Injectable

Injectable Preparation

1. Parenteral Preparation Prepared By: Roshni S. Vora PhD Research Scholar Guided By: Dr. Yamini D. Shah Associate Professor 1 L. M. College Of Pharmacy-Ahmedabad

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4. 4 Para-other than ; enteron -intestine According to I.P " parenterals are injectable preparations, sterile products intended for administration by injection, infusion or implantation in to the body." Parenteral products are unique from any other type of pharmaceutical dosage form for the following reasons: Parenterals should be free of physical, chemical and biological contamination. Parenteral preparations are sterile, pyrogen free liquids (solutions, emulsions, or suspensions) or solid dosage forms packaged in either single-dose or multi dose containers. These preparations are administered through the skin or mucus membranes into internal body compartments. These includes any method of administration that does not involve passage through the digestive tract. Injectable solutions must be free from visible particulate matter. This includes reconstituted sterile powders.

5. Products should be isotonic, although strictness of isotonicity depends on the route of administration. Ophthalmic products, although not parenteral, must also be isotonic. Products to be administered by bolus injection by routes other than intravenous (IV) should be isotonic, or at least very close to isotonicity . IV infusions must be isotonic. All products must be stable, not only chemically and physically like all other dosage forms, but also ‘stable’ microbiologically (i.e., sterility, freedom from pyrogenic and visible particulate contamination must be maintained throughout the shelf life of the product). Products must be compatible, if applicable, with IV diluents, delivery systems, and other drug products co administered. a small-volume therapeutic injection (SVI), such as an antibiotic, to large-volume injections (LVIs), such as 1000 mL of 0.9% sodium chloride solution, to avoid the discomfort, for the patient, of a separate injection. 5

6. 6 During hospital pharmacy practice a pharmacist have been undertaken and more information has been gained, it has been shown that knowledge of variable factors, such as pH and the ionic character of the active constituents, aids substantially in understanding and predicting potential incompatibilities. Kinetic studies of reaction rates may be used to describe or predict the extent of degradation. a thorough study should be undertaken of each therapeutic agent in combination with other drugs and IV fluids, not only of generic, but also of commercial preparations, from the physical, chemical, and therapeutic aspects. Types of Processes Small scale dispensing - usually one unit at a time : Hospital Pharmacy Large scale - manufacturing in which hundreds of thousands of units may constitute one lot of product : In pharmaceutical Industry

7. 7 Parenteral products from non-sterile components in the highly technologically advanced plants of the pharmaceutical industry, using cGMP principles includes following Ensuring that the personnel responsible for assigned duties are capable and qualified to perform them. Ensuring that ingredients used in compounding the product have the required identity, quality, and purity. Validating critical processes to be sure that the equipment used and the processes followed ensure that the finished product has the qualities expected. Maintaining a production environment suitable for performing the critical processes required, addressing such matters as orderliness, cleanliness, asepsis, and avoidance of cross contamination. Confirming, through adequate quality-control procedures, that the finished products have the required potency, purity, and quality.

8. 8 Establishing, through appropriate stability evaluation, that the drug products retain their intended potency, purity, and quality, until the established expiration date. Ensuring that processes are always carried out in accord with established, written procedures. Providing adequate conditions and procedures for the prevention of mix-ups. Establishing adequate procedures, with supporting documentation, for investigating and correcting failures or problems in production or quality control. Providing adequate separation of quality-control responsibilities from those of production to ensure independent decision making. General Manufacturing Process The preparation of a parenteral product may encompass four general areas: Procurement and accumulation of all components in a warehouse area, until released to manufacturing; Processing the dosage form in appropriately designed and operated facilities; Packaging and labeling in a quarantine area, to ensure integrity and completion of the product; and Controlling the quality of the product throughout the process.

9. 9 Procurement:- selecting and testing according to specifications of the raw-material ingredients and the containers and closures for the primary and secondary packages Processing :- cleaning containers and equipment to validated specifications, compounding the solution (or other dosage form), filtering the solution, sanitizing or sterilizing the containers and equipment, filling measured quantities of product into the sterile containers, stoppering (either completely or partially for products to be freeze-dried), freeze-drying, terminal sterilization (if possible), and final sealing of the final primary container. Packaging normally consists of the labeling and cartoning of filled and sealed primary containers. Control of quality :- the incoming supplies, being sure that specifications are met. Each step of the process involves checks and tests to ensure the required specifications at the respective step are being met. The quality control unit is responsible for reviewing the batch history and performing the release testing required to clear the product for shipment to users.

10. Typical water storage and distribution schematic Water must be kept circulating Spray ball Cartridge filter 1 µm Air break to drain Outlets Hygienic pump Optional in-line filter 0,2 µm UV light Feed Water from DI or RO Heat Exchanger Ozone Generator Hydrophobic air filter & burst disc Water for Pharmaceutical Use 10

11. On site inspection : Walk through the system, verifying the parts of the system as indicated in the drawing Review procedures and "on site" records, logs, results Verify components, sensors, instruments Start with source water supply – follow whole system "loop“ Water treatment system inspection Dead Legs Filters Pipes And Fittings Ionic Beds Storage Tanks By-pass Lines Pumps UV Lights Sample Points R everse Osmosis Valves Heat Exchangers 11

12. Additional documentation to review Qualification protocols and reports Change control request (where applicable) Requalification (where applicable) QC and microbiology laboratory SOP for sampling Sampling There must be a sampling procedure Sample integrity must be assured Sample point and Sample size Testing Chemical testing Microbiological testing Test method Types of media used Incubation time and temperature Objectionable and indicator organisms 12

13. Suggested bacterial limits (CFU / mL ) Sampling location Target   Alert Action Raw water 200 300 500 Post multimedia filter 100 300 500 Post softener 100 300 500 Post activated carbon filter 50 300 500 Feed to RO 20 200 500 RO permeate 10 50 100 Points of Use 1 10 100 13

14. Water systems Water systems (water used for product compounding or final rinsing of surfaces which will contact the product), are typically operated in the temperate ranges hot, ambient and cold: Hot systems are operated above 70 °C and Cold systems are operated in the range between 2°C and 10°C Ambient systems are operated in the range of the environment in which the system is located. Purified water systems can be operated at any temperature. WFI systems are preferably operated hot and with continuous recirculation to control microbial growth. When WFI is stored and distributed at cold or ambient temperatures, special precautions are taken to prevent the ingress and proliferation of microbial contaminants, as e.g. appropriate sanitization cycles which are defined as part of the system qualification 14

15. Pharmaceutical water - used for product compounding or final rinsing of surfaces - exists in different (compendia) qualities such as: Preparation of the different types of water must be performed according to current USP and/or European Pharmacopoeia requirements and - if applicable - according to other pharmacopoeias (e.g. Japanese) and local requirements. http://www.nayagara.net/ 15

16. Monitoring – General Requirements Water systems undergo periodic monitoring of the specified required characteristics . The monitoring program is based on the results of the qualification work and/or according to the results of a risk assessment. Monitoring is performed according to written procedures , describing in sufficient detail the responsibilities for sampling, the sampling sites, and the sampling frequencies. Typical minimum sampling frequencies for process systems are described in slide 11-12. Higher or lower sampling frequencies for specific processes or products are justified according to the results of a risk assessment. 16

17. Sampling Sampling sites must be selected based on a risk evaluation and / or as result of the initial qualification . Samples have to be taken from representative locations within the distribution and processing system . Selection of sampling sites must not compromise the quality (e.g.: microbiological status) of the system being monitored. The sampling plan has to be dynamic allowing for adjustments to sampling frequency and locations based on system performance trends. When routine monitoring points are reduced or increased, the reason for the change has to be documented. Sampling practice must simulate the use of a process system during manufacturing, for example where water for manufacturing is delivered through a hose, sampling has to be performed through this hose. 17

18. Monitoring – Typical Minimum Sampling & Testing Frequencies 1 18

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20. 20 Water For Injection (WFI) The source water contaminated with natural suspended mineral and organic substances, dissolved mineral salts, colloidal material, viable bacteria, bacterial endotoxins , industrial or agricultural chemicals, and other particulate matter. The degree of contamination varies with the source and will be markedly different, whether obtained from a well or from surface sources, such as a stream or lake. The source water must be pretreated by one or a combination of the following treatments: chemical softening, filtration, deionization, reverse osmosis, purification.

21. 21 A conventional still consists of a boiler (evaporator), containing feed water ( distilland ); a source of heat to vaporize the water in the evaporator; a headspace above the level of distilland , with condensing surfaces for refluxing the vapor, thereby returning nonvolatile impurities to the distilland ; a means for eliminating volatile impurities (demister/separation device) before the hot water vapor is condensed; and a condenser for removing the heat of vaporization, thereby converting the water vapor to a liquid distillate. The specific construction features of a still and the process specifications have a marked effect on the quality of distillate obtained from a still. Several factors must be considered in selecting a still to produce WFI: The quality of the feed water will affect the quality of the distillate. For example, chlorine in water, especially, can cause or exacerbate corrosion in distillation units, and silica causes scaling within. Controlling the quality of the feed water is essential for meeting the required specifications for the distillate.

22. 22 The size of the evaporator will affect the efficiency. It should be large enough to provide a low vapor velocity, thus, reducing the entrainment of the distilland either as a film on vapor bubbles or as separate droplets. The baffles (condensing surfaces) determine the effectiveness of refluxing. They should be designed for efficient removal of the entrainment at optimal vapor velocity, collecting and returning the heavier droplets contaminated with the distill and. Redissolving volatile impurities in the distillate reduces its purity. Therefore, they should be separated efficiently from the hot water vapor and eliminated by aspirating them to the drain or venting them to the atmosphere. Contamination of the vapor and distillate from the metal parts of the still can occur. Present standards for high-purity stills are that all parts contacted by the vapor or distillate should be constructed of metal coated with pure tin, 304 or 316 stainless-steel, or chemically resistant glass.

23. 23 There are two basic types of WFI distillation units—the vapor compression still and the multiple effect still. Compression Distillation The vapor-compression still, primarily designed for the production of large volumes of high-purity distillate with low consumption of energy and water. To start, the feed water is heated from an external source in the evaporator to boiling. The vapor produced in the tubes is separated from the entrained distilland in the separator and conveyed to a compressor that compresses the vapor and raises its temperature to approximately 107°C. It then flows to the steam chest, where it condenses on the outer surfaces of the tubes containing the distilland ; the vapor is, thus, condensed and drawn off as a distillate, while giving up its heat to bring the distilland in the tubes to the boiling point. Vapor-compression stills are available in capacities from 50 to 2800 gal/hr.

24. 24 vapor-compression still

25. 25 Multiple-Effect Stills The multiple-effect still is also designed to conserve energy and water usage. In principle, it is simply a series of single-effect stills or columns running at differing pressures where phase changes of water take place. A series of up to seven effects may be used, with the first effect operated at the highest pressure and the last effect at atmospheric pressure. Steam from an external source is used in the first effect to generate steam under pressure from feed water; it is used as the power source to drive the second effect. The steam used to drive the second effect condenses as it gives up its heat of vaporization and forms a distillate. This process continues until the last effect, when the steam is at atmospheric pressure and must be condensed in a heat exchanger.

