AN AUDIT APPROACH

TO ADDRESS MICROBIAL
CONTAMINATION IN
PROCESS EQUIPMENT
Paul Lopolito and Elizabeth Rivera

Reprinted from Contamination Control in Healthcare Product Manufacturing

Copyright © 2013, co-published by PDA and DHI. All rights reserved.
420-600-0056
 15

AN AUDIT APPROACH TO
ADDRESS MICROBIAL CONTAMINATION
IN PROCESS EQUIPMENT

Paul Lopolito and Elizabeth Rivera

STERIS Corporation
Mentor, OH
USA

INTRODUCTION
Regulatory agencies require that drug manufacturing processes be
designed to ensure that end-products are safe for the user.
Responsibility lies with the manufacturer to evaluate the hazards of
drug preparations per the intended use, the bioburden that the
product may contain, and the suitability of testing to ensure a safe
product. The emphasis of this regulatory expectation is to
investigate, identify and correct contamination events that may
compromise product quality, safety and efficacy, as well as to
establish controls that prevent future contamination.

“21 CFR 211.113 Control of Microbiological Contamination

(a) Appropriate written procedures, designed to prevent
objectionable microorganisms in drug products not required to
be sterile, shall be established and followed.

(b) Appropriate written procedures, designed to prevent

microbiological contamination of drug products purporting to be

323
324 Contamination Control in Healthcare Product Manufacturing
sterile, shall be established and followed. Such procedures shall
include validation of all aseptic and sterilization processes.”

The above statements indicate that, in the eyes of the United States
Food and Drug Administration (US FDA), microbial contamination
is a concern for both sterile and non-sterile drug products. This
concern may also be extended to other regulated industries and
international regulatory agencies. For example, the statements
below may imply that a substance that threatens end-user safety
may include microbial contamination.

21 CFR 111.365 What precautions must you take to prevent

contamination?
“You must take all the necessary precautions during the manufacture
of a dietary supplement to prevent contamination of components or
dietary supplements. These precautions include: (a) Performing
manufacturing operations under conditions and controls that protect
against the potential for growth of microorganisms and the potential
for contamination;”

21 CFR 820.70 Production and Process Control

“(e) Contamination control. Each manufacturer shall establish and
maintain procedures to prevent contamination of equipment or
product by substances that could reasonably be expected to have
an adverse effect on product quality.”

21 U.S.C. 361 — Adulterated Cosmetics

A cosmetic shall be deemed to be adulterated ...
(a) If it bears or contains any poisonous or deleterious substance
which may render it injurious to users under the conditions of
use prescribed in the labeling thereof, or under such conditions of
use as are customary or usual, ...”

Microbial contamination in the pharmaceutical, biopharmaceutical,

medical device, cosmetic, and dietary supplement industries is
recognized as a source of risk to the consumer. The presence of
objectionable microorganisms in non-sterile products, or any type
of microorganism in sterile products, denotes inadequate process
controls. The control of microbial contaminants in Good
Manufacturing Practice (GMP)-regulated facilities requires the
 An Audit Approach to Contamination in Process Equipment 325

interaction of a multidisciplinary group that includes, at a

minimum, microbiology, engineering, operations, and quality
assurance, to identify root causes and to implement corrective
actions that prevent reoccurrence.

To better assess microbial control issues in processing

equipment, it is necessary to have a plan that allows the evaluation
of various factors that can contribute to the problem. This chapter
assembles important information to assist in this investigational
process and describes a holistic approach to remediate microbial
contamination in process equipment. An auditing model is used to
help this reader develop a new plan or improve current methods for
addressing microbial contamination events.

PART 1: PRE-INSPECTION WORK

Pre-inspection work is vital to a successful internal or external audit.
Information assembled and delivered in advance of the audit allows
the auditor to ask more relevant questions and gain a better
understanding of the manufacturing process and contamination
control practices. Information such as the identification of micro-
organism (genus and species), history of microorganism on site,
process, personnel and waste flow diagrams, equipment drawings,
equipment cleaning procedures, and sanitization practices should be
reviewed prior to the audit. This information provides the auditor
with an idea of where to focus attention during the inspection. The
ultimate goal of an auditor is to focus on plausible root causes and
provide input for the corrective and preventative action plan by the
end of the audit. For the purpose of this chapter a person
investigating the cause(s) of microbial contamination is called an
auditor.

Different types of microbial contamination

Microbial contamination can include Gram-negative bacteria from
process water, Gram-positive bacteria from personnel, or spore-
forming microbes from raw materials. Bacterial spores are a concern
due to their high resistance to destruction (McDonnell, 2007).
Gram-negative bacteria are often associated with biofilm in process
326 Contamination Control in Healthcare Product Manufacturing

systems or utilities and they are more resistant to destruction when

in a biofilm. Gram-negative microbes also contain endotoxins in the
cell membrane, which can build up on equipment surfaces and
eventually lead to product failure. This is a particular concern for
medical implants, parenterals and ophthalmic products.

Case Study 1: Endotoxin levels related to process change

A small biomedical device company scaled up its raw material protein
extraction vessels from two-liter high density polyethylene (HDPE) roller
bottles to 200-liter HDPE top-mixed bioreactor vessels. This scale-up
reduced manufacturing process time and increased protein yield. However,
the endotoxin level in the final product was gradually increasing from batch
to batch. A review of the manufacturing process change revealed that the
cleaning procedure changed from automated washer cleaning using an
alkaline detergent and hot water rinses before steam sterilization, to a
manual cleaning step with an alkaline cleaning agent and warm water rinses
before steam sterilization. High endotoxin levels were detected on the
surfaces of the bioreactor and the company implemented a minimum
overnight alkaline cleaning agent soak of all tanks and processing aids used
in the manufacturing process. The endotoxin levels in the final product
immediately returned to that of baseline values after the cleaning process
was adjusted to address endotoxin removal.

The presence of biofilm or endotoxins often requires a modification

of the cleaning procedure to effectively remove the residue
(extracellular polymeric substances or lipid polysaccharides)
associated with microbial contamination. Mycobacteria,
mycoplasma, viruses and prions from animal derived materials, as
well as fungi (mold and yeast) from the environment or raw
materials, can also contaminate products. Microbial contamination
can be introduced from raw materials including active and non-
active components, personnel, the environment, utilities, product,
equipment surfaces and packaging materials. Procedures and
systems should be in place to reduce the risk of exposure of the
product to direct or indirect contact with microbes.

