8 IMPLEMENTING A CLEANING AND DISINFECTION PROGRAM IN PHARMACEUTICAL AND BIOTECHNOLOGY CLEAN ROOM ENVIRONMENTS

Arthur L. Vellutato, Jr.
Veltek Associates, Inc.

INTRODUCTION
The implementation of an effective cleaning and disinfection program in a GMP facility is a multi-phased process that requires the harnessing of a multitude of critical disciplines in order to achieve success. Like many compliant GMP systems, the design of a cleaning and disinfection regime utilizes the talents of the quality assurance, production and validation departments. This compilation of skills is mandatory to completely address a system that will net continually acceptable environmental conditions and prove competence during regulatory inspections. In looking at the basics surrounding the formation of a compliant system, the ultimate goal is to clean and disinfect the surface. This seems to be very simple; however, it is slightly more complex than just cleaning and disinfecting surfaces. By definition, cleaning a surface would require the removal of any contaminant that is not part of the material surface itself. Such contaminants may be in the form of spilled product from a previous batch, chemicals used in manufacturing product, disinfectants, particulates and residues. When cleaning surfaces in the clean room, the removal of all particulates and residues is virtually impossible. Thus, the clean room will, once cleaned, have an existent level of particulates and residues that may or may not affect the next batch of product. Likewise, disinfecting the surface would require one to ensure that the bioburden of the surface has been eliminated or reduced to a safe 1

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and contained level which will not affect final product. However, one could not render such surfaces “sterile”, as technically defined. Sterilizing a surface would require it to be exposed to a terminal sterilization process such as autoclaving, heat, gamma radiation, ethylene oxide or extended chemical sterilization. The only fitting method for use in a clean room from the above list would be chemical sterilization. Chemical sterilization of a clean room surface using an EPA registered sporicidal agent may take up to five hours of soaking of all surfaces. This is not possible in the clean room environment. Thus, an existent bioburden, even after disinfection, may exist in the area. Comprehending that the surfaces may not be perfectly clean and sterilized is critical to an understanding of what may and may not be accomplished. In many phases of life, “perfect” performance is impossible to achieve, and cleaning and disinfection are no exception. If a clean room were cleaned, filled with ultra-clean water, then the water drained and its chemical content evaluated for non-viable matter, one would find a multitude of spikes in an IR spectrum analysis; and, complete identification of all contaminants would be unlikely. In a viable sense, if a clean room was cleaned and disinfected, then subsequently filled with a growth medium like trypticase soy broth (TSB), and a reasonable growth period allowed, one would see growth. These two examples, at extreme ends of the spectrum, show that cleaning and disinfecting of a controlled environment will not be perfect. But despite the complexity, the goal is to near as close to perfection as is humanly possible.

DESIGNING A COMPLIANT GMP SYSTEM
The first step in developing a compliant cleaning and disinfection system is to evaluate what is presently being done in the operation. The author's personal experience as a consultant involved in cleaning and disinfection in this industry for many years is that Quality Assurance (QA) departments may not know absolutely everything that is done in a production department; likewise, production may not completely understand what quality assurance's recommendation may be. This is understandable, as QA and production are two separate functions with differing agendas but their combination of skills and disciplines makes for the development of a very intricate and effective system. But first, QA and production professionals need to understand what is done on a daily basis, and what should be done on a daily basis. The next step to achieving near-perfection in our cleaning and disinfection systems is to put the basics into perspective. The criteria for implementing a cleaning and disinfection program within a GMP facility require companies to tailor their system to address their operations' specialized needs. Development of a systematic approach to addressing and controlling of contamination will serve organizations for

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years to come, with attention to detail the key to success. And success is measured by consistently attaining acceptable environmental conditions, making use of the available disinfectants and application devices, and utilizing environmental monitoring data to address the specifics of the system. In turn, regulatory auditors will routinely scrutinize the system with a fine-tooth comb, and dissatisfaction can result in possible 483 observations or the shut-down of manufacturing processes. For this reason, systems need to be complete in their content, and assumptions that situations may not arise could be fatal. And most importantly, the design must be documented within the constraints of a quality system. To complicate things further, there is no common methodology in the industry; nor will companies receive a specific requirement list of expectations from the regulatory agencies. It requires the use of a multitude of separate scientifically-based methodologies and disciplines, combined to render the final infrastructure. For years, the pharmaceutical and biotechnology industries have been two of the most regulated in the world. However, within the spectrum of manufacturing a product that will enter or come in contact with the body, there are several critical areas where there is a lack of written directive from regulatory agencies. Two of these critical areas are environmental monitoring, and disinfection of a controlled area. Together these two critical capacities serve as the basis for determining and maintaining the environmental conditions during the time of manufacture of a drug product. Initially, one would assume that there is a specific need for regulatory specifications in these critical areas. However, after careful evaluation, it appears that these functions require a very unique design for each manufacturing setting and for each product that is produced. Specifications from regulatory agencies do, in certain circumstances, standardize the function or testing requirements throughout the industry. However, in the areas of environmental monitoring and disinfection, they may place possibly unattainable criteria to very process-specific functions. Despite the lack of written regulatory specifications, this does not mean that regulatory agencies do not have specific expectations. In fact, their expectations may be greater than if specifications were in place. In the present day, regulatory agencies have placed the responsibility for system design, implementation, documentation and justification on the manufacturers themselves. The overall theme is for a drug manufacturer to "figure out how they are going do it, do it, document it, and be prepared to justify their methodology upon request." An example of this can be found in the 1987 FDA Guideline on Sterile Drug Products Produced by Aseptic Processing, 2 (page 9) . At that time, regulatory specified what they expected for viable airborne conditions: "Air should also be of a high microbial quality. An incidence of no more than one colony forming unit per 10 cubic feet is considered as attainable and desirable." Within this text, regulatory said "here is what we expect; now you figure out how you need to accomplish and document this capacity." However, within this short statement, technical problems arise.

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The first is that humans, by nature, try to interpret hidden meanings in what is written. Personnel in pharmaceutical and biotechnology varied in their interpretation of the guidance. At the same time, regulatory inspectors and reviewers also varied amongst each other understanding and enforcement policies. In short, the guidance text was not complete. It did not specify many details such as the required total volume to be sampled, the number of locations to be sampled, and the frequency in which sampling should be done. This causes confusion as all then develop and implement their opinions and commonality of task to be performed and the method by which it will be inspected varies from firm to firm and regulatory inspector to regulatory inspector. This is just one example of problematic situations that arise with certain guidance documents. Earlier in the chapter, the probable imperfection of cleaning and disinfecting was discussed. In terms of guidance documents, the same is also true. In terms of environmental monitoring, cleaning and disinfection, no one organization will ever be able to create the perfect guideline. Guidelines will always lack the critical uniqueness that exists for implementation in each and every manufacturing and testing operation. This concept must be understood. Thus, the problem with developing an environmental monitoring and disinfection program is one is not able to pick up a book and follow its content. In reality, one may need to view many books, many guidelines or more effectively, rely on direct experience. Direct experience is this venue is invaluable and without it in one's arsenal, there is less of a chance for success. Experienced professionals design environmental monitoring and disinfection systems that work. The utilization of inexperienced personnel to design the systems is a way of ensuring failure and possibly future regulatory complications. It is necessary to look at a few guideline documents that may assist in understanding what needs to incorporate in the design of a system. First and foremost, one needs to look as the specific tone from the US FDA. Presently the 1987 FDA Guideline on Sterile Drug Products Produced by Aseptic Processing is the current document. However a revision 3 from FDA, entitled Sterile Drug Products Produced by Aseptic Processing Draft has 4 been recently released. A second source is the PDA's Technical Report #13 . the third source would be the United States Pharmacopeia (USP), Chapter <1116> (Environmental 5 6 Monitoring) and Chapter <1072> (Disinfectants and Antiseptics) . A fourth source looks to review ISO 146447 and 14698 , and the fifth source reviews current and past industry publications listed in the bibliography of this chapter. The last, and most important, source relays on what has been learned internally as an organization and externally from colleagues in the industry. While this list of written guidance is not complete, coupled together they provide us with the basic concepts needed to design our systems.
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While we review the existent guidelines, publications and past experience data, we must understand that all differ in their content. The design of a complete and compliant cleaning and disinfection system uses these critical documents only as tools to write a master plan; they could never be as complete for one's operations as a system one could develop ourselves. So where does one start? Cognizant of the multitude of issues that will require review, the GMP facility needs to look at the entire picture in a full circle approach. Prior to design or re-design we need to put some basic perspectives into context. While incomplete in lengthy discussion for each item noted, this short guide will hopefully identify the important areas where firms will need to focus their attention.

MAINTAINING CONTROL OF THE ENVIRONMENT
Maintaining control of the environment is critical. It is the most important aspect surrounding a cleaning a disinfection program. We need to understanding that control of the environment does not come from using a chemical agent that will destroy all microorganisms. Rather, control of the environment is the assurance level we establish to reduce or possibly eliminate the contamination from ever entering our controlled areas. Once a chemical agent has been applied to the surface and subsequently dries, its destructive capabilities to a microorganism are complete. Disinfection (or sporicidal) characteristics of a chemical agent require the organism to be wetted by the agent. While certain residues from chemical agents may have some remaining antimicrobial properties, the destructive capabilities of the residue are minimal. Control over what enters the controlled area now takes over. In reality, control of the environment has nothing to do with disinfection. Disinfection is complete when control takes over. Manufacturing operations will not disinfect surfaces while production is occurring. Disinfection is done prior to manufacturing and the environment released to produce a product. Our success or failure in control is measured each and every time as environmental monitoring is conducted during our filling operations. Addressing contamination prior to its entry will ensure that we will not have to contend with its presence. While disinfection of surfaces always take the lead in structuring our procedures, control over the environment should remain one of the main focuses. Criteria for reducing the bioburden that enters the controlled area require us to evaluate the entry of personnel, components, water, tanks, carts, and even disinfectants to name a few. This is done to ensure that one do not undermine disinfection efforts by introducing high levels of contamination after disinfection is complete. We must ensure each item's appropriateness in the room classification to which it enters. In

sterility and appropriate fit of garments for personnel. saturation and penetration of the cell wall. And second. we must ensure that such environmental conditions do not adversely affect the Class 100 area. the dirtied solution on the floor is collected and removed from the area (normally by a squeegee). Procedurally. contact time. however. Residues. existent soil load. the key to disinfection in the clean room is keeping the surface wetted for 5–10 minutes. cleaning is confused with disinfection. we can use a toothbrush and mouthwash as examples. cleaning prepares the surface for disinfection. They are not the same.000–100. while our concern is less.000. In short. disinfection efforts become simpler. By lessening the level of particulates. Cleaning tries to remove contamination from the surface while disinfection attempts to destroy what viable cells exist on the surface. First there are fewer organisms to destroy as most have been removed from the area. If we were to discuss the options of not using a toothbrush anymore and only utilizing a daily mouthwash rinse with our dentist. microbes and possibly existent residues from surfaces. particulates and microbials will build up on the surface of the teeth and the teeth would eventually deteriorate. as well as assuring the cleanliness. Cleaning characterizes the removal of particulates. surface and bioburden of the surface. contaminates and residues are loosened and rinsed to the floor. Subsequently. It further requires that an organism remain wetted for a specified contact time with a chemical agent capable of killing the organism in question. he/she would inform us that we would soon have no teeth. Disinfection relates to the saturation and penetration of the cell wall of an organism by a chemical agent. We forget to brush and just try to kill anything that exists on the . one needs to carefully evaluate each item as clean and sterile prior to entry. as the movement of air via laminar flow tends to dry surfaces quicker. As discussed. there are significant difference between cleaning and disinfection. a room can be monitored as having very little if any bioburden at rest. In Class 10. This scenario depicts what occurs too often in the pharmaceutical and biotechnology industry. the possible obstructions blocking the chemical agent from contacting the organism are minimized.6 Laboratory Validation Class 100 aseptic operations. Disinfection depends upon temperature. concentration of the chemical agent and pH. In short. as bioburden and residues are lower. In a realistic example. CLEANING THE ENVIRONMENT Too often. microbes and residues on the surface. Provided the appropriate chemical agent is utilized. Cleaning requires a non-destructive mechanical action be applied that loosens and removes contaminants from the area. in a static condition we can easily corrupt our efforts previously achieve through poor control. This is difficult.

