ANATEL Clean-In-Place
(CIP) Application Note
On-line Total Organic Carbon (TOC) & Conductivity
Analyzers for Verification of CIP Processes
ANATEL Clean-In-Place (CIP) Application Note
•••
ANATEL Clean-In-
Place (CIP) Highlights
•••
Application Note • The use of on-line
TOC & Conductivity
Analyzers to verify
On-line Total Organic Carbon (TOC) & Conductivity CIP can provide a
Analyzers for Verification of CIP Processes return on investment
of <1 year through
savings in time and
Introduction reagents used for
Frequently, CIP processes are validated and verified using laboratory grab-
laboratory grab sample analysis. The disadvantages of using this sample analysis.
on an ongoing basis is that the number of grab samples required
• TOC and conductivity
can be costly in terms of time and laboratory instrument reagents.
combined can be used
In addition, the time to collect grab samples and carry out the tests
to detect the process
can delay returning the process vessel to use, thus putting a vessel is clean of all
restriction on the capacity of the production facility, reducing inorganic and organic
throughput and economies of scale. matter down to WFI
levels.
The use of on-line direct conductivity TOC & Conductivity
verification analyzers located on the CIP skids can eliminate the • TOC & Conductivity
verification analyzers
need for grab-sample analysis. These analyzers use no reagents
employing direct
and can be stable in calibration for up to one year, thus reducing conductivity and
analysis cost and increasing production up-time. In some cases needing only small
pay-back for the analyzer can be <1year. sample aliquots are
best suited to this
In the sterile manufacturing environment, there are a lot of application.
advantages of cleaning the process vessel in place, not least of
• Direct conductivity
which is the advantage that keeping it in the cleanroom area means
TOC instruments offer
that it is far less likely to be contaminated. Traditional cleaning an advantage over
methods would involve removing the process equipment from the membrane-based
sterile manufacturing environment in order to clean and sterilize it analyzers because
before moving the equipment back into the sterile environment. infrequent use may
allow the wetted path
This type of process greatly increases the risk of contamination as
to dry, which could
the equipment is moved in and out of the controlled environment. damage membranes.
Introduction 1
ANATEL Clean-In-Place (CIP) Application Note
•••
The Aim of CIP
CIP aims to carry out all necessary cleaning steps to ensure that the process equipment is fit for use for
the next batch. In bulk sterile manufacturing using process vessels, the vessels can be cleaned at a
cleaning station located inside the cleanroom. In many cases, the CIP cleaning stations are on wheels
and portable (see Fig. 1). In other cases the process vessels themselves are on wheels and portable (see
Fig. 2) and they are moved to a fixed cleaning station within the cleanroom and this is sometimes
referred to as Clean Out-of Place (COP), but the intention and net effect is the same. All CIP/COP
cleaning steps are carried out with the process equipment in the normal production environment
where the equipment is used. The cleaning process is determined according to the cleaning
requirement of the process equipment and the risk of contamination to the next batch of product. In
some cases the risk of cross-contamination from one batch to another is too high to allow CIP and
multi-use of process equipment, one example being the manufacture of biologically active products
such as vaccines where typically separate process areas and equipment are dedicated to each product
being manufactured.
Where the risk assessment shows that production areas and process equipment can safely be used for
the manufacture of different products, the CIP process must be validated to demonstrate that the
resultant cleaning is sufficient to not represent a risk to the next batch.
The Aim of CIP 2
ANATEL Clean-In-Place (CIP) Application Note
•••
The Role of Conductivity and Total Organic Carbon
The attraction of Total Organic Carbon (TOC) and conductivity measurements for determining the
effectiveness of cleaning processes is that, between the two measurements, they will detect most
organic and inorganic contaminants, including the drug product itself and any cleaning agents used in
the CIP process. Conductivity tends to detect inorganic materials and, of course, TOC will detect trace
amounts of organic compounds that conductivity will not detect. Conductivity can be used to detect
gross contaminants present as the wash-down process commences and then, as the process equipment
cleans down, TOC can be used to verify the end-point of the cleaning process, i.e. the point at which the
process equipment is cleaned to the pre-determined ‘acceptably clean’ state and can be released for use
for the next production batch.