26. 26 The capacity of a multiple-effect still can be increased by adding effects. The quantity of the distillate will also be affected by the inlet steam pressure; thus, a 600-gal/hr unit designed to operate at 115 psig steam pressure could be run at approximately 55 psig and would deliver about 400 gal/hr. These stills have no moving parts and operate quietly. They are available in capacities from about 50 to 7000 gal/hr. Multiple-Effect Stills

27. 27 Reverse Osmosis (RO) It is a natural process of selective permeation of molecules through a semi permeable membrane separating two aqueous solutions of different concentrations is Reverse osmosis. Pressure, usually between 200 and 400 psig, is applied to overcome osmotic pressure and force pure water to permeate through the membrane. Membranes, usually composed of cellulose esters or polyamides, are selected to provide an efficient rejection of contaminant molecules in raw water. The molecules most difficult to remove are small inorganic molecules, such as sodium chloride. Passage through two membranes in series is sometimes used to increase the efficiency of removal of these small molecules and decrease the risk of structural failure of a membrane to remove other contaminants, such as bacteria and pyrogens .

28. 28 Several WFI installations utilize both RO and distillation systems for generation of the highest quality water. Since feedwater to distillation units can be heavily contaminated and, thus, affect the operation of the still, water is first run through RO units to eliminate contaminants. Whichever system is used for the preparation of WFI, validation is required to be sure that the system, consistently and reliably, produces the chemical, physical, and microbiological quality of water required. Such validation should start with the determined characteristics of the source water and include the pretreatment, production, storage, and distribution systems. Storage and Distribution The rate of production of WFI is not sufficient to meet processing demands; therefore, it is collected in a holding tank for subsequent use. In large operations, the holding tanks may have a capacity of several thousand gallons and be a part of a continuously operating system. In such instances, the USP requires that the WFI be held at a temperature too high for microbial growth, normally a constant 80°C.

29. 29 Such a system requires frequent sanitization to minimize the risk of viable microorganisms being present. The stainless-steel storage tanks in such systems are usually connected to a welded stainless steel distribution loop, supplying the various use sites with a continuously circulating water supply. The tank is provided with a hydrophobic membrane vent filter capable of excluding bacteria and nonviable particulate matter. Such a vent filter is necessary to permit changes in pressure during filling and emptying. The construction material for the tank and connecting lines is usually electro polished 316L stainless steel with welded pipe. The tanks also may be lined with glass or a coating of pure tin. Such systems are very carefully designed and constructed and often constitute the most costly installation within the plant. When the water cannot be used at 80°C, heat exchangers must be installed to reduce the temperature at the point of use. Bacterial retentive filters should not be installed in such systems, due to the risk of bacterial buildup on the filters and the consequent release of pyrogenic substances.

30. 30 Purity Although certain purity requirements have been alluded to, the USP and EP monographs provide the official standards of purity for WFI and Sterile Water for Injection (SWFI). The chemical and physical standards for WFI have changed in the past few years. The only physical/chemical tests remaining are the new total organic carbon (TOC), with a limit of 500 ppb (0.5 mg/L), and conductivity , with a limit of 1.3 μS /cm at 25°C or 1.1 μS /cm at 20°C. The pH requirement of 5 to 7 in previous revisions has been eliminated. The SWFI requirements differ in that, since it is a final product, it must pass the USP Sterility Test. WFI and SWFI may not contain added substances. Bacteriostatic Water for Injection (BWFI) may contain one or more suitable antimicrobial agents in containers of 30 mL or less.

31. Sampling Point & Point of Use Preparation Vessel point of use point of use = sampling point sampling point sampling point 31

32. UV Disinfection unit Points of Use Return Storage Tank Mixed ion exchange bed Particle Filter Pump Ventilation Filter Particle Filter UV disinfection unit Feed Water Inflow Monitoring – Typical Minimum Sampling & Testing Frequencies 32

33. 33 System-specific sampling points Depend on the construction and the technical conditions of the installation or system (e.g. Begin and end [=return] of the distribution system). Relevant sampling points (API/potable water) Evenly distributed throughout the plant (e.g. One sampling point per floor). Critical points of use Depend on the individual manufacturing process: - For cleaning product contacting surfaces - During final crystallization of APIs - for final rinsing of product contacting - For aqueous granulation processes) Selected sampling points Not directly relevant for the production (e.g. In cleaning/washing areas) Selected sampling points for endotoxin testing Where purified water is ultra-filtered to meet the endotoxin specification) Monitoring – Typical Minimum Sampling & Testing Frequencies

34. Return Inflow Monitoring – Typical Minimum Sampling & Testing Frequencies 34

35. 35 System-specific sampling points Depend on the construction and the technical conditions of the installation or system (e.g. Begin and end [=return] of the distribution system). Relevant sampling points (API/potable water) Evenly distributed throughout the plant (e.g. One sampling point per floor). Critical points of use Depend on the individual manufacturing process: - For cleaning product contacting surfaces - During final crystallization of APIs - for final rinsing of product contacting - For aqueous granulation processes) Selected sampling points Not directly relevant for the production (e.g. In cleaning/washing areas) Selected sampling points for endotoxin testing Where purified water is ultra-filtered to meet the endotoxin specification) Monitoring – Typical Minimum Sampling & Testing Frequencies

36. process-relevant but not critical: organic coating critical: aqueous coating Monitoring – Typical Minimum Sampling & Testing Frequencies 8 36

37. 37 System-specific sampling points Depend on the construction and the technical conditions of the installation or system (e.g. Begin and end [=return] of the distribution system). Relevant sampling points (API/potable water) Evenly distributed throughout the plant (e.g. One sampling point per floor). Critical points of use Depend on the individual manufacturing process: - For cleaning product contacting surfaces - During final crystallization of APIs - for final rinsing of product contacting - For aqueous granulation processes) Selected sampling points Not directly relevant for the production (e.g. In cleaning/washing areas) Selected sampling points for endotoxin testing Where purified water is ultra-filtered to meet the endotoxin specification) Monitoring – Typical Minimum Sampling & Testing Frequencies

38. selected: washing/cleaning Monitoring – Typical Minimum Sampling & Testing Frequencies 10 38

39. Physical, Chemical and Microbiological Testing Parameters TEST MODULE SPECIFICATIONS REFERENCE Appearance Clear, Colorless Liquid Ph. Eur. and USP Conductivity < 1.1 µS / cm (20 o C) or values as per Ph. Eur. Ph. Eur. and USP Ammonium Not more intense in color than reference, corresponding to < 0.05 ppm JP Chlorides Complies to the test JP Nitrates and Nitrites Not more intense in color than reference, corresponding to < 0.2 ppm Ph. Eur. and JP Total Organic Carbon (TOC) < 0.5 mg / L Ph. Eur. and USP Oxidizable Substances Complies to the test JP Acidity or Alkalinity Complies to the test JP Heavy Metals Not more intense in color than reference, corresponding to < 0.1 ppm Ph. Eur. and JP Sulfates Complies to the test JP Residue on Evaporation < 10 ppm JP Microbial Contamination max. 10 cfu / 100 ml Ph. Eur. and USP Bacterial Endotoxins < 0.25 EU / ml Ph. Eur. and JP 39

40. Test Methods and Method Requirements All methods must be performed according to current USP and/or European Pharmacopoeia and , if applicable other pharmacopoeia and/or local requirements (e.g. in case of potable water). All methods or culture media have to be suitable to detect microorganisms that may be present. The cultivation conditions, are selected to be appropriate for the specific growth requirements of microorganisms to be detected, for example: Total aerobic count can be obtained by incubating at 30 to 35 °C for not less than three days Suitable culture media (low nutrient medium) is used for monitoring of water systems (30 to 35°C, at least 5 days). Testing of viable monitoring samples is performed under aerobic conditions unless there are indications that the process is at risk for contamination with anaerobic microorganisms. It must be assured that cleaning or disinfection agents remaining on surfaces sampled does not interfere with microbial recovery when methods using culture media are applied. 40

41. Alert and Action Level in Microbiological Monitoring An Alert level in microbiological monitoring is that level of microorganisms that shows significant differences from normal operating conditions. Alert levels are usually based upon historical information gained from the routine operation of the process in a specific controlled environment . In a new facility, these levels are based on prior experience from similar facilities/ processes . Alert levels are re-examined and – if necessary – re-set at an established frequency. Trends that show a deterioration of the environmental quality require respective CAPAs. An Action level is that specification level of microorganisms or particles that when exceeded requires immediate follow-up and, if necessary, corrective action. Common procedure of setting alter level based on a set of at least 12 months data: 95 % of all results < alert level AND 5 % of all results ≥ alert level Typically, the initial alert level is set to… 50 % of the action level 41

42. Procedures when an Alert level is exceeded Exceeding the Alert level does not necessarily require a definitive corrective action, but it prompts at least documented follow-up measures, as established in a local procedure. These measures include but are not limited to the following: Comparison with results obtained concurrently with other related sampling points. Comparison with historical data from the same sampling point. If possible re-sampling of the affected sampling point; routine sample(s) taken from the affected point(s) within this period can be considered as resample. no further Alert level => no additional action again Alert level exceeding => repetition of re-sampling according to the procedure described above consecutive Alert level exceeding => escalation of measures (e.g. following the procedures of exceeding an Action level) 42

43. Procedures when an Action level is exceeded As soon as an Action level excursion is reported, “immediate corrective actions” and an investigation have to be performed as described in a local procedure. An evaluation of the potential impact this exceeding has on manufactured products has to be made. When a definitive cause for the excursion can be determined immediately, specific corrective actions are performed before re-sampling starts. Re-sampling of the affected points has to be performed immediately after the implementation of “immediate / specific corrective actions”. Monitoring critical sampling points includes routine identification of microorganisms to the species (or, where appropriate , genus) level at least when Alert and Action Levels are exceeded . 43

44. Documentation and Trending of Monitoring Data All monitoring activities are documented properly (typically on form sheets which are laid down in SOPs). The results from critical sampling locations must be assignable to the respective activity at the time of sampling (important in case of batch-related monitoring, i.e. the environmental monitoring data must have a formal linkage to product release as defined by procedures). Monitoring data must be summarized on a periodic basis and issued to the responsible senior management on a periodic basis (e.g. via Product Quality Review). Based on this summary, trends have to be evaluated and corrective action to be defined, if appropriate. 44

45. Q & A Purified water systems have to be sampled (monitored) daily for microbiological testing. For chemical/physical testing of water systems, it is highly recommended to define the last point of use (return) in the system as a routine sampling point. Any Alert Level excursion initiates an immediate procedure . Purified water with endotoxin limit is required for the final purification of a non-sterile API to be used in a sterile parenteral Drug Product. 45

46. Process Systems – General Qualification Provisions Qualification is required for any process system (e.g. Water, Nitrogen, Clean Steam, Compressed Air) … that is involved in the manufacture of APIs (beginning with the regulatory starting materials), Drug Products or intermediates that may affect testing results of an API, Drug Product or intermediate, that is involved in final cleaning processes , where the utility supplied directly contacts an API, Drug Product or intermediate , where the utility supplied comes in contact with surfaces that have direct contact with APIs, Drug Products or intermediates , … and, therefore, could have an impact on the quality of the API, Drug Product or intermediate. 46

47. Before beginning the qualification of a process system, the following documentation has to be available: 47

48. Test Items for Qualification of Process Systems Following table outlines parameters and aspects to be checked, evaluated and tested within the qualification study of a process system, provided that these are relevant for the particular qualification (see following slide) . 48