Importance of microbial identification

Understanding the microbial species and genus of the
contamination allows the auditor to investigate intrinsic or extrinsic
 An Audit Approach to Contamination in Process Equipment 327

sources. Intrinsic sources are those integral to the manufacturing

process or equipment. Examples of intrinsic microbial
contamination would be mycoplasma or viral contamination from
bovine serum albumin or Gram-negative bacterial contamination
associated with a biofilm in the process equipment or water system.
Case Study 2 is an example of intrinsic contamination.

Case Study 2: Intrinsic source of microbial contamination

An oral solid dose manufacturer was performing a continuous coating
process of a single product over six days using a high viscosity coating.
During manufacturing the firm would routinely sample the coating for
bioburden, and throughout the six-day coating process it remained below its
microbial alert and action levels. However, when the firm changed to a new
low-viscosity coating, the in-process microbial counts exceeded the alert
levels with Pseudomonas aeruginosa by day two or three of a six-day coating
process.When it returned to the high viscosity coating there were no issues
in running the process. In this case, Pseudomonas aeruginosa was part of the
native flora in the coater and in the low viscosity coating the microbe was
able to proliferate, resulting in out-of-specification results, shorter
production runs, discarded coating solution, wasted coated product, and lost
revenue. The corrective action was a thorough cleaning of the coating
equipment with an alkaline cleaning agent, and an increase in the
concentration of the quaternary ammonium sanitizer to “broad spectrum
disinfectant” concentration, validated as effective against Gram-negative
bacteria such as Pseudomonas aeruginosa.

Extrinsic sources are those associated with a transfer of microbial

contamination into the product via air, liquid or surface contact.
Examples of extrinsic microbial contamination are Gram-positive
bacterial contamination resulting from poor gowning practices or
fungal spore contamination from open process containers. Case
Study 3 is an example of extrinsic contamination.

Case Study 3: Extrinsic source of microbial contamination

A non active ingredient used in pharmaceutical, biopharmaceutical, dietary
supplement, personal care and cosmetic products had high levels of a Gram-
positive bacteria. The identified microbial contamination was Bacillus
halodurans, a spore-forming microbe that prefers alkaline growth conditions
and is commonly found in soil. The manufacturing process consisted of a
blending step, storage, nanofiltration and distillation. The raw materials
and manufacturing process intermediates are easy to clean. Inspection of the
328 Contamination Control in Healthcare Product Manufacturing
manufacturing process and material traffic flow revealed that there was a
high probability of introducing soil microbes from raw material received on
the property. The traffic flow of forklifts with incoming raw materials and
intermediate products was in close proximity to open process containers
used for production. The level of detection was low at time of release due
to the just-in-time manufacturing process.The product’s pH was around 9.0.
The high pH condition supported growth of the Bacillus halodurans during
shipping and storage. The corrective actions included the use of closed
containers for storage of in-process and finished product.

Historical information regarding prior microbial contamination can

also be helpful in understanding whether the contamination is a
single incident or an indication of insufficient practices or controls.

Understanding historical significance and microbial

resistance
Identification of the microbial contamination and prior history
provides valuable information related to the source of the microbial
contamination, conditions for optimum growth of the microbe, and
relative resistance to destruction. The relative resistance to destruct-
ion of the microbe is critical because if it is not removed or killed as
part of the corrective action plan, it can cause future problems.
Prions and bacterial spores are the most resistant microorganisms
(Figure 15.1). They would require sterilization cycles or high level
disinfection to effectively inactivate or destroy the microorganisms.
For organisms with less resistance, a broad spectrum disinfectant or
sanitizer may be effective at destroying them.

All hard-surface germicidal cleaners in the United States fall

under the Federal Insecticide, Fungicide and Rodenticide Act
(FIFRA) as amended in 1988 and administered by the
Environmental Protection Agency (EPA). Exceptions to this are
germicides used to reprocess medical devices, or used as a sterilant
for medical devices, which are regulated by the FDA per the Food
Quality Protection Act of 1996. These two agencies govern
germicidal product safety, use, disposal and efficacy through
clearance of each product label. Therefore, it is always important to
review each germicidal product label for efficacy claims before use.
 An Audit Approach to Contamination in Process Equipment 329
Figure 15.1 Hierarchy of microorganisms resistance

Microorganism Examples

More
Resistant Prions Creutzfield-Jacob Disease (CJD), Bovine
Spongiform Encephalopathy (BSE),
Chronic Wasting Disease (CWD), Scrapie

Bacterial spores Bacillus, Geobacillus, Clostridium, Paenibacillus

Mycobacteria Mycobacterium tuberculosis, M. bovis, M. terrae,

M. chelonea

Small, non-enveloped Poliovirus, Parvovirus

viruses

Fungal spores Aspergillus, Penicillium, Stachybotrys

Mycoplasma Acholeplasma laidlawi, Mycoplasma

gallisepticum, M. pneumoniae

Gram-negative bacteria Pseudomonas, Salmonella, Burkholderia,

Escherichia

Vegetative fungi Apergillus, Trichophyton, Candida

and algae

Large, non-enveloped Adenovirus, Rotaviruses

viruses

Gram-positive bacteria Staphylococcus, Streptococcus,

Enterococcus, Actinobacteria,
Corynebacterium

Less Enveloped viruses Influenza A Virus, HIV, Hepatitis B Virus

Resistant

(McDonnell, 2007)
330 Contamination Control in Healthcare Product Manufacturing

Material flow diagrams and equipment drawings

Diagrams displaying how product, personnel and waste flow
through the facility provide useful information for understanding
the manufacturing process and contamination control practices
already in place. With an understanding of how materials flow
within the process and the identification of microorganism(s)
present, one can assess potential microbial risks from intrinsic and
extrinsic sources prior to or during the inspection. Equipment
drawings can help identify areas exposed to microbial
contamination from extrinsic sources, as well as areas that would be
difficult to clean and disinfect.

Cleaning process design control

Cleaning procedures are designed to remove process residues. If
cleaning procedures are not robust enough, then process residues can
affect sanitization and sterilization practices. If the cleaning
procedure is not validated or designed to remove the specific process
residue, then this should be investigated through laboratory studies
in parallel with the microbial contamination remediation. A robust
cleaning procedure is critical to ensure successful sanitization and
sterilization procedures and to reduce equipment maintenance.
When microbial contamination is present, cleaning procedures need
to be robust enough to clean the microbial residue as well as the
process residue. In Case Study 3, a laboratory cleaning evaluation
was performed prior to a site audit. The laboratory study revealed
that water at ambient temperature was effective at removing the
process residue. Therefore their current cleaning procedure with an
alkaline cleaning agent and hot water rinses should be effective at
removing the process and microbial residue. This information, along
with identification of the microbe as native to soil, allowed the
auditor to focus on personnel and environmental contamination
control practices as the root cause.
 An Audit Approach to Contamination in Process Equipment 331

PART 2: INSPECTION
Time is critical during a microbial contamination investigation, and
an internal or external auditor is a valuable resource for identifying
plausible root causes and the formulation of a corrective and
preventative action plan. The auditor provides an experienced eye
and has advanced knowledge of critical factors including
microorganism identification, historical information, material flow
diagrams, equipment drawings, and cleaning procedures, which
can help reduce the scope of the investigation. A reduction in scope
saves both internal resources and time in correcting the issues and
getting back to manufacturing product.

