Environmental Monitoring and Media Fill Observation

Microbiological monitoring is a potential source of contamination that must be simulated in media fills, as one of the best ways of diagnosing the source of contamination arising in media fills. It should always be assumed that management is anxious to know what, where and why media fill contamination has arisen, in order to decide on appropriate corrective and preventive actions and improve their processes. For this reason it is advisable to have intensive microbiological monitoring over the...

Media Fills Microbiological Considerations And Controls

The ownership of media fills should properly lie with the management of the aseptic process. Properly, media fills should be scheduled into the manufacturing program in the same way as a routine filling activity except that the product is units filled with media. In practice, this ownership tends to be held jointly between production and microbiological QA. Numerous microbiological considerations and controls must be complied with for a media fill to be fit for its intended purpose within the...

Contamination from People

People are a significant source and the most unpredictable vector of microbiological contamination. Microorganisms are always present on hair and skin, which are shed into the surrounding environment. With more movement, more microorganisms are shed. Concentrations of microorganisms are found in the nose, throat, mouth, anal and genital regions, and may be dispersed by breathing, coughing, sneezing, talking, flatulence and hand contact. The most likely types of microorganism traceable to...

Facility

The author does not know of any regulatory-defined microbiological standards for the manufacture of nonsterile liquids, ointments and semisolids. Standards for sterile manufacture are, on the other hand, well defined. At first it might seem that the direction for defining microbiological standards for nonsterile manufacturing environments would be to scale down those already defined for sterile manufacture. However, this is not as easy as it would appear. Environmental microbiology standards...

Direct Epifluorescent Filtration Technique DEFT

This technique has also been extensively used in the food industry. It utilizes the observation that viable cells, when exposed to acridine orange, can be visualized under a fluorescent microscope, appearing bright orange, while nonviable cells appear green. By sample filtration, the viable cells can be stained in situ and immediately counted, giving a result within one hour or less. The most obvious drawback is that the technique is limited to filterable samples and its sensitivity, to some...

Diluents Media and Incubation Conditions

Samples are typically diluted 10-fold prior to counting, and the diluting fluid should be of such a pH and osmolarity that there is no possibility of viability loss before the sample reaches the Petri dish. It is therefore necessary to confirm this as part of the validation program. Water is unsuitable as a diluent, not only because its pH can be outside the range of 6 to 8 normally considered acceptable, but also because some sensitive organisms can suffer osmotic shock and die rapidly....

Mathematical Models

Roark (1972) developed the following model to describe the microbiological contamination of spacecraft. Its principles are applicable to any form of contamination, including the manufacture of sterile pharmaceutical products. Number of contaminants after time t A . X(t) . (t) . q-, where A the surface area exposed to contamination. The larger the area the greater the probability of microbiological contamination X(t) the deposition rate of microbial contaminants on the item of surface A. The...

Process Design and Mode of Operation

Many process-related aspects of manufacture of pharmaceutical preparations contribute to contamination, or conversely, to contamination control. These are generally product-specific, but some have intrinsic similarities amongst groups of products. Figure 4.4 shows a process flow diagram for creams manufacture, given as an example for creams, lotions, gels and ointments. The major items of equipment are the fats vessel, the manufacturing vessel and the holding vessel. These three vessel types...

Site Environmental Monitoring Program Document

The policy document should be very stable. If found otherwise in practice, it merits serious review. Conversely, the site program document is intended to reflect actual practice, and may be subject to more frequent revision. First of all the program document should contain a floor plan of the aseptic facility with the areas within the area clearly identified by their grades. On this floor plan the locations for environmental monitoring should be marked. The locations should be identified versus...

Definitions And Scope

The word bioburden has been ascribed subtly different meanings, not only in scientific texts, but also in official documents. Many authorities use the word simply in a quantitative sense, i.e., in a way suggesting merely a determination of numbers, with little or no specific mention of types of organisms present. ISO 111341, for example, defines bioburden as Population of viable microorganisms on a raw material, component, a finished product and or a package. More commonly, however, the word...

Media Fills Incubation

Filled units must be incubated as soon as possible after filling. Regulators, the FDA in particular, are anxious that all units are incubated (with the exception of those without caps, obvious cracks, etc.). This is intended to include those perfect units that may in practice never be released, e.g., units cleared off the line after a stoppage or similar event. Regulators prefer incubation for information purposes, with indirect rather than direct impact on the success or failure of the trial....

