Equipment Cleaning Validation Within a Multi-Product Manufacturing Facility

Currently, there are multiple publications, as well as guidelines from regulatory agencies that
make the critical process of equipment cleaning validation easier. These sources provide in-depth
information for the validation specialist, making the development and implementation of a robust
cleaning validation program possible within any particular facility developing or manufacturing
parenteral, biological, or sterile ophthalmic products.

Extremely important, specific, and above all, mandatory, are the requirements established by
regulatory agencies such as the US Food and Drug Administration (FDA), the European
Medicinal Evaluation Agency (EMEA), Australia's Therapeutic Goods Administration (TGA),
etc. For example, the 2004 Code of Federal Regulations (CFR) Title 21, Volume 4, Section
211.67, states:

"Equipment and utensils shall be cleaned, maintained, and sanitized at appropriate intervals to
prevent malfunctions or contamination that would alter the safety, identity, strength, quality or
purity of the drug product beyond the official or other established requirements."

Additionally, Section 211.182 requires that cleaning procedures must be documented

appropriately, and that a cleaning and use log should be established.

This article provides the reader with cleaning validation information enhanced by the author's
thirteen years of hands-on experience working in equipment cleaning validation.

SCOPE

This article focuses on manual cleaning procedures because these are considered the worst-case
scenario. It applies to parenterals, ophthalmic, and biologic presentations and is intended to cover
equipment validation for raw materials, contaminants, cleaning agents, as well as the control of
potential microbial contaminants associated with those products.

The flowchart in Figure 1 graphically shows the different aspects that should be considered when
developing a cleaning validation program. Understanding each aspect of the process, the
relationships among these actions, and the sequence in which they should take place will make
the development of a cleaning validation program a successful experience.

PROCESS FLOW

After all applicable cleaning information sources and regulatory guidelines have been consulted,
the first item to consider when establishing a cleaning validation program is the raw material and
final product flow. By following the flow of the product, one can identify the equipment that
comes in contact with it, such as utensils (scoops, spatulas, funnels, pipettes, etc.), tanks, filter
housings, pressure vessels, syringes, and others. Equipment such as this is considered critical
equipment because it comes in direct contact with the product.
Other areas where raw materials or products are processed, which might be considered non-
critical because they are not in direct contact with the product, should also be considered. From
the point of view of microbial load, inappropriate cleaning and sanitation of these areas may
contribute to cross-contamination. Some examples of these areas include: sampling and weighing
rooms, as well as formulation and filling rooms.

Typical steps to follow in process flow are as follows:

Raw Materials Sampling: Raw materials include both active and inactive ingredients. Many
active ingredients are potent compounds, such as steroids, cortisone, antibiotics, proteins, and
therefore it is important to demonstrate their removal. But be aware that some inactive
ingredients have poor solubility in water and their residues may be more difficult to remove than
those of an active drug.

Utensils used during the sampling process of raw materials require cleaning validation unless
they are disposable. Typically, use of disposable utensils is the preferred practice for parenteral
and biological products. The sampling of raw materials should be performed within a controlled
environment (classified as 1,000, 10,000, 100,000, etc.,) in order to reduce the introduction of
non-intrinsic contamination to the process.

Some firms use extraction hoods equipped with high efficiency particulate air (HEPA) filters and
room air conditions that provide an acceptable environment for raw materials sampling. In
addition, sampling area and rooms should be cleaned with sanitizing agents. Cleaning
effectiveness on those surfaces should be challenged using microbiological tests for verification
of bioburden reduction of non-sterilized parts, such as tables, walls, ceiling, fillers, etc.

Weighing of Raw Materials: Some rules followed during raw materials sampling also apply to
the raw materials weighing process. Some ingredients will require being weighed inside a glove
box due to special environmental requirements (e.g., under nitrogen, etc.). Because glove boxes
are usually shared, they too will require cleaning validation.

Product Compounding: This is one of the most critical steps because the equipment used will
have direct impact on the finished product. Compounding a finished product requires equipment
with large product contact surface areas. The handling of ingredients requires utensils with less
product contact surface area. This significant difference is important because the more complex
the equipment, the more samples must be taken to demonstrate effective cleaning.

