THORNTON

Leading Pure Water Analytics

Pharmacopeia Requirements for

White Paper
Pure Steam and Final Rinse Verification of CIP

Drinking Water

Validation of Pure Steam and the verification of the

final aqueous rinse in a CIP process are defined by
regulatory standards, which change periodically, and
challenge system designers and owners to know the
latest requirements. This white paper contains an
overview of current pharmacopeia requirements,
which will assist in the design and monitoring of
compliant systems for Pure Steam production
and CIP.
The first section of the paper details the requirements
for Pure Steam. The second section concerns the
requirements for validating the final rinse of a CIP
process which may have been performed using Pure
Steam, pharmaceutical grade water, or by chemical
cleaning. Regardless of the method used, all require
an aqueous final rinse and verification.
Pure Steam – pharmacopeia requirements
In 2006, the United States Pharmacopeia (USP) published a
monograph in USP 29 Supplement 1, detailing the testing
requirements for Pure Steam. The monograph details the
regulations, and it also explains the lethal properties of steam,
which addresses the absence of microbial testing. For testing
purposes, the condensate of the steam must be tested by the
end-user for the specific parameters listed. Pure Steam, in
simpler terms, is vaporized water with condensate equivalent to
WFI. In USP 38 (official May 1, 2015), the same requirements
are found.
Purified Water Water for Pure Steam
Injection
CIP
Final
Rinse
Figure 1 - Acceptable water sources for Pure Steam production.
Feedwater for Pure steam is drinking water quality, at minimum.
The requirements for production of Pure Steam:
The requirements are clearly defined as follows:
1. P ure Steam is produced from source water that meets the same
source water requirements for Purified Water and Water for
Injection, i.e., drinking water that meets the standards
established by the United States Environmental Protection
Agency National Primary Drinking Water Regulations (US EPA
NPDWR), or the drinking water regulations of the European
Union, Japan or World Health Organization (WHO). (See Figure
1) Please note that Pure Steam may be prepared from drinking
water but the quality, specifically the hardness of the drinking
water, may increase the maintenance of the Pure Steam
generator; therefore, Purified Water or Water for Injection can
be used as the source water for Pure Steam also.

Pure Steam and CIP
2 METTLER TOLEDO White
2. Pure Steam contains no added substances. This so-called
“no added substances” rule requires that if you add any
chemical or compound to the water during purification
you must show you have removed it before it is used. In
this case, the “no added substances” is intended to prevent
the use of corrosion inhibitors and other additives that are
often used in “plant steam”.
3. P ure Steam is water that has been heated above 100°C and
vaporized in a manner which prevents source water
entrainment. Entrainment occurs when the boiling rate of
the water is violent. This water is still in the liquid phase,
i.e., it has not undergone the phase change to vapor and is
pulled over with the steam to the condensate.
4. T he level of steam saturation or dryness, and the amount
of non-condensable gases need to be determined by the
Pure Steam application.
The testing and validation of Pure Steam:
USP 29, Supplement 1 continues: “Pure Steam quality is
difficult to assess in its vapor state; therefore, the attributes
of its condensate are used to indirectly test its quality. The
process used to create and collect the condensate for analysis
must not adversely impact these quality attributes.” The tests
for Pure Steam are:
1. B acterial Endotoxins <85>: condensate contains less than
0.25 Endotoxin Unit EU/mL.
2. Total Organic Carbon <643>: condensate meets the
requirement.
3. W ater Conductivity <645>: condensate meets the
requirement.
There is no microbial test required for Pure Steam because
any water borne bacteria will have been killed at the elevated
temperatures that are used to prepare steam. Water raised to
temperatures above 65˚C is lethal to water borne bacteria.
This is described in USP general information chapter
<1231> Water for Pharmaceutical Purposes: “Owing to the
lethal properties of Pure Steam, monitoring of microbial
control within a steam system is unnecessary. Therefore,
microbial analysis of the steam condensate is unnecessary.”
Paper
In addition, the USP Pure Steam monograph discusses other
characteristics of Pure Steam and its use and application, which
are determined and defined by the user. There is no regulation
for the degree of saturation or moistness of the steam, but the
monograph does explain some of the issues with steam that may
be superheated or too dry. USP <1231> continues: “Steam
attributes not detailed in the monograph, in particular, the
presence of even small quantities of non-condensable gases or
the existence of a superheated or dry state, may also be important
for applications such as sterilization.” Again, in USP <1231>,
“The large release of energy (latent heat of condensation) as
water changes from the gaseous to the liquid state is the key to
steam’s sterilization efficacy and its efficiency, in general, as a
heat transfer agent. If this phase change (condensation) is not
allowed to happen because the steam is extremely hot and in a
persistent superheated, dry state, then its usefulness could be
seriously compromised.”
The monograph defines the test methods but permits the user to
determine the degree of saturation or moistness of the steam for
their specific application. This is important because ‘dry’ steam
or superheated steam will not have the same lethal properties as
moist steam when used for sterilization of distribution systems,
especially the valves. If the steam is dry when it condenses it will
not effectively sterilize the backside of valves or manifolds.
This USP monograph for Pure Steam was made official to clarify
industry questions and finally define Pure Steam and how to
validate its purity. Pure Steam is used in a pharmaceutical
facility in a number of applications, including sterilization of
vessels, containers, filling equipment and piping; and the
manufacture of certain products.
With the definition of the Pure Steam test limits and the source
water mandate to produce the Pure Steam, pharmaceutical
facilities can now establish an SOP for the verification of the
quality. The only test that cannot be performed on-line or at-line
is the bacterial endotoxin test, which still requires an off-line
Limulus Amebocyte Lysate (LAL) test to be conducted. The
conductivity and TOC tests can be performed on-line and at-line,
saving the end-user time, money and eliminating sampling
errors and delays. Pure Steam conductivity can be tested directly
 in the condensate to validate that it meets USP <645> limits and terms of strength and time and temperature. This ensures the
Pure Steam and CIP

