 300
ISO 9 35 200 000 8 320 000 293 000
QUALITATIVE LAYOUT OF PARENTERAL MANUFACTURING (circular flow)

QUALITATIVE LAYOUT OF PARENTERAL MANUFACTURING (parallel flow)

LAYOUT FOR TERMINAL STERILIZATION

Pdt. Material
Exit Entry
Soln
Unidirectional Prepn
Clean Zone Area

Aseptic
Filling zone
Clean
Changing
Entry Room
Oven Equipment &
Component
Auto Prepn
clave Area
Aseptic
Receiving
Area
Hatch Comp.
Entry
3. LAMINAR AIR FLOW UNIT
o HEPA (HIGH EFFICIENCY PARTICULATE AIR filtration)
HEPA Filter

 HEPA filters are composed of a mat of randomly arranged fibers (poly-

vinylidene fluoride -PVDF)
 Key metrics affecting function are fiber density and diameter, and filter
thickness

 There are four basic mechanism in which fibrous air filter remove contamination
from the airstream.

1. Straining or Sieving 2. Impaction 3. Interception 4. Diffusion

Laminar flow hoods: These are clean air work benches are specially designed to
ensure the aseptic preparation of sterile products. Laminar air flow hoods are generally
used in conjunction with clean rooms.
o For laminar air flow work station the air flow rates shall be 0.3 meter per second
(vertical) and 0.45 (horizontal)
o Introduction of personnel, equipment, and material into the work area provides
sources of particulate matter which may contaminate the product.
o Very small particles are not heavy enough to settle due only to the force of gravity,
but instead are carried and directed by air currents.and if there is turbulent air,
particles may be driven into product.
o Laminar air flow velocity satisfactorily sweeps the area yet does not create
unacceptable turbulence.

 LIST OF EQUIPMENTS( as per schedule-M)

The following equipment's is recommended:

a) Manufacturing area: -
1. Storage equipment for ampoules, vials bottles and closures.
2. Washing and drying equipment.
3. Dust proof storage cabinet
4. Water still.
5. Mixing and preparation tanks or other containers.
6. Mixing equipment where necessary.
7. Filtering equipment.
8. Hot air sterilizer.
b) Aseptic filling and sealing rooms -
9. Benches for filling and sealing.
10. Bacteriological filters.
11. Filling and sealing unit under laminar flow work station.
c) General Room.
12. Inspection table.
13. Leak testing table.
14. Labeling and packing benches.
15. Storage of equipment including cold storage and refrigerators if necessary.

An area of minimum sixty square meters partitioned into suitable sized cubicles with air
lock arrangement, is recommended for the basic installation.

Types of containers:

1. Ampoules: They are intended for single use only, ampoules are opened by breaking
the glass at a score line on the neck. Because glass particles may become
dislodged during ampoule opening, the product must be filtered before it
administered. Because of their unsuitability for multiple-dose use, the need to filter
solutions before use and other safety considerations have markedly reduced
ampoule use.

2. Vials are glass or plastic containers are closed with a rubber stopper and sealed
with an aluminum crimp.
Advantages over ampoules.
 They can be designed to hold multiple doses (if prepared with a
bacteriostatic agent).
 It is easier to remove the product.
 They eliminate the risk of glass particle contamination during opening.
Drawbacks
 The rubber stopper may become cored.
 Multiple withdrawals(as with multiple-dose vials)may result in microbial
contamination.
Some drugs that are unstable in solution are packaged in vials in powder form and
must be reconstituted with sterile sodium chloride for injection before use.
Some of this drugs come in vials that contain a double chamber.(a) The top chamber
containing sterile water for injection is separated from the unreconstituted drug by a
rubber closure. (b) To dislodge the inner closure and mix the contents of the
compartments, external pressure is applied to the outer rubber closure. This system
eliminates the need to enter the vial twice, thereby reducing the risk or microbial
contamination.

3. Prefilled syringes -These designed for quickest administration and maximum

convenience. Drugs administered in an emergency (e.g.,atropine,epinephrine) may
be available for immediate injection when packaged in prefilled syringes.