49. Based on this table, the qualification team determines by means of a risk-based approach … the sampling points, e.g. by answering the following questions… Which points of use are critical ? Which points of use are system-specific ? Is it necessary to realize a particular sampling point (due to the unattainability of the point of use) ? Usually, selected sampling points include… significant points of use return loop points prior to and after each significant treatment step storage tank 49

50. Requalification of Process Systems For-Cause Requalification Generally, in case of changes or modifications, the same test items apply for requalification as for initial qualification . However, based on a risk evaluation, the extent of a requalification may be reduced in comparison to the initial qualification. Periodic Requalification The following periodic requalification intervals apply: However , t he regular evaluation of the existing documentation such as… monitoring data, quarterly reports, change documentation, logbooks, maintenance/servicing documentation, technical reports … equates to periodic requalification, provided that relevant requalification item are appropriately covered. “streamlined” requalification approach 50

51. In case of water systems, the qualification process entails a three-phase approach in order to satisfy the objective of demonstrating the reliability and robustness of the system in service over an extended period. 51

52. Phase 1: Initial phase, usually taking 2 to 4 weeks, serves to establish operating parameters and procedures, Does not end until the system operates stable and within the required ranges, Might be shortened in case of modifications to a water system already in use . Phase 2: Short-term control phase usually taking 2 to 4 weeks . Before water is permitted to be used for pharmaceutical purposes, an interim qualification report is required, documenting the successful completion of Phase 2. However, water can also be used for pharmaceutical purposes during this phase, provided that the respective batches are not released until the interim qualification report has been finalized . Phase 3: Long-term control phase usually taking 1 year, serves to demonstrate continuous and consistent operation irrespectively of external and seasonal variations. Physico-chemical properties, microbial counts (as well as endotoxin where required) are monitored and evaluated at close intervals, Where the season affects the quality of the feed water (e.g. potable water), sampling should be increased. Phase 3 ends with the preparation of the final Qualification Report. 52

53. 53 Water-Miscible Vehicles These solvents are used to solubilize certain drugs in an aqueous vehicle and to reduce hydrolysis. The most important solvents in this group are ethyl alcohol, liquid polyethylene glycol, and propylene glycol. Ethyl alcohol is used in the preparation of solutions of cardiac glycosides and the glycols in solutions of barbiturates, certain alkaloids, and certain antibiotics. Such preparations are given intramuscularly. There are limitations with the amount of these co-solvents that can be administered, due to toxicity concerns, greater potential for hemolysis , and potential for drug precipitation at the site of injection.

54. 54 Non-Aqueous Vehicles The most important group of non-aqueous vehicles is the fixed oils. The USP provides specifications for such vehicles, indicating that the fixed oils must be of vegetable origin so they will metabolize, will be liquid at room temperature, and will not become rancid readily. The USP also specifies limits for the free fatty acid content, iodine value, and saponification value (oil heated with alkali to produce soap, i.e., alcohol plus acid salt). The oils most commonly used are corn oil, cottonseed oil, peanut oil, and sesame oil. Fixed oils are used as vehicles for certain hormone (e.g., progesterone, testosterone, deoxycorticicosterone) and vitamin (e.g., Vitamin K, Vitamin E) preparations. The label must state the name of the vehicle, so the user may beware in case of known sensitivity or other reactions to it.

55. 55 Solutes Care must be taken in selecting active pharmaceutical ingredients and excipients to ensure their quality is suitable for parenteral administration. A low microbial level will enhance the effectiveness of either the aseptic or the terminal sterilization process used for the drug product. It is now a common GMP procedure to establish microbial and endotoxin limits on active pharmaceutical ingredients and most excipients. Chemical impurities should be virtually nonexistent in active pharmaceutical ingredients for parenterals , because impurities are not likely to be removed by the processing of the product. Depending on the chemical involved, even trace residues may be harmful to the patient or cause stability problems in the product. Therefore, manufacturers should use the best grade of chemicals obtainable and use its analytical profile to determine that each lot of chemical used in the formulation meets the required specifications.

56. 56 Reputable chemical manufacturers accept the stringent quality requirements for parenteral products and, accordingly, apply good manufacturing practices to their chemical manufacturing. Examples of critical bulk manufacturing precautions include: Using dedicated equipment or properly validated cleaning to prevent cross-contamination and transfer of impurities; Using WFI for rinsing equipment; Using closed systems, wherever possible, for bulk manufacturing steps not followed by further purification; and Adhering to specified endotoxin and bioburden testing limits for the substance. Added Substances The USP includes in this category all substances added to a preparation to improve or safeguard its quality. An added substance may: Increase and maintain drug solubility. Examples include complexing agents and surface active agents. The most commonly used complexing agents are the cyclodextrins , including Captisol . The most commonly used surface active agents are polyoxyethylene sorbitan monolaurate ( Tween 20) and polyoxyethylene sorbitan monooleate ( Tween 80).

57. 57 Provide patient comfort by reducing pain and tissue irritation, as do substances added to make a solution isotonic or near physiological pH. Common tonicity adjusters are sodium chloride, dextrose, and glycerin. Enhance the chemical stability of a solution, as do antioxidants, inert gases, chelating agents, and buffers. Enhance the chemical and physical stability of a freezedried product, as do cryoprotectants and lyoprotectants . Common protectants include sugars, such as sucrose and trehalose , and amino acids, such as glycine . Enhance the physical stability of proteins by minimizing self-aggregation or interfacial induced aggregation. Surface active agents serve nicely in this capacity. Minimize protein interaction with inert surfaces, such as glass and rubber and plastic. Competitive binders, such as albumin, and surface active agents are the best examples.

58. 58 Protect a preparation against the growth of microorganisms. The term ‘preservative’ is sometimes applied only to those substances that prevent the growth of microorganisms in a preparation. However, such limited use is inappropriate, being better used for all substances that act to retard or prevent the chemical, physical, or biological degradation of a preparation. Although not covered in this chapter, other reasons for adding solutes to parenteral formulations include sustaining and/or controlling drug release (polymers), maintaining the drug in a suspension dosage form (suspending agents, usually polymers and surface active agents), establishing emulsified dosage forms (emulsifying agents, usually amphiphilic polymers and surface active agents), and preparation of liposomes (hydrated phospholipids). Although added substances may prevent a certain reaction from taking place, they may induce others. Not only may visible incompatibilities occur, but hydrolysis, complexation , oxidation, and other invisible reactions may decompose or otherwise inactivate the therapeutic agent or other added substances. Therefore, added substances must be selected with due consideration and investigation of their effect on the total formulation and the container-closure system.

59. 59 Antimicrobial Agents The USP states that antimicrobial agents in bacteriostatic or fungistatic concentrations must be added to preparations contained in multiple-dose containers. The European Pharmacopeia requires multiple-dose products to be bacteriocidal and fungicidal. They must be present in adequate concentration at the time of use to prevent the multiplication of microorganisms inadvertently introduced into the preparation, while withdrawing a portion of the contents with a hypodermic needle and syringe. The USP provides a test for Antimicrobial Preservative Effectiveness to determine that an antimicrobial substance or combination adequately inhibits the growth of microorganisms in a parenteral product.

60. 60 Large-volume, single-dose containers may not contain an added antimicrobial preservative. Therefore, special care must be exercised in storing such products after the containers have been opened to prepare an admixture, particularly those that support the growth of microorganisms, such as total parenteral nutrition (TPN) solutions and emulsions. It should be noted that, although refrigeration slows the growth of most microorganisms, it does not prevent their growth. Buffers are used to stabilize a solution against chemical degradation or, especially for proteins, physical degradation (i.e., aggregation and precipitation) which might occur if the pH changes appreciably. Buffer systems should have as low a buffering capacity as feasible, so as not to significantly disturb the body’s buffering systems when injected. In addition, the buffer type and concentration on the activity of the active ingredient must be evaluated carefully. Buffer components are known to catalyze degradation of drugs. The acid salts most frequently employed as buffers are citrates, acetates, and phosphates. Amino acid buffers, especially histidine , have become buffer systems of choice for controlling solution pH of monoclonal antibody solutions.

61. 61 Because antimicrobials may have inherent toxicity for the patient, the USP prescribes maximum volume and concentration limits for those commonly used in parenteral products (e.g., phenylmercuric nitrate and thimerosal 0.01%, benzethonium chloride and benzalkonium chloride 0.01%, phenol or cresol 0.5%, and chlorobutanol 0.5%). Benzyl alcohol, phenol, and the parabens are the most widely used antimicrobial preservative agents used in injectable products. In oleaginous preparations, no antibacterial agent commonly employed appears to be effective. However, it has been reported that hexylresorcinol 0.5% and phenylmercuric benzoate 0.1% are moderately bactericidal. A physical reaction encountered is that bacteriostatic agents are sometimes removed from solution by rubber closures. Protein pharmaceuticals, because of their cost and/or frequency of use, are preferred to be available as multiple dose formulations (e.g., Human Insulin, Human Growth Hormone, Interferons , Vaccines, etc.). Phenoxyethanol is the most frequently used preservative in vaccine products. Single-dose containers and pharmacy bulk packs that do not contain antimicrobial agents are expected to be used promptly after opening or discarded.

62. 62 Anitoxidants are frequently required to preserve products, due to the ease with which many drugs, including proteins with methionine or cysteine amino acids conformationally exposed, are oxidized. Sodium bisulfite and other sulfurous acid salts are used most frequently. Ascorbic acid and its salts are also good antioxidants. The sodium salt of ethylenediaminetetraacetic acid (EDTA) has been found to enhance the activity of antioxidants, in some cases, by chelating metallic ions that would otherwise catalyze the oxidation reaction. Displacing the air (oxygen) in and above the solution, by purging with an inert gas, such as nitrogen, can also be used as a means to control oxidation of a sensitive drug. Process control is required for assurance that every container is deaerated adequately and uniformly. However, conventional processes for removing oxygen from liquids and containers do not absolutely remove all oxygen. The only approach for completely removing oxygen is to employ isolator technology, where the entire atmosphere can be recirculating nitrogen or another non-oxygen gas. Tonicity Agents are used in many parenteral and ophthalmic products to adjust the tonicity of the solution.

63. The water is contaminated as it passes through the valve Bacteria can grow when the valve is closed Stagnant water inside valve Bio contamination control techniques There should be no dead legs Water scours dead leg If D=25mm & distance X is greater than 50mm, we have a dead leg that is too long Dead leg section >1.5D Flow direction arrows on pipes are important Sanitary Valve D X Ball valves are unacceptable 63

64. Bio contamination control techniques Pressure gauges separated from system membranes Pipe work laid to fall (slope) – allows drainage Maintain system at high temperature (above 70 degrees Celsius) Use UV radiation Flow rate, life-cycle of the lamp Suitable construction material Periodic sanitization with hot water Periodic sanitization with super-heated hot water or clean steam Reliable Monitoring temperature during cycle Routine chemical sanitization using, e.g. ozone Removal of agent before use of water important 64

65. 65 Although it is the goal for every injectable product to be isotonic with physiologic fluids, this is not an essential requirement for small volume injectables administered intravenously. However, products administered by all other routes, especially into the eye or spinal fluid, must be isotonic. Injections into the subcutaneous tissue and muscles should also be isotonic to minimize pain and tissue irritation. The agents most commonly used are electrolytes and mono- or disaccharides. Cryoprotectants and Lyoprotectants are additives that servem to protect biopharmaceuticals from adverse effects, due tofreezing and/or drying of the product during freeze-dry processing. Sugars (non-reducing), such as sucrose or trehalose , amino acids, such as glycine or lysine, polymers, such as liquid polyethylene glycol or dextran , and polyols , such as mannitol or sorbitol , all are possible cryo - or lyoprotectants .