At 45 days. bags of USP Water for Injection. Grade A. Such cleaning and was conducted in a Class 100. In the report. Manufacturing operations ran for four hours per day and upon completion of manufacturing. Such surfaces trap residues and other contaminants and make the surface more difficult to disinfect. particulates and possibly microbials is also aided by the surface itself. ISO 5.000 units of 500 mL. demonstrated what affect cleaning the surfaces has on the level of microbial contamination found on the surface. Vellutato. Normality is to clean surfaces either bi-weekly or on a monthly basis. Grade A. Jr. test reports have shown the effect that cleaning the surface has on the level of microbial levels in controlled areas. The cleaning mechanism utilized a mop and two-bucket system. all surfaces were cleaned with a sodium lauryl sulfate detergent and mechanical cleaning action on a daily basis. the phrase. "But that's an additional cleaning operation we need to do!" The correct response is "That is correct. a sprayer and a squeegee. Environmental Monitoring was routinely conducted on air. The effect of the buildup of residues. the clean room was completely cleaned. Many publications purport this concept. Vellutato. a test report conducted and published in 9 1989 by A.Implementing a Cleaning and Disinfection Program 7 surface. Sr and A. the chemical was then sprayed on to the surface and all excess liquid on the surfaces pulled downward by squeegee to the floor. And unfortunately. ISO 5 area. Upon completion of the mopping. The manufacturing operations filled an average of 4. “simple common sense” is not the title of any GMP. CFR or guidance document. Within our note to technical brilliance we sometimes forget simple common sense. control was lost. The final conclusion was one to two uses of a sporicidal agent with cleaning regime controlled the environment. The limits returned to acceptable levels for 31 days (day 76 of the study). Some may say. The remaining liquid was then collected on the floor and removed. Cleaning is based on a few physical factors: • the surface to be cleaned . the results met industry limits for Class 100. The study focused on the concept that cleaning alone would remove most of the existent microbial contamination. Inc." Within the healthcare setting and most commonly reported in the hospital setting. results exceeded limits and a sporicidal agent was used once on day 45. In the pharmaceutical and biotechnology setting. Clean room surfaces are irregular in nature as depicted in the Scanning Electron Microscope (SEM) photographs (Figures 1–8) seen later in this chapter. Eventually our surfaces become residue-laden and more difficult to disinfect and eventually deteriorates. For 30 days. it needs to be done. Sooner or later we have to clean. The procedure for cleaning was to apply the sodium lauryl sulfate detergent (DECON-Clean®) to the surface utilizing a top to bottom approach. The frequency of cleaning can vary from a daily function to a monthly function. surface and personnel. of Veltek Associates.

cleaning becomes more difficult. The lack of available products forces the clean room professional to adapt a nonclean room product to address this specific need. the possibility for contamination to be vested underneath the particulate and residue may be probable. . residues and even microbial contamination to vest itself within the rutted or porous areas of the surface. The surface irregularity may compromise cleaning and disinfection efforts for two basic reasons. This is a subject that cleaning apparatus manufacturers need to address more closely. disinfection becomes harder as the chemical agent cannot contact all the surfaces for the required wetted time period (as obstructions may exist). a clean room brush is not a commonly-available item in the industry. First. so a multitude of contamination sources exist on the surface to be cleaned.8 Laboratory Validation • • • • • the contamination vested on the surface the chemical used to clean the surface the effect of the chemical agent on the surface to be cleaned the level of surfactants in the chemical agent the effect of the chemical on the contamination. Unfortunately. The lack of such an abrasive cleaning action bypasses the opportunity to loosen and remove these contaminants. surface irregularity varies as composite materials change. For example. Most irregular surfaces are commonly so across the span of the material. we are disinfecting the surface of the particulates and residues and never disinfecting the actual surface. but a toothbrush. Cleaning becomes more difficult as the surface's irregular nature may allow particulates. that exists on the surface Knowledge of the type of surface to be cleaned is very important. Thus. Cleaning an irregular surface requires one to use a cleaning device that can penetrate into the nooks and crannies and make contact with the existent contaminants and residues. These nooks and crannies are very hard to clean with most clean room apparatus designed for cleaning. Nor would requiring a controlled area to be cleaned with it be a reasonable request to make of production personnel. The second basic complication is that if particulates and residues exist in such nooks and crannies. Without such assurance for cleaning and disinfecting the actual surface. the possibility for a disinfecting agent to contact the surface within the nooks and crannies is improbable. we would not use a wiper or a pad to try to clean our teeth. Most clean room mops and wipes and flat and do not allow the penetration of the fibers or surfaces of the cleaning mechanism to reach into the crevices. As depicted in Figures 1–5 (further in the chapter). Second. moreover residue.

When we attempt to use such data in the field. Upon completion of the soaking. during such validation testing we look to inoculate a surface with a known enumeration of microorganisms and soak such surfaces in a disinfecting agent for a predetermined time period. giving an understanding that. However. We will have the same trouble. the opposite occurs and we find it more difficult to disinfect the irregular or porous surfaces. The smooth surfaces are more easily rinsed and existent microorganisms removed and can grow in our growth medium. if not a more complicated problem. and due to this. we find the rinsing of the irregular surfaces more difficult as microorganisms may vest and cling within the irregular areas of the surface. This means they were never rinsed to the filter and plated for growth. In our test. First. in disinfecting and rinsing the microorganisms from the irregular or porous surfaces in our manufacturing and testing areas. after we have soaked the surfaces.Implementing a Cleaning and Disinfection Program 9 In other sections of this chapter. In the manufacturing or testing areas. antimicrobial effectiveness testing will be discussed. we will then test the surface to account for the possibly remaining viable contamination. Some surfaces are more irregular than others. microorganisms from the irregular or porous surface may have never been removed from the surface itself. Understanding this concept is critical to successful disinfection. However. we could conclude smooth surfaces are harder to disinfect than irregular or porous surfaces in our manufacturing and testing areas. clean room material grades can be separated into six basic categories: • • • • aluminum stainless steel epoxy-coated finishes plastics . Thus. The effectiveness report may show higher remaining colony forming units (CFUs) for the smooth surfaces than the irregular or porous surfaces. Care needs to be taken when disinfecting irregular or porous surfaces. our testing inoculates a variety of surfaces with a multitude of microorganisms. In general. we may leave viable contamination on such surfaces. This means the possibility to rinse free a microorganism from a smooth surface is easier than from an irregular surface. This assumption would be incorrect. as they are harder to clean and disinfect than smoother surfaces. we will find our testing skewed for two basic factors. What we may learn from this testing is that it may seem that our disinfecting agent is more effective on the irregular or porous surface but this is due to our inability to rinse free all of the existent microorganisms. we need to rinse clean the surfaces of any possible contamination to a filter which is then plated to a growth medium.

deterioration of the surface from chemical exposure normally shows as a turquoise bluish gray bubbling on the metal. Figure 2: 316 L Stainless Steel surface . we see an aluminum surface. of most clean room surfaces. quaternary ammonium. Aluminum is also easily stained or discolored from the residues from by phenol. However. This surface is a metal grade that is soft and easily scratches and is deteriorated by chlorine solutions. aluminum is normally easy to clean and disinfect.10 Laboratory Validation Figure 1: Aluminum surface • • vinyl glass In Figure 1. gulteraldehyde and peroxide and peracetic acid and hydrogen peroxide solutions (from a list of basic disinfecting agents). and iodine to name a few. While aluminum is a commonly-used metal.

however this metal grade cannot be used for every component due to its brittle nature. of most clean room surfaces. Epoxy-type surfaces are very numerous in types and materials. Some perfect examples of this would be a spring or a solenoid mechanism. The irregular surface appearance of epoxy-coated surfaces challenges the craft of the disinfection professional. quaternary ammonium. many metal grades exist from of 302 to 402 stainless. Surface rust is the rusting of airborne heavy metals that deposit atop the surface. Within the clean room operation. Most clean room walls and floors are made of an epoxy material or similar material so understanding cleaning and disinfection of this surface is critical. stainless steel is normally one of the easiest surfaces to clean and disinfect. Rusting of the stainless itself is from chemicals or water oxidizing and/or reacting with impurities in the metal. Problematic situation can arise in the crevices of the surfaces as air pockets can form and once disinfection is compete. This surface is very smooth and contains few impurities in the metal that may be deteriorated by a disinfecting agent. deterioration of the surface is in the form of a rusting that pits the surface or surface rust. They are a coating that is applied in a liquid form and then hardens. we see a 316L stainless steel surface. In Figure 3.Implementing a Cleaning and Disinfection Program 11 Figure 3: Epoxy-oated surface In Figure 2. and iodine to name a few. we see an epoxy-coated surface. Common build up of particulates and moreover residues in the crevices complicate cleaning and disinfection. be broken an . However. Stainless is also easily stained or discolored from the residues from by phenol. This is where disinfection becomes complicated. Utilizing too hard a metal will cause the spring or moving part to routinely break as friction of movement on the metal will cause stress and fracture. Many in the industry demand the use of 316L stainless. Stainless comes in grade based on the level of impurities in the metal. gulteraldehyde and peroxide and peracetic acid and hydrogen peroxide solutions (from a list of basic disinfecting agents). Normally. Most impurities in the metal grade are in the carbon family and are deteriorated by chlorine solutions.

and iodine to name a few. The normal deterioration occurs in the form of over drying and cracking of the material finish. isopropyl alcohol. The most difficult task with the epoxy-coated surface is to clean the contaminants from the surface so that the disinfectant can be applied and have the ability to address the surface itself.12 Laboratory Validation Figure 4: Clean Room Curtains #1 possibly release existent contamination to the environment. quaternary ammonium. ethanol and peracetic acid and hydrogen peroxide solutions (from a list of basic disinfecting agents). Figure 5: Clean Room Curtains #2 . Epoxy-coated surfaces need routine replacement or refinishing and as such remain continuously on a preventative maintenance schedule. peroxide. The epoxy-coated finish is also easily stained or discolored from the residues from by phenol. Normally epoxy-coated surfaces are deteriorated by chlorine solutions. The material becomes powder-like and orange to blue in color (dependent upon the material).

A yellowing. The normal deterioration occurs in the form of over-drying and cracking of the material itself. Replacement is costly but necessary. Figure 7: Vinyl surface . and other similar type products. These materials need replacing every two or three years on average. The porous material is very easily stained or discolored from the residues of phenol. peroxide. In Figures 4. and should be viewed as a cost of doing business. ethanol and peracetic acid and hydrogen peroxide solutions (from a list of basic disinfecting agents). Cleaning and disinfection are very difficult with these materials. isopropyl alcohol. porous surfaces are deteriorated by chlorine solutions. plexiglass. or change of color to an orange or brown. Characteristically they have pore openings that are rather deep.Implementing a Cleaning and Disinfection Program 13 Figure 6: Phenolic Residue The next basic clean room surface type is a collage or porous material such as plastic. 5 and 7 we can see what these surfaces may look like when magnified through an SEM microscope. are common symptoms of the drying process. Normally. vinyl. Normally these type of surfaces need to be replaced over time. delron.

If such materials were as clear as glass. the residue. Cleaning these surfaces is normally done frequently. However. The last category is glass. We have seen from this section the importance of cleaning. the effect of cleaning surfaces ensures the best possible opportunity to disinfect the surface as the microbial levels and the possibly existent residues and particulates will be lower. However. Items such as plastic curtains are one of the most widely used materials in the clean room environment. and provides a constant example of how difficult all surfaces are to clean. such residues and/or stains only increase the deterioration process. . In general. Glass is very difficult to clean. first we must understand the chemical agents that cause the main problem associated with a dirtied surface. glass does not stain easily from disinfectants or sporicides. residues build up and it is discolored from the residues of phenol. The reason glass seems harder to clean is that one can see through it. Glass is a relatively smooth which does not deteriorate. and iodine to name a few. All other materials discussed to this point are not clear. and is a very difficult material to keep clean. quaternary ammonium. the appearance of these surfaces would look horrifically dirty in comparison to glass. Simply. This over-cleaning shortens the life of the material. However. Most notably these materials are use as curtains that separate process control areas. as they represent the second-closest nonproduct contact surface next to the filling line itself. if not daily.14 Laboratory Validation Figure 8: Sodium Hypochlorite Residue quaternary ammonium. this surface is used more and more in clean room operations as it allows viewing of the operation by supervisors and visitors. Glass is very easily dirtied. and iodine to name a few. Later in this chapter we will discuss the mechanisms to clean such surfaces.

Once developed. CFR Title 40 governs and registers by United States of America law antimicrobial effectiveness claims for hard surfaces disinfection. while having a common theme. the revision to the EU Annex I and several others. In very simple terms. This ensures that we use a scientific rationale to control contamination in our operations. The guidelines. one should use the guidelines as informational sources. While structuring one's environmental monitoring program. are not harmonized. Doing so would be a claim. ISO 14698. Some of the existing guidelines include. the FDA's 1987 Aseptic Processing Guidelines (in revision). the United States Environmental Protection Agency (EPA) under FIFRA. Through one's environmental monitoring program. We test our environment and subsequently address the contamination noticed with a proven and validated efficacious disinfection solution. one can determine what contamination exists in the facility and appropriately design their cleaning and disinfection program surrounding this scientific knowledge. Gain what knowledge can be gleaned from the writings. By review of one's environmental isolate list. a disinfectant and a sporicide. In choosing the appropriate chemical agents.Implementing a Cleaning and Disinfection Program 15 TESTING THE ENVIRONMENT AND DEVELOPING DATA Environmental monitoring is a subject warranting its own presentation. Each guideline's numerical values and recommendations slightly differ from each other. we need to review what the difference between a sanitizer. a sanitizer has the lowest ability to destroy vegetative cells of the three listed. In the world today. one can develop a list of environmental isolates that have been noticed in their operations. CHOOSING THE APPROPRIATE DISINFECTANTS AND SPORICIDES The utilization of environmental monitoring data to implement a cleaning and disinfection program in controlled environments uses science to justify our ends. the key is to successfully integrate and document a plan for the assured demise of these organisms. Rather. USP Chapter <1116> (in revision). The disinfectant destroys a broader spectrum of vegetative cells than the does a sanitizer and incorporates some log reduction of spores. one will find it necessary to test for viable cells both in the air and on the surface. Some contradict each other's content. . and specifically following the guideline's parameters may be required. one should cautiously approach the claim of adherence to one particular guideline. ISO 14644. then design an environmental monitoring program based on what information is applicable for one's specific operations. the existent guidelines are numerous. PDA Technical Report # 13. While there is no commonality in the guidelines. Specifically. ISO TC209. And the sporicide destroys all vegetative cells and spores.