Product formulations involve the use of tanks and ancillary equipment, such as gaskets, pipes,
hoses, mixers, and filter housings. Gaskets and hoses are disposable in many pharmaceutical
processes, so they do not generally require cleaning validation.

Product Filling: Filling of parenteral, ophthalmic, and biologic products is usually performed
within areas of controlled bioburden, ranging in scale from clean to aseptic rooms. Drug products
are capable of being contaminated in many ways. Contamination may occur via filling
components (tips, caps, bottles, or stoppers); when coming in contact with processing equipment
(tanks, manifolds, fillers, machine-syringes, pistons, and blocks); the manufacturing
environment, or manufacturing operators.
Some equipment may require cleaning revalidation if components come in contact with the
product. Stoppers are siliconized and then are placed in a hopper during filling. Small quantities
of silicone are accumulated in the lower part of the hopper where it can degrade over time. If this
silicone were to come in contact with the product, it would probably cause product
contamination.

Requirements for aseptic processing include cleanable floors, walls, ceilings, particulate,
temperature, humidity, cleaning, and disinfecting procedures. When disinfectants are used in the
manufacturing area, care must be taken to prevent the product from becoming contaminated with
chemical disinfectants. The selection of suitable disinfectants, verification of their effectiveness,
and a surface challenge are critical in developing a cleaning and sanitization program. Written
procedures for cleaning, maintenance, and sanitization of manufacturing equipment and
appropriate areas of the facility are required. Removal of residual disinfectants should be
monitored as a precaution against the possibility of product contamination.

Know Product Ingredients and Intended Use of the Final Product

Previous to designing the cleaning procedures, it is necessary to know all physical and chemical
characteristics of the product ingredients. Characteristics such as appearance, solubility, potency,
and toxicity play an important part in the design strategy of a cleaning validation program. These
characteristics will indicate whether solvents or detergents are needed for removal of product
residues. Avoid the use of detergents or solvents whenever possible because their use demands
added controls.

Regarding the intended use of the product, cleaning procedures related to the production of
parenterals are the most critical. Great precautions should be taken with the cleaning procedures
surrounding these products, because they will be intravenously administrated to patients and any
adverse reaction could cause serious damage to patient health.

Developing Standard Operating Procedures (SOPs) for Cleaning Processes

Once we know and understand the product process flow, the product's ingredients, and the
product's intended use, standard operating procedures associated with the cleaning process
should be established. These procedures should clearly address the specific method chosen, the
cleaning process itself, any detergents, and the allotted cleaning time.

There are three types of cleaning procedures for process equipment: automated Clean in Place
(CIP), Clean out of Place (COP), and the manual process. However, the major concern of
regulatory agencies has targeted pieces of equipment that will be cleaned manually, and where
the primary responsibility for the removal of product residues lies with the cleaning operator. In
the case of manual cleaning methods, the effectiveness of cleaning depends upon the design of
the procedure and the commitment of the operators to follow that procedure. This requires a
well-explained SOP, personnel training, and operator commitment. With all three elements
present, reproducibility of the results in terms of removal of product residues from equipment
surfaces can be achieved.
Cleaning procedures should cover every type of equipment to be cleaned. They should specify
all critical parameters including water flow, temperature, and pressure as well as washing and
rinsing times. Usually these will have been previously set through engineering studies. Cleaning
procedures should also include specific details regarding disassembly of equipment, where the
small parts should be placed, the quantity of detergent to be prepared and its concentration, and
the type of brush or piece of cloth used to apply the detergent. The size (length, and internal
diameter) and the type of hose used to perform the cleaning process should also be specified.
This is important because, if the hose diameter is small, less water is used and a higher pressure
is achieved during the operation. Finally, cleaning procedures should be challenged and validated
to demonstrate the reproducibility of results for all cleaned parts.

Validated Analytical Methods

Appropriate validated analytical methods for analyzing cleaning validation samples will be used.
A validated method is rugged and robust enough to measure the residue limit established. The
method used should be based upon previously established residue limits of the active, cleaning
agents, and excipients. The method must be appropriate for measuring the analyte at and below
the acceptance limit for residue.

Two important parameters in an analytical method used for cleaning validation are: the Limit of
Detection (LOD) and the Limit of Quantitation (LOQ). The LOD is the lowest amount of a
compound that can be detected. The LOQ is the lowest amount of a compound that can be
quantified. The LOD is usually lower than the LOQ, but is never higher. The LOD should never
be used to establish residue acceptance limits. Residue acceptance limit should be established
based on LOQ for accurate measurement.

