Designing_Disinfection_Systems_Chapt_8

IMPLEMENTING A CLEANING ANDDISINFECTION PROGRAM IN PHARMACEUTICAL

AND BIOTECHNOLOGY CLEAN ROOMENVIRONMENTS

Arthur L. Vellutato, Jr.

Veltek Associates, Inc.

INTRODUCTION

The implementation of an effective cleaning and disinfection program in a GMPfacility is a multi-phased process that requires the harnessing of a multitude of criticaldisciplines in order to achieve success. Like many compliant GMP systems, the designof a cleaning and disinfection regime utilizes the talents of the quality assurance,production and validation departments. This compilation of skills is mandatory tocompletely address a system that will net continually acceptable environmentalconditions and prove competence during regulatory inspections.

In looking at the basics surrounding the formation of a compliant system, theultimate 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 contaminantthat is not part of the material surface itself. Such contaminants may be in the form ofspilled product from a previous batch, chemicals used in manufacturing product,disinfectants, particulates and residues. When cleaning surfaces in the clean room, theremoval of all particulates and residues is virtually impossible. Thus, the clean roomwill, once cleaned, have an existent level of particulates and residues that may or maynot affect the next batch of product. Likewise, disinfecting the surface would requireone to ensure that the bioburden of the surface has been eliminated or reduced to a safe

1

8

and contained level which will not affect final product. However, one could not rendersuch surfaces “sterile”, as technically defined. Sterilizing a surface would require it tobe exposed to a terminal sterilization process such as autoclaving, heat, gammaradiation, ethylene oxide or extended chemical sterilization.

The only fitting method for use in a clean room from the above list would bechemical sterilization. Chemical sterilization of a clean room surface using an EPAregistered sporicidal agent may take up to five hours of soaking of all surfaces. This isnot possible in the clean room environment. Thus, an existent bioburden, even afterdisinfection, may exist in the area.

Comprehending that the surfaces may not be perfectly clean and sterilized iscritical to an understanding of what may and may not be accomplished. In manyphases of life, “perfect” performance is impossible to achieve, and cleaning anddisinfection are no exception. If a clean room were cleaned, filled with ultra-cleanwater, 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, completeidentification of all contaminants would be unlikely. In a viable sense, if a clean roomwas cleaned and disinfected, then subsequently filled with a growth medium liketrypticase soy broth (TSB), and a reasonable growth period allowed, one would seegrowth. These two examples, at extreme ends of the spectrum, show that cleaning anddisinfecting of a controlled environment will not be perfect. But despite thecomplexity, 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 evaluatewhat is presently being done in the operation. The author's personal experience as aconsultant involved in cleaning and disinfection in this industry for many years is thatQuality Assurance (QA) departments may not know absolutely everything that is donein a production department; likewise, production may not completely understand whatquality assurance's recommendation may be. This is understandable, as QA andproduction are two separate functions with differing agendas but their combination ofskills and disciplines makes for the development of a very intricate and effectivesystem. But first, QA and production professionals need to understand what is done ona daily basis, and what should be done on a daily basis.

The next step to achieving near-perfection in our cleaning and disinfectionsystems is to put the basics into perspective. The criteria for implementing a cleaningand disinfection program within a GMP facility require companies to tailor theirsystem to address their operations' specialized needs. Development of a systematicapproach to addressing and controlling of contamination will serve organizations for

Laboratory Validation2

years to come, with attention to detail the key to success. And success is measured byconsistently attaining acceptable environmental conditions, making use of theavailable disinfectants and application devices, and utilizing environmentalmonitoring data to address the specifics of the system. In turn, regulatory auditors willroutinely scrutinize the system with a fine-tooth comb, and dissatisfaction can resultin possible 483 observations or the shut-down of manufacturing processes. For thisreason, systems need to be complete in their content, and assumptions that situationsmay not arise could be fatal. And most importantly, the design must be documentedwithin 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 theregulatory agencies. It requires the use of a multitude of separate scientifically-basedmethodologies and disciplines, combined to render the final infrastructure.

For years, the pharmaceutical and biotechnology industries have been two of themost regulated in the world. However, within the spectrum of manufacturing a productthat will enter or come in contact with the body, there are several critical areas wherethere is a lack of written directive from regulatory agencies. Two of these critical areasare environmental monitoring, and disinfection of a controlled area. Together thesetwo critical capacities serve as the basis for determining and maintaining theenvironmental conditions during the time of manufacture of a drug product. Initially,one would assume that there is a specific need for regulatory specifications in thesecritical areas. However, after careful evaluation, it appears that these functions requirea very unique design for each manufacturing setting and for each product that isproduced. Specifications from regulatory agencies do, in certain circumstances,standardize the function or testing requirements throughout the industry. However, inthe areas of environmental monitoring and disinfection, they may place possiblyunattainable criteria to very process-specific functions.

Despite the lack of written regulatory specifications, this does not mean thatregulatory agencies do not have specific expectations. In fact, their expectations maybe greater than if specifications were in place. In the present day, regulatory agencieshave placed the responsibility for system design, implementation, documentation andjustification on the manufacturers themselves. The overall theme is for a drugmanufacturer to "figure out how they are going do it, do it, document it, and beprepared to justify their methodology upon request." An example of this can be foundin the 1987 FDA Guideline on Sterile Drug Products Produced by Aseptic Processing,(page 9)

2. 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 morethan one colony forming unit per 10 cubic feet is considered as attainable anddesirable." Within this text, regulatory said "here is what we expect; now you figureout how you need to accomplish and document this capacity." However, within thisshort statement, technical problems arise.

Implementing a Cleaning and Disinfection Program 3

The first is that humans, by nature, try to interpret hidden meanings in what iswritten. Personnel in pharmaceutical and biotechnology varied in their interpretationof the guidance. At the same time, regulatory inspectors and reviewers also variedamongst each other understanding and enforcement policies. In short, the guidancetext was not complete. It did not specify many details such as the required total volumeto be sampled, the number of locations to be sampled, and the frequency in whichsampling should be done. This causes confusion as all then develop and implementtheir opinions and commonality of task to be performed and the method by which itwill be inspected varies from firm to firm and regulatory inspector to regulatoryinspector.

This is just one example of problematic situations that arise with certain guidancedocuments. Earlier in the chapter, the probable imperfection of cleaning anddisinfecting was discussed. In terms of guidance documents, the same is also true. Interms of environmental monitoring, cleaning and disinfection, no one organizationwill ever be able to create the perfect guideline. Guidelines will always lack the criticaluniqueness that exists for implementation in each and every manufacturing and testingoperation. This concept must be understood.

Thus, the problem with developing an environmental monitoring and disinfectionprogram is one is not able to pick up a book and follow its content. In reality, one mayneed to view many books, many guidelines or more effectively, rely on directexperience. 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 disinfectionsystems that work. The utilization of inexperienced personnel to design the systems isa way of ensuring failure and possibly future regulatory complications.

It is necessary to look at a few guideline documents that may assist in understandingwhat needs to incorporate in the design of a system. First and foremost, one needs to lookas the specific tone from the US FDA. Presently the 1987 FDA Guideline on Sterile DrugProducts Produced by Aseptic Processing is the current document. However a revisionfrom FDA, entitled Sterile Drug Products Produced by Aseptic Processing Draft

3has

been recently released. A second source is the PDA's Technical Report #134. the third

source would be the United States Pharmacopeia (USP), Chapter <1116> (EnvironmentalMonitoring)

5and Chapter <1072> (Disinfectants and Antiseptics)

6.

A fourth source looks to review ISO 146447 and 146988, 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 anorganization and externally from colleagues in the industry. While this list of writtenguidance is not complete, coupled together they provide us with the basic conceptsneeded to design our systems.


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 andcompliant cleaning and disinfection system uses these critical documents only as toolsto write a master plan; they could never be as complete for one's operations as a systemone could develop ourselves.

So where does one start? Cognizant of the multitude of issues that will requirereview, 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. Whileincomplete in lengthy discussion for each item noted, this short guide will hopefullyidentify 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 aspectsurrounding a cleaning a disinfection program. We need to understanding that controlof the environment does not come from using a chemical agent that will destroy allmicroorganisms. Rather, control of the environment is the assurance level we establishto reduce or possibly eliminate the contamination from ever entering our controlledareas. Once a chemical agent has been applied to the surface and subsequently dries,its destructive capabilities to a microorganism are complete. Disinfection (orsporicidal) characteristics of a chemical agent require the organism to be wetted by theagent. While certain residues from chemical agents may have some remainingantimicrobial properties, the destructive capabilities of the residue are minimal.Control over what enters the controlled area now takes over. In reality, control of theenvironment has nothing to do with disinfection. Disinfection is complete whencontrol takes over. Manufacturing operations will not disinfect surfaces whileproduction is occurring. Disinfection is done prior to manufacturing and theenvironment released to produce a product. Our success or failure in control ismeasured each and every time as environmental monitoring is conducted during ourfilling operations.

Addressing contamination prior to its entry will ensure that we will not have tocontend with its presence. While disinfection of surfaces always take the lead instructuring our procedures, control over the environment should remain one of themain focuses.

Criteria for reducing the bioburden that enters the controlled area require us toevaluate the entry of personnel, components, water, tanks, carts, and even disinfectantsto name a few. This is done to ensure that one do not undermine disinfection efforts byintroducing high levels of contamination after disinfection is complete. We mustensure each item's appropriateness in the room classification to which it enters. In


Class 100 aseptic operations, one needs to carefully evaluate each item as clean andsterile prior to entry, as well as assuring the cleanliness, sterility and appropriate fit ofgarments for personnel. In Class 10,000–100,000, while our concern is less, we mustensure that such environmental conditions do not adversely affect the Class 100 area.

In short, a room can be monitored as having very little if any bioburden at rest;however, in a static condition we can easily corrupt our efforts previously achievethrough poor control.

CLEANING THE ENVIRONMENT

Too often, cleaning is confused with disinfection. They are not the same. Cleaningcharacterizes the removal of particulates, microbes and possibly existent residues fromsurfaces. Cleaning requires a non-destructive mechanical action be applied thatloosens and removes contaminants from the area. Procedurally, contaminates andresidues are loosened and rinsed to the floor. Subsequently, the dirtied solution on thefloor is collected and removed from the area (normally by a squeegee). By lesseningthe level of particulates, microbes and residues on the surface, disinfection effortsbecome simpler. First there are fewer organisms to destroy as most have been removedfrom the area. And second, as bioburden and residues are lower, the possibleobstructions blocking the chemical agent from contacting the organism are minimized.In short, cleaning prepares the surface for disinfection.

Disinfection relates to the saturation and penetration of the cell wall of anorganism by a chemical agent. It further requires that an organism remain wetted fora specified contact time with a chemical agent capable of killing the organism inquestion. Disinfection depends upon temperature, saturation and penetration of thecell wall, contact time, surface and bioburden of the surface, existent soil load,concentration of the chemical agent and pH. Provided the appropriate chemical agentis utilized, the key to disinfection in the clean room is keeping the surface wetted for5–10 minutes. This is difficult, as the movement of air via laminar flow tends to drysurfaces quicker.

As discussed, there are significant difference between cleaning and disinfection.Cleaning tries to remove contamination from the surface while disinfection attempts todestroy what viable cells exist on the surface. In a realistic example, we can use atoothbrush and mouthwash as examples. If we were to discuss the options of not usinga toothbrush anymore and only utilizing a daily mouthwash rinse with our dentist,he/she would inform us that we would soon have no teeth. Residues, particulates andmicrobials will build up on the surface of the teeth and the teeth would eventuallydeteriorate. This scenario depicts what occurs too often in the pharmaceutical andbiotechnology industry. We forget to brush and just try to kill anything that exists on the


surface. Eventually our surfaces become residue-laden and more difficult to disinfectand eventually deteriorates. Within our note to technical brilliance we sometimes forgetsimple common sense. And unfortunately, the phrase, “simple common sense” is notthe title of any GMP, CFR or guidance document. The effect of the buildup of residues,particulates and possibly microbials is also aided by the surface itself. Clean roomsurfaces are irregular in nature as depicted in the Scanning Electron Microscope (SEM)photographs (Figures 1–8) seen later in this chapter. Such surfaces trap residues andother contaminants and make the surface more difficult to disinfect.

Sooner or later we have to clean. The frequency of cleaning can vary from a dailyfunction to a monthly function. Normality is to clean surfaces either bi-weekly or ona monthly basis. Some may say, "But that's an additional cleaning operation we needto do!" The correct response is "That is correct, it needs to be done."

Within the healthcare setting and most commonly reported in the hospital setting,test reports have shown the effect that cleaning the surface has on the level ofmicrobial levels in controlled areas. Many publications purport this concept. In thepharmaceutical and biotechnology setting, a test report conducted and published in1989 by A. Vellutato, Sr and A. Vellutato, Jr. of Veltek Associates, Inc.

9demonstrated

what affect cleaning the surfaces has on the level of microbial contamination foundon the surface. The study focused on the concept that cleaning alone would removemost of the existent microbial contamination. In the report, all surfaces were cleanedwith a sodium lauryl sulfate detergent and mechanical cleaning action on a dailybasis. Such cleaning and was conducted in a Class 100, Grade A, ISO 5 area.Environmental Monitoring was routinely conducted on air, surface and personnel.The manufacturing operations filled an average of 4,000 units of 500 mL. bags ofUSP Water for Injection. Manufacturing operations ran for four hours per day andupon completion of manufacturing, the clean room was completely cleaned.