A multitude of categorizations of chemical agents exist in the world. In Table 1. Staphylococcus aureus. we identified the EPA's Table 1: EPA-Approved Disinfectant Test Methods (for hard surfaces) Types of Claims Sterilizer AOAC Sporicidal Test Tuberculocide AOAC Tuberculocidal Activity Method Modified AOAC Tuberculocidal Activity Method Quantitative Tuberculocidal Activity Test Method Hospital Disinfectant AOAC Use-Dilution Method AOAC Germicidal Spray Products Test General Disinfectant AOAC Use-Dilution Method AOAC Germicidal Spray Products Test Limited Disinfectant Fungicide AOAC Fungicidal Test AOAC Use-Dilution Method AOAC Germicidal Spray Products Test Virucide EPA Virucidal Test Parameters Sanitizing Rinse (food contact surfaces) AOAC-available chlorine germicidal equivalent concentration method AOAC germicidal detergent sanitizer method Sanitizer (inanimate. and Pseudomonas aeruginosa Salmonella cholerasuis and Staphylococcus aureus Same as Above Trichophyton mentagrophytes Mycobacterium tuberculosis Test organism(s) Bacillus subtilis and Clostridium sporogenes 8 . nonfood contact surfaces) EPA Sanitizer Test Parameters Staphylococcus aureus and Klebsiella pneumoniae or Enterobacteraer aerogenes Escherlchia coli and Salmonella typhi or Staphylococcus aureus Specific Virus Claimed Salmonella cholerasuis. Understanding disinfectant categories is essential.16 Laboratory Validation Table 1 gives is a basic overview of what is required by a disinfectant manufacturing company by the US EPA to make the following claims on marketed products.

In development and validating of a cleaning and disinfection system. . high level disinfectants.25% Hydrogen peroxide above 6% (sporicidal reduction at 6%. Sanitizers: Isopropyl Alcohol 70% Ethanol at 70% Ethyl Alcohol at 70% Iodine (not normally used in clean room operations) Disinfectants: Phenols Quarternary Ammonium Hydrogen Peroxide at 3% or below Sodium hypochlorite below 0. Defined in the categories below are chemicals agents normally used in pharmaceutical and biotechnology operations. a sterilant at 10–35% depending on the wetted time) Peracetic Acid & Hydrogen Peroxide Glutaraldehyde Formaldehyde In reviewing the basic types of chemical agents used in pharmaceutical and biotechnology operations we can come to understand their basic differences and applicability in our operations. one or two disinfectants (depending on our belief in resistance) and one sporicide/sterilant. Others have categorized disinfectants as sanitizers. We need to choose one sanitizer. disinfectants. intermediate level disinfectants.Implementing a Cleaning and Disinfection Program 17 classifications for antimicrobial effectiveness claims that based on their passing of particular AOAC test parameters 10.10% Sporicides/Sterilants: Sodium Hypochlorite above 0. we need to identify three categories. Sanitizer: a germicidal product that reduces the population of viable microorganisms Disinfectant: a chemical agent that completely destroys pathogenic organisms but does not destroy all microbal forms such as bacterial endospores. Simplifying this list to three basic categories allows us to determine what we will require for our operations and subsequently be required to validate. Such chemical can render a product free of viable organisms including all bacterial endospores. sporicides and sterilants. Sterilants/Sporicides: a chemical agent that is capable of sterilizing. low level disinfectants.

ISOPROPYL ALCOHOL. 12 13 known to inhibit sporulation and spore germination . surfaces. However. When used its dry times are significantly less. its destructive power substandard to that of the liquid itself on the surface. the summaries are brief so we can develop a basic understanding of each chemical. . More than 90% of industry operation will utilize a 70% isopropyl alcohol solution to address clean room organisms as it has been proven more efficacious than ethyl alcohol (190–200 proof diluted to 70%) or ethanol 9a mixture with the base as ethyl alcohol [63%] and spiked with methyl [3%] and isopropyl [4%]) at a small percentage (rendering it undrinkable). but this effect is reversible . and thus. While the mechanism of wiping destroys cells in its action. The most used forms in the clean room operation are isopropyl alcohol and ethyl alcohol (ethanol and alcohol). This should not be confused in any way with the product's effectiveness if used in a saturated wipe. As alcohols are not sporicidal. Alcohols demonstrate rapid broad-spectrum antimicrobial activity against vegetative bacteria 11 (including mycobacteria). and fungi but are not sporicidal . As a complete chapter could be written on each. Alcohols come in a variety of forms. a 70% solution has far superior efficacy performance than the higher or lower concentrations. for viruses. A saturated wipe contains a limited amount of the chemical agent. Published alcohol effectiveness or results of alcohol effectiveness are presented when sprayed to the surface and allowed to air dry. viruses. ETHYL ALCOHOL.18 Laboratory Validation To follow is a brief description of each of the most used chemical agents in the industry. ETHANOL SOLUTIONS: Alcohols have been used for years in pharmaceutical and biotechnology operations for three basic purposes: • • as a sanitizing spray for gloves. they are not recommended for sterilization and are widely used for hard surface disinfection and skin antisepsis. as a cleaning or wipe down agent to remove possible existent residues from critical non-product contact surfaces as a product contact cleaning agent (ethyl alcohol only) • In testing antimicrobial products' effectiveness. As has been previously stated. the dry time of the alcohol is significantly faster. ethyl alcohol or ethanol seems to be slightly more effective and is used basically within the confines of the laboratory environment. etc. They are however. of the existent alcohols. carts. isopropyl alcohols demonstrate superior effectiveness against clean room organisms.

phenolics demonstrate superior antimicrobial effectiveness in an acidic base as opposed to an alkaline base. both residue-cleaning products eventually give . their ability to soak up residues and clean the surface is minimal. This will remove the majority of contaminants on the surface. PHENOLS Phenolics have been used for years as a disinfecting agent. or sodium para-tertiaryamylphenate. Overall. ISO 7) areas. During isopropyl alcohol wipe downs of non-product contact but critical surfaces. Sterilization of disinfection agents is discussed in depth in the section on Sterility of Disinfecting Agents. ISO 5) and adjacent Class 10. Phenolics are normally an amber (light tan) or light tan color when manufactured. we need to employ the cleaner of the two methodologies. phenols exhibit better antimicrobial effectiveness against gram-positive organisms than they do against gram-negative organisms. Here also such products pose a problem. Type 2. The soaking of the liquid with contaminants in suspension from the surface into a dry wipe is a superior methodology. they are used to wipe a surface in a cleaning operation (IPA wipe down) to remove existent disinfectant residues. While somewhat effective. we would spray the solution to the surface and wipe. However. and look as though the dirt was just moved around. Rendering of the products as sterile is a must prior to use in a Class 100 (Grade A and B. If one were to conduct the same experiment with a dry wipe and spray isopropyl alcohol. One drawback is the horrific residues that are noticed from long-term use of the products. The use of 70% isopropyl alcohol or certain residue removers can remove such residues in their early existence on surfaces.Implementing a Cleaning and Disinfection Program 19 Isopropyl alcohol wipes carry few if any claims. As they are saturated with isopropyl alcohol. the window will have streaks or swirls on it. sodium ortho-phenylphenate. They have limited activity against fungi and certain virus strains such as HIV-1 (AIDS virus) and Herpes Simplex.000 (Grade C. the surface would appear much cleaner. Phenolics normally are available in an alkaline and acid base version. A superior methodology would be the use of an isopropyl alcohol and a dry wipe. Normally. Like cleaning a window. This can be proven in a home experiment by using a saturated wipe to clean a window. Residues start as a “dripping droplet” that is not easily removed. Some of the most common chemical compounds in phenolic germicidal detergents are in a low pH phenolic: an ortho-phenyl-phenol and ortho-benzyl-benzyl-para-chlorophenol and in a high pH phenolic: a sodium orthobenzyl-para-chlorophenate. While phenols provide good broad-spectrum disinfection they are not sporicidal and have major drawbacks in their use. This color darkens with age and exposure to light (especially fluorescent). Phenolics are effective against gram-positive and gram-negative organisms. Most of the time these products just move around the contamination of the surface. The theory surrounding the rotation of these two compounds is described later in this section of this chapter. When completed.

The formulation in a closed container is relatively stable and should remain stable for a 30-day period in a closed container. As with phenols. Their spectrum of use is very broad and ranges throughout the industrial world. through the hospital and institutional setting and even can be found in home use. The . phenols exhibit better antimicrobial effectiveness against gram-positive organisms than they do against gram-negative organisms. ISO 7) areas. quarternary ammonium compounds have the most organisms registered as label claims with the US EPA. and less if the solution is in an open container (such as a bucket) and should be discarded each day. However. Quarternary ammonium compounds are effective against gram-positive and gram-negative organisms. Quarternary ammoniums have excellent detergency. however. Expiration of a formulated phenol also carries some concern. the positively charged molecule attracts to the negatively charged microorganism's cell wall. Compatibility with most chemicals is normally very good. Quaternary ammonium compounds normally utilize an alkyl dimethyl benzyl ammonium chloride or a dimethyl ethyl benzyl ammonium chloride. Their mechanism of antimicrobial effectiveness is related to their positively charged molecule. quaternary ammoniums do leave sticky residues that become problematic over time. While a multitude of formulations exist in the industry. Sterilization of disinfection agents is discussed in depth in the section on Sterility of Disinfecting Agents. Expiration of a formulated (from concentrate) quarternary ammonium also carries some concern. these are two of the most popular components. However. Rendering of these products as sterile is a must prior to use in a Class 100 (Grade A and B. They are one of the best cleaners among the spectrum of disinfecting agents. These cationic solutions also have excellent deodorizing capabilities. The formulation in a closed container should remain stable for a period of 7–30 days (dependent upon storage). Quarternary ammoniums come are available in both alkaline and acid based compounds. Transfer of such residues to an unwanted location is a concern and precautionary measures need to be implemented to ensure minimization of this scenario. Simply. The normality in the industry is 7 days. the effect of anionic characteristic of the chemical in relation to applications in conjunction with a cationic surfactants such as a quarternary ammonium have been reported as problematic. Formulated solutions should be marked accordingly. They have limited activity against fungi and certain virus strains such as HIV-1 (AIDS virus) and Herpes Simplex. ISO 5) and adjacent Class 10. The cycle of kill is complete when the chemical agent is absorbed into the cell and spread throughout the organism. Formulated solutions should be marked accordingly.20 Laboratory Validation way to the darkening phenolic that stains the surface with a dark brown color. QUARTERNARY AMMONIUM COMPOUNDS Quarternary ammonium products are used in more disinfection applications than phenolic germicidal detergents. they are not of the scale of phenolic residues. In fact and due to competition in this arena. Type 2.000 (Grade C.

in recent years such formulations from the Clorox Corporation® have been increase to a near 7% solution to ensure continued stability of the active percentage. However. acidification causes rapid degradation of the active elements in the solution and use of the product is limited to approximately two hours.25% and 0. ISO 7) areas. In formulation of a sodium hypochlorite solution. expiration of the solution can become a problem.25 or 0. Sterility of Disinfecting Agents. Sterilization of disinfection agents is discussed in depth in this chapter in section. However.000 (Grade C. many choose to acidify the solution.0 x 106 ) and there is no soil load. One of the main problems with the use of sodium hypochlorite is the industry is that it is used at too strong an active percentage. ISO 5) and adjacent Class 10. Sodium hypochlorite is used throughout the heath care setting and normally diluted to concentrations of 0. which makes it a more potent mixture when focusing on bacterial endospores. viruses.52% (or a 1-10 dilution or 500 ppm) sodium hypochlorite is effective against gram positive and gram negative organisms. Rendering of these products as sterile is a must prior to use in a Class 100 (Grade A and B. Some companies have .25% and 10% solutions.52% solution. however. In reality. Some formulate to parts per million.25% concentrate sodium hypochlorite normally carries a one-year expiration for an unopened container. Bleach as we know it is normally found in a 5.25% (or a 1-20 dilution or 250 ppm) sodium hypochlorite is effective against gram positive and gram negative organisms. A normal 5.Implementing a Cleaning and Disinfection Program 21 normality in the industry is 7–30 days and less if the solution is in an open container (such as a bucket) and should be discarded each day. viruses. SODIUM HYPOCHLORITE Sodium Hypochlorite solutions are one of the oldest known disinfectants and 14 sporicidal agents . The product is available in many forms including ready to use 0. to powders that are mixed with water to formulate a variety of solutions. One of the problems with sodium hypochlorite formulations is the method of formulation designation which varies from firm to firm. acidification may not be necessary as the product demonstrates excellent sporicidal characteristics in its neutral state and bacterial endospore levels in clean rooms are not exuberant in numbers (above 1. A formulation of 250 or 500 ppm has excellent sporicidal activity. in recent years. quarternary ammoniums have been made in ready-to-use formulas that are very stable. At a slightly increase use-dilution of 0. At a use-dilution of 0. fungi and more effective against a wider range of bacterial endospores.25% concentration. Both formulations are used throughout the pharmaceutical and biotechnology industry. However.52%. fungi and bacterial endospores. to concentrate 5. some to a percentage of a solution and some to a dilution such as 1–10.

