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 o o o o o Previously sterilized container are taken. Filters having nominal pore size 0.22 m or less are used for filtration Remove bacteria and moulds but Not viruses & Mycoplasmas Double filter layer or second filtration No fibre shedding or asbestos filters 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 its 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:

1). Type of production

Depends on

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 Square meter Production Warehouse Utility Quality control Administration Maintenance Employee services Security Total 11,094 7,606 1,716 1,716 1,018 1,014 1,014 39 24,607 Area Percentage 45.1 30.9 4.1 7.0 4.1 4.5 4.1 0.9 100.0

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: Zone 7:- Filling line Zone 6:- Filling area Zone 5:- Weighing, mixing & transfer area. Zone 4:- Clean area Zone3:-General production Zone 2:- Warehouse Zone 1:- Exterior

1st. Zones as per Gazette of India

WHITE BLACK GRAY

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. Adequate sterilization procedure Barrier 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 Adequate material control and selection Material Adequate sterilization and filtration procedure Adequate air filtration system Air Adequate monitoring of air cleanliness level. Adequate air system validation procedure.
AIR HANDLING SYSTEM (AHU)

CRITICAL AREAS Aseptic feeling area Sterilized component unloading area Change room

GRADES B C 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 0.5m 3520 35,200 3,52,000 35,20,000 5m 29 293 2,930 29,300 0.5m 3500 3,52,000 35,20,000 Not (c) defined 5m 29 2,930 29,300 Not defined (c)

A B (a) C (a) D (a)

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

TABLE III Types of operations to be carried out in the various Grades for terminally sterilized products. Grade A C Types of operations for terminally sterilized products. Filling of products, which are usually at risk Placement of filling and sealing machines, preparation of solutions when usually at risk. Filling of product when unusually at risk. 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: >0.5 m Clean Area ISO 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 0.5m 5.0m 0.5m 5.0m 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 Microbiological Limit Clean Area <0.5 m <0.5 m 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 A (LAF) B C D 0.5m 3,500 3,500 3,50,000 3,500,000 5.0m 0 0 2,000 20,000 Microorganisms <1 5 100 500

4. Air Classifications as per ISO Grade ISO Class A 5 100 B 6 1000 C 7 10000 D 8 100000

Particle/cum 3.5 35 350 3500

Class(SI) M 3.5(filling) M 4.5 M 5.5 (preparation) 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 nonporous, 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

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

Selected ISO 209 airborne particulate cleanliness classes for cleanrooms and clean zones.
numbers (N) ISO ISO ISO ISO ISO ISO ISO ISO ISO 1 2 3 4 5 6 7 8 9 Maximum concentration equal to and larger than 0.1m m 0.2m m 10 2 100 24 1 000 237 10 000 2 370 100 000 23 700 1 000 000 237 000 limits (particles/m^3 of air) for particles the considered sizes shown below 0.3m m 0.5m m 1m m 5.0m m 10 102 1 020 10 200 102 000 4 35 352 520 200 000 000 000

3 35 352 3 520 35 200

8 83 832 8 320

8 83 832 320 200 000 000

29 293 2 930 29 300 293 000

QUALITATIVE

LAYOUT

OF

PARENTERAL

MANUFACTURING

(circular

flow)

QUALITATIVE LAYOUT OF PARENTERAL MANUFACTURING (parallel flow)

LAYOUT FOR TERMINAL STERILIZATION

Pdt. Exit Unidirectional Clean Zone

Soln Prepn Area

Material Entry

Aseptic Filling zone Clean Changing Room Oven Auto clave Aseptic Receiving Area Equipment & Component Prepn Area

Entry

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 (polyvinylidene 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 Millipores 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. Millipores 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 Pyrogen test Particulate test

Sterility test. Uniformity of weight. 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. Doesnt 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 10m 5 particulates of 25 m 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 Temp. Humidity (in substances) case of hygroscopic

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 no Ingredients Item code Standards ATR no Label claim Qty required Qty 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 started

oven

Desired temp. attained

Temp.

Time when oven 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 started

Filling completed

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.

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=t 10 (T-T0)/Z T0;REFERENCE TEMP T;PRODUCT TEMP Where t 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

Z=(T2-T1/LOGD1-LOGD2)

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 Steam Dry heat Gas Species B. stearothermophilus Cl. Sporogenes B.Subtills var. niger 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. BROWNS 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