Tests for Absence of Specific Indicator Microorganisms

Both pharmacopoeias describe methods for determining if particular microorganisms are present or absent from specified weights or volumes (10g or 10 ml in USP, 1 g or 1 ml in PhEur) of pharmaceutical preparations. These methods involve incubation of the pharmaceutical preparation in enrichment media, which encourages growth of one particular microorganism at the expense of others. This is followed by surface spread plating on media (selective and differential) upon which the particular...

Active Air Sampling

Active air sampling is intended to provide an index of the number of microorganisms per unit volume of air space in clean rooms. If the clean room is served by a good air-handling system, with integral HEPA filters in place, airborne microbial contamination arises from personnel operating within the clean room. Most active air sampling should be done when the clean room is operational. However, if a clean room has been nonoperational for a few days (e.g., a long weekend) or a few weeks (e.g., a...

Choice of Method

Having prepared the sample in a suitable form for a TVC, the choice of counting method is the next consideration. The methods described in the pharmacopoeias are listed in Table 5.3, which also identifies their relative merits. Other methods e.g., surface drop (Miles and Misra method) and semiautomated techniques (e.g., spiral plating) may be used, if validated, and are considered elsewhere. 9,10 The EP and USP differ in the guidance they offer on choice of method. The EP directs that membrane...

Media Fills Purposes

Media fills, broth fills, simulation trials and so on, are all synonymous names for an exercise undertaken as part of the validation of a new aseptic process, and as a frequent validation review thereafter. Regulatory pressure to repeat media fills at six- and even three-month intervals, is gradually merging media fills into routine environmental monitoring programs. The purpose of the media fill is to provide a measure of the likelihood of microbiological contamination arising in particular...

Validation

In addition to the minor aspects of validation, a bioburden validation programme has principally to demonstrate that the procedures routinely used are capable of Accurately and reproducibly enumerating low concentrations of organisms contained in, or, in the case of devices, on, the surfaces of samples or products Detecting low levels of specific objectionable organisms in products Adequately neutralizing any antimicrobial activity associated with the product and that any chemical inactivator...

Simulation of Lyophilization Processes

Fiber Bonding Scaffold

Those sterile dosage forms that are stable only for a short time in solution are frequently marketed in lyophilized presentations (see Figure 3.3). Figure 3.3. Simplified representation of aseptic filling and lyophilization. Figure 3.3. Simplified representation of aseptic filling and lyophilization. The process is more complicated than standard vial filling, although it may involve many items of common equipment. Vials are aseptically filled in the normal way, but the closures (which are of a...

Sources and Vectors for Contamination

There are several broad areas to which contamination may be traced in the manufacture of all pharmaceutical preparations. Figure 4.3 shows a schematic representation of the sources of contamination in any nonsterile manufacturing facility. The main sources are Incoming raw materials and ingredient water Facilities, services and cleaning materials Figure 4.3. Generalized sources of microbiological contamination. Figure 4.3. Generalized sources of microbiological contamination. Raw materials are...

Whytes Analyses

Throughout the 1980s and 1990s William Whyte and his co-workers attempted a more ambitious model of contamination than the experimentally based views on deposition published by Bradley et al. (1991). In 1986 Whyte listed five mechanisms by which airborne particles can be deposited on surfaces. He analyzed in some detail the significance of each of these mechanisms to contamination in practice in pharmaceutical clean rooms. Whyte's analysis is based on common sense, observation and experience,...

Growth Support and Sterility Controls

The first responsibility in any microbiological exercise that is expected to produce no growth results, and for which no growth is the favorable condition, is to ensure that the medium is capable of supporting growth. Maintenance of aseptic clean rooms must ensure that only materials that can be safely presumed to be sterile should be permitted entry. Growth supportiveness of the media should be verified before use. It should also be checked after it has been in contact with the filling...

Passive Air Sampling

Passive air sampling is done by means of settle plates agar plates are left open and exposed in clean rooms for defined periods. They are used widely in Europe where they have been strongly advocated over many years in the work of Whyte 1986 , but less so in the U.S., except in facilities manufacturing for export to Europe. The principles of the settle plate were empirically demonstrated by Whyte 1986 . Most airborne microorganisms are associated with physical particles of 12- m diameter or...

Media Fills in Validation of Aseptic Processes

The Guideline on Sterile Drug Products Produced by Aseptic Processing FDA, 1987 refers to media fills as an acceptable method of validating the aseptic assembly process. By 1994, the Guideline to Industry for the Submission Documentation for Sterilization Process Validation in Applications for Human and Veterinary Drug Products FDA, 1994 said that specifications for media fills should be among the information submitted in support of sterility assurance for products manufactured by aseptic...