4. Infusion solutions are divided into two categories : small volume parenterals
(SVP), those having a volume of 100 ml;and large volume parenterals (LVP), those
 having a volume of 100 ml or greater. Infusion solutions are used for the intermittent
or continuous infusion of fluids or drugs.

 EQUIPMENTS
 Sterile Garment Cabinet
 Made up of Stainless steel.
 Ensure a clean storage space by making use of UV disinfectant and heating
through IR lamps.
 These cabinets may be designed in horizontal air flow system and clean air
through HEPA filters
 Syringe Filling Machine
Characteristics
• Barrier isolators
• In-process check weighing
• Filling : rotary piston pumps.
• Volume: 0.2 to 29 ml
• All types of syringe including glass, plastic can be filled.
• Filling Rate: 300 to 600 syringes in a minute.
 Ampoule Washing Machine
PROCESS

 Water is sprayed onto the ampoules.

 Turned to an angle of 180 degree with their mouth
downward to remove water.
 Finally the ampoules are filled with compressed
air to remove residual water.
 Certain machines have a high temperature zone
meant for killing any bacteria.

 Vial Filling Machine

• Fill vials and bottles
• liquids, viscous material and suspensions and powders.
PROCESS
 The machine comprises of an intake section which
loads the vials.
 Transferred through an intermittent transport section.
 liquid filling section which fill the vials with predetermined
quantity.
 Finally the filled and rubber stoppered vials are released and discharged.
 SIP System
• For in-line sterilization of various processing equipments.
• Handling various biological solutions and mixtures requires cleaning and sterilizing
these equipments from time to time as they are susceptible to contamination.
• Proper SIP integration with pharmaceutical equipment is very important for the
overall success of the operation.
 ANTIMICROBIAL EFFICACY OF A SILVER-ZEOLITE MATRIX COATING ON
STAINLESS STEEL
• A silver and zinc-containing zeolite matrix (AgION) used as a coating for stainless
steel.
• Test against- e.coli, s.aereus, p.aeroginosa etc.
• Result:- The silver-zeolite mixture reduced microbial colony-forming units upto
84.536 – 99.999% after 4 h exposure, and upto 99.992-100% after 24 h in all
cases.

 FILTERS IN FILTRATION STERILIZATION

Millipore’s Airvent filters
• Constructed with a PTFE membrane.
• These filters have been qualified to withstand at least 40 SIP cycles at 135 °C for
30 minutes.

Millipore’s Durapore filters

• Constructed with a PVDF membrane
• These filters have been qualified to withstand 5 to 30 SIP cycles at 135 °C for 30
minutes

Verification of integrity of filter

• Bubble point method
• Diffusive flow
• Pressure hold test
Bubble Point Test
Test Method

1. Record the filter part number(s), lot number, and product information. Also include
physical observations.
2. Wet the filter to be tested with the appropriate solvent (water for hydrophilic filters,
alcohol for hydrophobic filters).
3. Place the wetted filter in the appropriate housing.
4. Connect the outlet fitting from the compressed air pressure regulator to the upstream
side of the test filter.
5. Connect a piece of flexible tubing from the downstream port of the test filter into a
beaker filled with water.
6. Starting from zero pressure, gradually increase the pressure to the test filter using
the pressure regulator.
7. Observe the submerged end of the tubing for the production of bubbles as the
upstream pressure is slowly increased in 0.5 psig increments. Note the rate that the
bubbles appear for the end of the submerged tube.
8. The bubble point of the test filter is reached when bubbles are produced from the
tube at a steady rate. Record the pressure to the nearest 0.5 psig as indicated on
the pressure gauge.

 STANDARAD OPERATION PROCEDURE

For aseptic filling:-

 Check all sterilized material has indicator and expiration date.

 Open sterilized container, filling assembly and tubing on LAF bench.
 Connect the tubing of filling lines.
 Connect solution tank to the inlet of the filling assembly.
 Connect the nitrogen over lay in tank for pre and post flushing.
 Pump the solution in filling tubing up to the filling nozzle (remove any air bubble)
 After that wipe the filling nozzle with 70%alcohol.
 Switch on the machine.