The cleaning mechanism utilized a mop and two-bucket system, a sprayer and asqueegee. The procedure for cleaning was to apply the sodium lauryl sulfate detergent(DECON-Clean®) to the surface utilizing a top to bottom approach. Upon completionof the mopping, the chemical was then sprayed on to the surface and all excess liquidon the surfaces pulled downward by squeegee to the floor. The remaining liquid wasthen collected on the floor and removed. For 30 days, the results met industry limits forClass 100, Grade A, ISO 5. At 45 days, control was lost, results exceeded limits and asporicidal agent was used once on day 45. The limits returned to acceptable levels for31 days (day 76 of the study). The final conclusion was one to two uses of a sporicidalagent with cleaning regime controlled the environment.

Cleaning is based on a few physical factors:

• the surface to be cleaned


• 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, moreover residue, that exists onthe surface

Knowledge of the type of surface to be cleaned is very important. As depictedin Figures 1–5 (further in the chapter), surface irregularity varies as compositematerials change. The surface irregularity may compromise cleaning anddisinfection efforts for two basic reasons. First, cleaning becomes more difficult.Second, disinfection becomes harder as the chemical agent cannot contact all thesurfaces for the required wetted time period (as obstructions may exist). Cleaningbecomes more difficult as the surface's irregular nature may allow particulates,residues and even microbial contamination to vest itself within the rutted or porousareas of the surface. These nooks and crannies are very hard to clean with most cleanroom apparatus designed for cleaning. Most clean room mops and wipes and flat anddo not allow the penetration of the fibers or surfaces of the cleaning mechanism toreach into the crevices. The lack of such an abrasive cleaning action bypasses theopportunity to loosen and remove these contaminants. Most irregular surfaces arecommonly so across the span of the material, so a multitude of contaminationsources exist on the surface to be cleaned. Cleaning an irregular surface requires oneto use a cleaning device that can penetrate into the nooks and crannies and makecontact with the existent contaminants and residues. For example, we would not usea wiper or a pad to try to clean our teeth, but a toothbrush. Unfortunately, a cleanroom brush is not a commonly-available item in the industry. Nor would requiring acontrolled area to be cleaned with it be a reasonable request to make of productionpersonnel. This is a subject that cleaning apparatus manufacturers need to addressmore closely.

The lack of available products forces the clean room professional to adapt a non-clean room product to address this specific need.

The second basic complication is that if particulates and residues exist in suchnooks and crannies, the possibility for a disinfecting agent to contact the surfacewithin the nooks and crannies is improbable. Thus, we are disinfecting the surface ofthe particulates and residues and never disinfecting the actual surface. Without suchassurance for cleaning and disinfecting the actual surface, the possibility forcontamination to be vested underneath the particulate and residue may be probable.


In other sections of this chapter, antimicrobial effectiveness testing will bediscussed, giving an understanding that, during such validation testing we look toinoculate a surface with a known enumeration of microorganisms and soak suchsurfaces in a disinfecting agent for a predetermined time period. Upon completion ofthe soaking, we will then test the surface to account for the possibly remaining viablecontamination.

When we attempt to use such data in the field, we will find our testing skewed fortwo basic factors. First, our testing inoculates a variety of surfaces with a multitude ofmicroorganisms. Some surfaces are more irregular than others. In our test, after wehave soaked the surfaces, we need to rinse clean the surfaces of any possiblecontamination to a filter which is then plated to a growth medium. However, we findthe rinsing of the irregular surfaces more difficult as microorganisms may vest andcling within the irregular areas of the surface. This means the possibility to rinse freea microorganism from a smooth surface is easier than from an irregular surface. Whatwe may learn from this testing is that it may seem that our disinfecting agent is moreeffective on the irregular or porous surface but this is due to our inability to rinse freeall of the existent microorganisms. The smooth surfaces are more easily rinsed andexistent microorganisms removed and can grow in our growth medium. However,microorganisms from the irregular or porous surface may have never been removedfrom the surface itself. This means they were never rinsed to the filter and plated forgrowth. Thus, we could conclude smooth surfaces are harder to disinfect than irregularor porous surfaces in our manufacturing and testing areas. The effectiveness reportmay show higher remaining colony forming units (CFUs) for the smooth surfaces thanthe irregular or porous surfaces. This assumption would be incorrect.

In the manufacturing or testing areas, the opposite occurs and we find it moredifficult to disinfect the irregular or porous surfaces. Understanding this concept iscritical to successful disinfection. We will have the same trouble, if not a morecomplicated problem, in disinfecting and rinsing the microorganisms from theirregular or porous surfaces in our manufacturing and testing areas, and due to this,we may leave viable contamination on such surfaces. Care needs to be taken whendisinfecting irregular or porous surfaces, as they are harder to clean and disinfect thansmoother surfaces.

In general, clean room material grades can be separated into six basic categories:

• aluminum

• stainless steel

• epoxy-coated finishes

• plastics


• vinyl

• glass

In Figure 1, we see an aluminum surface. This surface is a metal grade that is softand easily scratches and is deteriorated by chlorine solutions, gulteraldehyde andperoxide and peracetic acid and hydrogen peroxide solutions (from a list of basicdisinfecting agents). While aluminum is a commonly-used metal, deterioration of thesurface from chemical exposure normally shows as a turquoise bluish gray bubblingon the metal. Aluminum is also easily stained or discolored from the residues from byphenol, quaternary ammonium, and iodine to name a few. However, of most cleanroom surfaces, aluminum is normally easy to clean and disinfect.


Figure 1: Aluminum surface

Figure 2: 316 L Stainless Steel surface

In Figure 2, we see a 316L stainless steel surface. This surface is very smooth andcontains few impurities in the metal that may be deteriorated by a disinfecting agent.Most impurities in the metal grade are in the carbon family and are deteriorated bychlorine solutions, gulteraldehyde and peroxide and peracetic acid and hydrogenperoxide solutions (from a list of basic disinfecting agents). Stainless comes in gradebased on the level of impurities in the metal. Normally, deterioration of the surface isin the form of a rusting that pits the surface or surface rust. Rusting of the stainlessitself is from chemicals or water oxidizing and/or reacting with impurities in the metal.Surface rust is the rusting of airborne heavy metals that deposit atop the surface.Within the clean room operation, many metal grades exist from of 302 to 402 stainless.Many in the industry demand the use of 316L stainless; however this metal gradecannot be used for every component due to its brittle nature. Some perfect examplesof this would be a spring or a solenoid mechanism. Utilizing too hard a metal willcause the spring or moving part to routinely break as friction of movement on themetal will cause stress and fracture. Stainless is also easily stained or discolored fromthe residues from by phenol, quaternary ammonium, and iodine to name a few.However, of most clean room surfaces, stainless steel is normally one of the easiestsurfaces to clean and disinfect.

In Figure 3, we see an epoxy-coated surface. Epoxy-type surfaces are verynumerous in types and materials. They are a coating that is applied in a liquid formand then hardens. This is where disinfection becomes complicated. The irregularsurface appearance of epoxy-coated surfaces challenges the craft of the disinfectionprofessional. Most clean room walls and floors are made of an epoxy material orsimilar material so understanding cleaning and disinfection of this surface is critical.Common build up of particulates and moreover residues in the crevices complicatecleaning and disinfection. Problematic situation can arise in the crevices of thesurfaces as air pockets can form and once disinfection is compete, be broken an


Figure 3: Epoxy-oated surface

possibly release existent contamination to the environment. The most difficult taskwith the epoxy-coated surface is to clean the contaminants from the surface so that thedisinfectant can be applied and have the ability to address the surface itself. Normallyepoxy-coated surfaces are deteriorated by chlorine solutions, peroxide, isopropylalcohol, ethanol and peracetic acid and hydrogen peroxide solutions (from a list ofbasic disinfecting agents). The normal deterioration occurs in the form of over dryingand cracking of the material finish. The material becomes powder-like and orange toblue in color (dependent upon the material).

Epoxy-coated surfaces need routine replacement or refinishing and as suchremain continuously on a preventative maintenance schedule. The epoxy-coated finishis also easily stained or discolored from the residues from by phenol, quaternaryammonium, and iodine to name a few.


Figure 4: Clean Room Curtains #1

Figure 5: Clean Room Curtains #2

The next basic clean room surface type is a collage or porous material such asplastic, vinyl, plexiglass, delron, and other similar type products. In Figures 4, 5 and7 we can see what these surfaces may look like when magnified through an SEMmicroscope. Characteristically they have pore openings that are rather deep. Cleaningand disinfection are very difficult with these materials. Normally these type ofsurfaces need to be replaced over time. These materials need replacing every two orthree years on average. Replacement is costly but necessary, and should be viewed asa cost of doing business. Normally, porous surfaces are deteriorated by chlorinesolutions, peroxide, isopropyl alcohol, ethanol and peracetic acid and hydrogenperoxide solutions (from a list of basic disinfecting agents). The normal deteriorationoccurs in the form of over-drying and cracking of the material itself. A yellowing, orchange of color to an orange or brown, are common symptoms of the drying process.The porous material is very easily stained or discolored from the residues of phenol,


Figure 6: Phenolic Residue

Figure 7: Vinyl surface

quaternary ammonium, and iodine to name a few; such residues and/or stains onlyincrease the deterioration process. Most notably these materials are use as curtainsthat separate process control areas. Items such as plastic curtains are one of the mostwidely used materials in the clean room environment. Cleaning these surfaces isnormally done frequently, if not daily, as they represent the second-closest non-product contact surface next to the filling line itself. This over-cleaning shortens thelife of the material.

The last category is glass. Glass is a relatively smooth which does not deteriorate.Glass is very difficult to clean, and provides a constant example of how difficult allsurfaces are to clean. The reason glass seems harder to clean is that one can see throughit. All other materials discussed to this point are not clear. If such materials were as clearas glass, the appearance of these surfaces would look horrifically dirty in comparisonto glass. Glass is very easily dirtied, and is a very difficult material to keep clean.However, this surface is used more and more in clean room operations as it allowsviewing of the operation by supervisors and visitors. In general, glass does not staineasily from disinfectants or sporicides. However, residues build up and it is discoloredfrom the residues of phenol, quaternary ammonium, and iodine to name a few.

We have seen from this section the importance of cleaning. Simply, the effect ofcleaning surfaces ensures the best possible opportunity to disinfect the surface as themicrobial levels and the possibly existent residues and particulates will be lower. Laterin this chapter we will discuss the mechanisms to clean such surfaces. However, firstwe must understand the chemical agents that cause the main problem associated witha dirtied surface, the residue.


Figure 8: Sodium Hypochlorite Residue

TESTING THE ENVIRONMENT AND DEVELOPING DATA

Environmental monitoring is a subject warranting its own presentation. In the worldtoday, the existent guidelines are numerous. The guidelines, while having a commontheme, are not harmonized. Each guideline's numerical values and recommendationsslightly differ from each other. Some contradict each other's content. Some of theexisting guidelines include, USP Chapter <1116> (in revision), ISO TC209, ISO14644, ISO 14698, PDA Technical Report # 13, the FDA's 1987 Aseptic ProcessingGuidelines (in revision), the revision to the EU Annex I and several others. While thereis no commonality in the guidelines, one should cautiously approach the claim ofadherence to one particular guideline. Doing so would be a claim, and specificallyfollowing the guideline's parameters may be required. Rather, one should use theguidelines as informational sources. Gain what knowledge can be gleaned from thewritings, then design an environmental monitoring program based on whatinformation is applicable for one's specific operations.

While structuring one's environmental monitoring program, one will find itnecessary to test for viable cells both in the air and on the surface. Through one'senvironmental monitoring program, one can develop a list of environmental isolatesthat have been noticed in their operations. By review of one's environmental isolatelist, one can determine what contamination exists in the facility and appropriatelydesign their cleaning and disinfection program surrounding this scientific knowledge.Once developed, the key is to successfully integrate and document a plan for theassured demise of these organisms.

CHOOSING THE APPROPRIATE DISINFECTANTS ANDSPORICIDES

The utilization of environmental monitoring data to implement a cleaning anddisinfection program in controlled environments uses science to justify our ends. Wetest our environment and subsequently address the contamination noticed with aproven and validated efficacious disinfection solution. This ensures that we use ascientific rationale to control contamination in our operations.

In choosing the appropriate chemical agents, we need to review what thedifference between a sanitizer, a disinfectant and a sporicide. In very simple terms, asanitizer has the lowest ability to destroy vegetative cells of the three listed. Thedisinfectant destroys a broader spectrum of vegetative cells than the does a sanitizerand incorporates some log reduction of spores. And the sporicide destroys allvegetative cells and spores. Specifically, the United States Environmental ProtectionAgency (EPA) under FIFRA, CFR Title 40 governs and registers by United States ofAmerica law antimicrobial effectiveness claims for hard surfaces disinfection.


Table 1 gives is a basic overview of what is required by a disinfectantmanufacturing company by the US EPA to make the following claims on marketedproducts.