The inspection process should consider potential causes of

microbial contamination and focus on the areas of high risk in
developing an interim corrective action plan to immediately
address the issue. These measures should be followed by
development of a long-term plan to reduce the likelihood of
another microbial contamination. Potential sources of microbial
contamination would be raw materials, packaging materials,
utilities, room environmental factors, process, personnel and waste
flow, cleanability and cleaning of equipment surfaces including
sterilization or disinfection practices. Figure 15.2 summarizes the
aforementioned sources and suggested controls.

A formal risk assessment is recommended by International

Organization of Standardization (ISO) 14698-1 (ISO, 2003), utilizing a
system for identifying and managing risk such as Fault Tree Analysis
(FTA), Hazard Analysis and Critical Control Point (HACCP), or
Failure Modes and Effect Analysis (FMEA) during the product
design phase and change control process. The FMEA risk assessment
method would calculate the microbial risk (priority number) by
multiplying the probability of contamination times the severity times
the likelihood of detection, and can be a valuable tool in assessing the
manufacturing process or change (Whyte, 2004). Cause and effect
diagrams for potential causes of contaminated products, such as the
fishbone and Ishikawa–6M, can also be used to investigate microbial
contamination (Sutton, 2012). If this has not been performed in
advance, then the inspection process can sort observations as low,
medium and high risk. For example, an open final product storage
container, as noted in Case Study 3, would be a high risk item.
332 Contamination Control in Healthcare Product Manufacturing
Figure 15.2 Potential sources of microbial contamination

How can microorganisms

get into finished products? Suggested controls

Raw materials • Establish microbial limits for incoming raw material

or processing aids
• Sterilize, filter or pasteurize raw materials on site
• Filter processing aids on site (e.g., cleaning agents,
sanitizers, etc.)

Packaging components • Establish microbial limits for packaging components

• Sterilize or disinfect packaging components on site

Personnel • Train personnel in good hygiene and behaviour

practices
• Train personnel in gowning procedures

Room environment • Establish housekeeping and cleanroom procedures

as applicable
• Control room humidity, temperature, air changes
and particulates
• Establish suitable traffic flow and material
segregation

Utilities • Qualify and maintain water generation system,

compressed air, nitrogen and any product contact
utility system

Equipment • Establish robust cleaning procedures

• Address equipment design issues
• Establish disinfection or sterilization parameters
as applicable

Raw materials, processing aids and packaging

components
Raw materials should be purchased from approved suppliers and a
microbial risk assessment of each raw material from each supplier
performed. Raw materials can be classified as synthetic, semi-
synthetic, plant derived, animal derived, and mineral derived
(Cundell, 2005). If microorganism contamination is a concern, the
raw material lots should be tested prior to use. Microbial reduction
 An Audit Approach to Contamination in Process Equipment 333

of the raw material through pasteurization, filtration or sterilization

can be performed before use, if warranted from the risk assessment.
Raw materials from plant or animal origins pose the greatest risk of
microorganism contamination. Liquid samples are generally a
greater concern than pastes or powders due to water activity levels.
A neutral pH raw material is a greater risk than acidic or basic
solution. All recent changes in raw materials or product
formulations (refer to Case Study 4) should be reviewed as part of
the microbial contamination investigation, and if needed, a new
microbial risk assessment performed.

Case Study 4: Change in formulation

A consumer product manufacturer revised its product formulations to
market more environmentally friendly, “greener” products. These new
product formulations consisted of more biodegradable components and
eco-friendly — but less effective — preservatives. After the new
formulations were implemented in production, continual product failures
due to microbial contamination resulted. The blending, storage and filling
equipment was not appropriately designed for the higher microbial risk
represented by the new formulations, and engineering, cleaning, and
sanitization procedures needed to be revised to reduce the finished
product’s microorganism level.

Raw material storage, sampling and transfer processes within a

manufacturing facility should also be reviewed. Raw materials
should be stored in closed containers under controlled conditions,
and expiration dates established for closed and open containers.
Raw materials that are lyophilized or frozen can be considered a
lower risk. Raw material sampling and dispensing activities should
be reviewed for steps that have the potential to introduce microbial
contamination. Any processing aids that are in direct or indirect
contact with the product during the manufacturing process should
also be assessed for potential microbial risk. Processing aids
include, but are not limited to, lubricants, antifoams, alcohols,
cleaners and germicidal agents. Antimicrobial Effectiveness Testing,
USP <51>, can be performed to assess potential microbial risk of
raw materials and processing aids used in the manufacturing
process. In addition, the bioburden of packaging components
should be assessed, and if needed, they should be cleaned and
depyrogenated and/or sterilized before use.
334 Contamination Control in Healthcare Product Manufacturing

Personnel and room environment controls

Personnel working in the manufacturing process area are the
largest contributors of viable and non-viable particles. A human
adult has approximately 1012 bacteria on the skin, 1010 bacteria in
the mouth and 1014 bacteria in the gastrointestinal tract (Todar,
1997; Martínez, 2002). In 2007, Whyte and Hejab tested the airborne
dispersion of particles from 55 people. The dispersion rate per
minute of microbe-containing particles was 2400 when wearing
indoor clothing and 177 when wearing cleanroom garments (Whyte
and Hejab , 2007). For this reason, access control, personnel attire
and gowning procedures and personnel practices are important
factors to evaluate, to reduce the likelihood of introducing viable
and non-viable particles from personnel. Access should be limited
to trained personnel, and procedures should be in place to allow
access to visitors, service personnel, auditors and inspectors.
Gowning attire should be selected based on the classification of the
area with a separate area for gowning. Personnel should be trained
in practices and attire that are not appropriate for the
manufacturing areas. For example, policies can be established
regarding jewelry, makeup, smoking, skin rashes, sunburns, eating,
drinking, excessive movement and hygiene. Refer to the Institute of
Environmental Sciences and Technology, Personnel Practices and
Procedures in Cleanrooms and Scottish Society for Contamination
Control, Cleanroom Disciplines and Entry for detailed examples of
acceptable and non-acceptable behavior when working in
cleanrooms and controlled environments (IEST, 2006).