In the pharmaceutical and biotechnology industry. and bacterial spores. Precautions should be vested in this arena. colorless liquid that is environmentally friendly as it can rapidly degrade into water and oxygen. Open containers (such as buckets. Sterilization of disinfection agents is discussed in depth in this chapter in section. ISO 7) areas. viruses. Depending upon the concentration used. viruses and bacterial endospores in lower enumerations. Some of the positive features of the product are its mild (if any) odor and its low residue characteristics. whether in a ready to use formula or a concentrate product. Destruction of spores is greatly 15 increased both with a rise in temperature and increase in concentration . hydrogen peroxide is effective against bacteria. hydrogen peroxide is used at 35% as a sterilant in isolators and at 3–10% for surface disinfection. yeasts. Sterility of Disinfecting Agents. the chlorine in the solution begins to burn off leaving only the sodium chloride. most hydrogen peroxide formulations contain a preservative to prevent decomposition. Hydrogen Peroxide Hydrogen peroxide is one of the most common disinfectants in the industrial marketplace. While generally stable. Sterilization of disinfection agents is discussed in depth in the section. At lower concentrations (3–10%) the chemical is effective against gram-positive and gram-negative bacteria. Some drawbacks with sodium hypochlorite focus mainly on the residue and corrosiveness of the product. However. As previously stated. Sterility of Disinfecting Agents. This white crystal-like residue attacks the impurities in stainless steel (as an example) over a longer time frame. In hospital and consumer use at a 3% solution the product is commonly used as an antiseptic. Rendering of these products as sterile is a “must” prior to use in a Class 100 (Grade A and B. When using a sodium hypochlorite solution it is imperative to remove the sodium chloride residues frequently to minimize this corrosive action. and open bottles) have a shorter expiration as the chlorine in the solution begins to burn off leaving only the sodium chloride. at higher concentrations and longer contact times the product exhibits superior sporicial reduction of bacterial spores. the product needs to be used within a 30-day period.000 (Grade C. ISO 5) and adjacent Class 10.22 Laboratory Validation validated and increased this expiration with applicable assay data over time to 18 months. However. Once opened. the product also has some setbacks in exceeding OSHA exposure limits if used in confinement in too large a quantity. Open containers should be formulated and used within the same day. ISO 5) and adjacent Class 10. ISO 7) areas. . Rendering of these products as sterile is a must prior to use in a Class 100 (Grade A and B.000 (Grade C. Hydrogen peroxide is a clear.

The chemical is considered a more potent biocide than hydrogen peroxide being bactericidal. at 20°C. This will significantly reduce odors. bioburden is significantly lower.Implementing a Cleaning and Disinfection Program 23 ]PERACETIC ACID AND HYDROGEN PEROXIDE Peracetic acid and hydrogen peroxide mixtures have received much recent attention in the pharmaceutical and biotechnology industry in recent years. if not intolerable to many users.3% as a sterilant in both ready to use and concentrate solutions. if used in a clean room environment that may have 15–20% fresh air. The chemical destroys the cell by destroying vital membrane lipids. Active percentages of marketed products range from 0. this value drops to near 0. Due to the horrific smell and the characteristic drying of mucosal membranes.3% to 1. Safety precautions for end users should be ensured prior to its use. Peracetic acid and hydrogen peroxide mixtures decompose to acetic acid and oxygen. The sterilant claim on products/labels follows the AOAC sporicidal test. Thus end-users should look to validate a concentration of peracetic acid and hydrogen peroxide mixtures as well as other registered sporicides at realistic bioburden values as discussed later in this chapter in the section entitled Determining Antimicrobial Effectiveness.2%. DNA and denatures proteins and enzymes. peracetic acid and hydrogen peroxide mixtures have caused dissatisfaction among end-users.3%) . . One of the main misconceptions with the use of peracetic acid and hydrogen peroxide mixtures as well as most registered sporicides is that the product needs to be used at the sterilant label claim active percentage. not present in clean rooms. subtilis and C. 16 fungicidal and sporicidal at very low concentrations (<0. First we must understand the requirements set for by the US EPA. This test requires the complete reduction of 106 of B. virucidal. the active percentage needed to destroy the flora normally seen is significantly less. in a soil load. The US EPA requires all registrants making label claims to do so by following test methods outlined in by AOAC protocol.4%. The product as originally designed for the sterilization of medial devices. While concentrate products need smaller amounts of acetic acid in the formulation. cporogones in a 60carrier test. Peracetic acid and hydrogen peroxide mixtures have pungent vinegar smell that is offensive. Simply. registered sterilant label claims are too strong for what is noticed in a clean room. Facilities that have used sodium hypochlorite (bleach) for years find the transition to peracetic acid and hydrogen peroxide very difficult to handle in terms of worker satisfaction. Concentrate solutions incorporate approximately 8% acetic acid and upon formulation to a use dilution. In the clean room. The product. may easily exceed required levels when industrial hygiene testing is performed. Ready to use solutions require a very high level of acetic acid as a stabilizer nearing 5. Coupled with the high enumeration of the AOAC test parameters is also a soil load. deterioration of surfaces. Thus. problematic user situations and residues.

Many pharmaceutical and biotechnology organizations do not use a glutaraldehyde product in their operations. While formaldehyde is effective. The mechanism of action of formaldehyde is assumed to be due to the reaction with cell protein and DNA or RNA (Russell. epoxies and most clean room surfaces. GLUTARALDEHYDE Glutaraldehyde has been used for some time as a disinfectant and sterilant for endoscopes and surgical equipment. aluminums.24 Laboratory Validation While reports of the product deem it non-corrosive to metals.0% solution. industry reports have shown this product react adversely with most stainless steels. glutaraldehyde's use has been focused mainly on the hospital environment.5 hours exposure to 300 µ/L whereas at 250 µg/L only a 4-log reduction was obtained after six hours of exposure. viruses. subtilis spores was obtained after 1. Glutaraldehyde has broad spectrum activity against bacteria. bacterial spores. Glutaraldehyde is the only aldehyde to exhibit excellent sporicidal activity. 1976). Glutaraldehyde is a powerful biocidal agent having the advantage of continued activity 17 in the presence of organic material. FORMALDEHYDE Formaldehyde is widely known as a fumigant for rooms and buildings. a white cloudy residue is normally left that requires either an IPA wipe down or a WFI rinse to remove. After application. Acklund et al. (1980) showed that at 20°C and a relative humidity of approximately 100%. and fungi. In recent years. The mechanism of action involves the destruction of the outer layers of the cell. plastics. It has been 18 shown to be effective against bacteria and bacteria spores and vegetative bacteria . this chapter focuses on routine methods of cleaning and disinfection. The product is very toxic and specific handling precautions must be employed prior to its use. Formaldehyde is normally used in the pharmaceutical and the biotechnology industry as a means to bring back an area after shut down or major maintenance. Formaldehyde does not fit appropriately as a choice in this venue and would be used as a mechanism in opening of a new area or as a method . Glutaraldehyde is normally sold in a 2. a 6-log reduction of B. The product is usually supplied as an amber solution with an acid pH. Especially noted are the gaseous fumes and the possible absorption through human tissue (skin). During its implementation very stringent safety precautions are assured for personnel protection that include areas and building clearance and hold times of areas prior to release.

as some may be harmful to final product or incompatible with clean room surfaces. and whether the chemical is capable of killing the organism in question. The key to selection is answered in one's list of environmental isolates. The mechanisms by which chemical germicides and antibiotics work are completely different and there does not seem to be a relationship between antibiotic resistance and chemical germicide effectiveness. organisms do not develop and immunity or resistance to a chemical agent over time. Grade A or B area) is normality for the pharmaceutical and biotechnology industry. These are not found in the clean room environment. however. Resistance to antibiotics is usually acquired through modification of a single gene (or acquisition of a single gene) that blocks the very specific action of the antibiotic. antimicrobial effectiveness depends on a variety of factors. However. We need to also realize that “natural selection” is a slow process that requires high populations of cells.Implementing a Cleaning and Disinfection Program 25 when coming back from a shutdown period. a theory that has never been proven in the clean room environment. such theory was implemented into the clean room industry. The choice delineates the rotation parameters. This meaning that multiple mutations . the rotation to a more efficacious product such as sodium hypochloride at 0. published data. The antimicrobials we deal with do not act on a single enzyme. Process constraints may also affect one's choice of disinfectants. some may decide to rotate similar disinfectants while also utilizing a sporicide. Subsequently and without proven. he choice to use a phenol. Varying applications require various solutions to be in place.52% (a sporicide) may be warranted to destroy this organism. An example would be a phenol may not kill a b. The theory was originally borrowed from the medical industry where it was applicable to noticed resistance in the human body. Resistance is used to describe "not destroyed by" and incorrectly used to describe "the building of an immunity by the organism to the chemical agent". However. ISO 5. constant selective pressure and a rich growth environment. antibiotic resistant microorganisms are susceptible (or killed) to chemical germicides. The decision is based on the correlation between the organisms on the list versus the chemical agent that can be proven efficacious against such organisms. The basis for the rotation of disinfecting or sporicidal agents is to address an organism that may not be destroyed by a particular disinfectant with another that has proven efficacy performance against such organism. but have numerous effects on almost every aspect of cellular physiology. As previously discussed. This theory is exactly that. quaternary ammonium or hydrogen peroxide based solution on a daily basis (in a Class 100. The two most notable are the time period afforded to the disinfectant to saturate and penetrate the cell wall of the organism. Rotation systems are designed to address known or possibly existent contamination with proven efficacious disinfectant.subtilis spore in a 5–10 minute contact time and thus. This confusion arises by the multiple meaning of the word “resistance”. The choice of one disinfectant and a sporicide is completely appropriate.

However. aeruginosa (or resistant sub populations) became adapt or resistant to the alkaline phenolic disinfectant.000 per test of one organism against one disinfecting agent (in solution or on carrier surfaces). An subsequently. If organisms could develop a resistance to disinfectants at the low levels that they are seen in the clean room. nor ever was destroyed by. Again. And if it were proven so. It turned from "in this test we found that P. During this time period. The pharmaceutical and biotechnology industry demands thorough testing in all aspects of our operations. we needed to investigate the subject further. Therefore. the industry would be forced into reproducing antimicrobial effectiveness testing on a routine basis. one chemical agent with another that has proven efficacy performance against such organism. On the contrary. From the paper: "The testing method consisted of treating P. aeruginosa was not completely killed by a low pH phenolic and was subsequently destroyed by a low pH phenolic" to "all organisms can develop a resistance to disinfectants".500 to $2. given the environmental conditions and low cell numbers within the clean room 19 environment . this test report was conducted in a laboratory setting. many jumped to what may have been thought to be some applicable data that should be implemented across the board with all organisms. one such study that reported supported the resistance theory was published in article form in 1992. could develop a resistance to a phenolic germicidal detergent in a laboratory experiment. when this compound was applied to the culture on a rotational basis with the acid phenolic disinfectant during the same time period. Data indicated that the P. In this paper they examined whether it was possible for a Pseudomonas 6 aeruginosa (ATCC 15442) at a population of cells equal to 10 CFU/ml. this is not likely. the first question that would scientifically be asked is "when does this occur?" As this question could not be answered. aeruginosa on agar surfaces. During this time period." However. the industry would have a horrific problem. and then repeating the process. entitled Rotation of Phenolic Disinfectants (Connor 20 and Eckman ). such testing would be done for an unscientific reason that would burden firms with an extremely high overhead cost. the data as it was presented was bastardized throughout the industry. the basis for rotation is to address an organism that is not destroyed by. The average time contact kill study may cost a firm from $1. A total of 40 transfers per treatment per trial and two trials ere performed. Added to the cost of routinely reproducing antimicrobial effectiveness would be the question of how often should this data be reproduced. The published article attempted to prove that if such an occurrence existed that the "resistant microorganism" would be destroyed by either varying pH phenolic. no adaptation or selection occurred. Simply an unanswered question that would force firms into a greater .26 Laboratory Validation would be required to overcome their detrimental effects. the basis for implementing a rotation system may not be to address the building of immunity by an organism to a chemical germicide. subculturing (transferring) survival cells. This was not factually proven. But in reality as an industry. And.

what do we rotate other chemical compounds with? Rotation of two disinfecting agents may limit us to a phenol rotation that may or may not be the best choice to choose. the push for the rotation of two disinfecting agents is becoming less and less supported. we discussed the possible need to reproduce antimicrobial effectiveness on a routine basis if we believed the theory of resistance. P. Thus. are we not destroying the cell during the first month time period? Could it compromise our product? Further. Obviously the scale of flora in the clean room setting would be much broader. While some disinfectants may have a very small log reduction of spores. so the selective pressure . Likewise. if we do not rotate a high pH phenolic with a low pH phenolic. The rotation of disinfecting agents to combat microbial resistance incorporates yet another stumbling block when we review the subject of a routine sporicidal application to our controlled environments. if one believed that an organism could become resistant to the disinfectant used. aeruginosa. Would one species of Staphylococcus be affected like another? All of these questions could be pushed to the foreground when one utilizes an unproven beginning basis as our starting point. as disinfectants are more powerful biocidal agents than antibiotics and are applied in high concentrations against low populations of microorganisms. Disinfectants and Antiseptics which states: "The development of microbial resistance to antibiotics is a well-described phenomenon. The rotation of two disinfectants does not address the possible content of spore populations in our clean room environments. conclusive evidence that relates to our specific scenario should be the basis for us determining our starting ground. For the industry to base all disinfection systems on the testing of one organism would represent an incomplete analysis of pertinent data surrounding this subject. If we use a disinfecting agent that an organism may become resistant to in this month. However. if we believe in resistance then the list of questions continues. how will this resistant microorganism that was not killed by one disinfectant but destroyed by another survive through the application of a sporicide? There is no scientific theory or case study that would support the cell's ability to live through this scenario. As an industry. we will still incorporate. Simply stated. "What organisms do we test?" In completeness. A multitude of microorganisms would be noticed in normal clean room operation. The development of microbial resistance is less likely. Thus.Implementing a Cleaning and Disinfection Program 27 frequency as time continued. a sporicidal agent into our disinfection regime. and one that it is not resistant to the next month. what do we rotate with a quarternary ammonium? We cannot rotate a quarternary ammonium monthly with a phenol as they are not compatible and create a sticky residue on the surface. The Conner and Eckman article tested one organism. on a frequency. In the above paragraph. all environmental isolates could be suspect. We can see this in review of some of the current guidelines published in the industry such as USP 1072. coupled with the question of how frequently should we test would be the question.