66. What do you need to know about Injection Sites ? Safety Considerations: When preparing multiple injections, always label the syringe immediately . Keep the medication container with the syringe Do not rely on memory to determine which solution is in which syringe. Carefully monitor the patient for any adverse effects for at least 5 minutes after administration of any medication. Handle multi-dose vials carefully and with aseptic technique so that medicines are not wasted or contaminated. 66

67. Primary Parenteral Routes Routes Usual volume ( mL ) Needle commonly used Formulation constraints Types of medication administered SVP Sub cutaneous 0.5-2 5/8 in. , 23 gauge Need to be isotonic Insulin, vaccines Intra muscular 0.5-2 1.5 in. , 23 gauge Can be solutions, emulsions, oils or suspensions Isotonic preferably Nearly all drug classes Intra venous 1-100 Vein puncture 1.5 in. , 20-22 gauge Solutions, emulsions and liposomes Nearly all drug classes LVP 101 and larger (infusion unit) Venoclysis 1.5 in. , 18-19 gauge Solutions and some emulsions Nearly all drug classes 67

68. No. ADVANTAGES DISVANTAGES 1. Quick onset Wrong dose or over dose can be fatal 2. Vomiting and unconscious patients can take Pain at site 3. Prolonged action by modified formulation Trained person required 4. Nutritive fluids can be given Expensive 5. Drugs with poor absorption or instability from GIT Necessity Of Aseptic Conditions In Production, Compounding And Administration When to Aspirate (IM & SC injection) The reason for aspiration before injection a medication is to ensure that the needle is not in a blood vessel. If blood appears in the syringe, withdraw the needle, discard the syringe, and prepare a new injection. When Not To Aspirate When administering SC heparin/ insulin, it is recommended that you do NOT aspirate. Because of the anticoagulant properties of heparin, aspiration could damage surrounding tissue and cause bleeding and bursting. 68

69. 69

70. Intramuscular Administration Administered into a muscle or muscle group Onset : variable Volume : up to 4ml Equipment :1-5 ml syringe, needle (18-23 g, ⅝ to 3 inch needle), alcohol swab Identify site and Cleanse site with alcohol Pull skin taut and Hold needle like “dart” Insert quickly at a 90° angle and Stabilize needle Aspirate for blood, If no blood, instill medication slow and steady and Quickly remove needle. DO NOT RECAP . Activate safety feature. Place needle in sharps container uncapped. Massage site with alcohol swab 70

71. Injection Sites – Deltoid Location: upper arm Landmarks: Acromion Process, axillary fold Muscle mass: triangle apex at axillary line and base of triangle 2-3 finger breadths below acromion process. Injection area: in the middle of the triangle / into belly of the muscle mass. Avoid Brachial artery & Radial nerve (BARN) Should not be used in infants or children because of the muscle’s small size. Injection volume should not exceed 1m l in the adult Use a 23-28 gauge, 5/8 to 1 inch needle Rarely used for hospitalized patients. Primarily used for immunizations. 71

72. Injection Sites – Ventrogluteal Location: lateral (ventral) side of the hip Landmarks: Iliac crest, anterosuperior illiac spine, greater trochanter of femur Muscle mass: Gluteus medius and minimus Injection area: opposing palm of hand over greater trochanter , middle finger pointed toward the iliac crest, index finger toward anterosuperior iliac spine. Inject into the triangle created by these fingers. No major vessels / nerves. 72

73. Injection Sites – Vastus Lateralis Location: anterolateral aspect of the thigh Landmarks: greater trochanter , lateral femoral condyle Muscle mass: vastus lateralis muscle Injection area: between one handbreadth below the greater trochanter and one handbreadth above the knee. Width of area is from the midline on the anterior surface of the thigh to midline on the lateral thigh. Best to inject into outer middle third of the thigh. No major vessels or nerves to avoid. Identify the greater trochanter and the lateral femoral condyle . Select the site using the middle third and the anterior lateral aspect of the thigh. 73

74. Injection Sites – Dorsal Gluteal Location: Upper lateral aspect of the buttock Landmarks: Posterior superior iliac spine,greater trochanter Muscle mass: Gluteus maximus muscle Injection area: Draw an imaginary line between the anatomic landmarks listed above. Administer the injection lateral and slightly superior (2 inches) to the midpoint of this line. Avoid the sciatic nerve & superior gluteal artery Most dangerous site because of sciatic nerve location 74

75. Z – track Seals the medication into the muscle tissue. Minimizes subcutaneous tissue irritation from tracking of the medication as the needle is withdrawn. Used more frequently now to decrease discomfort and pain. Used for irritating medications ( Vistaril ) and tissue staining meds (iron dextran – Imferon ). Use in ventrogluteal or dorsogluteal sites 75

76. Intradermal Administration Used for allergy and tuberculin skin testing Site : inner forearm (may use back and upper chest) Volume : 0.01-0.05 ml Equipment : TB syringe (1ml, 25-27g, ⅝ or ½ inch needle), alcohol swab. Administration angle : 10-15° DO NOT massage . DO NOT RECAP . Subcutaneous Administration Site : deep into tissue Administration angle : 45-90 ° DO NOT ASPIRATE. DO NOT RECAP . Common drugs given SC: Anticoagulants Lovenox ( enoxaparin sodium ) Insulin Erythropoitic agents Some Analgesics (- caine type drugs) 76

77. Why there is requirement for Regulation for Pharmaceutical packaging materials?? “A container closure system refers to the sum of packaging components that together contain and protect the dosage form . This includes primary packaging components and secondary packaging components , if the latter are intended to provide additional protection to the drug product. A packaging system is equivalent to a container closure system.” FDA 1999 “The primary packaging components (e.g. bottles, vials, closures, blisters) are in direct physical contact with the product , whereas the secondary components are not (e.g. aluminium caps, cardboard boxes).” WHO guideline “Guidelines on packaging for pharmaceutical products, Annex 9” Selecting types of packaging is a critical point because packaging components are the major source of particulate matter; pyrogen and stability problems. 77

78. Packaging Materials – Ideal Requirements Protect the preparation from environmental conditions Non-reactive with the product and so does not alter the identity of the product Does not impart tastes or odors to the product Nontoxic and Protect the dosage form from damage or breakage Presentation & information Packaging is essential source of information on medicinal product. Information provided to patient may include: Identification no. for dispensing records. Direction for use and Name and address of dispensers. Name, strength, quantity and Storage instructions. Compliance Design should be such that product can be easily administered in safer manner to patient. 78

79. Packaging Components: Primary components: Syringes , Ampoules , Flexible Bags, Bottles And Closures Secondary components : Cartons and Overlaps Associated components : Dosing Droppers And Calibrated Spoon 79

80. 80 Containers Glass Plastic Rubber Highly Resistant Borosilicate Glass Treated Soda lime Glass Regular Soda Lime Glass N.P (Non- parenteral ) Glass Type 4 is not used for parenteral packaging, others all are used for parenteral packaging. Plastic containers are used but they face following problems Permeation Sorption Leaching Softening To provide closure for multiple dose vials, IV fluid bottles, plugs for disposable syringes and bulbs for ophthalmic pipettes, rubber is the material of choice. Problems associated with rubber closures are Incompatibility Chemical instability Physical instability

81. Advantages Economical Superior protective qualities Readily available in a wide variety of sizes & shapes Excellent barrier against every element except light. Colored glass, especially amber, can give protection against light Disadvantages: Fragility Heavy Weight Glass Containers 81

82. Types Of Glass Type I: Borosilicate Glass Highly resistant glass. Composed principally of silicone dioxide and boron oxide . It is used to contain strong acids & alkalies as well as all types of solvents. It is more chemically inert than the soda-lime glass . Type II: Treated Soda-Lime Glass When glassware is stored for several months, especially in a damp atmosphere or with extreme temperature variations, the wetting of the surface by condensed moisture (condensation) results in salts being dissolved out of the glass. This is called “ blooming” or “weathering” & it gives the appearance of fine crystals on the glass. Type II containers are made of commercial soda-lime glass that has been treated with sulfur dioxide or other dealkalizers to remove surface alkali. The de-alkalizing process is known as “ sulfur treatment” which increases the chemical resistant of the glass. 82

83. Type III – Regular Soda-Lime Glass Containers are untreated & made up of commercial soda-lime glass of average or better-than-average chemical resistance. Type NP – General Purpose Soda-Lime Glass Containers made up of soda-lime glass are supplied for non- parenteral products, those intended for oral or topical use. 83

84. QUALITY CONTROL TESTS FOR GLASSES Chemical Resistant Of Glass Containers A) Powdered Glass Test: It is done to estimate the amount of alkali leached from the powdered glass which usually happens at the elevated temperatures. When the glass is powdered, leaching of alkali is enhanced, which can be titrated with 0.02N sulphuric acid using methyl red as an indicator Step-1 : Preparation of glass specimen : Few containers are rinsed thoroughly with purified water and dried with stream of clean air. Grind the containers in a mortar to a fine powder and pass through sieve no.20 and 50. Step-2 : Washing the specimen : 10gm of the above specimen is taken into 250 ml conical flask and wash it with 30 ml acetone. Repeat the washing, decant the acetone and dried after which it is used within 48hr. 84

85. Procedure: 10gm sample is added with 50ml of high purity water in a 250ml flask. Place it in an autoclave at 121⁰C±2⁰C for 30min.Cool it under running water. Decant the solution into another flask, wash again with 15ml high purity water and again decant. Titrate immediately with 0.02N sulphuric acid using methyl red as an indicator and record the volume. B) Water Attack Test: Principle involved is whether the alkali leached or not from the surface of the container. Procedure: Fill each container to 90%of its overflow capacity with water and is autoclaved at 121⁰C for 30min then it is cooled and the liquid is decanted which is titrated with 0.02N sulphuric acid using methyl red as an indicator. The volume of sulfuric acid consumed is the measure of the amount of alkaline oxides present in the glass containers . 85

86. 3) Arsenic Test: This test is for glass containers intended for aqueous parenterals. Wash the inner and outer surface of container with fresh distilled water for 5 min. 50ml.pipette out 10ml solution from combined contents of all ampoules to the flask. Add 10ml of HNO3 to dryness on the water bath, dry the residue in an oven at 130⁰C for 30min cool and add 10ml hydrogen molybdate reagent. Swirl to dissolve and heat under water bath and reflux for 25min. Cool to room temp and determine the absorbance at 840nm.Do the blank with 10ml hydrogen molybdate. The absorbance of the test solution should not exceed the absorbance obtained by repeating the determination using 0.1ml of arsenic standard solution (10ppm) in place of test soln. 86

87. 4 ) Thermal Shock Test: Place the samples in upright position in a tray. Immerse the tray into a hot water for a given time and transfers to cold water bath, temp of both are closely controlled. Examine cracks or breaks before and after the test. The amount of thermal shock a bottle can withstand depends on its size, design and glass distribution. Small bottles withstand a temp differential of 60 to 80⁰C and 1 pint bottle 30 to 40⁰C.A typical test uses 45C temp difference between hot and cold water. 5 ) Internal Bursting Pressure Test: The test bottle is filled with water and placed inside the test chamber. A scaling head is applied and the internal pressure automatically raised by a series of increments each of which is held for a set of time. The bottle can be checked to a preselected pressure level and the test continues until the container finally bursts. 87

88. 88 TESTS CONTAINER Powdered glass test Type I Type II Type III Water attack test Type II(100ml or below) Type II(above 100ml) 6) Leakage Test: Drug filled container is placed in a container filled with coloured solution (due to the addition of dye)which is at high pressure compared to the pressure inside the glass container so that the coloured solution enters the container if any cracks or any breakage is present.