Other environmental controls include air quality, process

equipment, and cleaning and disinfection practices. The Heating,
Ventilation and Air Conditioning (HVAC) system is used to
maintain the room temperature, relative humidity, air flow and
differential pressure within the production areas. Typical settings
for relative humidity and temperature are between 30–60% and
17–23°C, respectively, for operator comfort; however, some
products might require warmer or cooler processing temperatures.
Warmer environments favor bacteria and yeast growth, while
cooler temperatures favor mold growth. The air exchanges per
hour, and air filtration through High-Efficiency Particulate Air
(HEPA) and ultra low penetration air (ULPA) filters, is dependent
 An Audit Approach to Contamination in Process Equipment 335

on the room classification. An ISO-5, Grade A area used for aseptic

manufacturing processes would have about 300 air exchanges per
hour, 100% HEPA filtration, a unidirectional air flow of about
90–100 feet per minute, an air makeup of about 75% re-circulated
and 25% fresh, and differential pressure between interior and
exterior rooms of 12.5 Pascals. Smoke studies during operational
conditions are an ideal way to evaluate product risk from airborne
particulates. These should be repeated on a regular basis or when
changes are made that could place the product at increased risk.

Process equipment should be selected based on functionality,

low particle generation, cleanability, and compatibility with cleaning
and disinfecting chemistries. The entire facility — walls, ceiling,
floors, grills, etc. — should be constructed of durable, washable, low
particle generating materials and compatible with cleaning and
disinfecting chemistries. The facility cleaning and disinfection
practices should be developed to address removal of gross process
soil, routine disinfection and sanitization of hard nonporous
surfaces, periodic sporicide application and disinfectant/sporicide
residue removal. Preparation, application and storage of containers
in-use of the sanitizer, disinfectant and sporicide should be defined.
The disinfectant and sporicide selection process should include a
detailed review of vendor literature and in-house testing
demonstrating microbial efficacy against native isolates on
representative surfaces (Bartnett et al., 2007; USP <1072>). Critical
parameters that can adversely affect microbial efficacy of germicides
are the concentration, temperature, exposure time, surface,
bioburden, process soil level and water quality (hardness) used in
preparing and applying the product. These critical parameters
should be defined in the cleaning and disinfection procedures.
When possible, disposable containers and application equipment
should be used. If this is not possible, cleaning and storage
procedures need to be established and validated.

Validation activities supporting environmental controls include

the HVAC system, facility validation, cleaning and disinfection
validation, and quality control environmental monitoring
procedures. Environmental monitoring procedures should include
testing methods, equipment procedures and sampling sites.
Environmental monitoring trends and recent excursions should be
336 Contamination Control in Healthcare Product Manufacturing

reviewed periodically, and adjustments made to the environmental

monitoring controls as needed.

Utilities (water, steam, compressed air and gases)

Any utilities used in direct contact with the product should be
inspected, and routine monitoring test results and contamination
control practices reviewed. Utilities commonly used include water,
steam, compressed air, and compressed gases such as helium or
nitrogen. For water and steam systems, this might include routine
microbial, Total Organic Carbon (TOC) and endotoxin testing. The
routine microbial test results (including identification) should
be reviewed as a possible source. Filters and ultraviolet (UV) bulbs
should be changed per manufacturer recommendations or more
frequently if operating conditions suggest it. TOC data should be
reviewed for any indication of upward trending before a microbial
contamination event, and prior to cleaning and sanitization activities.
Common bioburden reduction steps for water systems include
distillation, filtration and/or UV treatment to reduce micro-
organisms. The recent maintenance activities of the water system
should also be reviewed. If non-routine services were performed, a
microbial risk assessment should be performed and the services
scrutinized. The system piping, gaskets, valves and material surfaces
should be inspected for possible wear and visible residue. Routine
cleaning, derouging, passivation and sanitization practices should
also be reviewed. This may include an alkaline cleaning agent to
remove organic fouling, an acid cleaning agent to remove inorganic
fouling, and a sanitizing solution. Typical sanitizing solutions include
an oxidizing chemistry, such as sodium hypochlorite, ozone,
hydrogen peroxide or hydrogen peroxide/peracetic acid blends. The
concentration, temperature and time of the sanitizing solution should
be reviewed for its efficacy against the identified microorganism.
Some water systems may also have the capacity to increase the
temperature to 60–80°C or introduce steam to control microbial
contamination. In this case, steam traps and relief valves should be
inspected to ensure that they are in proper working order.

Compressed air and gases should be filtered prior to use to

prevent viable and non-viable particles, oil and condensation from
 An Audit Approach to Contamination in Process Equipment 337

entering the process system. Filters should be properly selected and

replaced on a preventative maintenance schedule. Non product
contact utilities, such as cooling or heating solutions, should be
inspected for possible incidents of cross-contamination with
product or product contact surfaces.

Walk-through of process and traffic flow of personnel

and waste
It is important to walk through the manufacturing process and
observe how each raw material, utility, and processing aid is
introduced or exposed to the product. From this walk-through the
auditor can assess possible intrinsic and extrinsic microbial risk and
verify in-place controls. The auditor can also integrate various
traffic flow patterns within the facility into the flow of the product.
Personnel from quality assurance, quality control, engineering and
operations should accompany the auditor to address questions or
discuss areas of microbial risk. Unless there is a safety risk to
personnel or product sampling of surfaces for process and
microbial residues and the taking of photographs should not be
discouraged. Photographs and additional sampling data outside of
routine environmental and product samples can be helpful for
getting support for changes to contamination control practices.

Processing equipment and elements affecting microbial

decontamination
The major elements that must be considered when addressing
microbial contamination in processing equipment are represented
in Figure 15.3 and are discussed in detail. It is important to notice
that these elements are linked by two-way arrows. These arrows
indicate the interaction of one element with the other, and they
should not be treated independently. Deficiencies found in one
element may have a significant impact on the other, and vice versa.
For example, during an investigation it is found that a certain
process residue is causing the microbial contamination because it is
not being removed from surfaces. While it is possible that the
cleaning procedure might not be robust enough for cleaning the
338 Contamination Control in Healthcare Product Manufacturing

residue, other factors might also be contributing to cleanliness

difficulties, including the type of surface to be cleaned.
Consequently, when a surface is not properly cleaned the current
sanitization parameters may not work successfully.

Figure 15.3 Elements affecting microbial control and

decontamination of processing equipment

Residue type and cleaning parameters

The cleaning of processing equipment is an essential part of all GMP
regulated manufacturing. The GMP regulations recognize process
cleaning to be a critical step in ensuring drug product quality,
efficacy, and safety (FDA, 1993). Usually, validation of cleaning
processes is mandatory in most countries for product contact surfaces
and equipment that could potentially contaminate drug products. To
date, there are many cleaning processes that are not validated. There
are others that lack scientific technical data and studies to support
full validation of the cleaning parameters being used.