Method
CIP methods are process-specific and generally share the aim of cleaning the process equipment down
to a level where the next batch will be successful and not contain product left over from the last batch.
In the sterile manufacturing environment, it is typical to have a follow-on stage of sterilize-in-place
(SIP) to ensure that the process equipment is sterile and suitable for use in the sterile manufacturing
process. In these sterile applications, it is considered attractive to use CIP as the process equipment
does not have to leave the sterile manufacturing environment. It is common to see CIP techniques
employed in factories manufacturing sterile bulk active pharmaceutical ingredients (API).
Depending on the aim of the clean, it may involve the use of commercially available cleaning
detergents, acids, or bases (e.g. CIP100 and CIP200); in other cases it may simply involve the use of
chemicals designed to inactivate the biological product being manufactured combined with simple
water for injection (WFI) for rinse down.
There are no published standards that define any of these processes, so users must determine an
appropriate CIP process according to their process goals.
An example of a CIP program for a fermentor / bio-reactor might look like this:
• Pre-rinse
Two pre-rinse cycles could be carried out with relative low- or ambient-temperature water (in
order to avoid denaturizing any protein-based residues that may be present). Typically the pre-
rinse cycles are sent straight to drain and the aim of this stage is to remove the majority of the
product residues from the vessel.
• Wash 1
The wash cycle for a bioreactor could consist of a detergent based on potassium hydroxide.
Typically this is recirculated through the vessel in order to allow the caustic detergent to clean the
vessel surfaces effectively. The temperature of this part of the cycle is typically elevated to boost
the detergent’s effectiveness.
The Role of Conductivity and Total Organic Carbon 3
ANATEL Clean-In-Place (CIP) Application Note
•••
• Post Wash 1 Rinse
Typically a post wash rinse is performed in once-through (single-pass to drain) aimed
exclusively at removing the caustic detergent residues.
• Wash 2
A second wash cycle using a formulated detergent based on phosphoric acid is typically carried
out with the aim of removing any caustic cleaning residues, again at an elevated temperature. A
secondary effect of using an acidic detergent is that it can help restore the stainless steel surfaces
of the bioreactor vessel.
• Post Wash 2 Rinse
After the acidic wash cycle a once-through (single-pass to drain) rinse with high temperature WFI
is typically carried out aimed exclusively at removing any acidic residues.
• Final rinse program
A final rinse program is then typically carried out using WFI with the expectation that the vessel
will be clean and free from any residues of the drug product, or cleaning agents at the end of this
program. It is on the final rinse where the cleanliness is verified by confirming that the conductivity
and TOC levels of the WFI as it is drained from the vessel meet target ‘clean’ values (conductivity is
primarily to detect any residual cleaning agents, while the TOC is primarily aimed at detecting any
drug product residuals . Figure 3 shows the Anatel PAT700 combined TOC and Conductivity
analyzer installed on a CIP system where it is used for measuring TOC and conductivity of the final
WFI rinse.
Fig. 3 Typical combined TOC & Conductivity verification analyzer installation
Method 4
ANATEL Clean-In-Place (CIP) Application Note
•••
During an automated CIP program, the controlling PLC/SCADA system will interact with the TOC &
Conductivity verification analyzer, typically as follows:
Ready to analyze
CIP PLC
Step 1
TOC & Conductivity
Verification Analyzer
Start analysis
CIP PLC (5 mins before
end of final rinse)
TOC & Conductivity
Verification Analyzer
Rinse sample line
Step 2
for 2 mins, then
take fixed sample
aliquot
TOC & Conductivity
Verification Analyzer
Step 3
Analyze fixed
sample aliquot for
TOC & Conductivity
Report TOC & Conductivity
Analysis complete
CIP PLC
Ready to analyze
TOC & Conductivity
Verification Analyzer
Fig. 4 Typical signal exchange between TOC & Conductivity analyzer and PLC
Challenges
Most CIP applications involve periodic cleaning. In some cases the CIP skid may be used several times
each day, especially in the case of a mobile CIP skid that is wheeled around the process area and used
to clean multiple pieces of process equipment each day. In other cases, the CIP equipment may have
prolonged periods of inactivity. This may make membrane-based TOC analyzers (indirect
Challenges 5
ANATEL Clean-In-Place (CIP) Application Note
•••
conductivity) unsuitable for applications where the CIP skid is unused for prolonged periods because
of the possibility that the membranes will dry out and need to be replaced.