Understanding disinfectant categories is essential. A multitude of categorizationsof chemical agents exist in the world. In Table 1, we identified the EPA's


Types of Claims Test organism(s)

Sterilizer Bacillus subtilis and Clostridium sporogenes

AOAC Sporicidal Test

Tuberculocide Mycobacterium tuberculosis

AOAC Tuberculocidal Activity MethodModified AOAC Tuberculocidal ActivityMethodQuantitative Tuberculocidal Activity TestMethod

Hospital Disinfectant Salmonella cholerasuis, Staphylococcusaureus, and Pseudomonas aeruginosa

AOAC Use-Dilution MethodAOAC Germicidal Spray Products Test

General Disinfectant Salmonella cholerasuis and Staphylococcus aureus

AOAC Use-Dilution MethodAOAC Germicidal Spray Products Test

Limited Disinfectant Same as Above

Fungicide Trichophyton mentagrophytes

AOAC Fungicidal TestAOAC Use-Dilution MethodAOAC Germicidal Spray Products Test

Virucide Specific Virus Claimed

EPA Virucidal Test Parameters

Sanitizing Rinse (food contact surfaces) Escherlchia coli and Salmonella typhi or Staphylococcus aureus

AOAC-available chlorine germicidalequivalent concentration methodAOAC germicidal detergent sanitizermethod

Sanitizer (inanimate, nonfood contact Staphylococcus aureus and Klebsiellasurfaces) pneumoniae or Enterobacteraer aerogenes

EPA Sanitizer Test Parameters

Table 1: EPA-Approved Disinfectant Test Methods8

(for hard surfaces)

classifications for antimicrobial effectiveness claims that based on their passing ofparticular AOAC test parameters 10. Others have categorized disinfectants assanitizers, disinfectants, high level disinfectants, intermediate level disinfectants, lowlevel disinfectants, sporicides and sterilants. In development and validating of acleaning and disinfection system, 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 butdoes not destroy all microbal forms such as bacterial endospores.

Sterilants/Sporicides: a chemical agent that is capable of sterilizing. Such chemicalcan render a product free of viable organisms including all bacterial endospores.

Simplifying this list to three basic categories allows us to determine what we willrequire for our operations and subsequently be required to validate. Defined in thecategories below are chemicals agents normally used in pharmaceutical andbiotechnology operations. We need to choose one sanitizer, one or two disinfectants(depending on our belief in resistance) and one sporicide/sterilant.

Sanitizers: Isopropyl Alcohol 70%Ethanol at 70%Ethyl Alcohol at 70% Iodine (not normally used in clean room operations)

Disinfectants: PhenolsQuarternary AmmoniumHydrogen Peroxide at 3% or belowSodium hypochlorite below 0.10%

Sporicides/Sterilants:Sodium Hypochlorite above 0.25%Hydrogen peroxide above 6% (sporicidal reduction at 6%, a sterilant at 10–35%depending on the wetted time)Peracetic Acid & Hydrogen PeroxideGlutaraldehydeFormaldehyde

In reviewing the basic types of chemical agents used in pharmaceutical andbiotechnology operations we can come to understand their basic differences andapplicability in our operations.


To follow is a brief description of each of the most used chemical agents in theindustry. As a complete chapter could be written on each, the summaries are brief sowe can develop a basic understanding of each chemical.

ISOPROPYL ALCOHOL, ETHYL ALCOHOL, ETHANOLSOLUTIONS:

Alcohols have been used for years in pharmaceutical and biotechnology operations forthree basic purposes:

• as a sanitizing spray for gloves, surfaces, carts, etc.

• as a cleaning or wipe down agent to remove possible existent residues fromcritical non-product contact surfaces

• as a product contact cleaning agent (ethyl alcohol only)

In testing antimicrobial products' effectiveness, a 70% solution has far superiorefficacy performance than the higher or lower concentrations. Alcohols come in avariety of forms. The most used forms in the clean room operation are isopropylalcohol and ethyl alcohol (ethanol and alcohol). More than 90% of industry operationwill utilize a 70% isopropyl alcohol solution to address clean room organisms as it hasbeen proven more efficacious than ethyl alcohol (190–200 proof diluted to 70%) orethanol 9a mixture with the base as ethyl alcohol [63%] and spiked with methyl [3%]and isopropyl [4%]) at a small percentage (rendering it undrinkable). Alcoholsdemonstrate rapid broad-spectrum antimicrobial activity against vegetative bacteria(including mycobacteria), viruses, and fungi but are not sporicidal

11. They are however,

known to inhibit sporulation and spore germination12, but this effect is reversible

13. As

alcohols are not sporicidal, they are not recommended for sterilization and are widelyused for hard surface disinfection and skin antisepsis. As has been previously stated,of the existent alcohols, isopropyl alcohols demonstrate superior effectiveness againstclean room organisms. However, for viruses, ethyl alcohol or ethanol seems to beslightly more effective and is used basically within the confines of the laboratoryenvironment.

Published alcohol effectiveness or results of alcohol effectiveness are presentedwhen sprayed to the surface and allowed to air dry. This should not be confused in anyway with the product's effectiveness if used in a saturated wipe. A saturated wipecontains a limited amount of the chemical agent. When used its dry times aresignificantly less, and thus, its destructive power substandard to that of the liquid itselfon the surface. While the mechanism of wiping destroys cells in its action, the dry timeof the alcohol is significantly faster.


Isopropyl alcohol wipes carry few if any claims. Normally, they are used to wipea surface in a cleaning operation (IPA wipe down) to remove existent disinfectantresidues. Here also such products pose a problem. As they are saturated with isopropylalcohol, their ability to soak up residues and clean the surface is minimal. Most of thetime these products just move around the contamination of the surface. A superiormethodology would be the use of an isopropyl alcohol and a dry wipe. Like cleaninga window, we would spray the solution to the surface and wipe. This will remove themajority of contaminants on the surface. This can be proven in a home experiment byusing a saturated wipe to clean a window. When completed, the window will havestreaks or swirls on it, and look as though the dirt was just moved around. If one wereto conduct the same experiment with a dry wipe and spray isopropyl alcohol, thesurface would appear much cleaner. The soaking of the liquid with contaminants insuspension from the surface into a dry wipe is a superior methodology. Duringisopropyl alcohol wipe downs of non-product contact but critical surfaces, we need toemploy the cleaner of the two methodologies. Rendering of the products as sterile is amust prior to use in a Class 100 (Grade A and B, ISO 5) and adjacent Class 10,000(Grade C, ISO 7) areas. Sterilization of disinfection agents is discussed in depth in thesection on Sterility of Disinfecting Agents.

PHENOLS

Phenolics have been used for years as a disinfecting agent. Phenolics are effectiveagainst gram-positive and gram-negative organisms. However, phenols exhibit betterantimicrobial effectiveness against gram-positive organisms than they do againstgram-negative organisms. They have limited activity against fungi and certain virusstrains such as HIV-1 (AIDS virus) and Herpes Simplex, Type 2. Phenolics normallyare available in an alkaline and acid base version. The theory surrounding the rotationof these two compounds is described later in this section of this chapter. Overall,phenolics demonstrate superior antimicrobial effectiveness in an acidic base asopposed to an alkaline base. Some of the most common chemical compounds inphenolic germicidal detergents are in a low pH phenolic: an ortho-phenyl-phenol andortho-benzyl-benzyl-para-chlorophenol and in a high pH phenolic: a sodium ortho-benzyl-para-chlorophenate, sodium ortho-phenylphenate, or sodium para-tertiary-amylphenate. While phenols provide good broad-spectrum disinfection they are notsporicidal and have major drawbacks in their use. One drawback is the horrificresidues that are noticed from long-term use of the products.

Phenolics are normally an amber (light tan) or light tan color when manufactured.This color darkens with age and exposure to light (especially fluorescent). Residuesstart as a “dripping droplet” that is not easily removed. The use of 70% isopropylalcohol or certain residue removers can remove such residues in their early existenceon surfaces. While somewhat effective, both residue-cleaning products eventually give


way to the darkening phenolic that stains the surface with a dark brown color. Transferof such residues to an unwanted location is a concern and precautionary measures needto be implemented to ensure minimization of this scenario. Compatibility with mostchemicals is normally very good, however, the effect of anionic characteristic of thechemical in relation to applications in conjunction with a cationic surfactants such asa quarternary ammonium have been reported as problematic. Expiration of aformulated phenol also carries some concern. The formulation in a closed containershould remain stable for a period of 7–30 days (dependent upon storage). Formulatedsolutions should be marked accordingly. The normality in the industry is 7 days, andless if the solution is in an open container (such as a bucket) and should be discardedeach day. Rendering of these products as sterile is a must prior to use in a Class 100(Grade A and B, ISO 5) and adjacent Class 10,000 (Grade C, ISO 7) areas.Sterilization of disinfection agents is discussed in depth in the section on Sterility ofDisinfecting Agents.

QUARTERNARY AMMONIUM COMPOUNDS

Quarternary ammonium products are used in more disinfection applications thanphenolic germicidal detergents. Their spectrum of use is very broad and rangesthroughout the industrial world, through the hospital and institutional setting and evencan be found in home use. Quarternary ammoniums have excellent detergency. Theyare one of the best cleaners among the spectrum of disinfecting agents. These cationicsolutions also have excellent deodorizing capabilities. Quarternary ammoniumcompounds are effective against gram-positive and gram-negative organisms.However, phenols exhibit better antimicrobial effectiveness against gram-positiveorganisms than they do against gram-negative organisms. They have limited activityagainst fungi and certain virus strains such as HIV-1 (AIDS virus) and HerpesSimplex, Type 2. In fact and due to competition in this arena, quarternary ammoniumcompounds have the most organisms registered as label claims with the US EPA. Theirmechanism of antimicrobial effectiveness is related to their positively chargedmolecule. Simply, the positively charged molecule attracts to the negatively chargedmicroorganism's cell wall. The cycle of kill is complete when the chemical agent isabsorbed into the cell and spread throughout the organism. Quarternary ammoniumscome are available in both alkaline and acid based compounds. Quaternary ammoniumcompounds normally utilize an alkyl dimethyl benzyl ammonium chloride or adimethyl ethyl benzyl ammonium chloride. While a multitude of formulations exist inthe industry, these are two of the most popular components. As with phenols,quaternary ammoniums do leave sticky residues that become problematic over time.However, they are not of the scale of phenolic residues. Expiration of a formulated(from concentrate) quarternary ammonium also carries some concern. The formulationin a closed container is relatively stable and should remain stable for a 30-day periodin a closed container. Formulated solutions should be marked accordingly. The


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. However, in recent years,quarternary ammoniums have been made in ready-to-use formulas that are very stable.Rendering of these products as sterile is a must prior to use in a Class 100 (Grade Aand B, ISO 5) and adjacent Class 10,000 (Grade C, ISO 7) areas. Sterilization ofdisinfection agents is discussed in depth in this chapter in section, Sterility ofDisinfecting Agents.

SODIUM HYPOCHLORITE

Sodium Hypochlorite solutions are one of the oldest known disinfectants andsporicidal agents

14. The product is available in many forms including ready to use

0.25% and 0,52% solution, to concentrate 5.25% and 10% solutions, to powders thatare mixed with water to formulate a variety of solutions. Bleach as we know it isnormally found in a 5.25% concentration, however, in recent years such formulationsfrom the Clorox Corporation® have been increase to a near 7% solution to ensurecontinued stability of the active percentage.

One of the main problems with the use of sodium hypochlorite is the industry isthat it is used at too strong an active percentage. Sodium hypochlorite is usedthroughout the heath care setting and normally diluted to concentrations of 0.25 or0.52%. One of the problems with sodium hypochlorite formulations is the method offormulation designation which varies from firm to firm. Some formulate to parts permillion, some to a percentage of a solution and some to a dilution such as 1–10. At ause-dilution of 0.25% (or a 1-20 dilution or 250 ppm) sodium hypochlorite is effectiveagainst gram positive and gram negative organisms, viruses, fungi and bacterialendospores. At a slightly increase use-dilution of 0.52% (or a 1-10 dilution or 500ppm) sodium hypochlorite is effective against gram positive and gram negativeorganisms, viruses, fungi and more effective against a wider range of bacterialendospores. Both formulations are used throughout the pharmaceutical andbiotechnology industry.

In formulation of a sodium hypochlorite solution, many choose to acidify thesolution, which makes it a more potent mixture when focusing on bacterialendospores. However, acidification causes rapid degradation of the active elements inthe solution and use of the product is limited to approximately two hours. In reality,acidification may not be necessary as the product demonstrates excellent sporicidalcharacteristics in its neutral state and bacterial endospore levels in clean rooms are notexuberant in numbers (above 1.0 x 106 ) and there is no soil load. A formulation of250 or 500 ppm has excellent sporicidal activity. However, expiration of the solutioncan become a problem. A normal 5.25% concentrate sodium hypochlorite normallycarries a one-year expiration for an unopened container. Some companies have


validated and increased this expiration with applicable assay data over time to 18months. Once opened, whether in a ready to use formula or a concentrate product, theproduct needs to be used within a 30-day period. Open containers (such as buckets,and open bottles) have a shorter expiration as the chlorine in the solution begins toburn off leaving only the sodium chloride. Open containers should be formulated andused within the same day.