Many cleaning agents specifically designed for GMP cleaning

applications are available (Verghese, 1998). Some cleaning agents
 An Audit Approach to Contamination in Process Equipment 339

may be effective against multiple residues, or more than one may be

necessary to clean one particular residue. Cleaning agents can be
categorized as commodity chemicals, organic solvents and
formulated detergents. Each type of cleaning agent may employ
one or more cleaning mechanisms, such as solubility, wetting,
emulsification, suspension, and more (Verghese and Kaiser, 2009).
In addition, cleaning agents are meant to be used at different
concentrations, temperatures, and contact times depending upon
the residues’ characteristics at the time of cleaning, the cleaning
method being used, and surface properties.

With this in mind, the evaluation of the right cleaning agent

and parameters should be determined through cleaning studies.
Laboratory scale evaluations are often used to provide guidance in
selecting the optimal cleaning parameters for the residues of
interest. Although this has been explained in multiple publications,
its significance for microbial control has not been fully discussed.
This lack of evaluation and understanding has led to microbial
contamination caused by inadequate cleaning in some
manufacturing operations. Case Study 5 below is an example of a
company that reported issues with a Gram-negative, biofilm
forming Burkholderia cepacia contamination, where the current
cleaning procedure was found to be ineffective at removing the
process residue.

Case Study 5: Inefficient cleaning procedure

A personal care product manufacturer of multiple products ranging from
shampoos, body washes, and lotions to topical products had repeated cases
of Burkholderia cepacia contamination in its finished products. Their current
cleaning practices included washing with a surfactant, followed by application
of a quaternary ammonium sanitizer. Personal care products typically contain
polymers, such as carbomers, which can be difficult to clean (Hadziselimovic
and Lopolito, 2012). Laboratory testing was performed and demonstrated
that the cleaning procedure left trace residues on the surface and that an
alkaline cleaning agent plus a detergent additive containing hydrogen
peroxide was effective at removing the process residue. The source of the
microbial contamination was an antifoam agent. Because of inadequate
cleaning, the microorganism was able to survive sanitization, and this led to
selective product failures.
340 Contamination Control in Healthcare Product Manufacturing

A successful cleaning process is necessary for microbial control.

Chemical residues left behind can provide a nutrient source for
microorganisms, especially when the raw materials are animal- or
plant-derived. Chemical residues can also serve as a physical trap in
which microorganisms hide and survive, even in the presence of
chemical sanitizers. Indeed, many biocidal agents (certain sterilants,
for example) require a clean, soil-free surface before application, or
their microbial efficacy is reduced. In theory, a good robust cleaning
procedure must achieve not only the removal of the chemical
residues, but also the effective control of microorganisms. In some
cases, a separate sanitization step might be necessary.

Some companies buy a cleaning agent and apply the general

“use directions” as described on the product label, data sheet, or
perhaps as suggested by the vendor, without additional laboratory
evaluation. This approach may work in some manual cleaning
applications, assuming the residues are relatively easy to clean, but
has its limitations because it does not provide knowledge on how
the cleaning parameters impact the cleanability of specific residues.
General use directions might work for one scenario but there is no
guarantee that the same directions would work under other
conditions. Process changes in manufacturing steps, batch
composition, dirty hold time, raw material sources, environmental
factors and others aspects may affect the soil condition at the time
of cleaning (refer to Figure 15.4). The cleaning parameters
previously set forth might not result in satisfactory results, leaving
minor residue spots or thin layers on the equipment surface that are
not easily detectable by the naked eye. Eventually, the residue spots
become food sites that allow microbes to proliferate.

A review of the current cleaning parameters and soiling effects

will provide the needed technical knowledge base for
understanding the effectiveness of the cleaning procedure and the
cleanability of the residue under current conditions. Laboratory
evaluations should then be performed to confirm the adequacy of
the current cleaning process by testing the four key parameters that
govern cleaning effectiveness. These include:

• the contact time of the cleaning solution with the dirty surface
 An Audit Approach to Contamination in Process Equipment 341
Figure 15.4 Effect of process related changes on cleanability

• the action or force acting on the surface

• the concentration of the cleaning agent

• the temperature of cleaning solution (Verghese and Lopolito,

2009).

The aforementioned parameters are commonly known as TACT

(time, action, concentration and temperature). In addition, if the
residue is dried, baked, autoclaved, denatured or polymerized onto
the surface, then it is important to mimic the actual conditions at
time of cleaning. Other characteristics include the amount of soil
present on the surface, the tenacity of the soil adhesion, and the
extent of dispersion of the soil in the cleaning solution during the
cleaning process. Figure 15.5 depicts a summary of a laboratory
evaluation for cleaning effectiveness.

Figure 15.5 Laboratory evaluation of cleaning effectiveness

342 Contamination Control in Healthcare Product Manufacturing

Laboratory evaluations should be conducted during a microbial

contamination investigation to examine the robustness of the
current cleaning process. As discussed earlier, a laboratory study
helps to clarify the difficulties in cleaning one or more residues and
identifies levels of chemical residues that can be contributing to the
contamination problem. Visual inspection of the cleaned coupons
and analytical and microbial tests should all be included in the
laboratory evaluation, to measure the magnitude of the problem. If
the current cleaning procedure does not produce the expected
cleanliness level, adjustments to the cleaning parameters are needed.

Surface and process equipment design

It is worth a brief discussion of rouge and scale origins to better
understand the following discussion and the significance of this
problem in GMP applications. Why is this important? Because
stainless steel is usually the preferred substrate in GMP industries.
Stainless steel constitutes the majority of the product contact
surface area for most applications. Stainless steel grades 304 and
316L have been used for many years due to their high quality and
corrosion resistance. Today, other grades are employed in certain
applications, such as medical device manufacturing.

Contrary to popular belief, stainless steel surfaces can corrode

when processing conditions are detrimental. Stainless steel
corrosion produces iron oxides that vary in color and solubility.
These oxides become particulates that eventually settle down onto
surfaces. Commonly called rouge, they can be composed of various
forms of iron oxides, with ferric oxide (Fe2O3) being predominant.