spp. the FDA released the Sterile Drug Products Produced by Aseptic Processing Draft." The use of theory may guide us to proof. Many common sanitizers are ineffective against spores. the writings make no mention of any concern that an organism may develop a resistance to a chemical agent over time. While this document may change." "After the initial assessment of sanitization procedures. and used for a limited time. disinfectants should be rendered sterile. ongoing sanitization efficacy should be frequently monitored through specific provisions in the environmental monitoring program. 2002. there are three types: . we think it could occur. Rarely in our internal scientific evaluations and our presentation of our validation to regulatory inspectors do we use the justification that "in theory. and limitations of sanitization agents should be assessed with their implementation for use in clean areas. Rotation is an applicable and a scientifically proven term." Resistance of microorganisms to disinfecting agents is a subject that is based on the theory that it could occur.e. 70% isopropyl alcohol is not effective against Bacillus. for example.28 Laboratory Validation for the development of resistance is less profound." On September 27. The effectiveness of these sanitization procedures should be measured by their ability to ensure that potential contaminants are adequately removed from surfaces (i. A sporicidal agent should be used regularly to prevent contamination of the manufacturing environment with otherwise difficult to eradicate spore forming bacteria or fungi. in several instances the guideline does mention that one should continually evaluate their systems. Sanitization Efficacy "The suitability. via obtaining samples before and after sanitization). Disinfectants should retain efficacy against the normal microbial flora and be effective against spore-forming microorganisms. An excerpt from the guideline is as follows. but never is a replacement for proof." "Upon preparation. Destroying contamination in a clean room operation requires addressing the known level of vegetative cells and the known level of spores. as specified by written procedures. While in the preliminary stages and not for implementation. If we define rotation of chemical agents as the "desire to assure a more broadspectrum kill in our disinfecting regime" then we have identified the key element for the reason why we would scientifically consider this subject. with a defined course of action in the event samples are found to exceed limits. However.. the rotation of two disinfectants is not applicable to address microorganisms we may see. In design of a rotation system. However. we should consider the basis for their thought process. efficacy. spores.

then apply the Sporicide Quaternary Ammonium Clean the surfaces.Implementing a Cleaning and Disinfection Program 29 • • • a single disinfectant rotated with a sporicide (Table 2) a two disinfectant system (rotated monthly) plus a sporicide (Table 3) the use of a sporicide alone (Table 4) A few suggested rotations of the chemical agents with frequency are presented here for use in a Class 100 and the adjacent Class 10. then apply the Sporicide Day 14 (if warranted Clean the surfaces. Table 2: Rotating One Disinfectant and a Sporicide DAY(S) Day 1–13 Phenolic Low pH Phenol Quaternary Ammonium Quaternary Ammonium Clean the surfaces. by EM data) allow to dry. allow to dry. allow to dry. Table 3: Rotating Two Disinfectants and a Sporicide DAY(S) Day 1–13 Phenolic High pH Phenol Quaternary Ammonium Quaternary Ammonium Clean the surfaces.000 environment. then apply the Sporicide Hydrogen Peroxide Clean the surfaces. when all surfaces are dry. allow to dry. when all surfaces are dry. then apply the Sporicide Day 15–29 Day 30 Low pH Phenol Clean the surfaces. allow to dry. allow to dry. all critical surfaces should be rinsed with hot WFI or an IPA wipe down performed. then apply the Sporicide After disinfection. allow to dry. then apply the Sporicide Hydrogen Peroxide Hydrogen Peroxide Clean the surfaces. by EM data) allow to dry. then apply the Sporicide Day 14 (if warranted Clean the surfaces. allow to dry. allow to dry. then apply the Sporicide Day 15–29 Day 30 Low pH Phenol Clean the surfaces. then apply the Sporicide Hydrogen Peroxide Hydrogen Peroxide Clean the surfaces. then apply the Sporicide Hydrogen Peroxide Clean the surfaces. allow to dry. allow to dry. . all critical surfaces should be rinsed with hot WFI or an IPA wipe down performed. then apply the Sporicide Quaternary Ammonium Clean the surfaces. then apply the Sporicide After disinfection.

as it is overkill for the known contamination level and will deteriorate surfaces over time. Validating one's sanitizing. allow to dry. we also need to address the cleaning aspect. The first two systems (Tables 2 and 3) require.52% Clean the surfaces. allow to dry. disinfecting and sporicidal agents requires them to delineate the organisms to be tested. then apply the Sporicide Day 15-29 Day 30 Sodium Hypochlorite at 0. As we have discussed previously.52% Peracetic Acid and Hydrogen Peroxide Peracetic Acid and Hydrogen Peroxide Clean the surfaces. Upon choosing 1 or 2 disinfecting agents and a sporicide. One could use a list of ATCC cultures. one can continue with the antimicrobial effectiveness studies.52% Sodium Hypochlorite at 0. or a Time Contact Kill Study (On User Surfaces) or an AOAC Protocol Study. however. then apply the Sporicide Hydrogen Peroxide Hydrogen Peroxide 6% Clean the surfaces. This may be increased or decrease in time frames and will be determined by the environmental conditions. then apply the Sporicide Hydrogen Peroxide 6% Clean the surfaces. . a monthly sporicidal application. by EM data) allow to dry. then apply the Sporicide Peracetic Acid and Hydrogen Peroxide Clean the surfaces.30 Laboratory Validation Table 4: Use of a Sporicide DAY(S) Day 1–13 Sodium Hypochlorite at 0. all critical surfaces should be rinsed with hot WFI or an IPA wipe down performed. when all surfaces are dry. These subjects will be completely discussed later in this chapter in the section entitled. allow to dry. then apply the Sporicide After disinfection. Industry standards look to a weekly application at the beginning and such application becoming lessened as control of the environment is consistently attained. The use of cleaner is considered an optional step in controlling existent residues and should be done at least once a month. cleaning and disinfecting are not the same. at minimum. After disinfection all critical surfaces should be rinsed with hot WFI or an IPA wipe down performed. allow to dry. DETERMINING ANTIMICROBIAL EFFECTIVENESS Determining what chemical agents will destroy a known level of one's environmental isolates or ATCC cultures is the next step. The third uses a sporicide daily and is the most effective but is not recommended. one needs to review the available disinfecting agents and determine which is initially appropriate for their operations. allow to dry. Within our cleaning and disinfection regime. Assuring Application of Our Disinfectants and Sporicides. then apply the Sporicide Day 14 (if warranted Clean the surfaces. Prior to conducting either a Time Contact Kill Study (Tube Dilution).

While this is the method used for registration. Pseudomonas aeruginosa. However. AOAC protocol testing is required by the US Environmental Protection Agency (EPA) to register a claim for a disinfecting agent. The rationales behind the choice of a carrier method are the beliefs that: • microorganisms that are dried are more difficult to chemically inactivate than those microorganisms in suspension that in the health care setting microorganisms are more often found in the dried 22 state than in suspension • Within the framework of antimicrobial effectiveness testing. normal dry time is 5 minutes at best. As the organism will be on the surface. The available surface area of the organism that can be contacted by the disinfectant that rests on the surface is 270°.0 x 10 CFUs. Candida albicans and Escherichia coli is acceptable. While our floors may remain . the surface test presents a more realistic scenario. a time contact kill study that confirms the destruction of a known enumeration of cells on an enduser's surface is more depictive than a time contact kill study done in a tube dilution (in suspension). Testing ATCC cultures such as Bacillus subtilis. This test requires the microorganism is to be dried on a non-porous carrier. The use of one's environmental isolates in a preferred methodology by most regulatory agencies. The EPA supports the use carrier methods for the evaluation of a disinfectant product's efficacy. one needs to review how one will address an organism in the clean room. It utilizes 60 carriers (a ceramic penicylinders) and requires a high enumeration value of equal to or greater 6 than 1.Implementing a Cleaning and Disinfection Program 31 utilization of one's environmental isolates nets a more exacting test for each unique manufacturing operations. Protocols use either AOAC Use-Dilution or AOAC Sporicidal tests procedures. Staphylococcus aureus. Normality is to test an enumeration greater 4 than or equal to 1. The reasoning for this is organisms dried on a surface better depict the situation of disinfecting an organism in the clean room. Our goal is to prove a 3-log reduction. Aspergillus niger. we also need to incorporate realistic contact times to depict representative dry times of the disinfectant on clean room surfaces. The available surface area of the organism that can be contacted by the disinfectant in the tube dilution study is 360°. Antimicrobial effectiveness studies need to be based on realistic bioburdens that may be noticed in the controlled areas.0 x 10 . Due to the significant movement of air from laminar flow in the clean room. 21 some guidelines like the British Standard BS EN 13697:2001 calls for a 4-log reduction as proof. Obviously. it may be too involved and expensive for pharmaceutical and biotechnology firms to utilize as a method for testing antimicrobial effectiveness. In determining which test to conduct. however not as exacting as testing a plane of one's known environmental isolates.

one should conduct their testing in a . one would open a bottle of disinfectant and age it for the time period that they plan to use it. For a concentrate. The use-dilution is then aged for the time period that it will be used (for example. Expiration for effectiveness can be determined by incorporating a simple variable in our test called aging of the disinfecting agent. Testing of a variety of times ensures one has data in the file to support a complete range of dry times (contact times) that may be noticed in their manufacturing or testing operations. Expiration dating of disinfectant effectiveness can also be tested by conducting an antimicrobial effectiveness test on a newly open bottle of disinfectant (ready to use or concentrate to use dilution) and then subsequently aging the solution for the expiry period and testing the active ingredients. the vertical surfaces will dry faster (3–5 minutes). ethyl alcohol or AAA ethanol at a concentration of 70%. the enumeration level of microorganisms. However. other critical factors also need to be determined. As time progresses. Upon completion. In performing antimicrobial effectives testing some other factors come to the fore that warrant attention. Thus antimicrobial effectiveness testing should incorporate a worse case scenario and utilize a 3–5 minute dry time. we need to continually updating our profile of organisms versus our chemical agents. At the expiration of the use-dilution it is then tested for antimicrobial effectiveness. While the type of test. and in 10 minutes excellent activity. for 7 days). Simply done. The correlation between the active ingredients at time of opening and their satisfactory stability at end of expiry provides the needed data to support the use of the agent for the time period. there must be the understanding and explanation of the relationship between the tested solution and the expiry active data that followed. Normally in 3 minutes the disinfectant shows average activity. many firms have begun to test three time frames to prove the disinfectant's activity over varying time periods. clean room operations do not have a soil load present and the most soil that exists would be that of disinfectant residues. In recent years. An exception to this rule would be sanitizing agents such as isopropyl alcohol. the aged concentrate solution is diluted to the prescribed use-dilution. in this test. A ready to use solution is tested at the 30-day period. 5 and 10 minutes be tested consecutively.32 Laboratory Validation wetted longer (possibly up to 10 minutes). and the contact time are some of the most critical factors to assess. say 30 days (concentrate and ready to use). The first is soil load. this testing will provide the justification for utilizing the chemical agents to destroy the known and possibly existent contamination in the facility. in 5 minutes good activity. This system of expiration proves that a solution. These products dry faster nearing 1–2 minutes and testing should be altered as such. in use for “X” time period has the ability to destroy an acceptable level of microorganisms that we have determined to be present in our operations. Normally. One of these is expiration of the disinfectant. The author supports this practice and would suggest the time period of 3. In the case of an existing soil load.