89. 89 GLASS PROPERTIES: 1. Chemical properties Sodium/alkali leaching Alkali/Acid resistance 4. Optical properties Refractive index Dispersion Absorption Transmission Reflectivity 2. Electrical properties Volume resistivity Surface resistivity Dielectric constant 5. Thermal properties Coefficient of thermal expansion (CTE) Thermal conductivity Specific heat 3. Mechanical properties Stress Density Specific gravity

90. PLASTIC CONTAINERS Advantage: Ease of manufacturing High quality Extremely resistant to breakage Limitations: Permeation Leaching and Sorption Chemical reactivity COMMONLY USED POLYMERS LESS COMMONLY USED POLYMERS Polyethylene Polypropylene Polyvinyl chloride (PVC) Polystyrene Polymethyl methacrylate Polyethylene terephthalate Polytrifluoroethylene Aminoformaldehydes Polyamides 90

91. QUALITY CONTROL TESTS FOR PLASTICS Leakage test for Injectable & Non- Injectable (IP 1996) Fill the 10 containers with water and fit the closure. Keep them inverted at RT for 24 hours. No sign of leakage from any container. 91

92. Water vapor permeability test for injectable preparation(IP 1996) Fill the 5 containers with nominal volume of water and seal. Weigh the each container. Allow to stand for 14 days at RH of 60 + 5% at 20 c to 25 c. Reweigh the container. Loss of the weight in each container should not be more than 0.2%. 92

93. Collapsibility test for Injectable and Non- Injectable preparation(IP 199 6) This test is applicable for those containers, which have to be squeezed for the withdrawal of product. A container by squeezing yields at least 90% of its nominal contents at require flow rate at ambient temperature. Measurement of diffusion coefficient through plastic Stopcock A is first opened allowing evacuation of the system and this gives rise to a barometric leg in the manometer . As soon as A is closed, gas will diffuse through the membrane and mercury level will increase. The height of mercury level indicates diffusion of gas through the plastic. 93

94. Physicochemical tests USP specifies the extracting medium; otherwise purified water is maintained at 70 c. After the extraction following tests are performed : Non-volatile residue which measures organic and inorganic residue soluble in extracting medium . Heavy metals: This detects the presence of metals such as lead, tin, zinc etc . Buffering capacity: It measures the alkalinity/acidity of the extract . Compatibility test Compatibility components will not interact with the dosage form and may not show leaching. Regular screening is done by liquid chromatography, mass spectrometry, GC-MS etc. Other changes like PH shift, precipitation, discoloration, which may cause the degradation of the product should be evaluated. 94

95. Specific Types Of Closures:- Tamper-evident closures Child resistant closures Closure Characteristics of Good Pharmaceutical rubbers Good ageing qualities Satisfactory hardness and elasticity Resistance to sterilization conditions Impermeable to moisture and air Examples Butyl Rubbers Natural Rubbers Neoprene Rubbers Polyisoprene rubbers Silicone Rubbers 95

96. Closures Most vulnerable & critical component of a container .. Resistant & compatible with the product & the product/air space. Should not lead to undesired interactions between contents and environment. If closure is re-closable, it should be readily openable & effectively resealed. Capable of high-speed application for automatic production by high speed machines without loss of seal efficiency. Offers additional functions: aid-pouring, metering, administration, child resistance, tamper evidence, etc. 96

97. Available in various designs like… Threaded screw cap Crown cap Roll on closures Pilferproof closures Closures 97

98. CLOSURE Closures are devices and techniques used to close or seal a bottle, jug, jar, tube, can, etc Closures can be a cap, cover, lid, plug, etc The closure is normally the most vulnerable and critical component of a container An effective closure must prevent the contents from escaping and allow no substance to enter the container 98

99. Function Of A Closure Provide a totally hermetic seal Provide an effective seal which is acceptable to the products Provide an effective microbiological seal Characteristics Of Closure It should be resistant and compatible with the product If closure is of re closable type, it should be readily operable and should be re-sealed effectively It should be capable of high speed application It should be decorative and of a shape that blends with the main containers 99

100. Types Of Closures Closures are available in five basic designs Screw-on , threaded, or lug Crimp-on (crowns ) Press-on ( snap) Roll-on Friction. Many variations of these basic types exist, including Tamperproof Child resistant Dispenser applicators 100

101. THREADED SCREW CAP The screw cap provides physical and chemical protection to content being sealed. The screw cap is commonly made of metal or plastics. The metal is usually tinplate or aluminum, and in plastics, both thermoplastic and thermosetting materials are used. 101

102. LUG CAP The lug cap is similar to the threaded screw cap and operates on the same principle It is simply an interrupted thread on the glass finish, instead of a continuous thread Unlike the threaded closure, it requires only a quarter turn The cap is widely used in the food industry 102

103. CROWN CAPS This style of cap is commonly used as a crimped closure for beverage bottles and has remained essentially unchanged for more than 50 years 103

104. ROLL-ON CLOSURES The aluminum roll-on cap can be sealed securely, opened easily, and resealed effectively It finds wide application in the packaging of food, beverages, chemicals, and pharmaceuticals The roll-on closure requires a material that is easy to form, such as aluminum or other light-gauge metal 104

105. PILFER PROOF CLOSURES The pilfer proof closure is similar to the standard roll-on closure except that it has a greater skirt length When the pilfer proof closure is removed, the bridges break, and the bank remains in place on the neck of the container The torque is necessary to remove the cap. 105

106. TAMPER RESISTANT Resistance to tampering is required for some types of products. 106

107. DISPENSING A wide variety of convenience dispensing features can be built in to closures. Spray bottles and cans with aerosol spray have special closure requirements. 107

108. CHILD-RESISTANT Child-resistant packaging or C-R packaging has special closures designed to reduce the risk of children ingesting dangerous items Tamper-evident 108

109. CLOSURE LINES A liner may be defined as any material that is inserted in a cap to effect a seal between the closure and the container. Liners are usually made of a resilient backing and a facing material. The backing material must be soft enough to take up any irregularities in the sealing surface and elastic enough to recover some of its original shape when removed and replaced. 109

110. FACTORS IN SELECTING A LINER The most important consideration is that the liner should be chemically inert with its product. Gas and vapor transmission rates are usually relative and depend chiefly on the shelf life required for the product. Homogenous Liner: These one piece liners are available as a disk or as a ring of rubber and plastic. Expensive & Complicated to apply Widely used in pharmaceuticals Uniform properties Can withstand high-temperature sterilization Heterogenous liner or composite liner: They are composed of layers of different materials. It consists of two parts: a) facing and b) backing 110

111. PLASTIC CLOSURES The two basic types of plastic generally used for closures are Thermosetting Thermoplastic resins Urea phenols 111

112. RUBBER CLOSURES Rubber is used in the pharmaceutical industry to make closures, cap liners and bulbs for dropper assemblies. The rubber stopper is used primarily for multiple dose vials and disposable syringes . Rubber closures for containers for aqueous parenteral P reparations for powders and for freeze-dried powders 112

113. GLASS CLOSURE 113

114. METAL CLOSURE 114

115. Packaging Evaluation An important step -- characterizing the materials and the chemicals that can migrate or extract from packaging components to the drug product . Figure shows the various types of chemicals that can migrate from polymeric materials. antioxidant stabilizer plasticizer monomer lubricant contaminant A number of tests can be used to establish initial qualification of the container closure system, and a quality control plan can help ensure compatibility and safety. 115

116. Test on Rubber closures  Closure efficiency Placing a desiccant in a packed stored under high RH . Putting liquid in side pack, storing at high temperature and low RH, detecting any moisture loss as reduction in weight . Checking of cap removal torque . Checking on compression ring seal in cap liner when a system contains a liners . Putting liquid in pack, inverting and applying a vaccum . A poor seal is detected by liquid seeping. 116

117. 2.   Fragmentation test   Place a volume of water corresponding to nominal volume minus 4 ml in each of 12 clean vials. Close the vial with closure and secure caps for 16 hours. Pierce the closures with 21 SWG hypodermic needle (bevel angle of 10 to 14) and inject 1 ml water and remove 1 ml air. Repeat the above operation 4 times for each closure (use new needle for each closure). Count the number of the fragments visible to the naked eye. Total numbers of the fragments should not be more than 10 except butyl rubber where the fragments should not exceed 15. 117

118. 3. Self – sealability This test is applicable to closures intended to be used with water close the vials with the ‘Prepared’ closures For each closure, use a new hypodermic needle with an external diameter of 0.8 mm & pierce the closure 10 times, each time at a different site. Immerse the vials upright in a 0.1% w/v solution of methylene blue & reduce the external pressure by 27KPa for 10 min. Restore the atmospheric pressure and leave the vials immersed for 30 minutes. Rinse the outside of the vials. None of the vials contains any trace of coloured solution. 118

119. 4 ) PH OF AQUEOUS EXTRACT: 20ml of solution A is added with 0.1ml bromothymol blue when it is added with a small amount of 0.01M NaOH which changes the colour from blue to yellow. The volume of NaOH required is NMT 0.3ml . 5 ) LIGHT ABSORPTION TEST: Solution is filtered through 0.5μ filter and its absorbance is measured at 220 to 360nm . Blank is done without closures and absorbance is NMT 2.0. 6 ) REDUCING SUBSTANCES: 20ml of solution A is added with 1ml of 1M H2SO4 and 20ml of 0.002M KMnO4 and boil for 3min then cool and add 1gm of potassium iodide which is titrated with sodium thio-sulphate using starch as an indicator. The difference between titration volumes is NMT 0.7ml. 7 ) RESIDUE ON EVAPORATION: 50ml of solution A is evaporated to dryness at 105⁰C.Then weigh the residue NMT 4mg. 119

120. Evaluations To establish suitability , evaluation of four attributes is required : protection, compatibility, safety, and performance/ drug delivery . Suitability refers to the tests used for the initial qualification of the container closure system with regard to its intended use . Which tests …….??? Suitability testing should be able to establish the following criteria: Materials of construction of container and closure components are safe for their intended use. Container components are compatible with the dosage form The container and closure system adequately protects the dosage form The entire system functions in the manner in which it is intended. 120

121. Dosage Form Condition Route Of Delivery Possible Package Form Solids Aseptic Inhalation Dry-Powder Inhaler Liquids Sterile Parenteral, Ophthalmic Glass Ampoules Glass Or Plastic Vial With Stopper Glass Or Plastic Vials With Applicators Pre-Filled Syringe Bag Pre-Filled Form-Fill-Seal Plastic Container Ointments Sterile Ophthalmic Collapsible Tube Glass Or Plastic Bottle And Cap Form-Fill-Seal Plastic Bottle Glass Of Plastic Jar Soft Gelatin Capsules Dosage forms and package forms 121