Rouge is a problem in water generation systems, whether they

produce Deionized Water (DI), Purified Water (PW), or Water for
Injection (WFI). It is also an issue for steam generators. It can be
found on tanks and pipelines that are constantly exposed to highly
corrosive solutions, such as those in the organic synthesis of small
molecule Active Pharmaceutical Ingredients (API) and in buffer
solution tanks used in biotechnology processes.
 An Audit Approach to Contamination in Process Equipment 343

Depending upon the type of rouge, some might be easy to wipe

off as shown in Figure 15.6a (Lopolito, 2010). The first photo is an
example of Type 1 rouge, which means that oxidized metal particles
generated from external sources are being deposited on this piece of
piping (Tverberg, 1999). This rouge tends to be yellow to orange in
color and it is very loose and fairly easy to remove. Type 2 rouge is
formed from in situ oxidation, is tightly adhered, and could have
underlying pits and damage (not illustrated in Figure 15.6). The
literature also mentions Type 3 rouge which is associated with high
temperature systems; it is mostly black in color, and very difficult to
remove from surfaces (illustrated in Figure 15.6b).

Figure 15.6 Examples of rouge and water scale build-up

(a) Cotton swab collecting Type 1 rouge (b) Steam trap with multiple in situ
from a piece of process piping dark spots typical of Type 3 rouge

(c) Washer compartment covered

with water scale
344 Contamination Control in Healthcare Product Manufacturing

Notice that Figure 15.6c illustrates the internal chamber of a parts

washer with scale build-up (not rouge) from water minerals. Hard
water exposed to high temperatures and alkaline solutions will
tend to precipitate the water minerals onto surfaces. Regular tap
water has been used for a long time in the washer in Figure 15.6c.
The washer was never treated with a descaling chemistry before
this picture was taken.

Rouge and scale particles rarely pose a contamination problem in

the manufactured product, but this depends on the type of product
being manufactured in the equipment. The main issue with rouge
and scale is that the accumulation of these particles forms a layer on
top of the metal surface. The layer is likely to have a higher roughness
than the stainless steel surface upon which it is forming. For optimal
cleaning, surfaces must be polished and free of irregularities. There
are standards suggesting a minimum surface roughness (Ra) of ≤1.6
µm for the biotech industry to ensure surface cleanability (EN13311-
1, 2001). Other standards are even more stringent, recommending
Ra between 0.38–0.76 µm (ASME-BPE, 2009).

As a consequence, process residues and microbes might adhere

more tenaciously to the rouged surface and become more difficult to
remove when compared to a highly polished surface. Residues and
microbes might also hide below the rouge or scale layers, making it
more challenging for the routine cleaner and sanitizer to reach the
chemical or microbial residue. In addition, once microbes attach to a
surface and proliferate, they may contribute to more rouge problems.
Some microorganisms are capable of consuming iron from a metal
surface in the absence of oxygen, and they use it as an energy source
for their metabolic processes (Beech et al., 2000; Videla et al., 2005).

“Written procedures are not followed for the maintenance of

equipment used in the manufacture, processing, packaging or holding
of drug substances. For example: a internal surfaces and manual valves
on the stainless steel chromatography columns used during drug
substance purification are not adequately maintained. Maintenance has
never been performed on the interior of columns to prevent adverse
impact on cell cultures due to metal contamination. Visible rouging was
observed on the exterior of the chromatography skid ...”
FDA Warning Letter (2009)
 An Audit Approach to Contamination in Process Equipment 345
Figure 15.7 Effects of rouge on microbial contamination

In other words, excessive rouging and scaling decreases the

cleanability of a stainless steel surface, promoting microbial
excursions in the processing equipment that eventually
contaminate the end product being manufactured (see Figure 15.7).
Rouge could also reduce the equipment’s useful life. It is important
to address rouge problems from a product contamination
perspective but also as part of a preventive maintenance procedure
to ensure proper long-term functioning of the equipment. Case
Study 6 summarizes the situation encountered by a company
reporting product contaminated with Enterobacter cloacae. Upon
inspection of its process piping, a serious rouge problem was
discovered that was never addressed in previous inspections.

Case Study 6: Rouge and microbial contamination

A manufacturer and bottler in Europe reported persistent issues with an
objectionable microbial contaminant identified as Enterobacter cloacae. The
company collected this microorganism inside the filling system, tanks,
transfer pipes, filling lines, and end-product. Enterobacter cloacae. is a human
pathogen and biofilm former that is typically associated with urinary tract
and respiratory infections.
346 Contamination Control in Healthcare Product Manufacturing
Several sanitization treatments were done in the process equipment without
success. After a thorough inspection of the process systems, it was noted
that there was rouge. A cleaning, derouging and disinfection procedure was
implemented to eradicate the contamination. First, an alkaline formulated
cleaner at a high temperature was recirculated through the equipment to
remove any organic deposits, including a possible biofilm. An alkaline
formulated cleaner is preferred over a commodity since it contains
surfactants that help lift organic residue from the surface. The second step
was an acid treatment using a formulated acid cleaner at high temperatures
to derouge and passivate the surfaces. At this point, most microorganisms
would be killed under such aggressive chemical and temperature conditions.
However, a third step was included to assure that all microorganisms were
eliminated. This additional step consisted of recirculating a formulated
sporicide of hydrogen peroxide and peracetic acid used at the temperature,
contact time and concentration indicated on its label. After performing this
procedure, the equipment was tested for microbes and none were found.
The company adopted this procedure as part of the equipment preventive
maintenance program.

In terms of equipment design, proper cleaning solution and

coverage must be taken in consideration. For manual cleaning
applications this is not a problem because the equipment is partially
or fully dismantled for cleaning. The person performing the
cleaning procedure has to apply the cleaning solution directly onto
the surface and exert some mechanical action to remove the soil.
However, the automatic system is different, since it involves the
distribution of the cleaning solution through spray devices that
exert almost no mechanical action in most cases. Hence, proper
coverage is important for successful cleaning.

Automatic processes are often validated. Once the validation is

done, very little attention is paid to the fact that automatic systems
might fail. Spray devices can get clogged, pump efficiency could be
compromised, controlled valves might not open, and pressure
could drop due to leaks — these are examples of issues that may
impact cleaning solution coverage. The worst part is that these
events can happen suddenly without any indication until there is a
significant cleaning failure.

The flow rate required for proper spray and cascading effect in
a vertical cylindrical tank can be estimated by multiplying 2.5 times
 An Audit Approach to Contamination in Process Equipment 347

the vessel perimeter in feet. This should result in a volumetric flow

rate in gallons per minute (gpm) for a vertical tank. For a horizontal
tank, it would be approximately 0.25 gpm per square foot of surface
area. It is important to inspect inserts such as baffles, dip tubes,
nozzles or agitators inside the tank because these may prevent the
sprayed fluid from striking the far side of the vessel walls. When
these mechanical items are present, additional spray devices may
be necessary to reach behind those potentially inaccessible areas.