Some drawbacks with sodium hypochlorite focus mainly on the residue andcorrosiveness of the product. As previously stated, the chlorine in the solution beginsto burn off leaving only the sodium chloride. This white crystal-like residue attacks theimpurities in stainless steel (as an example) over a longer time frame. When using asodium hypochlorite solution it is imperative to remove the sodium chloride residuesfrequently to minimize this corrosive action. Rendering of these products as sterile isa “must” prior to use in a Class 100 (Grade A and B, ISO 5) and adjacent Class 10,000(Grade C, ISO 7) areas. Sterilization of disinfection agents is discussed in depth in thesection, Sterility of Disinfecting Agents.

Hydrogen PeroxideHydrogen peroxide is one of the most common disinfectants in the industrialmarketplace. In hospital and consumer use at a 3% solution the product is commonlyused as an antiseptic. In the pharmaceutical and biotechnology industry, hydrogenperoxide is used at 35% as a sterilant in isolators and at 3–10% for surfacedisinfection. Depending upon the concentration used, hydrogen peroxide is effectiveagainst bacteria, yeasts, viruses, and bacterial spores. Destruction of spores is greatlyincreased both with a rise in temperature and increase in concentration

15. Hydrogen

peroxide is a clear, colorless liquid that is environmentally friendly as it can rapidlydegrade into water and oxygen. While generally stable, most hydrogen peroxideformulations contain a preservative to prevent decomposition. At lower concentrations(3–10%) the chemical is effective against gram-positive and gram-negative bacteria,viruses and bacterial endospores in lower enumerations. However, at higherconcentrations and longer contact times the product exhibits superior sporicialreduction of bacterial spores. Some of the positive features of the product are its mild(if any) odor and its low residue characteristics. However, the product also has somesetbacks in exceeding OSHA exposure limits if used in confinement in too large aquantity. Precautions should be vested in this arena. Rendering of these products assterile is a must prior to use in a Class 100 (Grade A and B, ISO 5) and adjacent Class10,000 (Grade C, ISO 7) areas. Sterilization of disinfection agents is discussed indepth in this chapter in section, Sterility of Disinfecting Agents.


]PERACETIC ACID AND HYDROGEN PEROXIDE

Peracetic acid and hydrogen peroxide mixtures have received much recent attention inthe pharmaceutical and biotechnology industry in recent years. The chemical isconsidered a more potent biocide than hydrogen peroxide being bactericidal, virucidal,fungicidal and sporicidal at very low concentrations (<0.3%)

16. The product as

originally designed for the sterilization of medial devices. The chemical destroys thecell by destroying vital membrane lipids, DNA and denatures proteins and enzymes.Peracetic acid and hydrogen peroxide mixtures decompose to acetic acid and oxygen.Active percentages of marketed products range from 0.3% to 1.3% as a sterilant inboth ready to use and concentrate solutions. Ready to use solutions require a very highlevel of acetic acid as a stabilizer nearing 5.2%. While concentrate products needsmaller amounts of acetic acid in the formulation. Concentrate solutions incorporateapproximately 8% acetic acid and upon formulation to a use dilution, this value dropsto near 0.4%.

One of the main misconceptions with the use of peracetic acid and hydrogenperoxide mixtures as well as most registered sporicides is that the product needs tobe used at the sterilant label claim active percentage. First we must understand therequirements set for by the US EPA. The US EPA requires all registrants makinglabel claims to do so by following test methods outlined in by AOAC protocol. Thesterilant claim on products/labels follows the AOAC sporicidal test. This testrequires the complete reduction of 106 of B. subtilis and C. cporogones in a 60-carrier test, at 20°C, in a soil load. In the clean room, bioburden is significantlylower. Thus, the active percentage needed to destroy the flora normally seen issignificantly less. Simply, registered sterilant label claims are too strong for what isnoticed in a clean room. Coupled with the high enumeration of the AOAC testparameters is also a soil load, not present in clean rooms. Thus end-users shouldlook to validate a concentration of peracetic acid and hydrogen peroxide mixturesas well as other registered sporicides at realistic bioburden values as discussed laterin this chapter in the section entitled Determining Antimicrobial Effectiveness. Thiswill significantly reduce odors, deterioration of surfaces, problematic usersituations and residues.

Peracetic acid and hydrogen peroxide mixtures have pungent vinegar smell thatis offensive, if not intolerable to many users. Due to the horrific smell and thecharacteristic drying of mucosal membranes, peracetic acid and hydrogen peroxidemixtures have caused dissatisfaction among end-users. Facilities that have usedsodium hypochlorite (bleach) for years find the transition to peracetic acid andhydrogen peroxide very difficult to handle in terms of worker satisfaction. Theproduct, if used in a clean room environment that may have 15–20% fresh air, mayeasily exceed required levels when industrial hygiene testing is performed. Safetyprecautions for end users should be ensured prior to its use.


While reports of the product deem it non-corrosive to metals, industry reports haveshown this product react adversely with most stainless steels, aluminums, plastics,epoxies and most clean room surfaces. After application, a white cloudy residue isnormally left that requires either an IPA wipe down or a WFI rinse to remove.

GLUTARALDEHYDE

Glutaraldehyde has been used for some time as a disinfectant and sterilant forendoscopes and surgical equipment. Glutaraldehyde is normally sold in a 2.0%solution. The product is usually supplied as an amber solution with an acid pH.Glutaraldehyde is a powerful biocidal agent having the advantage of continued activityin the presence of organic material.

17Glutaraldehyde has broad spectrum activity

against bacteria, bacterial spores, viruses, and fungi. The mechanism of actioninvolves the destruction of the outer layers of the cell. Glutaraldehyde is the onlyaldehyde to exhibit excellent sporicidal activity. In recent years, glutaraldehyde's usehas been focused mainly on the hospital environment. Many pharmaceutical andbiotechnology organizations do not use a glutaraldehyde product in their operations.The product is very toxic and specific handling precautions must be employed prior toits use. Especially noted are the gaseous fumes and the possible absorption throughhuman tissue (skin).

FORMALDEHYDE

Formaldehyde is widely known as a fumigant for rooms and buildings. It has beenshown to be effective against bacteria and bacteria spores and vegetative bacteria

18.

Acklund et al. (1980) showed that at 20°C and a relative humidity of approximately100%, a 6-log reduction of B. subtilis spores was obtained after 1.5 hours exposure to300 µ/L whereas at 250 µg/L only a 4-log reduction was obtained after six hours ofexposure. The mechanism of action of formaldehyde is assumed to be due to thereaction with cell protein and DNA or RNA (Russell, 1976).

Formaldehyde is normally used in the pharmaceutical and the biotechnologyindustry as a means to bring back an area after shut down or major maintenance.During its implementation very stringent safety precautions are assured for personnelprotection that include areas and building clearance and hold times of areas prior torelease.

While formaldehyde is effective, this chapter focuses on routine methods ofcleaning and disinfection. Formaldehyde does not fit appropriately as a choice in thisvenue and would be used as a mechanism in opening of a new area or as a method


when coming back from a shutdown period.

As previously discussed, antimicrobial effectiveness depends on a variety offactors. The two most notable are the time period afforded to the disinfectant tosaturate and penetrate the cell wall of the organism, and whether the chemical iscapable of killing the organism in question. The key to selection is answered in one'slist of environmental isolates. The decision is based on the correlation between theorganisms on the list versus the chemical agent that can be proven efficacious againstsuch organisms. Process constraints may also affect one's choice of disinfectants, assome may be harmful to final product or incompatible with clean room surfaces.Varying applications require various solutions to be in place. he choice to use a phenol,quaternary ammonium or hydrogen peroxide based solution on a daily basis (in a Class100, ISO 5, Grade A or B area) is normality for the pharmaceutical and biotechnologyindustry. The choice delineates the rotation parameters. The choice of one disinfectantand a sporicide is completely appropriate; however, some may decide to rotate similardisinfectants while also utilizing a sporicide.

Rotation systems are designed to address known or possibly existentcontamination with proven efficacious disinfectant. The basis for the rotation ofdisinfecting or sporicidal agents is to address an organism that may not be destroyedby a particular disinfectant with another that has proven efficacy performance againstsuch organism. An example would be a phenol may not kill a b.subtilis spore in a 5–10minute contact time and thus, the rotation to a more efficacious product such assodium hypochloride at 0.52% (a sporicide) may be warranted to destroy thisorganism.

However, organisms do not develop and immunity or resistance to a chemicalagent over time. This theory is exactly that, a theory that has never been proven in theclean room environment. This confusion arises by the multiple meaning of the word“resistance”. Resistance is used to describe "not destroyed by" and incorrectly used todescribe "the building of an immunity by the organism to the chemical agent". Thetheory was originally borrowed from the medical industry where it was applicable tonoticed resistance in the human body. Subsequently and without proven, publisheddata, such theory was implemented into the clean room industry. However, antibioticresistant microorganisms are susceptible (or killed) to chemical germicides. Themechanisms by which chemical germicides and antibiotics work are completelydifferent and there does not seem to be a relationship between antibiotic resistance andchemical germicide effectiveness. We need to also realize that “natural selection” is aslow process that requires high populations of cells, constant selective pressure and arich growth environment. These are not found in the clean room environment.Resistance to antibiotics is usually acquired through modification of a single gene (oracquisition of a single gene) that blocks the very specific action of the antibiotic. Theantimicrobials we deal with do not act on a single enzyme, but have numerous effectson almost every aspect of cellular physiology. This meaning that multiple mutations


would be required to overcome their detrimental effects. Again, this is not likely, giventhe environmental conditions and low cell numbers within the clean roomenvironment

19. Therefore, the basis for rotation is to address an organism that is not

destroyed by, nor ever was destroyed by, one chemical agent with another that hasproven efficacy performance against such organism. And, the basis for implementinga rotation system may not be to address the building of immunity by an organism to achemical germicide.

On the contrary, one such study that reported supported the resistance theory waspublished in article form in 1992, entitled Rotation of Phenolic Disinfectants (Connorand Eckman

20). In this paper they examined whether it was possible for a Pseudomonas

aeruginosa (ATCC 15442) at a population of cells equal to 106CFU/ml. could develop

a resistance to a phenolic germicidal detergent in a laboratory experiment. From thepaper: "The testing method consisted of treating P. aeruginosa on agar surfaces,subculturing (transferring) survival cells, and then repeating the process. A total of 40transfers per treatment per trial and two trials ere performed. Data indicated that the P.aeruginosa (or resistant sub populations) became adapt or resistant to the alkalinephenolic disinfectant. However, when this compound was applied to the culture on arotational basis with the acid phenolic disinfectant during the same time period, noadaptation or selection occurred."

However, this test report was conducted in a laboratory setting. The publishedarticle attempted to prove that if such an occurrence existed that the "resistantmicroorganism" would be destroyed by either varying pH phenolic.

The pharmaceutical and biotechnology industry demands thorough testing in allaspects of our operations. During this time period, many jumped to what may havebeen thought to be some applicable data that should be implemented across the boardwith all organisms. But in reality as an industry, we needed to investigate the subjectfurther. During this time period, the data as it was presented was bastardizedthroughout the industry. It turned from "in this test we found that P. aeruginosa wasnot completely killed by a low pH phenolic and was subsequently destroyed by a lowpH phenolic" to "all organisms can develop a resistance to disinfectants". This was notfactually proven. And if it were proven so, the industry would have a horrific problem.If organisms could develop a resistance to disinfectants at the low levels that they areseen in the clean room, the first question that would scientifically be asked is "whendoes this occur?" As this question could not be answered, the industry would be forcedinto reproducing antimicrobial effectiveness testing on a routine basis. Ansubsequently, such testing would be done for an unscientific reason that would burdenfirms with an extremely high overhead cost. The average time contact kill study maycost a firm from $1,500 to $2,000 per test of one organism against one disinfectingagent (in solution or on carrier surfaces). Added to the cost of routinely reproducingantimicrobial effectiveness would be the question of how often should this data bereproduced. Simply an unanswered question that would force firms into a greater


frequency as time continued. The Conner and Eckman article tested one organism, P.aeruginosa. Obviously the scale of flora in the clean room setting would be muchbroader. A multitude of microorganisms would be noticed in normal clean roomoperation. For the industry to base all disinfection systems on the testing of oneorganism would represent an incomplete analysis of pertinent data surrounding thissubject. Thus, coupled with the question of how frequently should we test would bethe question, "What organisms do we test?" In completeness, if one believed that anorganism could become resistant to the disinfectant used, all environmental isolatescould be suspect. Would one species of Staphylococcus be affected like another? Allof these questions could be pushed to the foreground when one utilizes an unprovenbeginning basis as our starting point. Simply stated, conclusive evidence that relates toour specific scenario should be the basis for us determining our starting ground.

In the above paragraph, we discussed the possible need to reproduce antimicrobialeffectiveness on a routine basis if we believed the theory of resistance. However, if webelieve in resistance then the list of questions continues. If we use a disinfecting agentthat an organism may become resistant to in this month, and one that it is not resistantto the next month, are we not destroying the cell during the first month time period?Could it compromise our product?