122. OVERVIEW OF MANUFACTURING PROCESS OF PARENTERALS documentation Planning & scheduling Material management -Raw material & API -Packaging material Warehousing Equipment & facility Manufacturing requirement personal Finishing Manufacturing Bulk analysis Sterilization Q.C. Testing Aseptic filling Visual inspection Labeling & packing 122

123. FLOW OF MATERIALS :- Ingredients vehicle solute Processing equipment Container component Compounding of product Cleaning Cleaning Filtration of solutes Sterilization Sterilization Filling Packaging Sealing Product storage Diagram of flow of materials through the production department 123

124. [QUALITATIVE 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 24,607 100.0 124

125. 1.AREA PLANING AND ENVIRONMENTAL CONTROL:- Area planning may be addressed by functional groups ground this critical area with particular attention given to maintaining cleanliness. Functional groupings :- Warehousing :- The storage of spare parts, air filters, change parts, water treatment chemicals, office supplier, janitorial supplies, uniforms, an so on may be handled as central storage or individually by department. Finished product and certain raw materials need special environmental storage conditions, such as, temperature and humidity control. Administrative areas :- Administrative area planning requires careful analysis of the direct and indirect administrative requirements of a particular plant. Successively higher levels of supervision are usually provided successively larger office areas. Some offices are individual, while some are grouped in an “open area concept”, 125

126. ZONES AS PER GAZZETE OF INDIA : 1 st .zones as per gazette of India White zone:- final step (filling of parenteral) Grey zone:- weighing, dissolution & filtration. Black zone:- storage, worst area from contamination view point. BLACK GRAY WHITE 126

127. ENVIRONMENTAL CONTROL ZONE GROUPING :- 1 st .zones as per the c GMP:- Zone 1:- Exterior Zone 7:- Filling line Zone 6:- Filling area Zone 5:- Weighing, mixing & transfer area Zone 4:- Clean area Zone 3:- General production Zone 2:- Warehouse 127

128. Zone 7:- filling line:- The walls of the filling area are the last physical barrier to the ingress of contamination, but within the filling area a technique of contamination control known as laminar flow may be considered as the barrier to contamination. For aseptic filling process Sterilization and Depyrogenation of containers before filling, normally hot air oven or autoclave. Provision must be made for Filling requires An aseptic environment with the attendant support rooms Inspection and packaging 128

129. Zone 6:- filling area:- Zone 6 is a distinct zone of the controlled environment area for an aseptic filling process but may not be distinct zone for non-aseptic filling process. Zone 5:- weighing, mixing, and transfer area:- Zone 5 encompasses activities of “ weighing, mixing, filling or transfer operations ” addressed by c GMP section 212.81 which are not handled as zone 6 but which require a controlled environment. Zone 4:- clean area:- Activities in this may include washing and preparations of equipment or accumulation and sampling of filled product. Zone 3:- general production and administration area:- The third zone of environmental controls is formed by the periphery of the general production area. Only essential materials-handling equipment and personnel. Zone 2:- plant exterior:- It is a base point from which to work in determining the requirements for the various control barriers. Control zone 1 might include the maintenance of sterile areas around the facility. 129

130. 2. WALL & FLOOR TREATMENT: The design of filling areas or more generally, controlled environment areas involves attention to many seemingly minor details. The basic cleanlability requirement includes smooth, celanable walls, floors, ceilings, fixtures, and partition exposed columns, wall studs, bracing, pipes, and so on are unacceptable. The need for cleanability also eliminates the open floor system commonly used in the microelectronics industry for laminar airflow rooms. The goal of the designer, when creating the details for the architectural finishes and joining methods, is to eliminate all edges or surfaces within the room where dirt may accumulate. All inside walls must be finished; Example: common methods of finish are block, plaster, or gypsum board. 130

131. 3. LIGHTNING FIXTURES : Lighting fixtures should be reduced flush with the ceiling. Areas having a full HEPA ceiling obviously cannot accommodate recessed lighting fixtures. In these areas, fixtures are of a special “ tear drop ” shape which minimizes disruption to the laminar airflow pattern. 4. CHANGE ROOMS : Personnel access to all controlled areas should be through change rooms. Change rooms concepts and layouts vary from single closet size rooms to expensive multi-room complexes. Entrance to a change area is normally through vestibules whose doors are electrically interlocked so that both cannot be opened simultaneously, thus maintaining the necessary air pressure differential to prevent the entry of airborne contamination. Upon entry into the change room wash skins are provided for scrubbing hands and forearms. Further control may be achieved by using filtered and heated compressed air for drying to reduce further particular potential. After hands are dry, garments are taken from dispensers and donned while moving across a dressing bench. As a final growing step, aseptic gloves are put on and sanitized. Exit from the change room to the controlled area is, like entrance, through an interlocked vestibule. 131

132. Clothing dispenser hand dryer hand wash Exit Glove dispenser Change bench entrance Change room 132

133. 5. Personnel flow :- The movement of personnel should be planned during the design of individual plant areas. Each individual production area may have a smooth and efficient personnel flow pattern, a discontinuous or crowded pattern may develop when several individual production area plants are combined. The flow of material and personnel through corridors are inefficient and unsafe paths for moving materials, particularly if heavy forklifts are required. Discontinuous and crowed flow patterns can decrease production efficiency, increase security problems, and increase the problem of maintaining a clean environment. x Design \ / Design 3 1 4 2 1 2 4 3 133

134. 6. UTILITES AND UTILITY EQUIPMENT LOCATION :- Utilities :- Piping system in particular, must be initially and often periodically cleaned and serviced. Exposed overhead piping is not acceptable from a cleanliness or contamination stand point since it collects dirt, is difficult to clean and may leak. Buried or concealed pipe may require unacceptable demolition for cleaning or repair. Utilities equipment location :- Public utilities require space for metering. In addition to meeting, electrical power system require for switchgear and transformer. Water systems usually require treatment to ensure consistent quality. Plant generated utilities typically require steam boilers, air compressors, and distillation, the typical “ boiler room ” approach. Proper equipment maintenance is difficult in foul weather, especially winter. Heavy equipment may damage the roof-structure, particularly if the equipment location requires numerous penetrations through the roof which, coupled with equipment vibration, will invariable lead to leakage. 134

135. 7.Engineering and maintenance :- From an engineering stand point, even a location outside the plant can serve well if access to the production area by engineers for field work is not too difficult often particularly in small or less complex plants, maintenance or other plant service functions such as utilities or combined with engineering, making an in-plant location desirable. Maintenance responsibilities cover all areas of the plant and can generally be grouped into two categories: plant maintenance and production maintenance. production maintenance is a direct production support function and all the routine and recurring operating maintenance work. Production maintenance facilities are usually minimal, often only a place to store a tool box, and seldom have more than a small workbench. plant maintenance operations, in contrast, are more diverse. They vary from heavy maintenance on production equipment to cosmetic work on the building exterior and often include plant service functions such as sanitation, ground sweeping, or waste disposal. 135

136. LIST OF EQUIPMENTS (as per schedule- M ) : The following equipments is recommended: a)Manufacturing area :- Storage equipment for ampoules, vials bottles and closures. Washing and drying equipment. Dust proof storage cabinet. Water still. Mixing and preparation tanks or other containers. Mixing equipment where necessary. Filtering equipment. Hot air sterilizer. b) Aseptic filling and sealing room:- 9. Benches for filling and sealing. Bacteriological filters. Filling and sealing unit under laminar flow work station. C) General room:- 12. Inspection table Leak testing table. Labeling and packing benches. Storage of equipment including cold storage and refrigerators if necessary. 136

137. Production facilities of parenterals The production area where the parenteral preparation are manufactured can be divided into five sections: Clean-up area: All the parenteral products must be free from foreign particles & microorganism. Clean-up area should be withstand moisture, dust & detergent. This area should be kept clean so that contaminants may not be carried out into aseptic area . Preparation area: In this area the ingredients of the parenteral preparation are mixed & preparation is made for filling operation. It is not essentially aseptic area but strict precautions are required to prevent any contamination from outside. 137

138. Aseptic area: The parenteral preparations are filtered, filled into final container & sealed should be in aseptic area. The entry of personnel into aseptic area should be limited & through an air lock . Ceiling, wall & floor of that area should be sealed & painted. The air in the aseptic area should be free from fibers, dust and microorganism. The High efficiency particulate air filters (HEPA) is used for air. UV lamps are fitted in order to maintain sterility. 138

139. Quarantine area: After filling, sealing & sterilization the parenteral product are held up in quarantine area. Randomly samples were kept for evaluation. The batch or product pass the evaluation tests are transfer in to finishing or packaging area. Finishing & packaging area: Parenteral products are properly labelled and packed. Properly packing is essential to provide protection against physical damage. The labelled container should be packed in cardboard or plastic container. Ampoules should be packed in partitioned boxes 139

140. Aseptic Processing Certain pharmaceutical products must be sterile injections, ophthalmic preparations, irrigations solutions, haemodialysis solutions Two categories of sterile products those that can be sterilized in final container (terminally sterilized) those that cannot be terminally sterilized and must be aseptically prepared Objective is to maintain the sterility of a product, assembled from sterile components Operating conditions so as to prevent microbial contamination To review specific issues relating to the manufacture of aseptically prepared products 140

141. Manufacturing Environment Classification of Clean Areas “ At rest” - production equipment installed and operating “In operation” - Installed equipment functioning in defined operating mode and specified number of personnel present 141

142. Grade D (equivalent to Class 100,000, ISO 8): Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg . handling of components after washing. Grade C (equivalent to Class 10,000, ISO 7): Clean area for carrying out less critical stages in manufacture of aseptically prepared products eg . preparation of solutions to be filtered. Grade B (equivalent to Class 100, ISO 5): Background environment for Grade A zone, eg . cleanroom in which laminar flow workstation is housed . Grade A (equivalent to Class 100 (US Federal Standard 209E), ISO 5 Local zone for high risk operations eg . product filling, stopper bowls, open vials, handling sterile materials, aseptic connections, transfer of partially stoppered containers to be lyophilized. Conditions usually provided by laminar air flow workstation. Each grade of cleanroom has specifications for viable and non-viable particles Non-viable particles are defined by the air classification 142

143. Limits for viable particles (microbiological contamination) These are average values Individual settle plates may be exposed for less than 4 hours Values are for guidance only - not intended to represent specifications Levels (limits) of detection of microbiological contamination should be established for alert and action purposes and for monitoring trends of air quality in the facility 143

144. Environmental Monitoring - Physical Particulate matter Particles significant because they can contaminate and also carry organisms Critical environment should be measured not more than 30cm from worksite, within airflow and during filling/closing operations Preferably a remote probe that monitors continuously Difficulties when process itself generates particles (e.g. powder filling) Appropriate alert and action limits should be set and corrective actions defined if limits exceeded Differential pressures Positive pressure differential of 10-15 Pascals should be maintained between adjacent rooms of different classification (with door closed) Most critical area should have the highest pressure Pressures should be continuously monitored and frequently recorded. Alarms should sound if pressures deviate 144