A nice tool for testing coverage deficiencies in an automatic

system is a riboflavin test. A coverage study using riboflavin is a
simple test with many benefits:

• It helps identify shadowed areas that can lead to cleaning

failures

• It helps identify some of the hard-to-reach locations within the

vessel

• It serves as a training opportunity for personnel directly

involved in cleaning execution

• It assists in the understanding of the proper function of a spray

device and helps identify issues during routine visual
inspections

The basic steps of the coverage test are:

• Prepare a riboflavin solution of approximately 0.2 g/L and

spray the solution on the surfaces, preferably using a short
droplet sprayer. Drops should stick to the surfaces without
dripping down

• Inspect the surfaces while wet with a UV lamp to ensure that

everything was covered with riboflavin

• Run a short water rinse and recirculate through the system,

especially the spray devices to be tested. Drain the water from
the equipment

• Inspect the surfaces again while wet with a UV lamp and check
for residual riboflavin
348 Contamination Control in Healthcare Product Manufacturing

The presence of the riboflavin after the water rinse in locations

where the cleaning solution should reach the surface may be an
indication of insufficient coverage. The interpretation of the results
varies, depending upon the severity of the problem. Whatever the
outcome, it must be addressed to ensure proper coverage and
reduce cleaning failures. This procedure should be periodically
performed to check the proper functioning of spray devices,
especially if a microbial contamination event might be related to
improper cleaning. It is important to note that this is a coverage test
and not a cleaning test. Assurance of coverage increases the chances
of a successful cleaning procedure. Even so, additional laboratory
testing would be necessary to determine proper cleaning
parameters for the removal of the process residues.

There are a variety of valves available on the market for multiple

uses and applications. For cGMP applications, with more stringent
requirements, very few valves are considered to be intrinsically
sanitary, so diaphragm and pinch valves, which are generally
considered to be sanitary, are most often recommended. Their
mechanisms consist of a screw device that pushes a diaphragm
down to interrupt the fluid flow. When properly installed and
oriented, they are self-drainable. Other types of valves (ball valves,
gate valves and butterfly valves, for example) may carry some
contamination risk by accumulating soil and debris at gasket
spindles and crevices, so they become more difficult to clean in
place. Specific instructions should be included in the cleaning
procedure for dismantling and visually inspecting ball valves once
cleaning is completed. Additional manual cleaning steps and
sampling for cleanliness must also be considered.

Fluid flow can be described in two ways: laminar, which means

that a particle flow in the middle would travel at maximum speed
while a particle located at the wall is practically stagnant; and
turbulent, in which eddying motions can be very effective at mixing
and can quickly transport energy and heat from one place to another
(Perry and Green, 2008). As a result, velocity differences are
smoothed out more effectively than in laminar flow and the time-
averaged velocity profile is much more uniform. A fluid velocity of
approximately five feet per second (1.5 meters per second) for water-
based solutions is optimal in order to displace gases, penetrate dead-
 An Audit Approach to Contamination in Process Equipment 349

legs and fill vertical downward piping. This fluid velocity is quite
enough for pipe diameters of four inches or less.

Dead legs are sites in a piping system where liquid or gases can
become stagnant and where cleaning agents and water are not
properly exchanged during flushing. For obvious reasons, dead legs
should be minimized or, if possible, avoided. Bacteria and soil in
dead legs become sheltered from cleaning, flushing, and sanitization
procedures. Dead ends in piping are unacceptable unless they can be
physically inspected and sampled for cleanliness. Branches and/or
tees used for instrumentation should be horizontally positioned,
with their length limited to 1.5 times the pipe diameter.

Dead legs should not be positioned up as they can trap air

bubbles, or down because residues and microbes may be trapped
there. Figure 15.8 is an illustration of a dead leg with recommended
orientation.

Figure 15.8 Optimal dead leg orientation

Whenever possible, welding should be the preferred method for

connecting piping used to handle product and transfer cleaning
solutions. A properly welded connection should not have excessive
surface deformities such as cracks and crevices that will later
contribute to corrosion. Non-permanent joints used for
maintenance or multi-process reconfigurations can be clamp-type
connections which are accepted as a sanitary option for joining two
similar end pieces with a centered elastomeric gasket.
350 Contamination Control in Healthcare Product Manufacturing

Processing tanks should have a round bottom and a port that

facilitates draining. Flat bottom tanks must be pitched to facilitate
draining. The round or curved shape also applies to sanitary
connections, fittings, and nozzles. Overall, everything that has a
curved design is easier to clean and drain off than squared designs,
which tend to accumulate residues and debris between connecting
corners.

Horizontal piping must be inclined at 1/16 to 1/8 inches per

foot of piping (0.5 to 1 cm per meter of piping) to facilitate
drainability. This inclination must be towards drain points.

In the case of manual cleaning, repeated assembly and

disassembly procedures may result in surface damage if done
incorrectly. Corners, scratches, and crevices are often missed. The
cleaning or preventive maintenance procedure must specify surface
damage that is not acceptable, and how to fix it. It should never
be left to personnel to guess what is acceptable or not. It is common
to notice surface damage that should not be there, and to have
cleaning personnel say, “that has always been there.” A careful
visual inspection is not only about looking for process residues; it
also must include identify and report unacceptable surface damage.

The design of some processing equipment, for example, ribbon

blenders, tends to puddle residue at the bottom. If process
equipment does not drain properly after cleaning, additional
drying steps must be included in the cleaning procedure, and hard-
to-reach locations that may pose a challenge to cleaning must be
identified. If a portion of equipment is not easily accessible,
cleaning personnel have been known to simply not clean it. Any
portion left uncleaned is almost akin to no cleaning at all. As
mentioned earlier, residues left behind will serve as food and home
for microbes. Refer to Case Study 7, which provides an example.

Case Study 7: Hard-to-reach location

An oral solid dose manufacturer was having mold contamination in its
finished product. After a thorough investigation, it was discovered that the
source of the contamination was located in a V-blender. Figure 15.9
illustrates a V-blender positioned upside down. The bottom of the blender
connects to a solids charge chute located on the top floor. The cleaning
 An Audit Approach to Contamination in Process Equipment 351
operators were able to clean the charge chute as well as the blender, but
there was no easy access to the piece of tubing and piping connection
between the blender and charge chute.This was considered a hard-to-reach
location since it was 20 feet above the floor. Basically, there was product
build-up on the gasket of that connection where mold had proliferated,
contaminating select products that were manufactured in that blender.

