Excerpt from Cleaning Validation: An Exclusive Publication

Using Swabs for Cleaning Validation: A Review

By Douglas W. Cooper, Ph.D.
The Texwipe Company

Cleaning and cleaning validation are crucial to many industries. Clean swabs
applied skillfully to areas that have been cleaned can be used to show the
effectiveness of that cleaning. The swab, usually wet, is wiped across a cleaned
region. The swab, or an extract from the swab, is then analyzed physically,
chemically or biologically for contamination. The levels of contamination found
are compared against the "blanks," the levels in swabs of the same type that
were not rubbed on the surface being tested, and against a limit. Sufficient
replication is done to make a statistically valid statement that cleaning was
effective. The major choices - swabs, cleaning liquid, areas to be sampled,
extraction, analytical method and statistical analysis - are discussed.

Introduction

Pharmaceutical, bio-medical device and even food preparation industries are

concerned about the cleanliness - physical, chemical and especially biological -
of their products. These industries have materials that need regular careful
cleaning, and some have requirements for validating such cleaning,
demonstrating that required levels of cleanliness have been met. They want to
prove the efficacy of their cleaning methods. For example, "Cleaning and
validation of cleaning are among the most critical issues facing producers of
recombinant DNA protein products, monoclonal antibodies (MAbs), and
oligonucleotide therapeutics," according to Adner and Sofer (1994). As noted
below, the Food and Drug Administration (FDA) has taken validation of cleaning
very seriously.
It is widely recognized that contamination control is crucial to these industries.
Here we deal with the cleaning of surfaces, rather than the cleaning of gases or
liquids, generally done by filtration. This article discusses the use of swabs
(fabrics on handles or shafts) for cleaning validation.

Contaminants

Contamination comes from the environment, from the materials in use, the
processes and the people. Viable and non-viable particles and various organic
and inorganic materials often contaminate the surfaces of interest. Biological
contamination in clean rooms, such as those used for aseptic filling, has been
discussed in a monograph by the IES (Institute of Environmental Sciences,
1993).
Autoclaved pharmaceutical residues can be particularly hard to clean due to
their proteinaceous nature, probably requiring an acidic cleaner, but only
somewhat less hard are product residues, such as lipids, sugars and salts
(Parenteral Drug Association (PDA), 1996), requiring different solvents. Solvents
and cleaning compounds generally are relatively easy to remove, and product
residues may or may not be.

Validation

Validation is proof or demonstration that something is what it claims to be.

Carlberg (1995) defined it as "Full, detailed documentation that all processes and
procedures are functioning in the manner they were designed for. Required by
the FDA." Byers (in Carleton and Agalloco, 1985) gives several definitions,
including, "A scientifically designed program to prove that a process consistently
does what it is designed (intended) to do." The FDA (U.S. Food and Drug
Administration, 1993) stated, "In the end, the test of any validation process is
whether scientific data shows that the system consistently does as expected and
produces a result that consistently meets predetermined specifications." The
essence of all these definitions seems to be: documented, scientific proof of
consistent successful performance.
The FDA Guide to Inspections (U.S. Food and Drug Administration, 1993),
"intended to cover equipment cleaning for chemical residues only," includes:

1. "FDA expects firms to have written procedures (SOPs) detailing the

cleaning processes...."
2. "FDA expects firms to have written general procedures on how cleaning
processes will be validated."
3. These procedures will "address who is responsible for performing and
approving the validation study, the acceptance criteria, and when
revalidation will be required."
4. "FDA expects firms to conduct the validation studies in accordance with
the protocols and to document the results of studies."
5. Besides assuring chemical cleanliness, "the microbiological aspects of
equipment cleaning should be considered. This consists largely of
preventive measures...."
6. Sterilization is not part of the scope of the document, except that reduction
of the biological material will assist in successful sterilization and
minimization of pyrogens.
7. "Determine the specificity and sensitivity of the analytical method used to
detect residuals."
8. The sampling and analysis combination should be challenged to
determine what fraction of the target material is actually sampled and then
detected.
9. " Direct sampling (e.g., with swabs) is "most desirable," although rinse
sampling may be satisfactory.