One of the important considerations in choosing an analytical method is the type of residue to be
analyzed. Residues can be active drugs, cleaning agents, or organic compounds. The
methodologies available are either specific or nonspecific. A specific methodology detects a
unique compound in the presence of potential contaminants. Some examples of specific methods
are high performance liquid chromatography (HPLC), ion chromatography, atomic absorption,
capillary electrophoresis, and other chromatographic methods. Nonspecific methods detect any
compound that produces a certain response. Some examples of nonspecific methods include total
organic carbon (TOC), titrations, pH, and conductivity. Currently, methods that are
complimentary to each other are applied to cleaning validation samples. That is, if a specific
method is used for active and cleaning agents, then a nonspecific method, such as TOC can be
used as a complementary methodology. Cleaning validation methodology includes other
important parameters such as linearity, ruggedness, method precision, and reproducibility.

A method used to analyze cleaning validation samples is properly designed if it identifies

possible interferences. Many possible sources of interferences include active drugs and
excipients, cleaning agents and compounds, swab materials, and solvents used in performing the
cleaning process. Still other sources of interference may be triggered when a sample is not
handled properly.

In addition, the method validation must contain sample stability for the reference analyte. Choice
of validation methods should guarantee that samples remain stable for the time specified for
analysis. This includes the stability of all residues that specifically determine the active and the
detergent residues.

Validated Swab Recovery Study

The swab recovery must be validated to determine the amount of analyte that can be recovered
from a surface. Previous to validating the equipment cleaning procedure, the types of equipment
surfaces for product manufacturing should be identified. This means if four types of surfaces are
identified such as glass, stainless steel, Teflon, or rubber; each of the four surfaces must be tested
for swab recovery.

The study can be performed on simulated product contact surfaces. The efficiency of the
swabbing procedure should be challenged at the acceptance criteria level for active ingredient,
excipients, and detergent.

Known amounts of active ingredient, excipients, and detergent will be added to each type of
surface. The surface is air-dried and swabbed. The nature of the swab extraction is dictated by
the analytical method, which has taken into consideration its dissolution properties as well as the
analytical technique used. The amount recovered by the swab is determined using a validated
trace level analytical method.

Recovery from the surface is calculated as follows:

A recovery factor of 70% is usually acceptable; but factors as low 50% may be obtained. In cases
where low results are obtained in a reproducible manner, the sample surface area may be
sampled again using a second swab,with the results obtained from both swabs added together.

Initial Cleaning of New Equipment

Another important aspect to consider when establishing a cleaning validation program is the
initial cleaning of new equipment. Most commonly, the equipment included in a cleaning
validation study is that which currently is in use for a specific operation. Nevertheless, it could
happen that new equipment purchased for any type of improvement or replacement is received
and installed at the site while the equipment validation study is being developed. In such case,
this is the best time to include the new equipment in the study.

Initial cleaning for new equipment requires some special considerations as noted here. This
cleaning should be performed to eliminate foreign matter or residues introduced through
maintenance, fabrication, or installation.

Description of this initial cleaning procedure is as follows:

Equipment should be passivated (if stainless steel material) as per current SOP.
Remove passivation residues from equipment surfaces as per existing cleaning SOP.
No detergent will be used during the cleaning process.
Let equipment dry, rub a lint free piece of cloth (preferably white) over equipment surface and
inspect the piece of cloth for visible residues.
Take rinse or swab samples to determine the absence of passivation agents by a validated
methodology. Additional samples can be taken for pH/conductivity or any organic reagent
determination by TOC technique. In some cases, a sample for total plate count determination is
also included.
The rinse sample for passivation agents will be collected in a 500 mL glass bottle and the sample
for organic test by TOC determination, in a 50 mL glass tube.
The microbiological samples will be collected in sterilized sampling bottles. When swabbing is
called for, a sterilized swab should be used.
Take samples as controls from the water source for chemical tests, TOC analysis, and micro-
biological analysis, respectively.
Submit the samples to the corresponding laboratories and analyze as per current SOP's.
Acceptable limits:
Passivation agents: ID test according to the chemical used in the passivation process
TOC: Three log reduction of passivation agent's formulation or USP limits
Conductivity: USP limits
Total Plate Counts: The most important consideration in setting any limit here is that it have a
scientific basis supported by historical data.