Figure 15.9 Mold growth on the gasket in between the tube

and pipe connection

There are multiple standards and references available elsewhere for

more details on sanitary options (P3-A; ASME). In general, the
surface conditions and the equipment design must be examined
during a microbial contamination investigation. Table 15.1 can be
used as a guide for inspecting surface and equipment parts that
could be associated with microbial contamination in the process
equipment. Please be aware that Table 15.1 is not an all-inclusive
list of possibilities.
352 Contamination Control in Healthcare Product Manufacturing
Table 15.1 Check list of surfaces and equipment design issues

What to look for?

Surface Rouge, scratches, roughness, crevices, welding

imperfections, porous substrates

Valves Non-sanitary designs, worn gaskets, operating

mechanism, orientation

Dead legs Orientation, size, quantity

Spray devices (automatic Spray device mechanism, clogged holes coverage

cleaning systems)

Piping Rouge, welding imperfections, misalignments, poor

drainability, non-sanitary fittings, worn seals and gasket

Vessels Flat bottoms, non-sanitary nozzles, poor drainability,

instrument or equipment inserts

Pumps Non-sanitary design, worn seals and gaskets, bearing

failure, shaft leakage, rouge

Sampling ports Orientation, drainability, operation, cleaning and flushing

procedure, non-sanitary fittings, environmental exposure
concerns, worn seals and gaskets

Flexible hoses Poor drainability and storage, non-sanitary fittings,

worn sidewall, seals and gaskets

Sterilization or high-level disinfection parameters

This section reviews alternatives for equipment high-level
disinfection or sterilization, whichever is applicable. Several tech-
nologies are available and they are summarized in Figure 15.10. The
technologies underlined in Figure 15.10 are currently suitable and
feasible for process equipment. High temperature sterilization
techniques would be steam-in-place. Gas/vapor would be vaporized
hydrogen peroxide, chlorine dioxide, and ozone technologies.
Chemical sterilization using commercially available sporicidal/
sterilant agents is also available.
 An Audit Approach to Contamination in Process Equipment 353
Figure 15.10 Disinfection and sterilization methods

The applicability of any of these technologies would depend on the

process requirements, the sensitivity of the end-product to any of
these technologies, and the safety requirements for performing any
of the above methods.

In the case of non-sterile pharmaceuticals, it is neither required

nor expected to sterilize processing equipment. Generally, a low and
controlled bioburden is acceptable. An effective cleaning procedure
followed by a high-level disinfection or sanitization step may be
sufficient, provided these can kill objectionable microorganisms.
References, such as USP <1111> provide assistance in defining and
setting microbial limits for finished products. Nonetheless, the
significance of other microorganisms must be established on a case-
by-case basis, as these are microbial contaminants that, depending
upon species, cell number, dosage form and intended use of the
drug, may adversely affect patient safety (USP <1111>). For
example, an article published by Jimenez identifies Burkholderia
cepacia as the most frequently found microbial specie with 34% of
non-sterile recalls reported from 2004 to 2011 (Sutton and Jimenez,
2012; Jimenez, 2007 ). While the recall data supports the inclusion of
354 Contamination Control in Healthcare Product Manufacturing

Burkholderia cepacia as an objectionable microorganism, none of the

current pharmacopeias specify it.

The aforementioned methods should be validated to ensure that

these will consistently meet pre-established criteria. Change in the
soil level, microbial load or species, and surface attributes can
impact the disinfection or sterilization method being used. Critical
parameters governing the effectiveness of any of these methods
must be periodically monitored to verify the validated state. Table
15.2 provides a checklist of critical parameters of typical disinfection
and sterilization technologies used on processing equipment. It is
recommended to review them during a contamination investigation.

Table 15.2 Critical parameters for disinfection and sterilization

methods used on processing equipment

Type Critical parameters

Steam Time, temperature and saturated steam

Hydrogen peroxide Vapor concentration, time, humidity and saturation

Chlorine dioxide Gas concentration, time, temperature, humidity and pressure

Ozone Gas concentration, time, humidity and pressure

Liquid chemical Concentration, wet contact time, temperature,

water hardness

CONCLUSION
Contamination of a product with an objectionable microorganism is
a costly and time-consuming problem that needs to be addressed
quickly and efficiently by a cross-functional team. The cross-
functional team should consist of, but not be limited to, personnel
from quality, engineering, operations and microbiology. It is critical
to identify the microorganisms early in the investigation and
determine whether they are from an intrinsic (of the manufacturing
 An Audit Approach to Contamination in Process Equipment 355

process) or extrinsic (outside of the manufacturing process) source.

Based on the probable source, the investigation team can focus its
investigation. A careful walk-through of the process can be used to
flag areas of microbial risk and identify the presence or lack of
existing controls, understand when the product is at highest risk,
and to provide solutions to reduce the risk. A single root cause may
not always be determined. It is important to identify and correct
factors (within reasonable costs) that could have increased the
likelihood of microbial contamination within the process equipment.
A holistic approach is commonly performed to address the
contamination issues and get the manufacturing process running
again. As part of the holistic approach, assess the cleanability of the
system and make engineering changes to address contributing
factors such as coverage issues, drainability, etc., as needed. Perform
a robust high-temperature alkaline cleaning to remove process and
microbial residue. If rouge or water scale is present, perform a high-
temperature acid cleaning to remove the inorganic residue. When
possible, use a disinfection/sterilization procedure that is routinely
performed on-site and ensure that critical parameters are achieved
during the process. Once the manufacturing process is back up and
running, schedule regular meetings to review current microbial
control measures and internal plans to quickly and effectively
address the next microbial contamination event.

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ABOUT THE AUTHORS

Paul Lopolito is a technical services manager for the Life Sciences
Division of STERIS Corporation (Mentor, Ohio). He currently
provides global technical support related to process cleaning and
contamination control, which includes field support, site audits,
training presentations and educational seminars. Paul has more than
15 years of industry experience and has held positions as a technical
services manager, manufacturing manager and laboratory manager.
He has authored and published numerous articles on cleaning and
contamination control. He earned a B.A. in Biological Sciences from
Goucher College in Towson, MD.
360 Contamination Control in Healthcare Product Manufacturing

Elizabeth Rivera is a technical services specialist for the Life

Sciences Division of STERIS Corporation (Mentor, Ohio). Currently,
she provides technical support in the areas of formulated
detergents, disinfectants for critical environments, and sterility
assurance products. In addition, she specializes in cleaning
validation processes and lectures at technical educational forums,
such as IPA, Interphex, ExpoFYBI, IVT, and Executive Conferences.
Elizabeth has 10 years of industry experience and she has held
positions in pharmaceutical manufacturing companies. She has
published articles related to cleaning and cleaning validation. She
earned a Bachelor and Graduate degree in Chemical Engineering
from the University of Puerto Rico.