Further, if we do not rotate a high pH phenolic with a low pH phenolic, what dowe rotate with a quarternary ammonium? We cannot rotate a quarternary ammoniummonthly with a phenol as they are not compatible and create a sticky residue on thesurface. Likewise, what do we rotate other chemical compounds with? Rotation of twodisinfecting agents may limit us to a phenol rotation that may or may not be the bestchoice to choose.

The rotation of disinfecting agents to combat microbial resistance incorporates yetanother stumbling block when we review the subject of a routine sporicidal applicationto our controlled environments. The rotation of two disinfectants does not address thepossible content of spore populations in our clean room environments. While somedisinfectants may have a very small log reduction of spores, we will still incorporate,on a frequency, a sporicidal agent into our disinfection regime. Thus, how will thisresistant microorganism that was not killed by one disinfectant but destroyed byanother survive through the application of a sporicide? There is no scientific theory orcase study that would support the cell's ability to live through this scenario.

As an industry, the push for the rotation of two disinfecting agents is becomingless and less supported. We can see this in review of some of the current guidelinespublished in the industry such as USP 1072, Disinfectants and Antiseptics whichstates: "The development of microbial resistance to antibiotics is a well-describedphenomenon. The development of microbial resistance is less likely, as disinfectantsare more powerful biocidal agents than antibiotics and are applied in highconcentrations against low populations of microorganisms, so the selective pressure


for the development of resistance is less profound."

On September 27, 2002, the FDA released the Sterile Drug Products Produced byAseptic Processing Draft. While in the preliminary stages and not for implementation,the writings make no mention of any concern that an organism may develop aresistance to a chemical agent over time. However, in several instances the guidelinedoes mention that one should continually evaluate their systems. While this documentmay change, we should consider the basis for their thought process. An excerpt fromthe guideline is as follows.

Sanitization Efficacy"The suitability, efficacy, and limitations of sanitization agents should beassessed with their implementation for use in clean areas. Theeffectiveness of these sanitization procedures should be measured bytheir ability to ensure that potential contaminants are adequately removedfrom surfaces (i.e., via obtaining samples before and after sanitization)."

"Upon preparation, disinfectants should be rendered sterile, and used fora limited time, as specified by written procedures. Disinfectants shouldretain efficacy against the normal microbial flora and be effective againstspore-forming microorganisms. Many common sanitizers are ineffectiveagainst spores, for example, 70% isopropyl alcohol is not effectiveagainst Bacillus, spp. spores. A sporicidal agent should be used regularlyto prevent contamination of the manufacturing environment withotherwise difficult to eradicate spore forming bacteria or fungi."

"After the initial assessment of sanitization procedures, ongoingsanitization efficacy should be frequently monitored through specificprovisions in the environmental monitoring program, with a definedcourse of action in the event samples are found to exceed limits."

Resistance of microorganisms to disinfecting agents is a subject that is based onthe theory that it could occur. Rarely in our internal scientific evaluations and ourpresentation of our validation to regulatory inspectors do we use the justification that"in theory, we think it could occur." The use of theory may guide us to proof, but neveris a replacement for proof.

If we define rotation of chemical agents as the "desire to assure a more broad-spectrum kill in our disinfecting regime" then we have identified the key element forthe reason why we would scientifically consider this subject. Rotation is an applicableand a scientifically proven term. However, the rotation of two disinfectants is notapplicable to address microorganisms we may see. Destroying contamination in aclean room operation requires addressing the known level of vegetative cells and theknown level of spores. In design of a rotation system, there are three types:


• 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 presentedhere for use in a Class 100 and the adjacent Class 10,000 environment.


DAY(S) Phenolic Quaternary Hydrogen Ammonium Peroxide

Day 1–13 High pH Phenol Quaternary HydrogenAmmonium Peroxide

Day 14 (if warranted Clean the surfaces, Clean the surfaces, Clean the surfaces,by EM data) allow to dry, then allow to dry, then allow to dry, then

apply the Sporicide apply the Sporicide apply the Sporicide

Day 15–29 Low pH Phenol Hydrogen QuaternaryPeroxide Ammonium

Day 30 Clean the surfaces, Clean the surfaces, Clean the surfaces,allow to dry, then allow to dry, then allow to dry, thenapply the Sporicide apply the Sporicide apply the Sporicide

After disinfection, when all surfaces are dry, all critical surfaces should be rinsed with hotWFI or an IPA wipe down performed.

Table 3: Rotating Two Disinfectants and a Sporicide

DAY(S) Phenolic Quaternary Hydrogen Ammonium Peroxide

Day 1–13 Low pH Phenol Quaternary HydrogenAmmonium Peroxide



Day 15–29 Low pH Phenol Quaternary HydrogenAmmonium Peroxide



Table 2: Rotating One Disinfectant and a Sporicide

The first two systems (Tables 2 and 3) require, at minimum, a monthly sporicidalapplication. This may be increased or decrease in time frames and will be determinedby the environmental conditions. Industry standards look to a weekly application at thebeginning and such application becoming lessened as control of the environment isconsistently attained. The third uses a sporicide daily and is the most effective but is notrecommended, as it is overkill for the known contamination level and will deterioratesurfaces over time. Within our cleaning and disinfection regime, we also need toaddress the cleaning aspect. As we have discussed previously, cleaning and disinfectingare not the same. The use of cleaner is considered an optional step in controllingexistent residues and should be done at least once a month. After disinfection all criticalsurfaces should be rinsed with hot WFI or an IPA wipe down performed. These subjectswill be completely discussed later in this chapter in the section entitled, AssuringApplication of Our Disinfectants and Sporicides.

DETERMINING ANTIMICROBIAL EFFECTIVENESS

Determining what chemical agents will destroy a known level of one's environmentalisolates or ATCC cultures is the next step. Prior to conducting either a Time Contact KillStudy (Tube Dilution), or a Time Contact Kill Study (On User Surfaces) or an AOACProtocol Study, one needs to review the available disinfecting agents and determinewhich is initially appropriate for their operations. Upon choosing 1 or 2 disinfectingagents and a sporicide, one can continue with the antimicrobial effectiveness studies.

Validating one's sanitizing, disinfecting and sporicidal agents requires them todelineate the organisms to be tested. One could use a list of ATCC cultures, however,


DAY(S) Sodium Hypochlorite Peracetic Acid and Hydrogen at 0.52% Hydrogen Peroxide Peroxide

Day 1–13 Sodium Hypochlorite Peracetic Acid and Hydrogenat 0.52% Hydrogen Peroxide Peroxide 6%



Day 15-29 Sodium Hypochlorite Peracetic Acid and Hydrogenat 0.52% Hydrogen Peroxide Peroxide 6%



Table 4: Use of a Sporicide

utilization of one's environmental isolates nets a more exacting test for each uniquemanufacturing operations. Testing ATCC cultures such as Bacillus subtilis, Aspergillusniger, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans andEscherichia coli is acceptable, however not as exacting as testing a plane of one'sknown environmental isolates. The use of one's environmental isolates in a preferredmethodology by most regulatory agencies.

Antimicrobial effectiveness studies need to be based on realistic bioburdens thatmay be noticed in the controlled areas. Normality is to test an enumeration greaterthan or equal to 1.0 x 10

4CFUs. Our goal is to prove a 3-log reduction. However,

some guidelines like the British Standard BS EN 13697:200121

calls for a 4-logreduction as proof.

In determining which test to conduct, one needs to review how one will addressan organism in the clean room. As the organism will be on the surface, a time contactkill study that confirms the destruction of a known enumeration of cells on an end-user's surface is more depictive than a time contact kill study done in a tube dilution(in suspension). The reasoning for this is organisms dried on a surface better depict thesituation of disinfecting an organism in the clean room. The available surface area ofthe organism that can be contacted by the disinfectant that rests on the surface is 270°.The available surface area of the organism that can be contacted by the disinfectant inthe tube dilution study is 360°. Obviously, the surface test presents a more realisticscenario. AOAC protocol testing is required by the US Environmental ProtectionAgency (EPA) to register a claim for a disinfecting agent. It utilizes 60 carriers (aceramic penicylinders) and requires a high enumeration value of equal to or greaterthan 1.0 x 10

6. Protocols use either AOAC Use-Dilution or AOAC Sporicidal tests

procedures. While this is the method used for registration, it may be too involved andexpensive for pharmaceutical and biotechnology firms to utilize as a method fortesting antimicrobial effectiveness.

The EPA supports the use carrier methods for the evaluation of a disinfectantproduct's efficacy. This test requires the microorganism is to be dried on a non-porouscarrier. The rationales behind the choice of a carrier method are the beliefs that:

• microorganisms that are dried are more difficult to chemically inactivate thanthose microorganisms in suspension

• that in the health care setting microorganisms are more often found in the driedstate than in suspension

22

Within the framework of antimicrobial effectiveness testing, we also need toincorporate realistic contact times to depict representative dry times of the disinfectanton clean room surfaces. Due to the significant movement of air from laminar flow inthe clean room, normal dry time is 5 minutes at best. While our floors may remain


wetted longer (possibly up to 10 minutes), the vertical surfaces will dry faster (3–5minutes). Thus antimicrobial effectiveness testing should incorporate a worse casescenario and utilize a 3–5 minute dry time. An exception to this rule would besanitizing agents such as isopropyl alcohol, ethyl alcohol or AAA ethanol at aconcentration of 70%. These products dry faster nearing 1–2 minutes and testingshould be altered as such. In recent years, many firms have begun to test three timeframes to prove the disinfectant's activity over varying time periods. The authorsupports this practice and would suggest the time period of 3, 5 and 10 minutes betested consecutively. Normally in 3 minutes the disinfectant shows average activity, in5 minutes good activity, and in 10 minutes excellent activity. Testing of a variety oftimes ensures one has data in the file to support a complete range of dry times (contacttimes) that may be noticed in their manufacturing or testing operations. Uponcompletion, this testing will provide the justification for utilizing the chemical agentsto destroy the known and possibly existent contamination in the facility. As timeprogresses, we need to continually updating our profile of organisms versus ourchemical agents.

While the type of test, the enumeration level of microorganisms, and the contacttime are some of the most critical factors to assess, other critical factors also need tobe determined. One of these is expiration of the disinfectant. Expiration foreffectiveness can be determined by incorporating a simple variable in our test calledaging of the disinfecting agent. Simply done, one would open a bottle of disinfectantand age it for the time period that they plan to use it, say 30 days (concentrate andready to use). A ready to use solution is tested at the 30-day period. For a concentrate,the aged concentrate solution is diluted to the prescribed use-dilution. The use-dilutionis then aged for the time period that it will be used (for example, for 7 days). At theexpiration of the use-dilution it is then tested for antimicrobial effectiveness. Thissystem of expiration proves that a solution, in use for “X” time period has the abilityto destroy an acceptable level of microorganisms that we have determined to bepresent in our operations.

Expiration dating of disinfectant effectiveness can also be tested by conducting anantimicrobial effectiveness test on a newly open bottle of disinfectant (ready to use orconcentrate to use dilution) and then subsequently aging the solution for the expiryperiod and testing the active ingredients. The correlation between the activeingredients at time of opening and their satisfactory stability at end of expiry providesthe needed data to support the use of the agent for the time period. However, in thistest, there must be the understanding and explanation of the relationship between thetested solution and the expiry active data that followed.

In performing antimicrobial effectives testing some other factors come to the forethat warrant attention. The first is soil load. Normally, clean room operations do nothave a soil load present and the most soil that exists would be that of disinfectantresidues. In the case of an existing soil load, one should conduct their testing in a


similar situation. Commonly used as a soil load or an increase level of protein (a fetalbovine serum) at 5% v/v is added to the organism challenge to test the ability of thedisinfectant or sporicide under the circumstances of a soil load (dirtied) condition.

Another factor that may surface is hardness and temperature of water. Requiredby all registrants of disinfecting agents must state in their labeling their ability orinability to achieve antimicrobial effectiveness in the presence of hard water (400 ppmas CaCO3). At the same time the temperature of the solution may vary in its effectagainst microorganisms as elevated temperatures tend to increase the ability of thechemical agents performance.

While disinfectant validations are expensive and time consuming they areforemost in most regulatory agency's minds. And thus, are a required to assuring theeffectiveness of a cleaning and disinfection system that will prove an acceptableenvironment during the manufacture of product.

ASSAY OF DISINFECTING AGENTS

Analytical validation of our disinfecting agents tests that the required percentage ofthe chemical agent is present to ensure antimicrobial effectiveness. If the appropriateuse instructions are followed, a ready-to-use product or a formulation fromconcentrate is normally easy to prove as having a sufficient amount of the activeingredients to reconfirm the required percentage that was validated in ourantimicrobial effectiveness studies. Upon the use of this product past the first use iswhere we start to see the possibility that the percentage of the active ingredients maybegin to slowly become too low to warrant continued use. Varying products havevarying in-use time periods. Time periods range from 7–30 days. This scenario needsto be validated for each chemical agent and each container type. As an example of thesame chemical having varying in-use time periods, an isopropyl alcohol solution inan aerosol for can be used for a longer time period than a trigger spray bottle. Thereasoning for this difference is the aerosol container is sealed in a pressure vessel.The reduction in percentage of the active ingredient due to evaporation or thequestion of sterility over time is not founded in this container type. On the other hand,the trigger spray container may slowly aspirate room air to the master reservoir thatmay compromise the level of active ingredient and the sterility of the container overtime. The basic validation question is "what is the time frame that we can prove thechemical's active percentage and sterility to be valid for, once opened?"