S.O.P FOR OTHER MENUFACTURING PROCESSES IS SAME AS THAT OF NON

STERILE DOSAGE FORMS
 VALIDATION
Purpose: To minimize this reliance on end product sterility testing.
Three principle involved in validation process.
• To built sterility in the product.
• To demonstrate the maximum level of probability that the processing and
sterilization method have establish sterility to all units of product batch.
• To provide greater assurance and support to the result of the end product sterility.
It includes
a) Pre-processing quality control test
b) In process quality control test
c) Finished product quality control test
 Pre-processing quality control test
a) Raw material testing and assays
b) Packaging material test (glass, plastic, rubber etc)
c) sterility test and media fill (process simulation test)

 Tests for containers

(a). For Glass containers.
(i). Test for hydrolytic resistance.
(ii). Arsenic test.
(b). For Plastic container.
(i). Non volatile matter.
(ii). Sulphated ash.
(iii). Heavy metals.
(iv). Buffering capacity.
(v). Biological test. (Adverse reaction or toxicity)

 Media fill (process simulation test)

 Evaluation of the environment along with the process, the operator and the
equipment is the media fill.
Procedure

 Sterile Trypticase soy broth is filled into sterile container under condition
simulating as for a product.
 Entire lot at least 3000 units is incubated at suitable temp for 14 days .
 To pass the test not more than 0.1% of the unit may show growth.
 This is very stringent evaluation of an aseptic fill process and is considered to be
the most evaluative test available.

 In Process Quality Control Test

 Conductivity measurement
 Volume filled
 Temp for heat sterilized product
 Environmental control tests
 Visual inspection
 Finished product quality control test
 Leaker test  Sterility test.
 Pyrogen test  Uniformity of weight.
 Particulate test  Uniformity of content

 Leak test
• To detect incompletely sealed ampoules.
Principle
10% methylene blue or 0.1% FDC red one or red two.
Generally combined with autoclave.
Disadvantage
Leakage of 15 micron in diameter or smaller is not detected.
Vial and bottles are not subjected to this test.

 LAL test
Limulus Amoebocyte Lysate test or bacterial Endotoxin test for the validation of
depyrogenation process.
Reagent - LAL reagent (limulus Polyphemus)
Reaction - In presence of Endotoxin a firm gel is formed within 60 min when
incubated at 370 C.
o CHARACTERISTIC
• Test tube scale.
• Only pyrogen of gram negative bacteria detected.
• Semi quantitative test.
• Sensitivity in terms of Endotoxin unit.
• In-vitro test.
• Doesn‟t measure fever producing potential of Endotoxin.
• Sensitivity varies with different microbial source of LAL.

 Pyrogen test- Fever response of rabbit

• Sham test is performed to select the proper animals for the main tests.
• Rabbit test - Qualitative fever response test.
Procedure
• Test solution is injected into the vein of rabbit. Temperature elevation is seen for 3
hrs.
Disadvantage
 Biological variation
 Expensive
 Laborious
 Dose dependent.
 Not for anti pyretic drug.

 Particulate test USP

 Visually inspected- all (WHITE AND BLACK )
 Any with visible particle is discarded.
  Large volume parental
 50 particles of 10μm
 5 particulates of 25 μm per ml

Particulate count is done by:

1. Light obscuration particle count test
2. Microscopic particle count test

 USP requirements for packaging.

• Single dose container should not be more than 1 liter.
• Intra-spinal and intra-cisternal administered product must be in single dose
container.
• In case of multiple dose container dose should not be more than 30 ml.
 BFS Technology
Blow-fill-seal (BFS) technology is an automated process by which containers are
formed, filled, and sealed in a continuous operation. This manufacturing technology
includes economies in container closure processing and reduced human intervention
Most BFS machines operate using the following steps.

1. Extrusion
• An endless sterile plastic tube is continuously extruded from the melted granulate
in the filling cavity of the mould.
2. Blowing
• Final container is produced by sterile air pressure from Blow and Fill nozzle.
3. Filling
• After the container is formed inside the mould, sterile liquid product is introduced
into the container.
4. Sealing
• Final container is sealed in place by closing of the seal-mould form onto the
container top.
5. Mould opening
• Upon completion of filling and sealing steps, the mould is separated, producing
the sterile filled and sealed container.