CLEANING VALIDATION PROTOCOL

Once all the aforementioned aspects are considered, we are ready to develop the cleaning
protocol. A protocol used to validate the cleaning procedure contains the following basic
elements:
A. Purpose: This section describes the intention of the validation protocol.
B. Scope: This lists the boundaries of the project to be validated.
C. Responsibilities: Tasks are listed for each person involved in executing the protocol.
D. Strategy: The following steps should be included in the strategy section:

1. Description of the product to be challenged:

If two or more product presentations are used, the more concentrated should be challenged.
Bracketing may be considered acceptable for similar products and equipment, provided
appropriate scientific justification is present.

2. Holding time period for equipment being cleaned:

This is the time during which the equipment was soiled with the product.
Let the residue dry for a specified length of time (hours), followed by cleaning the equipment.

3. Holding time for cleaned equipment:

The rationale to establish this period of time should be based on microbiological challenges in
equipment areas identified as inaccessible locations or where the probability of stagnant water
can exist.
4. References
5. Cleaning effectiveness measures (rinse or swab)

Measuring and Test Equipment

Check calibrations of instruments used during protocol execution for proper compliance.

Procedure

1. Identify the procedure to be challenged.

2. List all safety precautions established for the affected area.
3. Indicate the number of runs that will be made
4. Describe the equipment to be cleaned, including:

Equipment pictures
Indicate equipment surface area calculations

5. List chemical tests (active, detergent, excipients, pH, and conductivity, as applicable to the
method)
6. Indicate the number of samples to be collected, the volume of rinse (mL), the container (e.g.,
glass) material, etc.

Selection of Acceptance Criteria

Chemical Limits

For any new product, the owner should be consulted for details concerning the therapeutic
potential and possible adverse reactions. A determination should be made as to the
appropriateness of the limits for that product. If the product is determined to be highly potent or
poses the strong possibility of adverse reaction, special limits must be determined. All special
limits should be determined on a case-by-case basis.

The lowest therapeutic dose (LTD) of 1/1000 is considered a safe level for residues of active
ingredients. The same approach will be adopted to calculate the contamination limit for the
cleaning agent, using LD50 instead of LTD. For the products that have excipients, (e.g.,
preservatives), the acceptable residual level should be set at not more than 1/1000 (three log
reduction) of TOC of the product formulation.

The maximum allowable carryover (MACO) limit for the active drug or cleaning agent in mg per
swab for a specific piece of equipment or for an equipment process train is as follows:

MACO (mg/swab) =

Where:
LTD = Lowest therapeutics dose (mg) or LD50 for cleaning agent (mg/kg)

D = Highest maximal daily dose (dose units)

W b = Smallest batch size (g)

W t = Highest unit dose weight (g)

S s = Swab area (cm2 or in2 )

S e = Equipment product contact surface area (cm2 or in2 )

R = Recovery factor of active ingredient or cleaning agent

LD50 = Lethal dose of 50% of animal population

Microbiological Limits

Depend on the type of product (e.g., injectable, etc.)

The most important consideration in setting any limit is that there be a scientific and historical
basis.

Discrepancies

Collect any unexpected situations that occurred during protocol execution.

Approvals

List approvers will vary with each firm. At the very least, the following should be included: a
validation specialist, head management of the affected area, appropriate representatives of the
microbiological and chemistry laboratories, and appropriate QA representatives. The protocol
must be approved before execution begins.

FINAL REPORT AFTER PROTOCOL EXECUTION

The final report will include the following:

1. Table of Contents

2. Introduction

Include a brief description of the cleaning process that was validated.

3. Data analysis and results

Summarize all validation outcome (use of tables is recommended)

4. Deviation

Any discrepancy found during protocol execution should be satisfactorily explained.

5. Conclusions

6. Approvals

The same personnel who approved the validation protocol should approve the final report. These
individuals included a validation specialist, head management of the affected area, appropriate
representatives of the microbiological and chemistry laboratories, and the appropriate QA
representatives.