STERILITY OF DISINFECTING AGENTS

Through our antimicrobial effectiveness studies, we realize that disinfectants do notkill all organisms. As all chemical agents may have an inherent bioburden (normallyspores), we must ensure that such bioburden is removed prior to their entry to ourcontrolled areas. The transfer of such organisms through our disinfectants to ourcontrolled areas, especially our aseptic filling areas, should be viewed as a catastrophicevent. To even further reinforce the issue, we spread the disinfecting agents all overour walls, ceiling and floors. Controlling the contamination from ever entering is mucheasier that subsequently removing it from the controlled area. If we review regulatoryexpectations in this area we will find the requirement to sterilize all disinfectants andsporicides prior to entry to the controlled environment.

The FDA has stated in its Sterile Drug Products Produced by Aseptic ProcessingDraft that, "Upon preparation, disinfectants should be rendered sterile, and used for alimited time, as specified by written procedures."

Purchasing a disinfectant or sporicidal product as sterile from an audited vendorrequires one to review the following critical items as a quality control measure toassure 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 0.2 microns

• Aseptically filling the product into pre-sterilized containers or exposing the entirecontents to a terminal sterilization process such as gamma irradiation.

• Requires a lot-by-lot sterility test per current USP or EP compendium (conductingsterility testing requires the completion of bacteriostasis and fungistasis (B/Ftesting. This proves that the sterility test is capable of growing organisms in thepresence of the chemical agent.)

• Requires the assessment for expiration dating of an unopened container


If the disinfectant or sporicide is to be processed sterile in-house, then the reviewof the following critical items as a quality control measure is suggested to assure whatone 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 providing microorganisms are retained by the filter

• Assay of the solution (RTU or concentrate) to an acceptable active percentage

• Filtering the solution at 0.2 microns

• Aseptically filling the product into pre-sterilized containers or exposing the entirecontents to a terminal sterilization process such as gamma irradiation.

• If the product is aseptically filtered, then the assurance that all that comes incontact with the product after the filter is rendered sterile.

• Requires validation to be performed using a lot by lot sterility test per current USPor EP compendium or a bioburden analysis for at least three lots at the beginningand subsequently routinely tested as a quality control check. Sterility testing on alot-by-lot basis need not be performed if sufficient validation testing is conducted.

• Conducting sterility testing requires the completion of bacteriostasis andfungistasis (B/F) testing. This proves that the sterility test is capable of growingorganisms in the presence of the chemical agent.

• Requires the assessment for expiration dating of an unopened container

Validation of a sterility claim for disinfectants should be a focus of one’svalidation efforts. Normally the testing of three processed lots for sterility providessufficient data. The pre-sterilization of our chemical agents prior to entry to thecontrolled area is simple common sense and common practice in the industry.

Container type is critical when using a sterile disinfectant. Container typesinclude aerosol, trigger spray, squeeze bottle, and larger closed containers (1–5 gallonand larger). Aerosol container is the most lucrative type, as the vessel does not aspiratethe room air to the master reservoir. However, the container and its contents must besterilized via gamma radiation or all the components pre-sterilized and subsequentlyfilled via a validated aseptic filling operation. For obvious reasons, gamma


sterilization of the entire contents is considered far superior methodology forachieving a sterile product. In 1992, the first sterile disinfectant, DECON-AHOL® (asterile USP IPA), was marketed under US patent 6,123,900, Method of Sterilization

23.

This patent and product showed the industry the effectiveness of this type of container.Thus, no viable or particulate contamination is returned to the solution contained. Apre-sterilized aerosol or pressurized vessel assures assay of the active ingredients (forthe time period they remain stable) and sterility for the expiration period designated bythe manufacturer.

Other smaller containers include the squeeze bottle and trigger spray bottles.While acceptable containers for all disinfectants and sporicides, the container itselfaspirates the room air back to the master reservoir. Thus, assay and sterility arecompromised after the initial use. The same is true for larger 1–5 gallon containers andlarger. Once opened sterility is compromised.

For varying disinfecting and sporicidal agents a variety of containers need to beutilized. For ready to use mixtures we have to decide how the product will be used.If in a smaller aerosol, trigger spray or squeeze bottle, we may want to utilize aproduct that is pre-sterilized in this smaller form rather than attempting to pour orfilter such solutions to a empty pre-sterilized container. We may also want to limitthat the capping of product “to be used later” as our assay and sterility may becompromised over time. For concentrate products that need to be diluted with aquality water grade, we may want to look to implement unit dose bottles that areincorporate a pre-measured dose and are sterile. This system is superior to that ofpouring a concentrate disinfectant or sporicide into a measuring cup and capping theremainder for later use. Questions may arise as to the assay and sterility of thisremaining solution over time.

Thus, no viable or particulate contamination is returned to the solution contained.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 perioddesignated by the manufacturer.

TYPES AND STERILITY OF CLEANING COMPONENTS

The sterility of disinfectants and sporicides is essential to assure the non-transfer oforganisms to the Class 100–10,000 areas. Of equal importance is the transfer oforganisms to the Class 100–10,000 areas via our cleaning components. Our cleaningcomponents are defines as spray-mop-fog pressure devices, buckets, mops sprayers,mop handles, mop frames, mop heads and other components used during the cleaningand disinfection operation.


In review of this subject we see problematic quality control situations that mayarise from these items not being rendered sterile. At the same time, manufacturingoperational difficulties may exist in implementing these items as sterile.

Let's review the quality assurance side of the subject: the utilization of a steriledisinfectant that is diluted in a sterile quality water grade (such as sterile water forinjection) and that is subsequently poured into a non-sterile bucket compromises oursolution's sterility and our environment. Moreover, if we then dip a non-sterile mophead into the solution, we again corrupt our solution and possibly the surfaces that wetouch with the mop head. If our handles and mop frames are not sterile they too canbring in unwanted contamination to the area. A typical quality assurance line may be"You need to sterilize each component prior to use to reduce the ingress ofcontamination." However, quality assurance may not be completely abreast of what itwill take to accomplish this scenario.

On the production side of the subject if we utilize sterile components we maypossibly need to purchase two to three times the numbers we presently have (due tosterilization schedules) which is very costly. Production will have to transfer thewrapped sterile components to the controlled areas, which can be awkward, and willbe required to decontaminate the exterior of the wrappings prior to entry to the Class100–10,000 area. Dependent upon the location of quality water that will be used todilute our concentrate disinfectants, we may need to open such components outside thecontrolled area. If this occurs, they will not be sterile any more. Incorporating apurchased or pre-made sterile water grade available in the area will also add significantcosts. And finally, of the components that enter the area during manufacturing, theseitems should be of little consequence, as they will have disinfectants in and on themduring the disinfecting operation. A typical statement from production may be "Well,we wheel in big tanks during manufacturing and they are not sterile."

Both sides have their valid points. All considerations need to be reviewed. Thegoal in the sterilization of the cleaning components is to reduce bioburden to the area.If this is done, chances are lessened for contamination entering. While not every itemcan be sterilized prior to entry to the aseptic manufacturing area, as much as can besterilized should be sterilized. Our cleaning components and our disinfectantsrepresent a common place where we can start anew each day with control. Prior tosterilization we must review the subject from the end of the cleaning operation andassure that these items are cleaned prior to sterilization. Otherwise, bioburden on theitems and deterioration to the items will increase over time.

But the real question is frequency. How often do we clean and sterilize thesecomponents? Spray-mop-fog devices, buckets, mop frames and mop handles need tobe cleaned after use. This includes the inside, the outside, underneath and the wheels.We can then start with a clean component. At this juncture we need to determine wherethe component will be taken and stored. If it is stored outside the Class 100 area, then


the item needs to be disinfected or sterilized again prior to entry. Most of the time, thisis the case. While we understand the need to clean the components after use,disinfection or sterilization prior to entry still looms as a unique question that needs tobe answered by each operation. An important factor in consideration is where thewater will be obtained to create the dilution. Of enormous expense would be theestablishment of new WFI water drop lines to serve this operation. And so the storyencounters more complication.

As an industry smaller firms or firms with only a few areas seem to be able tosterilize these components before use. As the scale increases to larger operations thisbecomes more difficult. As scientists we know that sterilizing the buckets is thesuperior methodology and we should make arrangements to move towards such ascenario. Sterilization of buckets on a weekly, monthly or quarterly basis does verylittle to control bioburden to the area on a daily basis and should not be considered. Atthe minimum level, we need to clean these devices each day and disinfect these itemsprior to entry to a controlled area. Disinfection should be done using the chemicalagent used in the cleaning operation. Science may say that we need to use a sporicidalon the items prior to entry; however, if a disinfectant is used in, say, the bucket duringthat cleaning shift, we may have a mixture of chemical agents. This should be avoided.

Mop handles and mop frames are a different story. They are small enough andinexpensive enough to have a variety on available, pre-sterilized and used per cleaningoperation. The mop handles and frames should also be cleaned prior to sterilization.The same is true for mop heads. Pre-sterilized mop heads (foam or string) should beused and changed daily in a class 100 and 10,000 operation. The frequency ofchanging a mop head used in class 100,000 could move forward to weekly, however,a better scenario is also to use this mop head and then discard. The reasoning forroutine discarding of the mop in Class 100,000 is it is dirtied and is normally air-dried.Bioburden levels build up on the mop head and may cause potential problems. Theseproblems may not be worth the cost of the mop head.

IN-USE EXPIRATION OF DISINFECTANTS AND SPORICIDES

Questions surrounding the expiration of an in-use disinfectant and sporicide will bequestioned by all involved internally and externally by regulatory agencies. Severalmethodologies were previously discussed in this chapter (in the AntimicrobialEffectiveness section) as to how to develop this data. The two main questions that ariseare:

• How long does the disinfectant retain its active percentage enabling it to destroymicroorganisms?


• How long does the container and the contents held within the container remainsterile?

Both are very important questions and, from the author's standpoint, two of themost frequently-asked questions in the industry surrounding disinfection. Simply anassay over time of use and an accompanying sterility or bioburden test will delineatethis time frame. The importance of reporting this information to cover all potentialsituations is critical. As an example, SOP's should look to address the various type ofuse situations as depicted in Table 5 and identified as concentrate bottles, ready to usebottles, unit dose concentrate bottles, open formulations (in tanks, etc.) and closedcontainers (such as squeeze bottles, trigger sprays and closed bottles).

CONDUCTING IN SITU FIELD STUDIES

Once a disinfection system has been chosen and antimicrobial effectiveness testing hasbeen completed, conducting an "in situation field study" is important to prove theeffectiveness of the combination of our cleaning SOPs (standard operating procedures)and our antimicrobial effectiveness testing. Simply, environmental monitoring, both airand surface, is conducted in a dirtied room. Upon completion of the monitoring, theroom is cleaned and disinfected per the current operating procedures. Upon completionand drying of all surfaces, the room is monitored again. Satisfactory results need to beobtained in three different and separate in situation field studies prior to acceptance ofthe disinfection system. This testing is the movement of lab knowledge to the real lifesituation. It tests our overall effectiveness in conducting a disinfection operation.

Coordination between the quality assurance group and the production services orcleaning group is sometimes a very difficult task. However, the magnitude of thistesting should outweigh the coordination of scheduling. As an alternative method,some organizations have taken cultures enumerated at 10–100 CFUs and inoculated asimilar production surface. Likewise, some have just monitored a dirtied surfacesimilarly used in production. After inoculation the surface is cleaned using a multitudeof methods that may be employed. Such methods are:


Approved Concentrate Ready to Use Formulations Formulations inDisinfectant Bottle Unit Dose in Open Closed Containersor Sporicide Containers such as Squeeze,

(Buckets and Trigger Spray orTanks) closed bottles

Chemical 30 days after 30 days after 1 day after 7 days afteragent opening opening formulation formulation

Table 5: In-Use Disinfectant Expiration

• mopping

• spraying

• wiping

• fogging

The surface is then monitored again after cleaning as a test for effectiveness of thecleaning operation in conjunction with the disinfecting agents. Inoculation of actualproduction services in controlled areas should never be done, as one would neverintroduce any microorganisms knowingly to the area that may compromisemanufacturing of product later in the scope. While the in situ or field study conductedon similar surfaces nets good information, it is still not a test of the actual environmentand the cleaning that will be performed. Thus, it behooves organizations to conduct theactual in situ or field study in their manufacturing areas.

Regulatory agencies such as the FDA, consider the in situ or field study extremelyimportant. In an excerpt from the FDA's Sterile Drug Products Produced by AsepticProcessing Draft 9/27/02

18guideline is as follows:

Sanitization Efficacy"The suitability, efficacy, and limitations of sanitization agents should beassessed with their implementation for use in clean areas. Theeffectiveness of these sanitization procedures should be measured bytheir ability to ensure that potential contaminants are adequately removedfrom surfaces (i.e., via obtaining samples before and after sanitization)."