ADVANTAGE OVER CONVECTIONAL ASEPTIC FILLING

• There is no need to purchase and stock a range of pre-fabricated container

and closures.
• Cleaning and sterilizing pre-fabricated container and closures are not required.
A clean sterile container is made with in the BFS machine.
• The cost of material transport, storage and inventory control is reduced.
• Validation requirement are reduced.
• There is a large choice of neck and opening device shapes.
• Saving floor space.
• Less labour intensive than conventional one.
• The code number and variables can be moulded into container it-self.
• With blow-fill-seal, you produce a one-piece, aseptically filled container with a
built-in safety seal.
• The blow-fill-seal process is suitable for heat-sensitive products.
List of equipments:- (mostly utilized in industry)

 Asep-Tech® Model 603 Blow/Fill/Seal Packaging Machine System

 PKV316 Vial and BFS container for Leak Detector
 Single chamber Autoclave Unit
 STERILIZER
 Sterilizing Tunnel

 Master formula records
Name of the product________________________________________

Name and Weight of API ____________________________________

Name and Weight of Ingredient _______________________________

Description of equipment ____________________________________

Statement of theoretical yield_________________________________

Process and packaging procedure_____________________________

A description of container____________________________________

closure and packaging material _______________________________

In process control during processing ___________________________

In process control during packaging____________________________

Precaution to be taken______________________________________

Environmental control
Product_______________________________ lot no.__________________________

Room________________________________ date exposed_____________________

Media__________________________

Date Time Incubation Humidity (in case of hygroscopic

substances)
Temp.

Plate no Duration Location No. of colonies

 Batch Manufacturing Records
Name of the company:-_______________________________________

Address:-___________________________________________________

Name of the product _________________________________________

Active pharmaceutical ingredient ______________________________

M F R No. __________________________________________________

Batch No._____________________ Batch size ____________________

Mfg. date _____________________Date of expiry_________________

Requisition slip

Sr Ingredients Item Standards ATR Label claim Qty Qty

no code no required issued

Preparation of equipment & containers

Description of containers _______________________________________

Q/C report of container ________________________________________

Date ________________________ Equipment used__________________

Cleaning agent used ___________________________________________

Cycle of washing: _____________________________________________

Sign. Of officer_______________________________________________

If sterilized by dry heat or autoclave:

Articles Date Time when oven Desired temp. Temp. Time when oven
started attained switched off

Filtration & filling:-

Equipments used for filtration ___________________________________

Date__________________________ Time_________________________

Result of test or analysis of solution_______________________________

Equipment used for filling_______________________________________

Date________________________________________________________

Sign. Of officer_______________________________________________

Time Filling Filling completed

started

IPQC records
1. Visual inspection:

Description ________________________________________________________

Total no of filled, sealed & sterilized containers rejected __________________

Nature of defects ____________________________________________________

Name of worker who examined:

 (i). ________________________________________________

(ii). _______________________________________________

(ii). _______________________________________________

Batch Packaging & Labeling Records

Product name_______________________ Batch no _______________________

Strength___________________________ batch size ______________________

Category___________________________ mfg date _______________________

MFG no____________________________ exp date _______________________

Batch Packaging & Labeling Records

Description of packaging______________________________________________

Pre-coding of labels & printed packaging materials,

examined & verified by _______________________________________________

No. of pre-coded ____________________________________________________

(ii). Printed packaging material received __________________________________

Result of bulk finished products ________________________________________

Sign. Of officer _____________________________________________________

Reconciliation of labeling and packaging material

Quantity of material received___________________________________________

Quantity of material destroyed__________________________________________

Quantity of material used _____________________________________________

Quantity of material returned___________________________________________

Date of release____________________ quantity release____________________

Signature of supervisor _______________

QAQC records
1.Visual inspection records

2. Uniformity of content

3. Uniformity of weight

4. Pyrogen test

 Sterilization

“The act or process, physical or chemical, that destroys or eliminates all viable
microbes including resistant bacterial spores from a fluid or a solid.”