Commonly used as a soil load or an increase level of protein (a fetal bovine serum) at 5% v/v is added to the organism challenge to test the ability of the disinfectant or sporicide under the circumstances of a soil load (dirtied) condition. As an example of the same chemical having varying in-use time periods. once opened?" . This scenario needs to be validated for each chemical agent and each container type. Another factor that may surface is hardness and temperature of water. At the same time the temperature of the solution may vary in its effect against microorganisms as elevated temperatures tend to increase the ability of the chemical agents performance. are a required to assuring the effectiveness of a cleaning and disinfection system that will prove an acceptable environment during the manufacture of product. The basic validation question is "what is the time frame that we can prove the chemical's active percentage and sterility to be valid for. While disinfectant validations are expensive and time consuming they are foremost in most regulatory agency's minds. Required by all registrants of disinfecting agents must state in their labeling their ability or inability to achieve antimicrobial effectiveness in the presence of hard water (400 ppm as CaCO3). an isopropyl alcohol solution in an aerosol for can be used for a longer time period than a trigger spray bottle. ASSAY OF DISINFECTING AGENTS Analytical validation of our disinfecting agents tests that the required percentage of the chemical agent is present to ensure antimicrobial effectiveness.Implementing a Cleaning and Disinfection Program 33 similar situation. The reasoning for this difference is the aerosol container is sealed in a pressure vessel. And thus. the trigger spray container may slowly aspirate room air to the master reservoir that may compromise the level of active ingredient and the sterility of the container over time. If the appropriate use instructions are followed. Upon the use of this product past the first use is where we start to see the possibility that the percentage of the active ingredients may begin to slowly become too low to warrant continued use. a ready-to-use product or a formulation from concentrate is normally easy to prove as having a sufficient amount of the active ingredients to reconfirm the required percentage that was validated in our antimicrobial effectiveness studies. The reduction in percentage of the active ingredient due to evaporation or the question of sterility over time is not founded in this container type. On the other hand. Varying products have varying in-use time periods. Time periods range from 7–30 days.

The FDA has stated in its Sterile Drug Products Produced by Aseptic Processing Draft that.34 Laboratory Validation STERILITY OF DISINFECTING AGENTS Through our antimicrobial effectiveness studies. As all chemical agents may have an inherent bioburden (normally spores). we realize that disinfectants do not kill all organisms. The transfer of such organisms through our disinfectants to our controlled areas. Controlling the contamination from ever entering is much easier that subsequently removing it from the controlled area. and used for a limited time. we must ensure that such bioburden is removed prior to their entry to our controlled areas. we spread the disinfecting agents all over our walls. disinfectants should be rendered sterile. should be viewed as a catastrophic event. especially our aseptic filling areas." Purchasing a disinfectant or sporicidal product as sterile from an audited vendor requires one to review the following critical items as a quality control measure to assure what one is using is sterile prior to use: • • • • • • • Assessment of the bioburden of the solution Assessment of the bioburden of the container that the solution is to be filled into Pre-washing of containers (cleanliness level) Requires a filter validation proving microorganisms are retained by the filter Assay of the solution (RTU or concentrate) to an acceptable active percentage Filtering the solution at 0. as specified by written procedures.) Requires the assessment for expiration dating of an unopened container • • . ceiling and floors. If we review regulatory expectations in this area we will find the requirement to sterilize all disinfectants and sporicides prior to entry to the controlled environment. "Upon preparation. This proves that the sterility test is capable of growing organisms in the presence of the chemical agent. To even further reinforce the issue.2 microns Aseptically filling the product into pre-sterilized containers or exposing the entire contents to a terminal sterilization process such as gamma irradiation. Requires a lot-by-lot sterility test per current USP or EP compendium (conducting sterility testing requires the completion of bacteriostasis and fungistasis (B/F testing.

Aerosol container is the most lucrative type. Normally the testing of three processed lots for sterility provides sufficient data. Container types include aerosol. Requires validation to be performed using a lot by lot sterility test per current USP or EP compendium or a bioburden analysis for at least three lots at the beginning and subsequently routinely tested as a quality control check. trigger spray. squeeze bottle.Implementing a Cleaning and Disinfection Program 35 If the disinfectant or sporicide is to be processed sterile in-house. For obvious reasons. Conducting sterility testing requires the completion of bacteriostasis and fungistasis (B/F) testing. Requires the assessment for expiration dating of an unopened container • • • • Validation of a sterility claim for disinfectants should be a focus of one’s validation efforts. gamma . The pre-sterilization of our chemical agents prior to entry to the controlled area is simple common sense and common practice in the industry. However. and larger closed containers (1–5 gallon and larger). Container type is critical when using a sterile disinfectant.2 microns Aseptically filling the product into pre-sterilized containers or exposing the entire contents to a terminal sterilization process such as gamma irradiation. then the assurance that all that comes in contact with the product after the filter is rendered sterile. If the product is aseptically filtered. This proves that the sterility test is capable of growing organisms in the presence of the chemical agent. the container and its contents must be sterilized via gamma radiation or all the components pre-sterilized and subsequently filled via a validated aseptic filling operation. • • • • • • • Assessment of the bioburden of the solution Assessment of the bioburden of the container that the solution is to be filled into Pre-washing of containers (cleanliness level) Requires a filter validation providing microorganisms are retained by the filter Assay of the solution (RTU or concentrate) to an acceptable active percentage Filtering the solution at 0. Sterility testing on a lot-by-lot basis need not be performed if sufficient validation testing is conducted. then the review of the following critical items as a quality control measure is suggested to assure what one is using is sterile prior to use. as the vessel does not aspirate the room air to the master reservoir.

TYPES AND STERILITY OF CLEANING COMPONENTS The sterility of disinfectants and sporicides is essential to assure the non-transfer of organisms to the Class 100–10. the container itself aspirates the room air back to the master reservoir. mop frames. mop heads and other components used during the cleaning and disinfection operation.900. Thus. If in a smaller aerosol. . We may also want to limit that the capping of product “to be used later” as our assay and sterility may be compromised over time. we may want to utilize a product that is pre-sterilized in this smaller form rather than attempting to pour or filter such solutions to a empty pre-sterilized container. Other smaller containers include the squeeze bottle and trigger spray bottles. A pre-sterilized aerosol or pressurized vessel assures assay of the active ingredients (for the time period they remain stable) and sterility for the expiration period designated by the manufacturer. trigger spray or squeeze bottle. assay and sterility are compromised after the initial use.000 areas. no viable or particulate contamination is returned to the solution contained. was marketed under US patent 6. the first sterile disinfectant. Method of Sterilization .000 areas via our cleaning components. mops sprayers. DECON-AHOL® (a 23 sterile USP IPA). Questions may arise as to the assay and sterility of this remaining solution over time. mop handles. This system is superior to that of pouring a concentrate disinfectant or sporicide into a measuring cup and capping the remainder for later use. Thus. In 1992. A pre-sterilized aerosol or pressurized vessel assures assay of the active ingredients (for the time period they remain stable) and sterility for the expiration period designated by the manufacturer. For varying disinfecting and sporicidal agents a variety of containers need to be utilized. The same is true for larger 1–5 gallon containers and larger. we may want to look to implement unit dose bottles that are incorporate a pre-measured dose and are sterile. buckets. While acceptable containers for all disinfectants and sporicides. Our cleaning components are defines as spray-mop-fog pressure devices. Of equal importance is the transfer of organisms to the Class 100–10. Thus. Once opened sterility is compromised.123. For ready to use mixtures we have to decide how the product will be used. no viable or particulate contamination is returned to the solution contained. For concentrate products that need to be diluted with a quality water grade.36 Laboratory Validation sterilization of the entire contents is considered far superior methodology for achieving a sterile product. This patent and product showed the industry the effectiveness of this type of container.

as much as can be sterilized should be sterilized. If it is stored outside the Class 100 area." Both sides have their valid points. A typical quality assurance line may be "You need to sterilize each component prior to use to reduce the ingress of contamination. We can then start with a clean component. If this is done. of the components that enter the area during manufacturing. buckets. quality assurance may not be completely abreast of what it will take to accomplish this scenario. the outside. But the real question is frequency. A typical statement from production may be "Well. How often do we clean and sterilize these components? Spray-mop-fog devices. Dependent upon the location of quality water that will be used to dilute our concentrate disinfectants. they will not be sterile any more. Prior to sterilization we must review the subject from the end of the cleaning operation and assure that these items are cleaned prior to sterilization." However. On the production side of the subject if we utilize sterile components we may possibly need to purchase two to three times the numbers we presently have (due to sterilization schedules) which is very costly. Moreover. and will be required to decontaminate the exterior of the wrappings prior to entry to the Class 100–10. underneath and the wheels. While not every item can be sterilized prior to entry to the aseptic manufacturing area. This includes the inside. The goal in the sterilization of the cleaning components is to reduce bioburden to the area. Our cleaning components and our disinfectants represent a common place where we can start anew each day with control. Production will have to transfer the wrapped sterile components to the controlled areas. If this occurs. Otherwise. we wheel in big tanks during manufacturing and they are not sterile. chances are lessened for contamination entering. All considerations need to be reviewed. these items should be of little consequence. At the same time. mop frames and mop handles need to be cleaned after use. we may need to open such components outside the controlled area. bioburden on the items and deterioration to the items will increase over time.Implementing a Cleaning and Disinfection Program 37 In review of this subject we see problematic quality control situations that may arise from these items not being rendered sterile. At this juncture we need to determine where the component will be taken and stored. we again corrupt our solution and possibly the surfaces that we touch with the mop head.000 area. if we then dip a non-sterile mop head into the solution. as they will have disinfectants in and on them during the disinfecting operation. And finally. Let's review the quality assurance side of the subject: the utilization of a sterile disinfectant that is diluted in a sterile quality water grade (such as sterile water for injection) and that is subsequently poured into a non-sterile bucket compromises our solution's sterility and our environment. then . which can be awkward. Incorporating a purchased or pre-made sterile water grade available in the area will also add significant costs. manufacturing operational difficulties may exist in implementing these items as sterile. If our handles and mop frames are not sterile they too can bring in unwanted contamination to the area.

IN-USE EXPIRATION OF DISINFECTANTS AND SPORICIDES Questions surrounding the expiration of an in-use disinfectant and sporicide will be questioned by all involved internally and externally by regulatory agencies. The same is true for mop heads.000 operation. we need to clean these devices each day and disinfect these items prior to entry to a controlled area. Most of the time. say. pre-sterilized and used per cleaning operation. Bioburden levels build up on the mop head and may cause potential problems. The mop handles and frames should also be cleaned prior to sterilization. Disinfection should be done using the chemical agent used in the cleaning operation. however. we may have a mixture of chemical agents. if a disinfectant is used in. They are small enough and inexpensive enough to have a variety on available. however. Science may say that we need to use a sporicidal on the items prior to entry.38 Laboratory Validation the item needs to be disinfected or sterilized again prior to entry. disinfection or sterilization prior to entry still looms as a unique question that needs to be answered by each operation.000 is it is dirtied and is normally air-dried. this is the case. As the scale increases to larger operations this becomes more difficult. These problems may not be worth the cost of the mop head.000 could move forward to weekly. The two main questions that arise are: • How long does the disinfectant retain its active percentage enabling it to destroy microorganisms? . An important factor in consideration is where the water will be obtained to create the dilution. The reasoning for routine discarding of the mop in Class 100. Pre-sterilized mop heads (foam or string) should be used and changed daily in a class 100 and 10. As scientists we know that sterilizing the buckets is the superior methodology and we should make arrangements to move towards such a scenario. While we understand the need to clean the components after use. Of enormous expense would be the establishment of new WFI water drop lines to serve this operation. The frequency of changing a mop head used in class 100. the bucket during that cleaning shift. monthly or quarterly basis does very little to control bioburden to the area on a daily basis and should not be considered. At the minimum level. This should be avoided. a better scenario is also to use this mop head and then discard. Mop handles and mop frames are a different story. Several methodologies were previously discussed in this chapter (in the Antimicrobial Effectiveness section) as to how to develop this data. Sterilization of buckets on a weekly. As an industry smaller firms or firms with only a few areas seem to be able to sterilize these components before use. And so the story encounters more complication.

Upon completion and drying of all surfaces. two of the most frequently-asked questions in the industry surrounding disinfection. Upon completion of the monitoring. the magnitude of this testing should outweigh the coordination of scheduling. some organizations have taken cultures enumerated at 10–100 CFUs and inoculated a similar production surface. CONDUCTING IN SITU FIELD STUDIES Once a disinfection system has been chosen and antimicrobial effectiveness testing has been completed. This testing is the movement of lab knowledge to the real life situation.Implementing a Cleaning and Disinfection Program 39 Table 5: In-Use Disinfectant Expiration Approved Disinfectant or Sporicide Chemical agent Concentrate Bottle Ready to Use Unit Dose Formulations in Open Containers (Buckets and Tanks) 1 day after formulation Formulations in Closed Containers such as Squeeze. the room is cleaned and disinfected per the current operating procedures.) and closed containers (such as squeeze bottles. As an alternative method. Simply an assay over time of use and an accompanying sterility or bioburden test will delineate this time frame. both air and surface. Satisfactory results need to be obtained in three different and separate in situation field studies prior to acceptance of the disinfection system. Coordination between the quality assurance group and the production services or cleaning group is sometimes a very difficult task. trigger sprays and closed bottles). unit dose concentrate bottles. As an example. environmental monitoring. SOP's should look to address the various type of use situations as depicted in Table 5 and identified as concentrate bottles. from the author's standpoint. Simply. the room is monitored again. It tests our overall effectiveness in conducting a disinfection operation. Such methods are: . is conducted in a dirtied room. Trigger Spray or closed bottles 7 days after formulation 30 days after opening 30 days after opening • How long does the container and the contents held within the container remain sterile? Both are very important questions and. ready to use bottles. conducting an "in situation field study" is important to prove the effectiveness of the combination of our cleaning SOPs (standard operating procedures) and our antimicrobial effectiveness testing. open formulations (in tanks. However. After inoculation the surface is cleaned using a multitude of methods that may be employed. The importance of reporting this information to cover all potential situations is critical. etc. some have just monitored a dirtied surface similarly used in production. Likewise.