One concern that is vested throughout the industry from regulatory guidance isthe need to routinely perform this task. In support of the industry, such routinetesting is a revalidation of a validated system. Once the system is in place and thetesting conducted thereof, the system, if performed to accompanying SOPs, shouldprove effective each time it is performed. As an industry, the concern needs to bevested in the absolute assurance that such cleaning and disinfection procedures arefollowed on a daily basis. If this is done, the system should satisfy the need for anacceptable environment. If the system fails, this will be noticed in ourenvironmental monitoring program and such corrective action must then be takenon an immediate basis.

The in situ or field study is a realistic test of the ability of our cleaning anddisinfection systems, as a system (chemical and method) to render a tested “dirtiedsurface” as free of microorganisms. Our lab work in our antimicrobial effectivenesstesting renders us the understanding of what our disinfectants and sporicides arecapable of destroying in the presence of high bioburden levels. While important, it is


still a laboratory test and never ventures to a real manufacturing setting. The in situ orfield study is a critical test of our systems.

ASSURING APPLICATION OF OUR DISINFECTANTS ANDSPORICIDES

The most important subject is the arena of disinfection is application of the chemicalagent to the surface. As an industry, we do not spend enough time on this extremelyimportant function. Throughout the industry, we hear tales of this chemical is noteffective against this species of an organism. In fact, we may spend too much timetrying to figure out which chemical agent is most appropriate to destroy all thepossible organisms that we may encounter. At the same time we are focusing onwhich chemical kills what organism, we need to assure that the chemical agent isappropriately applied to the surface. To do so, we need to view more than just awetted surface. We need to identify the factors, aside from the chemical agent, thatwill assist our disinfection efforts. Putting the basics into perspective can simplifyour understanding of this subject and allow us to refocus our efforts appropriately.

First and foremost, we need to assure the saturation and penetration of the cell wallover a lengthened contact time. We need to reproduce the contact times from ourantimicrobial effectives testing on all surfaces in the clean room. Second, we need to makeits existence in the area very difficult. One way we can do this is by removing theorganism's required food source, such as particulates and water. This requires routinecleaning of the area. Another way is to change the temperature it is exposed to (Hot WFI,if permitted by our safety group and by the characteristics of the disinfectant) or byassuring it must exist in an undesirable environment. In reality, the undesirableenvironment is the clean room itself. Sufficient support conditions for sustaining anorganism's existence are lacking in the clean room environment. In short, it's a difficultplace to survive. Some consider the disinfectant or sporicidal residue as an undesirableenvironment. This is true to some degree, and for a short time period. However, it is alsotrue that a residue on the surface can complicate our manufacturing for a variety ofreasons. This will be discussed in detail later in the chapter. And third, we can removeorganism from the area by our cleaning practices. We can wipe it, mop it or rinse it fromthe area; we can also fatally damage the cell wall by an abrasive friction type action as iscommon by wiping or mopping. All of these factors will assist our disinfection efforts.

Our design then relies on the cleaning and disinfecting procedures that we have inplace. Our procedures need to include elements such as: the cleaning of a surface, thesaturating of a surface, routine change of dirtied solutions, prior sterilization andcleaning of our buckets and mop handles, routine changing of our dirtied mop heads,cleaning and sterilization of our sprayers and foggers, and our procedures relating totraining of our personnel to accomplish these critical tasks.


Application is normally done by four methods: spraying, mopping, wiping andfogging.

Spray nets the best coverage of a surface and proves to be the best form forassuring efficacy against microorganisms on the surface as the dry time or contact timeis longer. Simply, the surface remains wetted for a longer time frame. Effectivespraying is done from a light spray and not a power or pressure spray. While pressurespraying does clean the surface better, the key to assuring saturation and penetrationof the cell wall for the specified contact time is though a light spray with a largerdroplet size. Spraying should be done from the top surfaces to the bottom of the walls.Spraying should be done by applying to the ceilings first (without contact to filters)and then be done to the walls. Spraying should not be done on floors and the correctoption would be to apply the chemical via mopping. While spraying is effective inefficacy performance, it does not clean the surface as well as mopping. Types of spraydevices include canister sprayers, spray-mop-fog devices (Core2Clean®), pressurizedcanisters, and piped facility pressure systems. Cleaning and sterilization of thesecomponents has been previously discussed, and arrangements need to be made toassure the reduction of bioburden prior to entry.

Mopping a disinfectant to the surface creates a non-destructive abrasive typeaction that loosens particulates, residues and microbial contamination. Moppingshould be done in order: ceilings, then walls then floors. The mopping action shouldbe done from the top of the wall to the bottom with overlapping stokes. Similarly, theceiling should incorporate the same method over overlapping stokes and start from thefurthest corner of the room. Mopping the floor is done by starting at the furthest cornerand working towards the exit door (similar to painting a floor). While mopping createsan excellent scenario for cleaning, it is less effective in addressing microorganisms asthe mop head and the wall are less wetted. Thus, the surface dries faster. However, itis a very critical step that needs to be accomplished on a routine basis. Methodologyfor cleaning a surface (not disinfecting) using a mop is enhanced by a quality waterrinse such as WFI. During the cleaning operation the non-destructive abrasive typeaction loosens particulates, residues and microbial contamination. After this has beendone, it is important to rinse the surface to remove the contaminants from the surface.Upon rinsing, the dirtied solution on the floor is collected and removed. Once allsurfaces dry, then a disinfectant or sporicide is applied.

Mopping requires a variety of components that include buckets, spray-mop-fogdevices, handles, mop frames and mop heads. Buckets come in a variety of makes andmodels. Some include the single, double and triple bucket systems. In the singlebucket system, the disinfectant or sporicide is placed into a bucket and the moppingprocedure calls for the dipping of the mop into the bucket, then to the wringer, then tothe surface where the chemical is to be applied. In this system, the wringer andapplication solutions are mixed in one chamber. Increase in soil load and bioburdenoccurs in the bucket and the solution is dirtied in a short time. The double and triple


bucket system attempts to separate the wringer solution from the application solutionby creating separate chambers. Procedurally (in a two-bucket system), the mop entersthe front chamber to be wetted and is wrung into the back chamber. This system issuppose to keep the solution cleaner as the rinse is collected in the back bucket andnever contacts the application solution in the front chambers. The three-bucket systemexemplifies the two-bucket system and adds one additional bucket as middle rinsebucket for the mop. While these systems attempt to reduce the soil and bioburden loadin each chamber, the mop head is the contaminating factor of the solutions andeventually corrupts or soils all cavities in the system. Material grades of buckets arenormally made of stainless steel, galvanized steel or some form of plastics orautoclavable plastics. Compatibility with the disinfecting agent and capability ofsterilization should be tested prior to use. Buckets should also remain chemicalspecific as the mixture of chemical agents and their accompanying residues should beavoided (especially in a liquid state).

In recent years spray-mop-fog systems have been designed and patented for usein controlled areas. These devices are made of stainless steel and completelyautoclavable. They provide a chamber for the disinfectant or sporicide and onceintroduced, the canister is pressurized with compressed air (ASME rated to 100 psi).Off the tank exists the ability to connect a trigger spray activated power sprayer or atrigger activated mop (sending liquid automatically to the mop head or a fogger(allowing 2–3 hours of fog time). These systems dispense a clean liquid continuouslyfrom the canister. Superior wetting to the surface occurs as the and cleaning time isreduced by nearly 50%

24.

The single, double and triple bucket system and the spray-mop-fog systems alluse mop heads. Mop heads come in disposable and reusable forms. Mop heads alsocome in foam flat mops for wall and string mops for floors. A single use principleshould be applied in all classified areas. The question of disposable or reusable is anoperation specific concern. Disposable means use once and throw away and the costof doing so is approximately $8.00 for each sterile mop head. Washing and re-sterilizing each mop head is an involved process and an operation should review thetotal costs and possible impact to the environmental conditions prior to implementingthis scenario. A mop is changed:

• after use in each class 100 room. The same mop head may be sequentially used inClass 10,000 or 100,000 areas. A mop is never used in a Class 10,000 and thenused in a class 100

• when it is noticeable dirtied

• when the mop head is damaged.


Mop heads for class 100 and 10,000 should be rendered sterile prior to use.

Wiping is the fourth way of application. As was expressed earlier in the chapter,a very wetted wipe is excellent for disinfection but very poor for cleaning. To cleanwith a wipe, one should use a chemical agent and a cleaner. The key is to wet andloosen the contaminants from the surface and then wipe them away with a dry wipe.Consider cleaning a window at home as an example of the action needing to beperformed in a cleaning operation in controlled environments. Likewise, fordisinfection, it is necessary to saturate the surface.

Fogging is the last method of application. While fogging a room can be veryeffective, there are many important aspects that need consideration. First is coverageand droplet size. The droplet size required to effectively fog an area and saturate allsurfaces is 25.0 µm

19. This droplet size accompanied with the ability of the fog or mist

to be circulated to all surfaces is the key to success. Fogging can produce very goodresults in destroying levels of vegetative bacteria and spores on the surface but isdependent upon the chemical agent that is used and the time frame of the foggingoperation. Carrier surface tests inoculated with b. subtilis at 1.6 x 10

5(11 carriers)

show the destruction of such spores using a peracetic acid and hydrogen peroxidemixture from a concentrate (DECON-SPORE 200 Plus® at a 0.4% use dilution) in a 12foot x 15 foot room, using two two-jet foggers for two hours

25. While this study shows

the ability of the method to destroy microorganisms on the surface, fogging does notclean the surface: it only disinfects the surface. Fogging also requires room releasetimes that may near 3–4 hours. This release time is to assure harmful vapors havesubsided to a safe level for personnel to enter per OSHA standards.

One of the most confusing subjects surrounding application is frequency.Frequency of application is dependent upon a variety of factors that include thebioburden of the area, contamination control capabilities, the classification of the area,the process or manufacturing operation, personnel, product changeover, and the typeof facility and its associated controls. While many other factors come into play, theseare the main culprits for contamination of a manufacturing facility.

Normality is frequency is dependent upon the classification of the environment. Aclass 100, 10,000 and 100,000 environment will all be cleaned and disinfected withvarying frequencies. Coupled with the classification is the review of the environmentaltest data from each area. The use of the environmental test data to classify thefrequency of an area is superior to any other methodology. In understanding how toaccomplish this we must understand that if we had an area that showed very lowbioburden, if any, and invoked a daily sporicidal application, we would destroy nomore microorganisms than existed in the area. Once disinfectants and sporicides havedried their ability to kill is complete. It is true that some chemical agent residues createan undesirable environment for an organism to live, however, such residue kill is short-term and should not be a part of our system design. By applying a disinfectant and/or


sporicide, we do not create a preventative measure for organisms that may enter theenvironment. From the experience of this author, the following charts cultivate aguideline structure to base our disinfection and sporicidal frequencies.

In discerning frequencies, we need to first identify the type of areas that may existin our facility. The can be classified as:

• Class 100 stand-alone (gowning room to a Class 100)

• Class 100 surrounded by a Class 10,000

• Class 100 surrounded by Class 100,000

• Class 10,000

• Class 100,000

• Not Classified Areas that may effect classified areas


Table 6: Frequency of Disinfectant Application

Classification Walls Floors Ceilings Curtains

Class 100 Daily spray Daily mop Weekly spray Daily or per shiftor mop or mop application followed

by 70% IPA wipe

Class 100 Daily spray Daily mop Weekly spray Daily or per shiftsurrounded by or mop or mop application followedClass 10,000 by 70% IPA wipe

Class 100 Daily spray Daily mop Weekly spray Daily or per shiftsurrounded by or mop or mop application followedClass 100,000 by 70% IPA wipe

Class 10,000 Daily spray Daily mop Weekly spray If applicable, daily or or mop or mop per shift application

followedby 70% IPAwipe

Class 100,000 Weekly spray Daily mop Weekly spray If applicable, daily or or mop or mop per shift application

followedby 70% IPAwipe

Not classified Monthly or Weekly mop Monthly or Not applicablequarterly spray quarterly sprayor mop or mop

RESIDUES

With any application of a disinfecting agent comes the accompanying residueproblem. Only a few disinfectants do not leave a residue. Isopropyl alcohol and ethyl(ethanol) alcohol are two of these disinfectants. In fact, both isopropyl and ethylalcohol can assist in the removal of disinfectant and sporicidal residues. In review ofthe level of residues attained from each chemical agent we see that if not addressed ona routine basis, such residue can develop into a critical problem in our operations.