145. Air Changes/Airflow patterns Air flow over critical areas should be uni -directional (laminar flow) at a velocity sufficient to sweep particles away from filling/closing area for B, C and D rooms at least 20 changes per hour are ususally required Clean up time/recovery Particulate levels for the Grade A “at rest” state should be achieved after a short “clean-up” period of 20 minutes after completion of operations (guidance value) Particle counts for Grade A “in operation” state should be maintained whenever product or open container is exposed Temperature and Relative Humidity Ambient temperature and humidity should not be uncomfortably high (could cause operators to generate particles) (18°C) Airflow velocity Laminar airflow workstation air speed of approx 0.45m/s ± 20% at working position (guidance value) 145

146. P ersonnel Minimum number of personnel in clean areas especially during aseptic processing Inspections and controls from outside Training to all including cleaning and maintenance staff initial and regular manufacturing, hygiene, microbiology should be formally validated and authorized to enter aseptic area Special cases supervision in case of outside staff decontamination procedures (e.g. staff who worked with animal tissue materials ) High standards of hygiene and cleanliness should not enter clean rooms if ill or with open wounds 146

147. Periodic health checks No shedding of particles, movement slow and controlled No introduction of microbiological hazards No outdoor clothing brought into clean areas, should be clad in factory clothing Changing and washing procedure No watches, jewellery and cosmetics Eye checks if involved in visual inspection Clothing of appropriate quality: Grade D hair, beard, moustache covered protective clothing and shoes Grade C hair, beard, moustache covered single or 2-piece suit (covering wrists, high neck), shoes/overshoes 147

148. Grade A and B headgear, beard and moustache covered, masks, gloves not shedding fibres, and retain particles shed by operators Outdoor clothing not in change rooms leading to Grade B and C rooms Change at every working session, or once a day (if supportive data) Change gloves and masks at every working session Frequent disinfection of gloves during operations Washing of garments – separate laundry facility No damage, and according to validated procedures (washing and sterilization) Regular microbiological monitoring of operators 148

149. Aseptic Processing In aseptic processing, each component is individually sterilised , or several components are combined with the resulting mixture sterilized. Most common is preparation of a solution which is filtered through a sterilizing filter then filled into sterile containers ( e.g active and excipients dissolved in Water for Injection) May involve aseptic compounding of previously sterilized components which is filled into sterile containers May involve filling of previously sterilized powder sterilized by dry heat/irradiation produced from a sterile filtered solution which is then aseptically crystallized and precipitated requires more handling and manipulation with higher potential for contamination during processing 149

150. Preparation and Filtration of Solutions Solutions to be sterile filtered prepared in a Grade C environment If not to be filtered, preparation should be prepared in a Grade A environment with Grade B background (e.g. ointments, creams, suspensions and emulsions) Prepared solutions filtered through a sterile 0.22μm (or less) membrane filter into a previously sterilized container filters remove bacteria and moulds do not remove all viruses or mycoplasmas filtration should be carried out under positive pressure consideration should be given to complementing filtration process with some form of heat treatment Double filter or second filter at point of fill advisable Same filter should not be used for more than one day unless validated If bulk product is stored in sealed vessels, pressure release outlets should have hydrophobic microbial retentive air filters 150

151. Preparation and Filtration of Solutions Time limits should be established for each phase of processing, e.g. maximum period between start of bulk product compounding and sterilization (filtration) maximum permitted holding time of bulk if held after filtration prior to filling product exposure on processing line storage of sterilized containers/components total time for product filtration to prevent organisms from penetrating filter maximum time for upstream filters used for clarification or particle removal (can support microbial attachment ) Filling of solution may be followed by lyophilization (freeze drying) stoppers partially seated, product transferred to lyophilizer (Grade A/B conditions) Release of air/nitrogen into lyophilizer chamber at completion of process should be through sterilizing filter 151

152. Prefiltration Bioburden (natural microbial load) Limits should be stated and testing should be carried out on each batch Frequency may be reduced after satisfactory history is established and biobuden testing performed on components Should include action and alert limits (usually differ by a factor of 10) and action taken if limits are exceeded Limits should reasonably reflect bioburden routinely achieved No defined “maximum” limit but the limit should not exceed the validated retention capability of the filter Bioburden controls should also be included in “in-process” controls particularly when product supports microbial growth and/or manufacturing process involves use of culture media Excessive bioburden can have adverse effect on the quality of the product and cause excessive levels of endotoxins / pyrogens 152

153. Filter integrity Filters of 0.22μm or less should be used for filtration of liquids and gasses (if applicable) filters for gasses that may be used for purging or overlaying of filled containers or to release vacuum in lyphilization chamber filter intergrity shoud be verified before filtration and confirmed after filtration bubble point pressure hold forward flow methods are defined by filter manufacturers and limits determined during filter validation Filter validation Filter must be validated to demonstrate ability to remove bacteria most common method is to show that filter can retain a microbiological challenge of 10 7 CFU of Brevundimonas diminuta per cm 2 of the filter surface a bioburden isolate may be more appropriate for filter retention studies than Brevundimonas diminuta 153

154. Challenge concentration is intended to provide a margin of safety well beyond what would be expected in production preferably the microbial challenge is added to the fully formulated product which is then passed through the filter if the product is bactericidal, product should be passed through the filter first followed by modified product containing the microbial challenge (after removing any bactericidal activity remaining on the filter) filter validation should be carried out under worst case conditions e.g. maximum allowed filtration time and maximum pressure integrity testing specification for routine filtration should correlate with that identified during filter validation 154

155. Equipment/container preparation and sterilization All equipment (including lyophilizers ) and product containers/closures should be sterilized using validated cycles same requirements apply for equipment sterilization that apply to terminally sterilized product particular attention to stoppers - should not be tightly packed as may clump together and affect air removal during vacuum stage of sterilization process equipment wrapped and loaded to facilitate air removal particular attention to filters, housings and tubing heat tunnels often used for sterilization/ depyrogenation of glass vials/bottles usually high temperature for short period of time need to consider speed of conveyor validation of depyrogenation (3 logs endotoxin units) worst case locations: tunnel supplied with HEPA filtered air 155

156. equipment should be designed to be easily assembled and disassembled, cleaned, sanitised and/or sterilized equipment should be appropriately cleaned - O-rings and gaskets should be removed to prevent build up of dirt or residues rinse water should be WFI grade equipment should be left dry unless sterilized immediately after cleaning (to prevent build up of pyrogens ) washing of glass containers and rubber stoppers should be validated for endotoxin removal should be defined storage period between sterilization and use (period should be justified) 156

157. Process Validation Not possible to define a sterility assurance level for aseptic processing Process is validated by simulating the manufacturing process using microbiological growth medium (media fill) Process simulation includes formulation (compounding), filtration and filling with suitable media using the same processes involved in manufacture of the product modifications must be made for different dosage formats e.g. lyophilized products, ointments, sterile bulks, eye drops filled into semi-transparent/opaque containers, biological products Media fill program should include worst case activities Factors associated with longest permitted run (e.g. operator fatigue ) 157

158. Representative number, type, and complexity of normal interventions, non-routine interventions and events (e.g. maintenance, stoppages, etc) Lyophilisation Aseptic equipment assembly Worst case activities (cont) No of personnel and their activities, shift changes, breaks, gown changes Representative number of aseptic additions (e.g. charging containers, closures, sterile ingredients) or transfers Aseptic equipment connections/disconnections Aseptic sample collections Line speed and configuration Weight checks Container closure systems Written batch record documenting conditions and activities Should not be used to justify risky practices 158

159. Duration Depends on type of operation BFS, Isolator processes - sufficient time to include manipulations and interventions For conventional operations should include the total filling time Size 5000 - 10000 generally acceptable or batch size if <5000 For manually intensive processes larger numbers should be filled Lower numbers can be filled for isolators Frequency and Number Three initial, consecutive per shift Subsequently semi-annual per shift and process All personnel should participate at least annually, consistent with routine duties Changes should be assessed and revalidation carried out as required 159

160. Line Speed: Speed depends on type of process Environmental conditions Representative of actual production conditions (no. of personnel, activity levels etc) - no special precautions (not including adjustment of HVAC), if nitrogen used for overlaying/purging need to substitute with air Media Anaerobic media should be considered under certain circumstances, should be tested for growth promoting properties (including factory isolates ) Incubation, Examination In the range 20-35ºC. All integral units should be incubated. Should be justification for any units not incubated. Units removed (and not incubated) should be consistent with routine practices (although incubation would give information regarding risk of intervention) Batch reconciliation 160

161. Interpretation of Results When filling fewer than 5000 units: no contaminated units should be detected One (1) contaminated unit is considered cause for revalidation, following an investigation When filling from 5000-10000 units One (1) contaminated unit should result in an investigation, including consideration of a repeat media fill Two (2) contaminated units are considered cause for revalidation, following investigation When filling more than 10000 units One (1) contaminated unit should result in an investigation Two (2) contaminated units are considered cause for revalidation, following investigation 161

162. Interpretation of Results Media fills should be observed by QC and contaminated units reconcilable with time and activity being simulated (Video may help) Ideally - no contamination. Any contamination should be investigated. Any organisms isolated should be identified to species level (genotypic identification) Invalidation of a media fill run should be rare Batch Record Review In-process and laboratory control results Environmental and personnel monitoring data Output from support systems(HEPA/HVAC, WFI, steam generator) Equipment function (batch alarm reports, filter integrity) Interventions, Deviations, Stoppages - duration and associated time Written instructions regarding need for line clearances Disruptions to power supply 162

163. Isolators Decontamination process requires a 4-6 log reduction of appropriate Biological Indicator (BI) Minimum 6 log reduction of BI if surface is to be free of viable organisms Significant focus on glove integrity - daily checks, second pair of gloves inside isolator glove Traditional aseptic vigilance should be maintained Blow-Fill-Seal (BFS) Located in a Grade D environment Critial zone should meet Grade A (microbiological) requirements (particle count requirements may be difficult to meet in operation) Operators meet Grade C garment requirements Validation of extrusion process should demonstrate destruction of endotoxin and spore challenges in the polymeric material Final inspection should be capable of detecting leakers 163

164. Issues relating to Aseptic Bulk Processing Applies to products which can not be filtered at point of fill and require aseptic processing throughout entire manufacturing process. Entire aseptic process should be subject to process simulation studies under worst case conditions (maximum duration of "open" operations, maximum no of operators) Process simulations should incorporate storage and transport of bulk. Multiple uses of the same bulk with storage in between should also be included in process simulations Assurance of bulk vessel integrity for specified holding times . Process simulation for formulation stage should be performed at least twice per year. Cellular therapies, cell derived products etc products released before results of sterility tests known should be manufactured in a closed system sterility testing of intermediates, microscopic examination (e.g. gram stain) endotoxin testing 164

165. Formulation:Methods of Sterilization Steam(autoclave ): Steam sterilization is conducted in an autoclave and employs steam under pressure. The usual temperature and the approximate length of time required is 121 °C for 15 to 30 minutes, depending on the penetration time of moist heat into the load. Dry heat: The transfer of energy from dry air to the object that is sterilized. The transfer occurs through conduction, convection and radiation, h igher temperature and longer time are required (250°C for two hours). Filtration: Sterilization by filtration depends on the physical removal of microorganisms by adsorption on the filter medium or by a sieving mechanism, for heat-sensitive solutions, membrane filters (0.22 μ m). Membrane filters are used exclusively for parenteral solutions, due to their particle-retention effectiveness, non-shedding property, non-reactivity, and disposable characteristics. 165