The effectiveness of these sanitization procedures should be measured by their ability to ensure that potential contaminants are adequately removed from surfaces (i.e. If the system fails. Regulatory agencies such as the FDA. it is still not a test of the actual environment and the cleaning that will be performed. If this is done.40 Laboratory Validation • • • • mopping spraying wiping fogging The surface is then monitored again after cleaning as a test for effectiveness of the cleaning operation in conjunction with the disinfecting agents. it behooves organizations to conduct the actual in situ or field study in their manufacturing areas. the system. efficacy. this will be noticed in our environmental monitoring program and such corrective action must then be taken on an immediate basis. should prove effective each time it is performed. consider the in situ or field study extremely important. While important. if performed to accompanying SOPs. the system should satisfy the need for an acceptable environment. Inoculation of actual production services in controlled areas should never be done. and limitations of sanitization agents should be assessed with their implementation for use in clean areas. it is . While the in situ or field study conducted on similar surfaces nets good information. Our lab work in our antimicrobial effectiveness testing renders us the understanding of what our disinfectants and sporicides are capable of destroying in the presence of high bioburden levels. Thus. In support of the industry. as a system (chemical and method) to render a tested “dirtied surface” as free of microorganisms." One concern that is vested throughout the industry from regulatory guidance is the need to routinely perform this task. As an industry.. as one would never introduce any microorganisms knowingly to the area that may compromise manufacturing of product later in the scope. via obtaining samples before and after sanitization). Once the system is in place and the testing conducted thereof. such routine testing is a revalidation of a validated system. In an excerpt from the FDA's Sterile Drug Products Produced by Aseptic 18 Processing Draft 9/27/02 guideline is as follows: Sanitization Efficacy "The suitability. The in situ or field study is a realistic test of the ability of our cleaning and disinfection systems. the concern needs to be vested in the absolute assurance that such cleaning and disinfection procedures are followed on a daily basis.

the undesirable environment is the clean room itself. and our procedures relating to training of our personnel to accomplish these critical tasks. As an industry. mop it or rinse it from the area. This requires routine cleaning of the area. Second. This is true to some degree. We can wipe it. such as particulates and water. we can also fatally damage the cell wall by an abrasive friction type action as is common by wiping or mopping. Putting the basics into perspective can simplify our understanding of this subject and allow us to refocus our efforts appropriately. And third. In reality. One way we can do this is by removing the organism's required food source. we may spend too much time trying to figure out which chemical agent is most appropriate to destroy all the possible organisms that we may encounter. we need to assure the saturation and penetration of the cell wall over a lengthened contact time. we need to make its existence in the area very difficult. Our procedures need to include elements such as: the cleaning of a surface. it is also true that a residue on the surface can complicate our manufacturing for a variety of reasons. the saturating of a surface. cleaning and sterilization of our sprayers and foggers. if permitted by our safety group and by the characteristics of the disinfectant) or by assuring it must exist in an undesirable environment. To do so. we can remove organism from the area by our cleaning practices. All of these factors will assist our disinfection efforts. Some consider the disinfectant or sporicidal residue as an undesirable environment. routine change of dirtied solutions. . First and foremost. we hear tales of this chemical is not effective against this species of an organism. However. we need to view more than just a wetted surface. We need to identify the factors.Implementing a Cleaning and Disinfection Program 41 still a laboratory test and never ventures to a real manufacturing setting. This will be discussed in detail later in the chapter. We need to reproduce the contact times from our antimicrobial effectives testing on all surfaces in the clean room. that will assist our disinfection efforts. and for a short time period. Throughout the industry. routine changing of our dirtied mop heads. In short. At the same time we are focusing on which chemical kills what organism. Our design then relies on the cleaning and disinfecting procedures that we have in place. we do not spend enough time on this extremely important function. Another way is to change the temperature it is exposed to (Hot WFI. we need to assure that the chemical agent is appropriately applied to the surface. it's a difficult place to survive. ASSURING APPLICATION OF OUR DISINFECTANTS AND SPORICIDES The most important subject is the arena of disinfection is application of the chemical agent to the surface. In fact. Sufficient support conditions for sustaining an organism's existence are lacking in the clean room environment. The in situ or field study is a critical test of our systems. prior sterilization and cleaning of our buckets and mop handles. aside from the chemical agent.

42 Laboratory Validation Application is normally done by four methods: spraying. Increase in soil load and bioburden occurs in the bucket and the solution is dirtied in a short time. In this system. and piped facility pressure systems. Buckets come in a variety of makes and models. Mopping a disinfectant to the surface creates a non-destructive abrasive type action that loosens particulates. handles. residues and microbial contamination. Thus. spray-mop-fog devices. then walls then floors. spray-mop-fog devices (Core2Clean®). it does not clean the surface as well as mopping. wiping and fogging. residues and microbial contamination. While spraying is effective in efficacy performance. the disinfectant or sporicide is placed into a bucket and the mopping procedure calls for the dipping of the mop into the bucket. the surface remains wetted for a longer time frame. In the single bucket system. While mopping creates an excellent scenario for cleaning. Types of spray devices include canister sprayers. it is important to rinse the surface to remove the contaminants from the surface. Upon rinsing. After this has been done. The mopping action should be done from the top of the wall to the bottom with overlapping stokes. Mopping the floor is done by starting at the furthest corner and working towards the exit door (similar to painting a floor). the key to assuring saturation and penetration of the cell wall for the specified contact time is though a light spray with a larger droplet size. mop frames and mop heads. Methodology for cleaning a surface (not disinfecting) using a mop is enhanced by a quality water rinse such as WFI. it is a very critical step that needs to be accomplished on a routine basis. the wringer and application solutions are mixed in one chamber. pressurized canisters. Cleaning and sterilization of these components has been previously discussed. mopping. Mopping requires a variety of components that include buckets. However. it is less effective in addressing microorganisms as the mop head and the wall are less wetted. the surface dries faster. Spraying should be done from the top surfaces to the bottom of the walls. Spraying should not be done on floors and the correct option would be to apply the chemical via mopping. then to the surface where the chemical is to be applied. double and triple bucket systems. Simply. Mopping should be done in order: ceilings. then a disinfectant or sporicide is applied. Once all surfaces dry. Similarly. While pressure spraying does clean the surface better. During the cleaning operation the non-destructive abrasive type action loosens particulates. and arrangements need to be made to assure the reduction of bioburden prior to entry. the ceiling should incorporate the same method over overlapping stokes and start from the furthest corner of the room. Some include the single. Spray nets the best coverage of a surface and proves to be the best form for assuring efficacy against microorganisms on the surface as the dry time or contact time is longer. The double and triple . Spraying should be done by applying to the ceilings first (without contact to filters) and then be done to the walls. then to the wringer. Effective spraying is done from a light spray and not a power or pressure spray. the dirtied solution on the floor is collected and removed.

A mop is changed: • after use in each class 100 room. Compatibility with the disinfecting agent and capability of sterilization should be tested prior to use. These devices are made of stainless steel and completely autoclavable.000 or 100. Mop heads also come in foam flat mops for wall and string mops for floors. the mop enters the front chamber to be wetted and is wrung into the back chamber. • • . Off the tank exists the ability to connect a trigger spray activated power sprayer or a trigger activated mop (sending liquid automatically to the mop head or a fogger (allowing 2–3 hours of fog time).00 for each sterile mop head. Buckets should also remain chemical specific as the mixture of chemical agents and their accompanying residues should be avoided (especially in a liquid state). The question of disposable or reusable is an operation specific concern. Disposable means use once and throw away and the cost of doing so is approximately $8. Procedurally (in a two-bucket system).Implementing a Cleaning and Disinfection Program 43 bucket system attempts to separate the wringer solution from the application solution by creating separate chambers.000 areas. Mop heads come in disposable and reusable forms. The three-bucket system exemplifies the two-bucket system and adds one additional bucket as middle rinse bucket for the mop. galvanized steel or some form of plastics or autoclavable plastics. A mop is never used in a Class 10. The same mop head may be sequentially used in Class 10. This system is suppose to keep the solution cleaner as the rinse is collected in the back bucket and never contacts the application solution in the front chambers. the canister is pressurized with compressed air (ASME rated to 100 psi). Washing and resterilizing each mop head is an involved process and an operation should review the total costs and possible impact to the environmental conditions prior to implementing this scenario. While these systems attempt to reduce the soil and bioburden load in each chamber. A single use principle should be applied in all classified areas. These systems dispense a clean liquid continuously from the canister. In recent years spray-mop-fog systems have been designed and patented for use in controlled areas. They provide a chamber for the disinfectant or sporicide and once introduced. double and triple bucket system and the spray-mop-fog systems all use mop heads. the mop head is the contaminating factor of the solutions and eventually corrupts or soils all cavities in the system.000 and then used in a class 100 when it is noticeable dirtied when the mop head is damaged. Material grades of buckets are normally made of stainless steel. The single. Superior wetting to the surface occurs as the and cleaning time is 24 reduced by nearly 50% .

however. Fogging is the last method of application. fogging does not clean the surface: it only disinfects the surface. Consider cleaning a window at home as an example of the action needing to be performed in a cleaning operation in controlled environments. the process or manufacturing operation.000 and 100. Wiping is the fourth way of application. Likewise. personnel. One of the most confusing subjects surrounding application is frequency. The key is to wet and loosen the contaminants from the surface and then wipe them away with a dry wipe.4% use dilution) in a 12 25 foot x 15 foot room. Fogging also requires room release times that may near 3–4 hours. To clean with a wipe. It is true that some chemical agent residues create an undesirable environment for an organism to live. there are many important aspects that need consideration. contamination control capabilities. Coupled with the classification is the review of the environmental test data from each area. for disinfection.44 Laboratory Validation Mop heads for class 100 and 10.0 µm . one should use a chemical agent and a cleaner. In understanding how to accomplish this we must understand that if we had an area that showed very low bioburden. By applying a disinfectant and/or . the classification of the area. This release time is to assure harmful vapors have subsided to a safe level for personnel to enter per OSHA standards. While fogging a room can be very effective. Normality is frequency is dependent upon the classification of the environment. product changeover. it is necessary to saturate the surface. subtilis at 1. using two two-jet foggers for two hours . While many other factors come into play. This droplet size accompanied with the ability of the fog or mist to be circulated to all surfaces is the key to success. such residue kill is shortterm and should not be a part of our system design. we would destroy no more microorganisms than existed in the area. The droplet size required to effectively fog an area and saturate all 19 surfaces is 25. and the type of facility and its associated controls.000 environment will all be cleaned and disinfected with varying frequencies. a very wetted wipe is excellent for disinfection but very poor for cleaning.000 should be rendered sterile prior to use. these are the main culprits for contamination of a manufacturing facility. Once disinfectants and sporicides have dried their ability to kill is complete. First is coverage and droplet size. Fogging can produce very good results in destroying levels of vegetative bacteria and spores on the surface but is dependent upon the chemical agent that is used and the time frame of the fogging 5 operation. Carrier surface tests inoculated with b. The use of the environmental test data to classify the frequency of an area is superior to any other methodology. 10. and invoked a daily sporicidal application. if any. A class 100. While this study shows the ability of the method to destroy microorganisms on the surface.6 x 10 (11 carriers) show the destruction of such spores using a peracetic acid and hydrogen peroxide mixture from a concentrate (DECON-SPORE 200 Plus® at a 0. As was expressed earlier in the chapter. Frequency of application is dependent upon a variety of factors that include the bioburden of the area.

000 Class 100 surrounded by Class 100. the following charts cultivate a guideline structure to base our disinfection and sporicidal frequencies.000 Walls Daily spray or mop Daily spray or mop Daily spray or mop Daily spray or mop Weekly spray or mop Monthly or quarterly spray or mop Floors Daily mop Daily mop Daily mop Daily mop Ceilings Weekly spray or mop Weekly spray or mop Weekly spray or mop Weekly spray or mop Weekly spray or mop Monthly or quarterly spray or mop Curtains Daily or per shift application followed by 70% IPA wipe Daily or per shift application followed by 70% IPA wipe Daily or per shift application followed by 70% IPA wipe If applicable.000 Class 10.000 Class 100. From the experience of this author.000 Class 10. daily or per shift application followedby 70% IPA wipe If applicable.000 Not Classified Areas that may effect classified areas Table 6: Frequency of Disinfectant Application Classification Class 100 Class 100 surrounded by Class 10.000 Class 100 surrounded by Class 100. In discerning frequencies. we do not create a preventative measure for organisms that may enter the environment. The can be classified as: • • • • • • Class 100 stand-alone (gowning room to a Class 100) Class 100 surrounded by a Class 10.Implementing a Cleaning and Disinfection Program 45 sporicide. daily or per shift application followedby 70% IPA wipe Not applicable Class 100.000 Daily mop Not classified Weekly mop . we need to first identify the type of areas that may exist in our facility.