Media Fills Placebos

The most commonly used placebo for media fills is TSB used to simulate aqueous injections. It is a reasonably good all-round, general-purpose microbiological medium, which can support growth of aerobic bacteria when incubated at temperatures in the range 20-35 C. Equally, it is a reasonably good medium for supporting the growth of yeasts and fungi, when incubated at 20-25 C. It is the recommended test for sterility in all of the major pharmacopoeias. However, many microorganisms will not...

International Pharmacopoeial Guide Limits

In setting the limits for guides appropriate to particular pharmaceutical preparations, both USP and PhEur take account of The significance of microorganisms to different types of product The way in which the product is used The potential hazard to the patient. Although the pharmacopoeias describe tests for counting two different groups of microorganisms, and tests for detecting the presence or absence of four different selected indicator microorganisms, there is only one single guide in USP...

Objectionable Microorganisms

It is important to recognise that pharmacopoeial limits on microbiological contamination in pharmaceutical preparations are not all embracing. Contamination limits can never adequately specify everything present in a pharmaceutical preparation that might risk infection, or lead to patient refusal. Many microorganisms are not specifically restricted from pharmaceutical preparations, but if present even in numbers well within the quantitative limits would clearly constitute a risk to the patient....

Periodic Media Fills in Routine Operation

It is unlikely that any responsible regulatory body would tolerate a frequency of less than twice a year for periodic media fills. Media fills are probably the most sensitive method of detecting unexpected sources of process contamination. The regulatory standpoint coming from the principle of patient protection is that if unexpected process contamination occurs in a media fill, and is considered sufficient to compromise the sterility of past product, they would expect market withdrawal....

Surface Sampling

There are two methods available for sampling surfaces the contact plate and the swab. Only the contact plate is referenced in the E.U. requirements and in USP Chapter lt 1116 gt limits. Swabs are not thought, generally speaking, to produce quantitative data. In the contact plate, agar contained in a specially designed Petri dish, the Rodac plate, is pressed against the surface being sampled. Microorganisms are transferred from the surface to the agar and colonies develop on incubation. Devices...

Environmental Monitoring Applications And Limits

All pharmaceutical manufacturing environments merit a level of environmental monitoring. The greatest emphasis and the tightest limits are applied to sterile manufacturing facilities. When different areas within sterile manufacturing facilities serve different purposes, so the environmental monitoring programs differ. The question being so often asked is What limits should be applied in microbiological monitoring of sterile products manufacturing facilities In Europe the answer is easy....

Environmental Monitoring Microbiological Considerations And Controls

Tryptone Soy Agar TSA is the standard medium for microbial recovery in environmental monitoring programs. Incubation is at 30-35 C. Yeasts and moulds may also be specifically sought out. Sabouraud Agar SA incubated at 20-25 C is generally used for this purpose. Media used in antibiotic manufacturing facilities, particularly when they are solid dosage forms, should be reviewed for their capability to recover environmental microorganisms. Neutralizers may have to be included in the preparation of...

Personnel Monitoring

Personnel are probably the greatest source of microbial contamination in clean rooms. People are mobile, unpredictable and cannot be sterilized in the microbiological sense of sterilization . Some may be greater potential sources of contamination than others there may be sudden or periodic changes in their contamination potential for physical or even psychological reasons the garments provided to them may be inappropriate wrong fabric or fit and none of this may be obviously evident to...

Particulate Matter

Particulate matter has, for parenteral products, been defined as mobile undissolved substances which are unintentionally present. It is divided into subvisible and visible particles with the limit at 50 m. Intravenously administered pharmaceutical products enter the circulatory system and pass through the lungs, where the largest particles are filtered out before the product is pumped on through the arterial circulation. The potential for patient risk from nonviable particles was first reported...

Contamination of Sterile Products

Sterility is defined as freedom from all viable life forms. Two broad groupings of pharmaceutical products are required to be sterile parenteral and ophthalmic products. Such products must be free from all viable life forms, due to the potential consequential severity of the consequences of viable microorganisms present when the products are used in the manner intended or prescribed. Confirmed incidents of nonsterility in supposedly sterile parenteral and ophthalmic products have been...

Parenteral Products

Parenteral products are intended for administration by injection, by infusion, or by implantation into the human body. Products normally totally free from microbiological contamination or colonization are delivered to internal tissues, while the parenteral route of administration deliberately bypasses the body's external physical barriers to infection. No distinction can be made between microorganisms known to specifically cause infectious disease in humans, from those customarily thought to be...