166. Formulation:Methods of Sterilization Filtration: The most common membranes are composed of Cellulose esters, Nylon, Polysulfone, Polycarbonate, PVDF, Polyethersulfone (PES) or Polytetrafluoroethylene(Teflon). The integrity of the filters has to be proven . If the drug formulation content benzyl alcohol, it is recommend ed to use nylon filter instead of PES filter due to the incompatibility issue. Ionizing radiation: High-energy photons are emitted from an isotope source (Cobalt 60) producing ionization throughout a product.  It can be applied under safe, well-defined, and controlled operating parameters, and is not a heat- or moisture generating process . Most importantly, there is no residual radioactivity after irradiation (Gamma Radiation ) . 166

167. 167

168. EVALUATION OF PARENTERAL PREPARATIONS The finished parenteral products are subjected to the following tests, in order to maintain quality control: A) Sterility test B) Clarity test C) Leakage test D) Pyrogen test E) Assay 168

169. METHOD A: Membrane filtration METHOD B: Direct inoculation TEST FOR STERILITY 169 Sterility is defines as freedom from the presence of viable microorganism

170. 170 Fluid Thioglycolate Medium Soyabean-casein digest Medium

171. 171 M INIMUM QUANTITY TO BE USED FOR EACH MEDIUM Quantity per container Minimum quantity to be used for each medium Liquids 1. less than 1 ml The whole contents of each container 2. 1-40 ml Half the contents of each container but not less than 1 ml 3.Greater than 40 ml and not greater than 100 ml 20 ml 4. Greater than 100 ml 10 per cent of the contents of the container but not less than 20 ml Antibiotic liquids 1 ml

172. Membrane filtration method (METHOD 1 ): Membrane filtration Appropriate for : (advantage) Filterable aqueous preparations Alcoholic preparations Oily preparations Preparations miscible with or soluble in aqueous or oily (solvents with no antimicrobial effect) All steps of this procedure are performed aseptically in a Class 100 Laminar Flow Hood 172

173. Membrane filter 0.45 μ porosity Filter the test solution After filtration remove the filter Cut the filter in to two halves First halves (For Bacteria) Second halves (For Fungi) Transfer in 100 ml culture media (Fluid Thioglycollate medium) Incubate at 30-35 C for not less then 7 days Transfer in 100 ml culture media (Soyabeans-Casein Digest medium) Incubate at 20-25 C for not less then 7 days Observe the growth in the media Observe the growth in the media 173

174. Direct inoculation method (METHOD 2 ): Suitable for samples with small volumes volume of the product is not more than 10% of the volume of the medium suitable method for aqueous solutions, oily liquids, ointments and creams Direct inoculation of the culture medium suitable quantity of the preparation to be examined is transferred directly into the appropriate culture medium & incubate for not less than 14 days. 174

175. 175 INTERPRETATION OF RESULTS If the material being tested renders the medium turbid so that the presence or absence of microbial growth cannot be easily determined by visual inspection,14 days after the beginning of incubation , transfer portion (< 1 ml) of the medium to fresh vessels of the same medium and then incubate original and transfer vessel for not less than 4 days. If No evidence of microbial growth is found- complies with test for sterility. If evidence of microbial growth is found- does not complies with test for sterility.

176. B) Clarity test Particulate matter is defined as unwanted mobile insoluble matter other than gas bubble present in the product. If the particle size of foreign matter is larger than the size of R.B.C.. It can block the blood vessel. The permit limits of particulate matter as per I.P . are follows: 176

177. Methods for monitoring particulate matter contamination: Visual method Coulter counter method Filtration method Light blockage method 177

178. 178 C) LEAKAGE TEST Leakage test is employed to test the package integrity. Package integrity reflects its ability to keep the product in and to keep potential contamination out. Which is the flow of matter through the barrier itself. Leakage tests are 4 types V isual inspection B ubble test  D ye tests  V acuum ionization test

179. leakage test The sealed ampoules are subjected to small cracks which occur due to rapid temperature changes or due to mechanical shocks. Filled & sealed ampoules Dipped in 1% Methylene blue solution Under negative pressure in vacuum chamber Vacuum released colored solution enter into the ampoule Defective sealing Vials & bottles are not suitable for this test because the sealing material used is not rigid 179

180. 180 Leakage test apparatus High voltage leak detection

181. D ) Pyrogen test Pyrogen = “Pyro” (Greek = Fire ) + “gen” (Greek = beginning ). Fever producing , metabolic by-products of microbial growth and death. Bacterial pyrogens are called “ Endotoxins ”. Gram negative bacteria produce more potent endotoxins than gram + bacteria and fungi. Endotoxins are heat stable lipopolysaccharides (LPS) present in bacterial cell walls, not present in cell-free bacterial filtrates TEST FOR PYROGEN The test involves measurement of the rise in body temperature of rabbits following the IV injection of a sterile solution into ear vein of rabbit. Dose not exceeding 10 ml per kg injected intravenously within a period of not more than 10 min Test animals: Use healthy, adult rabbits of either sex, preferably of the same variety. Recording of temperature: Clinical thermometer 181

182. 182 PRELIMINARY TEST(SHAM TEST) If animals are used for the first time in a pyrogen test or have not been used during the 2 previous weeks condition them 1 to 3 days before testing the substance by injecting IV 10ml per kg pyrogen free saline solution warmed to about 38.5° Record the temperature of the animals beginning at least 90 min before injection and continuing for 3 hours after injection. Any animal showing a temperature variation of 0.6° or more must not be used in main test MAIN TEST Carry out the test using a group of 3 rabbits. Dissolve the substance in or dilute with pyrogen free saline solution . Warm the liquid to approximately 38.5° before injection. Inject the solution under examination slowly into the marginal veins of the ear of each rabbit over a period not exceeding 4 min. Record the temperature of each animal at half-hourly intervals for 3 hours after injection. The highest temperature recorded for a rabbit is taken to be its response.

183. 183 INTERPRETATION OF RESULT

184. E ) Assay Assay is performed according to method given In the monograph of that parental preparation in the pharmacopoeia Assay is done to check the quantity of medicament present in the parenteral preparation UNIFORMITY OF WEIGHT Remove the labels& wash the container & dry Weigh the container along with content Empty the container completely Rinse with water & ethanol,dry at 100°C to a constant weight Cool& weigh Net weight shout be calculated 184

185. 185 UNIFORMITY OF CONTENT 30 sterile units are selected from each batch. The weight of 10 individual sterile units is noted and the content is removed from them and empty individual sterile unit is weighed accurately again. Then net weight is calculated by subtracting empty sterile unit weight from gross weight. The dose uniformity is met if the amount of active ingredient is within the range of 85-115.0% of label claim. Relative standard deviation is equal to or less than 6.0%. If one unit is outside the range of 85-115.0%, and none of the sterile unit is outside the range of 75-125.0% or if the relative standard deviation of the resultant is greater than 6.0% ,or if both condition prevail, an additional 20 sterile unit should be tested. The sterile units meet the requirements if not more than one unit is out side the range of 85-115%, no unit is outside the range of 75-125.0% and the calculated relative standard deviation is 7.8%.

186. 186 PARTICULATE MATTER TEST Particulate matter refers to the extraneous, mobile, undissolved particles, other than gas bubbles, unintentionally present in the solutions. Two methods are used: Light obstruction Particle Count Test Microscopic particle count test LIGHT OBSTRACTION PARTICLE COUNT TEST Use a suitable apparatus based on the principle of light blockage which allows an automatic determination of the size of particles and the number of particles according to size.

187. 187 Sample Particle size in μ m Maximum no. of particles. LVP ≥ 100 ml 10 25 Average in the units tested 25 per ml 3 per ml SVP – 100 ml and less than 100 ml 10 25 6000 per container 600 per container Limits

188. MICROSCOPIC PARTICLE COUNT TEST Wet the inside of the filter holder fitted with the membrane filter with several milliliter of particle-free water . Transfer the total volume of a solution pool or of a single unit to the filtration funnel, and apply vacuum. Place the filter in a Petri dish and allow the filter to air-dry. After the filter has been dried, place the Petri dish on the stage of the microscope, scan the entire membrane filter under the reflected light from the illuminating device, and count the number of particles Sample Particle size in μ m Maximum no. of particles. LVP ≥ 100 ml 10 25 Average in the units tested 12 per ml 2 per ml SVP – 100 ml and less than 100 ml 10 25 3000 per container 300 per container Limits : 188

189. TEST FOR BACTERIAL ENDOTOXIN Measures the concenration of bacterial endotoxin Test is using lysate derived from hemolymph cells or amoebocytes of horse shoe crab Endotoxin limit calculated by K/M K  maximum no.of endotoxin which receive the patient without suffering toxic reaction M maximum dose administered to a patient/kg/hr Procedure Equal volume of LAL reagent and test solution (usually 0.1 ml of each) are mixed in a depyrogenated test-tube Incubation at 37 o C, for1 hour Remove the tube – invert in one smooth motion (180 o ) Observe the result 189

190. 190 Mechanism of LAL Test The test is based on the primitive blood-clotting mechanism of the horseshoe crabLimulus amebocyte lysate [LAL] test LAL reagent Bleeding adult crabs blood into an anticlotting solution Washing and centrifuging to collect the amoebocytes Lysing in 3% NaCl Lysate is washed and lyophilized for storage

191. 191 Different Techniques Three different techniques: The gel-clot technique – gel formation The turbidimetric technique – the development of turbidity after cleavage of an endogenous substrate The chromogenic technique – the development of color after cleavage of a synthetic peptide – chromogen complex Chromogenic Technique This is based on the measurement of color change which is caused by the release of the chromogenic chemical p - nitroanilide The quantity of the p - nitroanilide produced is directly proportional to the endotoxin concentration

192. 192 Gel Clot Technique A solid gel is formed in the presence of endotoxins This technique requires positive and negative controls Positive controls – a known concentration of endotoxin added to the lysate solution Negative controls – water, free from endotoxins, added to the lysate solution Turbidimetric Technique The test is based on the measurement of opacity change due to the formation of insoluble coagulin Opacity is directly proportional to the endotoxin concentration This technique is used for water systems and simple pharmaceutical products

193. References Donald C. Liebe , Packaging of Pharmaceutical Dosage Form, Modern Pharmaceutics by G.S.Banker , Marcel Dekker, p 681-725. C.P.Croce , A.Fischer & R.L.Thomas , Packaging material Science, The theory & Practice of Industrial Pharmacy by Leon Lachman , Third edition, p 711-732 Plastic Packaging , Remington: The Science and Practice of Pharmacy , 19 th edition, Volume II, p 1487 Indian Pharmacopoeia, 2007, Government of Indian ministry of health and family welfare, The Indian pharmacopoeia commission, Ghaziabad, volume-1, 599-609. Dean D. A., Evans E. R. and Hall I. H.: Pharmaceutical Packaging Technology, Taylor and Francis, London and New York, First Indian reprint, 2006, 5 and 73. Carter S.J., “Packaging”; Cooper and Gunn’s Tutorial Pharmacy, sixth edition, CBS publicashers and distributors, New Delhi, 2005, 133-136 and 139-140. Pharmaceutical Dosage Forms. Vol. 3 : Parenteral Medications, Pub Informa Healthcare, Edi. Avis, Kenneth E, Vol I, pg173-180 Encyclopedia of pharmaceutical technology by James Swarbrick pg.no.1266-1299 193