MONITORING SYSTEM

The purpose of monitoring is to assure the adequacy of the equipment cleaning process. It is
important to verify that the cleaned equipment performs as it was intended and that it remains in
a validated state. Monitoring may be achieved through taking representative samples, and
evaluation of product non-conformances; by following through on quality alert notifications and
complaints and by conscientiously completing "Annual Products Reviews." Any or all of these
should be followed by revisions to change controls as needed.

Failures in any of these areas might indicate the need for revalidation. Revalidation is also
required when there is a significant change in product formulation, change, or modification to a
process or equipment and change of cleaning agents.

COMMON ERRORS AND RECOMMENDATIONS

The section is based on the author's experience during his 13 years of dealing with cleaning
validations.

Common Errors

1. Failing to perform a good process flow, beginning with the raw material sampling process and
focusing on obviously important stages such as the compounding or filling process.

2. Failing to train operators well, and failure to instill a sense of high commitment:

Most common mistakes, such as not following procedures, not taking samples at the time
specified, improper handling of samples, not reporting discrepancies observed during the specific
process executed, etc., undermine high quality performance and contribute to poor results. It is
highly recommended to have representation from the validation department present throughout
all runs.
3. Missing or inaccurate documentation:

In addition, to the common errors noted above in #2, a lack of documentation also causes
difficult situations during future audits, because important key information will be missing
during investigations. This is often the basis for general misunderstanding of data.

4. Lack of good coordination and communication between the different areas involved in the
validation process:

Examples of poor coordination and communication include samples not being taken on time,
required materials not being available, disputes within departments, lack of cooperation, and bad
working environments based on placing blame rather than on cooperation.

5. Failing to understand the criticality of the sampling process:

It is very important to take the samples at the time(s) established and for the proper durations
(stability), taking correct quantities and volumes, attending to the proper handling and storage of
samples, using the correct equipment, and other special considerations for samples requiring
microbiological testing.

6. Failure to properly train persons in charge of sampling:

This can have a major impact on the accuracy of microbiological sampling.

7. Not having an effective change control program:

Once the validation process is completed, internal procedures impacted by the validation
outcomes must be revised accordingly. It is frequently observed that the communication and
follow up systems fail at some point and impacted procedures and activities are not modified.

8. Not having an adequate cleaning monitoring system:

It is frequently observed that changes are made to an already validated system, disregarding the
impact those changes might have on the validation state of the total system. Constant
communication between manufacturing and the validation department is critical.

9. Failure of manufacturing areas to consider the validation department in internal changes and
procedure approval:

Due to lack of technical knowledge in the validation area personnel in manufacturing might
overlook or be unaware of important aspects that have impact from a validation point of review.

10. Failure of plant personnel primarily focused on the manufacturing operation, to require
compliance with the cleaning validation procedures.
José A. Morales Sánchez is a technical services senior scientist at Janssen Ortho LCC a Johnson '
Johnson Company; 787.272.7463; jmoral12@janpr.jnj.com
.

REFERENCES

1. FDA. "Guide to Inspectors of Validation of Cleaning Procedures," 1993.

2. Technical Tip #3020. "Pharmaceutical Product Contact Surface Sampling." Steris Corporation
(Calgon Vestal Division), 410-200-3020. 4/97.

3. PDA Pharmaceutical Cleaning Validation Task Force, "Points to Consider for Cleaning
Validation," Journal of Pharmaceutical Science and Technology, Technical Report No. 29,
Volume 52, Number 6, 1998.

4. Maria J. Capote, "Documentation For Cleaning Validation: A Protocol Template," pgs 261-
271, Volume 2-Number 3, Journal of Validation Technology.

5. Gil Bismuth and Shosh Neumann, "Cleaning Validation: A Practical Approach," Interpharm
Press, 2000.

6. Health Products and Food Branch Inspectorate, "Good Manufacturing Practices - Cleaning
Validation Guidelines," Spring 2000.

7. Destin A. LeBlanc. "Equipment Cleaning Validation: Microbial Control Issues," Journal of

Validation Technology, Volume 8, Number 4, August 2002.

8. Herbert J. Kaiser and Maria Minowitz. "Analyzing Cleaning Validation Samples: What
Method?" (http://www.ivthome.com/free/analyzin...on_samples.htm, accessed July 16,2003).

9. Code of Federal Regulations Title 21, Volume 4, Section 211.67, April 2004.