References cited in this FDA document include Harder (1984), Smith (1992),
and Fourman and Mullen (1993).
Recently, Gold (1996) presented an extended discussion of validation, stating,
"Validation is necessary since testing of a product to assess its quality is not
enough." Rare failures can escape testing but might be prevented by improved
procedures. Table 1 shows the probability (binomial distribution) that a sample of
size "n" will find no defectives, when the true fraction of the population is "p,"
which probability is (1-p) to the nth power. The table shows, for example, that if
the true fraction defective is 1%, samples of size n=100 will find no defectives in
36.6% of the cases.

Sampling is clearly not enough. Validation requires checking the materials and
equipment and procedures that constitute the process. The essence of validation
is "establishing documented evidence that a process does what it purports to do."
Gold (1996) emphasized that what is needed is routine, documented application
of the scientific method. Documentation guidelines have been presented by
Capote (1996).
Sterilization validation is a specialty with its own extensive literature (see Bruch
in Morrissey and Phillips, 1993), and is not covered in detail here. Validation of
Aseptic Pharmaceutical Processes by Carleton and Agalloco explores this
subject in detail.

Cleaning Validation

Cleaning validation means proof that something has been cleaned, perhaps
that the contamination levels have been reduced by at least a certain ratio or,
more usefully, that they have been reduced below some target level. That means
there will be some target limit level of surface contamination, and sampling and
analysis will be carried out to assure that the area of interest almost certainly is
cleaner than the target limit. Often, the locations hardest to clean will have to
receive special attention to assure that they have been cleaned.
In an early paper on cleaning validation, Harder (1984) cited five crucial
elements:

1. A standard operating procedure (SOP) for cleaning (with a checklist)

2. A procedure for determining cleanliness (e.g., rinse or swab, then analyze)
3. An assay for testing for residual drug levels
4. Pre-set chemical and microbial limit to which the equipment must be
cleaned
5. A protocol for cleaning validation

Harder recommended that the procedure be tested by requiring it to be

successful on three successive cleanings. (See Figure 1) and that there should
be periodic revalidation, as well as revalidation after significant changes.
 Jenkins and Vanderwielen (1994) presented an overview of cleaning validation,
covering strategy, determination of residue limits, methods of sampling, and
analysis. They noted that "increased use of multipurpose equipment" has
produced increased interest in cleaning validation. The cleaning protocol must be
thorough and must be checked. Training is essential. A validation program
requires:

 Criteria for acceptance after cleaning

 Appropriate methods of sampling
 A maximum limit set for the residues
  The test methods must themselves be tested

Products to be tested may best be put into groups, rather than testing all of
them. The most important may not be the highest-volume products, but those
capable of causing the largest possible problems if contaminated or if they
contaminate other products. (Solubility of the drug is an important issue.)
Equipment may also be tested as groups.
Geigert et al. (1994) emphasized the importance of cleaning validation, as
making good business sense and being required by cGMP.
Table 2 has "points to consider" for cleaning validation, summarizing an
excellent article on the topic by Agalloco (1992), who stressed that not all of
these points need equal weight.
Recently a highly useful book has been published on cleaning and cleaning
validation (PDA, 1996), covering process equipment design, cleaning
mechanisms, automatic cleaning equipment, validation studies, sampling
methods, analytical methods, validation acceptance criteria and special
considerations for multi-product facilities. The following sampling methods
provide various levels of assurance concerning cleaning:

 Visual inspection
o Easy
o Qualitative
o Subjective
 Rinse water sampling and analysis.
o Sampling is easy
o Analysis can be quantitative, using pH, conductivity, particle count,
microbial count, total organic carbon determination,
spectrophotometry, bioassays, limulus amebocyte lysate for
pyrogens
o Recovery factor is uncertain; it involves dilution.
 Surface sampling and analysis
o Removes adherent materials
o Analysis can be quantitative
o Precise definition of the area sampled
 Surface sampling from coupons
o Quantitative
o Depends on whether coupons are equivalent to the surface of
interest
o Requires removing coupons from system...