The same applies to TOC analyzers designed for continuous flow through, especially those analyzers
that utilize automatic re-calibration/offset calculation of their multiple conductivity cells. With these
analyzers, the sporadic sample flow may force the analyzer to enter a re-calibration routine each time
the sample starts, thus extending the time before the analyzer can take a reading. In addition, as this is
not a continuous-flow application, the small sample aliquot typically provided for analysis does not
suit TOC analyzers designed for continuous flow as they require the sample to flow for a period of
minutes allowing the analyzer to stabilize before they start to report TOC results.
Another reason membrane-based TOC analyzers may not be well suited to the CIP application is that
the chemical residue from the cleaning process may foul the membranes affecting their permeability
and thus the calibration of the analyzer. Similar to laboratory TOC analyzers, membrane-based
analyzers used in this application will require a calibration check using TOC calibration reference
standards before each use to ensure that the membranes have not been fouled or dried out. Thus
membrane-based analyzers used in CIP applications may increase cost of ownership through frequent
re-calibrations and replacement of the membranes.
Ideally, an on-line, direct conductivity analyzer that is designed to capture a small aliquot and quickly
provide a TOC and Conductivity result is best-suited for the majority of CIP applications. Ideally, once
the CIP process has been validated, the CIP process should be automated and the TOC & Conductivity
verification analyzer simply used to verify the final rinse step has reached the desired ‘acceptably
clean’ state at the end of each and every CIP program.
Validating the Application
Typically CIP processes are validated and verified using laboratory grab sample analysis. The
challenge of moving to on-line analysis may be compounded by the fact that grab samples will most
likely be contaminated by both the process of collecting the grab sample and the vessel into which the
grab sample is collected. Laboratory TOC analysis will therefore typically report much higher results
than an on-line TOC analyzer. However, in order to build confidence in the on-line TOC analyzer
results during the validation process, a traceable TOC standard can be run at the start and end of each
validation run in order to bracket the validation run and provide confidence that the analyzer was
running correctly at the start of the validation run and still running correctly at the end of the run.
The advantage of the on-line TOC analyzer is that it will report a more accurate picture of the actual
TOC levels present in the CIP run-off and give an early warning if the process is starting to trend out of
control before product is put at risk of contamination.
Validating the Application 6
ANATEL Clean-In-Place (CIP) Application Note
•••
CIP applications generally require frequent re-qualification on a regular basis to ensure that nothing
has varied, either in the process or the interaction between the process products and the cleaning
process. The use of automated CIP and on-line TOC analysis can provide trend data to support an
extended period between these re-qualifications, or even be used to eliminate the requirement for
periodic re-qualification.
Conclusion
CIP can reduce the risk of process equipment contamination by keeping the process equipment within
the cleanroom environment. In most cases, automation of a validated CIP cycle with confirmation of
final rinse water quality will deliver the most robust process.
TOC and conductivity combined can be used to confirm the process vessel final rinse water is clean of
all inorganic and organic matter down to WFI quality levels. TOC analyzers capable of reporting both
TOC and conductivity and using direct conductivity and only requiring a small sample aliquot are best
suited to this application. Thus, following the trend in the industry to move from a focus on final
product quality testing to the use of on-line quality testing instrumentation, the use of on-line TOC &
Conductivity verification analyzers for CIP verification offers to both increase confidence in the CIP
process quality and at the same time provide a tool to drive process improvement.
Author Biography
Author: Tony Harrison is a subject matter expert to the UK British Standards Institute (BSI) cleanroom
mirror group and one of the UK subject matter experts on ISO TC209 Working Group 01 tasked with
revising ISO 14644-1 and -2. He is also Convenor of ISO TC209 Working Group 02 tasked with the
revision of ISO 14698-1 and -2 for the classification of the cleanroom environment by microbiological
cleanliness. Tony is employed by Hach Company as a Senior Product Manager.
Hach Company – World Headquarters
Particle Counting Business Unit
P.O. Box 608
Loveland, Colorado 80539 USA
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Conclusion 7