Table 7: Frequency of Sporicidal Application

Classification Walls Floors Ceilings Curtains

Class 100 Weekly or bi- Weekly or bi- Weekly or bi- Weekly or bi-weeklyweekly spray or weekly mop or weekly spray or spray or mop ormop or following EM mop or following EM atfollowing EM at at action following EM at action level. Followedaction level level action level by 70% IPA wipe

Class 100 Weekly or bi- Weekly or bi- Weekly or bi- Weekly or bi-weeklysurrounded by weekly spray or weekly mop or weekly spray or spray or mop orClass 10,000 mop or following EM mop or following EM at

following EM at at action following EM at action level. Followedaction level level action level by 70% IPA wipe

Class 100 Weekly or bi- Weekly or bi- Weekly or bi- Weekly or bi-weeklysurrounded by weekly spray or weekly mop or weekly spray or spray or mop orClass 100,000 mop or following EM mop or following EM at

following EM at at action following EM at action level. Followedaction level level action level by 70% IPA wipe

Class 10,000 Bi-weekly or Bi-weekly or Bi-weekly or If applicable, weeklymonthly spray monthly mop monthly spray spray or mop oror mop or or following or mop or following EM atfollowing EM at EM at action following EM at action level. Followedaction level level action level by 70% IPA wipe

Class 100,000 Monthly spray Monthly mop Monthly spray If applicable, weeklyor mop or or following or mop or spray or mop orfollowing EM at EM at action following EM at following EM ataction level level action level action level. Followed

by 70% IPA wipe

Not classified Upon Action EM Upon Action EM Upon Action EM Not applicabledetermination determination determinationof problem of problem of problem

Some problematic situations with residues relate to the complication of thesurface to be disinfected. The surface, if laden with chemical residues compromisesthe disinfectant and/or sporicide's ability to fully contact an organism. This is coupledwith the ability for air pockets to form underneath residues that may possibly breakand release contaminants to the environment. Most residues are unwanted. Personnelcan transfer them to a critical site. Such transfer may occur, as personnel would toucha surface and then make a critical aseptic connection. This may compromise theintegrity of the clean system in place.

Residues are most notably the cause of deterioration of surfaces in the clean room.Most notably is stainless steel. Deterioration occurs for two basic reasons:

• the chemical agent reacts adversely, or is incompatible with the surface andattacks the material composite material or impurities in the surface, or

• the chemical agent stains or discolors the surface

It is first important to conduct compatibility testing to assure the chemical agentdoes not harm the surface. Once this has been completed, routine cleaning can removeor reduce most residues.

DEVELOPING PROCEDURES AND TRAINING

Developing SOPs is critical in design and assuring correct implementation of oursystems. Likewise training our personnel on such procedures is critically important. Indeveloping the required applicable SOPs, a GMP facility needs to write a verycomplete “Use and Rotation” operating procedure. All other departmental operatingprocedures can then reference this particular operating procedure in their content.


Table 826

: Residues from Disinfectants

Chemical Tested When Applied After Rinse

High pH Phenol 759 ppm 61 ppm

Low pH Phenol 731 ppm 41 ppm

Quaternary Ammonium 133 ppm 11 ppm

Bleach @ 5.25% 929 ppm 66 ppm

Bleach @ 0.52% 144 ppm 14 ppm

Hydrogen Peroxide 0.067 ppm 0 ppm

Peracetic Acid/H202 RTU 123 ppm 16 ppm

Peracetic Acid/H202 Concentrate 44 ppm 6 ppm

Critical “Use and Rotation” SOP's should contain the following pertinent data. WhileSOP's will vary from firm to firm, the following is a general guideline list of theimportant areas.

Without the ability to prepare this critical standard operating procedure, wecannot assure personnel in varying departments will all follow a general companyscheme of disinfection. Standardizing disinfection plant-wide should be a mainobjective of the company. Only in specific instances where EM data warrants thealteration from the general master plan should such systems change in specific areas.Once we have established our SOPs, our next step is to assure that all is done to whathas been written. This is not an easy task and the main method of assuring this occursis through our training.

Training is without doubt one of the most critical elements in our spectrum ofdisinfection. Too often we train only our general workers and forget to train


Table 9: Standard Operating Procedures

Standard Operating Procedure DescriptionSection Title

Purpose The purpose of the SOP or why it is being done

Scope A scope of what is covered in the SOP

Responsibilities A listing of whom is responsible for each capacity in the SOP

Safety Precautions Internal corporate and product specific safetyprecautions for each product

Approved Disinfectants/Sporicides A listing of the approved disinfectants,and Cleaning Products sporicides, wipers, and saturated wipers

General Guidance in Preparing Specific guidance as to the general corporateDisinfectants and Sporicides issues surrounding the preparation of

disinfectants and sporicides such as approveddilution water types, dating of containers,storage, etc.

Specific Approved Disinfectant and Specific approved usage instructions for eachSporicide Usages disinfectant and sporicide. Including dilution

mixtures, container types, use instructions,expiration dating, storage of unopenedcontainers, storage of opened containers

Method of Application How are the disinfectants to be applied invarying classifications and on varying surfaces

Frequency Frequency of application of disinfectants andsporicides

Approved Equipment and Materials Approved mop heads, mop handles, mop frames,buckets, sprayers and foggers

Procedure How each specific are is to be cleaned and disinfected

management. An unknowledgeable management person in the area of disinfection canharm our efforts. They may lack the skills to train or supervise this important function.At the same time, they may lack the ability to notice a shift away from acceptability.It is important to train all involved in the disinfection process on a routine basis.

But “what is training and what content should be included” is a commonly askedquestion. First, people learn by doing. People learn very little in a classroom. Theirattention span is short within a classroom setting and "learning by doing" is a farsuperior method.

Second, know what you teach to be correct. The worst thing that can happen to anorganization is that the trainees learn and prove that the methods in place are notscientifically justified or flawed. Trust then becomes a concern.

Third, sometimes inside trainers become boring to trainees. An outside source oftraining may provide a fresh approach that one's employees may enjoy. It breaks upthe daily monotony.

Fourth, training must place in the foreground what is expected and if notaccomplished, what the organization will be required to do to assure the function'scompletion. People have to know what is expected.

And last, there are countless year 2000 techniques available to the trainer of today.Utilization of past training methods should be reevaluated to see if implementation ofa better methodology exists.

A FINAL SUMMARY AND CONCLUSION

Too often we just want to kill things. We forget the multitude of critical areas that needto be addressed to assure the continued and consistent attainment of acceptableenvironmental conditions in our controlled areas. If we wait for regulatory agencies tocomment before we change or if we wait for an excursion to exist before assuring acomplete system, we place ourselves into the situation of “hurried qualityimplementation”. This will cause us to fail. Hysteria during a crisis situation will causeus to place a band aide on the situation and not create a strong infrastructure that willassure our success for years to come. We will forget many critical aspects. Decisionsfor design are always made better without emotional turmoil.

As an industry we need to review the required time to adequately clean anddisinfect manufacturing areas. Too often time period are too short to complete this taskand shortcuts may be taken in order to run product sooner. However, when the productis in jeopardy as environmental levels have exceeded prescribed action levels, the first


scenario that is blamed is cleaning and disinfection. It must not have been donecorrectly. On the same note cleaning and disinfection is also the last word as too manytimes we use it as the corrective action procedure instead of delving more deeply intothe root cause.

We must also remember that the painting we have created will not gain value overtime as is common for artistic impressions. Each day our design becomes outdated andwe need to routinely reevaluate our systems to assure they include updates for processchanges, changes in technology and changes in our personnel structure.

BIBLIOGRAPHY

Vellutato, A. L., Utilizing environmental monitoring data to implement a cleaning anddisinfection program. CleanRooms Magazine, January 2001, p. S10–S20.

Center for Drugs and Biologics and Office of Regulatory Affairs, Food and DrugAdministration, Guidelines on Sterile Drug Products produced by AsepticProcessing, June 1987, p. 9.

Center for Drugs and Biologics and Office of Regulatory Affairs, Food and DrugAdministration, Sterile Drug Products Produced by Aseptic Processing Draft,Concept paper (Not for Implementation), September 27, 2002.

PDA TR #13

United States Pharmacopoeia, The National Formulary, USP 25 NF 20, Chapter 1116,United States Pharmacopeial Convention, Inc., National Publishing, Philadelphia,1999

United States Pharmacopoeia, The National Formulary, USP 25 NF 20, Chapter 1072,United States Pharmacopeial Convention, Inc., National Publishing, Philadelphia,1999

ISO 14644

ISO 14698

A. Vellutato Sr. and A. Vellutato, Jr., 1989. The Use of Cleaners and Disinfectants inGMP Controlled Environments, Technical update Notice Monthly. VeltekAssociates, Inc. p. 1–14

Official Methods of Analysis (1998) 16th Ed., 5th Revision, 1998, AOAC


INTERNATIONAL, Gaithersburg, MD

McDonnell, G. and R. Denver. Antiseptics and Disinfectants: Activity, Action, andResistance, Clinical Microbiological Reviews, Jan 1999, p. 151.

McDonnell, G. and R. Denver. Antiseptics and Disinfectants: Activity, Action, andResistance, Clinical Microbiological Reviews, Jan 1999, p. 151 (545)

McDonnell, G. and R. Denver. Antiseptics and Disinfectants: Activity, Action, andResistance, Clinical Microbiological Reviews, Jan 1999, p. 151 (513)

Chlorine and Chlorine Compounds. In Disinfection,, Sterilization and Preservation,4th Edition, edited by S.S. Block. Philadelphia: Lea and Febiger,

Peroxygen Compounds. In Disinfection, Sterilization and Preservation, 4th Edition,edited by S.S. Block. Philadelphia: Lea and Febiger,

McDonnell, G. and R. Denver. Antiseptics and Disinfectants: Activity, Action, andResistance, Clinical Microbiological Reviews, Jan 1999, Jan 1999, p. 156

Gluteraldehyde. In Disinfection, Sterilization and Preservation, 4th Edition, edited byS.S. Block. Philadelphia: Lea and Febiger.

Formaldehyde. In Disinfection, Sterilization and Preservation, 4th Edition, edited byS.S. Block. Philadelphia: Lea and Febiger.

Casey, W.M., Comment, The Pharmaceutical Microbiology Mail List, Rotation ofDisinfectants, Nov 17, 2000.

Connor, D.E. and M.K. Eckman, Rotation of Phenolic Disinfectants, PharmaceuticalTechnology, September 1992, 148–158.

European Committee for Standardization, Chemical Disinfectants and Antisepics –Quanitative non-porous surface test for the evaluation of bactericidal and/orfungicidal activity of chemical disinfectants used in food, industrial, domestic andinstitutional areas – Test methods and requirements without mechanical action(phase 2/step2), EN 13697 (2001) E, ICS 11.080.20; 71.100.35 Managementcenter: ue de Stassart 36, B-1050 Brussels .

Brady, C. S., A.T. Snyder and B.L. Baskin, 2001. Disinfectants Efficacy Testing. InMicrobiology in Pharmaceutical Manufacturing, edited by R.Prince. Godalming:Davis Horwood International Publishing.

Vellutato, Arthur, Sr., United States Patent 6, 123,900, Untites States patent Office,


1992

Vellutato, Arthur, Jr., Validation of the Core2Clean Spray Mop Fog Systems, InternalValidation Report, Veltek Associates, Inc. January 2000

Vellutato, Arthur, Jr., Validation of the Core2Clean Fog System, Internal ValidationReport, Veltek Associates, Inc. April 2000

Code of Federal Regulations. 1992. Title 21, Part 211, Good Manufacturing Practicesfor Finished Pharmaceuticals, 160, Washington, DC: US Government PrintingOffice

ADDITIONAL READING

Proceedings of the Second International Kilmer Memorial Conference on theSterilization of Medical Products, Washington, DC, 1980.

Sterilization of healthcare products – Requirements for validation and routine control– Radiation sterilization, ANSI/AAMI/ISO 11137–1994, Association for theAdvancement of Medical Instrumentation, Arlingto, VA, 1995.

Veltek Associates, Inc., DECON-AHOL Sterile Spray Validation Report, ValidationReport VL-101and VL 101B, Exton, PA, 1989.

Veltek Associates, Inc., DECON-AHOL WFI Sterile Spray Validation Report,Validation Report VL-201 and VL201B, Exton, PA, 1991.

Veltek Associates, Inc., Sterile Disinfectant and Sporicide Validation Reports,Validation Reports VL-301, VL401, VL-501, VL601, VL-701, VL-801, VL901,VL1011, VL2000, VL2001, Phoenixville, PA, 1993

ABOUT THE AUTHOR

Art Vellutato, Jr. is the vice-president of Technical Support Operations, vice-presidentof Sales & Marketing and one of the two founders of Veltek Associates, Inc. (an EPAand FDA registered facility) in 1981.

He is a frequent industry speaker with 21 industry publications. He lends over 18years of valuable experience that include his tenure as the Director of QualityAssurance at VAI for nine years and the Director of manufacturing for six years.


Art Vellutato, Jr. co-owns US Patent 6,123,900, Method of Sterilization, that wasused to market the first sterile disinfectant in the industry, DECON-AHOL®. He hasalso conducted disinfectant training for FDA's CDER and CBER divisions.

He is the President of the Delaware Valley Chapter of the Parenteral DrugAssociation and a faculty member teaching the Disinfection segment at PDA'sTraining and Research Institute for the Aseptic Processing Course. Art Vellutato, Jr. isalso the chairman of the PDA Special Interest Group on Cleaning and Disinfection(PDA Delaware Valley Chapter)

Combining experience of production, quality assurance and validation, ArtVellutato, Jr. has assisted many pharmaceutical manufacturing operations inimplementing their environmental monitoring and disinfection programs, worldwide.


Documents

Designing_Disinfection_Systems_Chapt_8