000 Class 100. In review of the level of residues attained from each chemical agent we see that if not addressed on a routine basis.000 Not classified Upon Action EM Upon Action EM Upon Action EM determination determination determination of problem of problem of problem RESIDUES With any application of a disinfecting agent comes the accompanying residue problem.000 Class 10. Followed by 70% IPA wipe If applicable.46 Laboratory Validation Table 7: Frequency of Sporicidal Application Classification Class 100 Walls Weekly or biweekly spray or mop or following EM at action level Weekly or biweekly spray or mop or following EM at action level Weekly or biweekly spray or mop or following EM at action level Bi-weekly or monthly spray or mop or following EM at action level Monthly spray or mop or following EM at action level Floors Weekly or biweekly mop or following EM at action level Weekly or biweekly mop or following EM at action level Weekly or biweekly mop or following EM at action level Bi-weekly or monthly mop or following EM at action level Monthly mop or following EM at action level Ceilings Weekly or biweekly spray or mop or following EM at action level Weekly or biweekly spray or mop or following EM at action level Weekly or biweekly spray or mop or following EM at action level Bi-weekly or monthly spray or mop or following EM at action level Monthly spray or mop or following EM at action level Curtains Weekly or bi-weekly spray or mop or following EM at action level. Followed by 70% IPA wipe If applicable. Followed by 70% IPA wipe Weekly or bi-weekly spray or mop or following EM at action level.000 Class 100 surrounded by Class 100. Followed by 70% IPA wipe Weekly or bi-weekly spray or mop or following EM at action level. . Isopropyl alcohol and ethyl (ethanol) alcohol are two of these disinfectants. Only a few disinfectants do not leave a residue. weekly spray or mop or following EM at action level. In fact. such residue can develop into a critical problem in our operations. weekly spray or mop or following EM at action level. Followed by 70% IPA wipe Not applicable Class 100 surrounded by Class 10. both isopropyl and ethyl alcohol can assist in the removal of disinfectant and sporicidal residues.

Implementing a Cleaning and Disinfection Program 47 Table 8 : Residues from Disinfectants Chemical Tested High pH Phenol Low pH Phenol Quaternary Ammonium Bleach @ 5. Likewise training our personnel on such procedures is critically important. This is coupled with the ability for air pockets to form underneath residues that may possibly break and release contaminants to the environment.52% Hydrogen Peroxide Peracetic Acid/H202 RTU Peracetic Acid/H202 Concentrate When Applied 759 ppm 731 ppm 133 ppm 929 ppm 144 ppm 0. Residues are most notably the cause of deterioration of surfaces in the clean room. or the chemical agent stains or discolors the surface • It is first important to conduct compatibility testing to assure the chemical agent does not harm the surface. Once this has been completed. DEVELOPING PROCEDURES AND TRAINING Developing SOPs is critical in design and assuring correct implementation of our systems. routine cleaning can remove or reduce most residues. Deterioration occurs for two basic reasons: • the chemical agent reacts adversely. The surface.067 ppm 123 ppm 44 ppm After Rinse 61 ppm 41 ppm 11 ppm 66 ppm 14 ppm 0 ppm 16 ppm 6 ppm 26 Some problematic situations with residues relate to the complication of the surface to be disinfected. or is incompatible with the surface and attacks the material composite material or impurities in the surface. Most notably is stainless steel. In developing the required applicable SOPs. . as personnel would touch a surface and then make a critical aseptic connection. if laden with chemical residues compromises the disinfectant and/or sporicide's ability to fully contact an organism. All other departmental operating procedures can then reference this particular operating procedure in their content. Personnel can transfer them to a critical site. This may compromise the integrity of the clean system in place. a GMP facility needs to write a very complete “Use and Rotation” operating procedure. Most residues are unwanted. Such transfer may occur.25% Bleach @ 0.

mop frames. etc. our next step is to assure that all is done to what has been written. we cannot assure personnel in varying departments will all follow a general company scheme of disinfection. wipers. Standardizing disinfection plant-wide should be a main objective of the company. expiration dating. storage. Including dilution mixtures. buckets. Without the ability to prepare this critical standard operating procedure. sporicides. mop handles. While SOP's will vary from firm to firm.48 Laboratory Validation Critical “Use and Rotation” SOP's should contain the following pertinent data. and saturated wipers Specific guidance as to the general corporate issues surrounding the preparation of disinfectants and sporicides such as approved dilution water types. use instructions. storage of unopened containers. Only in specific instances where EM data warrants the alteration from the general master plan should such systems change in specific areas. storage of opened containers How are the disinfectants to be applied in varying classifications and on varying surfaces Frequency of application of disinfectants and sporicides Approved mop heads. sprayers and foggers How each specific are is to be cleaned and disinfected Specific Approved Disinfectant and Sporicide Usages Method of Application Frequency Approved Equipment and Materials Procedure . container types. Too often we train only our general workers and forget to train Table 9: Standard Operating Procedures Standard Operating Procedure Section Title Purpose Scope Responsibilities Safety Precautions precautions for each product Approved Disinfectants/Sporicides and Cleaning Products General Guidance in Preparing Disinfectants and Sporicides Description The purpose of the SOP or why it is being done A scope of what is covered in the SOP A listing of whom is responsible for each capacity in the SOP Internal corporate and product specific safety A listing of the approved disinfectants. Once we have established our SOPs. Training is without doubt one of the most critical elements in our spectrum of disinfection. This is not an easy task and the main method of assuring this occurs is through our training. dating of containers. the following is a general guideline list of the important areas. Specific approved usage instructions for each disinfectant and sporicide.

what the organization will be required to do to assure the function's completion. Utilization of past training methods should be reevaluated to see if implementation of a better methodology exists. An unknowledgeable management person in the area of disinfection can harm our efforts. But “what is training and what content should be included” is a commonly asked question. Third. However. Hysteria during a crisis situation will cause us to place a band aide on the situation and not create a strong infrastructure that will assure our success for years to come. Their attention span is short within a classroom setting and "learning by doing" is a far superior method. The worst thing that can happen to an organization is that the trainees learn and prove that the methods in place are not scientifically justified or flawed. And last. It is important to train all involved in the disinfection process on a routine basis. Fourth. they may lack the ability to notice a shift away from acceptability. An outside source of training may provide a fresh approach that one's employees may enjoy. Trust then becomes a concern. Too often time period are too short to complete this task and shortcuts may be taken in order to run product sooner. People have to know what is expected. First. know what you teach to be correct. It breaks up the daily monotony. we place ourselves into the situation of “hurried quality implementation”.Implementing a Cleaning and Disinfection Program 49 management. people learn by doing. Decisions for design are always made better without emotional turmoil. Second. when the product is in jeopardy as environmental levels have exceeded prescribed action levels. the first . We forget the multitude of critical areas that need to be addressed to assure the continued and consistent attainment of acceptable environmental conditions in our controlled areas. sometimes inside trainers become boring to trainees. A FINAL SUMMARY AND CONCLUSION Too often we just want to kill things. there are countless year 2000 techniques available to the trainer of today. We will forget many critical aspects. At the same time. This will cause us to fail. As an industry we need to review the required time to adequately clean and disinfect manufacturing areas. training must place in the foreground what is expected and if not accomplished. They may lack the skills to train or supervise this important function. If we wait for regulatory agencies to comment before we change or if we wait for an excursion to exist before assuring a complete system. People learn very little in a classroom.

AOAC . Vellutato. 1989. Sterile Drug Products Produced by Aseptic Processing Draft. p. The National Formulary. Center for Drugs and Biologics and Office of Regulatory Affairs. Philadelphia. A. USP 25 NF 20. United States Pharmacopeial Convention. CleanRooms Magazine. and A. 2002. Vellutato Sr. National Publishing. We must also remember that the painting we have created will not gain value over time as is common for artistic impressions. USP 25 NF 20. The Use of Cleaners and Disinfectants in GMP Controlled Environments. Food and Drug Administration. Food and Drug Administration. The National Formulary. Inc. September 27. Chapter 1072. PDA TR #13 United States Pharmacopoeia. National Publishing. 1999 ISO 14644 ISO 14698 A. Inc. S10–S20. 1998. BIBLIOGRAPHY Vellutato. On the same note cleaning and disinfection is also the last word as too many times we use it as the corrective action procedure instead of delving more deeply into the root cause. Chapter 1116. p. p. 1999 United States Pharmacopoeia. 5th Revision. Veltek Associates.50 Laboratory Validation scenario that is blamed is cleaning and disinfection. L. changes in technology and changes in our personnel structure. Guidelines on Sterile Drug Products produced by Aseptic Processing.... Inc. Philadelphia. June 1987. Jr. January 2001. It must not have been done correctly. Each day our design becomes outdated and we need to routinely reevaluate our systems to assure they include updates for process changes. Utilizing environmental monitoring data to implement a cleaning and disinfection program. Center for Drugs and Biologics and Office of Regulatory Affairs.. 9. 1–14 Official Methods of Analysis (1998) 16th Ed. Concept paper (Not for Implementation). United States Pharmacopeial Convention. Technical update Notice Monthly..

Peroxygen Compounds..S. Philadelphia: Lea and Febiger. Denver.K. The Pharmaceutical Microbiology Mail List. Philadelphia: Lea and Febiger. Antiseptics and Disinfectants: Activity. . and Resistance. p.E. Block. Jan 1999. Antiseptics and Disinfectants: Activity. In Disinfection. Jan 1999. edited by S. 2000. 151.35 Management center: ue de Stassart 36. European Committee for Standardization. industrial. EN 13697 (2001) E. Clinical Microbiological Reviews. Sr. Jan 1999. and R. Casey. Brady. p. p.20. domestic and institutional areas – Test methods and requirements without mechanical action (phase 2/step2). edited by S. Clinical Microbiological Reviews. September 1992. Denver.. G. Block. Sterilization and Preservation. Disinfectants Efficacy Testing. McDonnell. 123. edited by S. Vellutato. 71. Gaithersburg. 151 (513) Chlorine and Chlorine Compounds. p. Nov 17. Action. Godalming: Davis Horwood International Publishing. and M. Rotation of Phenolic Disinfectants. Clinical Microbiological Reviews. 148–158. Comment. In Disinfection.Prince. Jan 1999. Block. 156 Gluteraldehyde. Chemical Disinfectants and Antisepics – Quanitative non-porous surface test for the evaluation of bactericidal and/or fungicidal activity of chemical disinfectants used in food. Philadelphia: Lea and Febiger. G.Implementing a Cleaning and Disinfection Program 51 INTERNATIONAL. Pharmaceutical Technology. Block. Sterilization and Preservation. Clinical Microbiological Reviews.S. Arthur. Action.S. edited by S. and R. Denver. Connor.T. Action. Antiseptics and Disinfectants: Activity. and R. G. A. In Disinfection.100. 2001. S. and Resistance. Sterilization and Preservation. Untites States patent Office. ICS 11. and Resistance.L. MD McDonnell. United States Patent 6. Jan 1999. 4th Edition. McDonnell. Sterilization and Preservation. Philadelphia: Lea and Febiger.080.S.. B-1050 Brussels .M. G.. 151 (545) McDonnell. Eckman. Antiseptics and Disinfectants: Activity. Rotation of Disinfectants. Formaldehyde. 4th Edition. Action. In Microbiology in Pharmaceutical Manufacturing. 4th Edition. In Disinfection. C. 4th Edition. Denver.900. Baskin. W. D. and R. and Resistance. Snyder and B. edited by R.

Validation Report VL-101and VL 101B. Veltek Associates. PA. Exton. Sterile Disinfectant and Sporicide Validation Reports. Arthur. Sterilization of healthcare products – Requirements for validation and routine control – Radiation sterilization. (an EPA and FDA registered facility) in 1981. Inc. 1980.. Title 21. DECON-AHOL Sterile Spray Validation Report. Washington. Inc. Veltek Associates. DECON-AHOL WFI Sterile Spray Validation Report. VA. He is a frequent industry speaker with 21 industry publications.52 Laboratory Validation 1992 Vellutato. DC. Veltek Associates. Inc. VL2001. Exton. Veltek Associates. January 2000 Vellutato. DC: US Government Printing Office ADDITIONAL READING Proceedings of the Second International Kilmer Memorial Conference on the Sterilization of Medical Products. Validation Reports VL-301. Inc. PA. Jr. vice-president of Sales & Marketing and one of the two founders of Veltek Associates.. VL401. 1993 ABOUT THE AUTHOR Art Vellutato. 160.. VL1011. Arlingto. VL601. Washington. Jr. Jr. Validation of the Core2Clean Spray Mop Fog Systems. Internal Validation Report. ANSI/AAMI/ISO 11137–1994. 1995. Inc.. Inc. Arthur. Association for the Advancement of Medical Instrumentation. VL2000. Veltek Associates. Part 211. 1989. is the vice-president of Technical Support Operations. VL-501. VL901. April 2000 Code of Federal Regulations. Phoenixville. Validation of the Core2Clean Fog System. VL-701. 1992. .. Validation Report VL-201 and VL201B. PA. Internal Validation Report. Good Manufacturing Practices for Finished Pharmaceuticals. 1991. He lends over 18 years of valuable experience that include his tenure as the Director of Quality Assurance at VAI for nine years and the Director of manufacturing for six years. VL-801.

is also the chairman of the PDA Special Interest Group on Cleaning and Disinfection (PDA Delaware Valley Chapter) Combining experience of production. .Implementing a Cleaning and Disinfection Program 53 Art Vellutato. Jr. He is the President of the Delaware Valley Chapter of the Parenteral Drug Association and a faculty member teaching the Disinfection segment at PDA's Training and Research Institute for the Aseptic Processing Course. quality assurance and validation. Jr. worldwide. Jr. Method of Sterilization. co-owns US Patent 6. Art Vellutato.900.123. He has also conducted disinfectant training for FDA's CDER and CBER divisions. that was used to market the first sterile disinfectant in the industry. DECON-AHOL®. Art Vellutato. has assisted many pharmaceutical manufacturing operations in implementing their environmental monitoring and disinfection programs.