Examples of sterilization methods are : steam treatment at 121℃, dry heat at

230℃, flushing with a sterilizing solution such as hydrogen peroxide (H 2O2) or ozone
(O3), irradiation, and filtration.

Sterility
“The reduction of anticipated levels of contamination in a load to the point where
the probability of survival is less than 10-6.”
EQUIPMENT

Single chamber Autoclave Unit

Steam Sterilize Item:

 Surgical Instruments
 Dressing materials,
 Injection liquids,
 Linen
----by means of
steam under pressure of 15 - 20 PSI (Adjustable)
HVAC Validation
Features of HVAC affect product quality (sterility).
1. HEPA integrity
a) Certification: by filter manufacturer indicates that filter is capable of removing all
particulate matter equal to or greater than 0.3 in size with an efficiency of
99.97%.
b) Installation: a certified filter if improperly installed will not perform its function &
provides a false sense of security.

C) Integrity testing: A popular method for certifying integrity of filter installation uses
polydisperse aerosol, created by blowing air through liquid Dioctyl phthalate,
introduced into upstream of HEPA filter followed by scanning the entire downstream
 of filter face with a probe nozzle of an aerosol photometer.
→This testing will indentify “leaks” caused by damage due to mishandling faulty
construction.
→ Small leaks can be repaired with a suitable silicone based compound without
removing filter.
d) Airflow resistance : Caused by dirty filter may reduce airflow volume, thereby
reducing the air change rate in critical areas.
 Airflow resistance is expressed as pressure differential between the air pressure
upstream of the filter and the downstream air filter.
 If the filters are not changed when they reach the maximum resistance as specified
by manufacturer, they may begin to lose their physical integrity or rupture, thereby
releasing some of the dust they have accumulated.

2.Airborne particle control

Particle count surveys should be performed at regular intervals.

Validation of the steam sterilizer

a) Validation on the basis of microbial death kinetic.

D- VALUE , F-VALUE AND Z-VALUE

1) D value: Quantitative expression of the rate of kill of the microorganism .The time
or dose required for the one log reduction in the microbial population.

SURVIVOR CURVE METHOD:- based upon plotting the log no of surviving organism
versus and independent variable such as time, gas concentration etc

Log N =A + Bt,

Where: - N is number of surviving organism, A is the y intercept.

B is the slope of the linear line in the graph
D value is of the linear slope: D=1/B
FRACTION NEGATIVE METHOD:- uses replicate samples containing identical spore
population treated in identical manner and determining the number of sample still
showing growth following treatment and incubation. Specially used for thermal
destruction processes.
-it is not applicable to ethylene oxide sterilization.

2 ) Z value:-for validation of heat sterilization process.Temp. Required for one log

reduction in D value. The Z value is the reciprocal of the slope resulting from the plot of
the logarithm of the D value was obtained.
 Z=(T2-T1/LOGD1-LOGD2) FOR SPORES;10-15
NON SPORES;4-5
3) F value:-The equivalent time at temp T delivered to a unit of product
calculated using a specified value of z. F0 is equivalent time at temp of 121oC delivered
to a unit of product calculated using a specified value of z equal to 10. .its current
application is limited to steam sterilization.

F0=Δt Σ10 (T-T0)/Z T0;REFERENCE TEMP T;PRODUCT TEMP

Where Δt time interval between product temp measurements T.

▶ II) Media fill (process simulation test)

Evaluation of the environment along with the process, the operator and the equipment is
the media fill.

METHODS USED FOR VALIDATION

1) PHYSICAL METHODS
2) BIOLOGICAL INDICATORS
3) CHEMICAL METHODS
1) PHYSICAL METHODS
A.MEASURING DEVICES FOR HEAT:
For heat sterilizers, including autoclaves and dry heat sterilizers, determination &
recording of heat distribution within the chamber is of prime importance. The most
commonly used equipment consists of the resistance temperature detectors or
thermocouples measuring systems.
B.PRESSURE SENSOR
They should be chosen to fit the purpose of the instrument
C.Dosimeters for radiation sterilization
Perspex (Polymethacrylate) strips,

2) BIO-INDICATORS FOR VALIDATION OF STERILIZER

 Bio-indicators are preparations of microorganisms innoculated into the product,
adsorbed onto paper strips or glass beads, or suspended in a liquid medium and
sealed in ampoules for steam sterilization.

Procedures Species
Steam B. stearothermophilus
Cl. Sporogenes
Dry heat B.Subtills var. niger
Gas B. subtilis var. niger
Radiation B. pumilus,
B. cereus,
B. Sphaericus

3) CHEMICAL METHODS
based on ability of steam,heat,etc. to alter the chemical &physical characteristic of
substances.
 BROWN‟S TUBE  KLINTEX PAPER
 BOWIE DICK TEST  TEST TABLET
 FILTER PAPER STRIP

VALIDATION PLANS
For all sterilization procedures a coherent validation plan should be developed.
This plan should include:
QUALIFICATION OF THE STERILIZER
Installation qualification
Operation qualification
PRODUCT VALIDATION
Compatibility of the sterilization process with the given equipment
Development of the sterilization cycle
PROCESS VALIDATION
Specification of the data to be routinely collected & evaluated
Revalidation, i.e. specification of frequency and types of operation
to be repeated at regular intervals.
VALIDATION OF STEAM STERILIZER
1. Mechanically checking, upgrading & qualifying the Sterilizer unit.
2. The autoclave must be checked for quality, Dependability, proper installation &
lack of Contamination.
3. All instruments used in studying the steam sterilizer such as
temperature & pressure instrumentations must be calibrated.

RESULTS SUMMARY SHEET

1. Autoclave identification number:___________
2. Location: Building_____________________
3. Validation date:_______________________
4. Revalidation date:_____________________
5. Description of process validated____________
6. Temperature set point for validation_________
7. Temperature range for validation___________
8. Cycle validated_______________________
9. Validation records stored in archives_________
10. Revalidation records stored in archives________

QUESTIONS BANK
1.Give qualitative and quantitative lay out, manufacturing steps with
suitable equipments, important IPQC parameter, and packaging records
and post marketing surveillance reports for sterile products. (JULY- 2004 )
2. Discuss the department layout, schedule „M‟ requirement, validation
parameters, and PMS report for sterile LVPs?(29th September, 2004 )
3. What is the importance of Bio film removal on product quality?( march 2005)
4. What are the facilities, environment control and air handling system with different
types of classification ?( march 2005)
5. Discuss clean room concept and level of protection in brief?(2005)
6. How will you evaluate the package for different sterile DF? Give the legal
requirement for keeping their records and reports? (sep 2006)
7. Discuss the qualities, national, international standard for clean room?
Discuss the pressure differential in the pharma. plant ?(sep 2006)
8. Validation of the steam sterilizer and importance of the D, Z, F value?
(sep 2006)
REFERENCES
▶ www.GMP.online.coms
▶ Pharmaceutical dosage forms (Parenteral Preparation) by Kenneth Avis, Leon
Lachman, Vol-1.
▶ Pharmaceutical dosage forms (Parenteral Preparation) by Kenneth Avis, Leon
Lachman, Vol-2
▶ Drugs & Cosmetics Act 1940.
▶ www.ispc.org
▶ www.whqlibdoc.who.org
▶ The theory and Industrial pharmacy by Leon Lachman, Third edition
▶ Aseptic Pharmaceutical Manufacturing by M.J.Groves
▶ Pharmaceutical science by Remington, 20th edition
▶ Pharmaceutical process Validation by Loftus & Nash: 29-90.
▶ Sterile Pharmaceutical Manufacturing by Groves Gisan.
▶ www.fda.gov.
▶ American Journal of Hospital Pharmacy, Vol. 38, Issue 8, 1144-1147
▶ Dispensing for pharmaceutical students; 10 th edition; by:-S J Carton
▶ www.dwscientific.co.uk
▶ www.pharmamachines.com
▶ www.bascotech.com
▶ www.getthatmag.com
▶ www.fabtecheng.com
▶ www.ahind.com
▶ www.nkambica.com