the TOC can be tested with an at-line connection meeting the
requirements for USP <643>.
Validation of the final rinse after the Clean in
Place process
There are several types of cleaning methods utilized for the
Clean-in-Place (CIP) process, including; water (WFI, Purified
Water, other water [often hot but not required], steam (SIP),
chemicals (caustic soak, acid soak, water rinse) and heat. In the
prior section of this paper, the control and verification of Pure
Steam was detailed, and for some cleaning applications Pure
Steam will be the cleaning agent. For other cleaning methods
utilizing aqueous or chemical cleaning, the final rinse must be
validated.
CIP is a vital procedure across the pharmaceutical industry. The
most critical step in the CIP process is the assurance that there is
cleaning agent accomplishes cleaning the system (bioreactors,
fermenters, vessels, filling equipment, etc.) of microbes and prior
chemicals. The next critical step is that the system has been
sufficiently rinsed. This is simply accomplished by measuring the
key chemical impurities in the quality of the water exiting the
system during the final rinse step. Along with the chemical (or
hot water filling process), this confirms that the previous steps in
the process have achieved the desired goals and allows equipment
to be returned to service. The pharmacopeias have issued
guidelines for the validation of the final rinse.
“In general, the final rinse used for equipment,
containers/closures should use the same quality of
water as used in the final stage of manufacture of the
API or used as an excipient in a medicinal product.”
Evaluation of Medicinal Products

proper contact of the cleaning agents (chemical and water) in (European Medicines Agency, EMA), See Table 1

Table 1

Guidance for CIP Final Rinse

Cleaning/Rinsing of Equipment, Product Type Minimum Acceptable
Containers, Closures Quality of Water
Use same quality of water as used in the
Final rinse API
API Manufacture
Purified Water or use the same quality of
Final rinse including CIP of
Pharmaceutical water as used in the manufacture of
equipment, containers and closures,
products – non sterile medicinal products, if higher quality than
if applicable
Purified Water
Initial rinse including CIP of
equipment, containers and closures, Sterile products Purified Water
if applicable
Purified Water or use the same quality of
Final rinse including CIP of
Sterile non-parental water as used in the manufacture of
equipment, containers and closures,
products medicinal product, if higher quality than
if applicable
Purified Water
Final rinse including CIP of
Sterile parental
equipment, containers and closures WFI
products
if applicable

METTLER TOLEDO White Paper 3

 A significant bottleneck for the pharmaceutical industry is the
Pure Steam and CIP
validation of the final rinse for CIP processes. The time involved
in off-line tests results in delays in restarting production (or re-
starting at risk) and therefore a significant loss of production
uptime. Indeed, more than 60% of a vessel’s downtime can be
attributed to cleaning and delays in validation testing. Lab
testing of grab samples is not simply time consuming, it also
carries risks of sample contamination and false test results that
can lead to additional production downtime and delays in
bringing the system back on-line.
In order to ensure a CIP process has achieved its aim, some
facilities may perform multiple rinse cycles using pharmaceutical
grade water. This not only contributes to lost production uptime,
it is also a waste of expensive water. Further, if a previous step in
the CIP cycle (e.g., an acid solution wash) has not been
conducted correctly, validation of the final rinse may not be
achieved.
On-line analytical measurements provide continuous, real-time
determination of not simply water exiting vessels, but ensure
each step in a CIP process will be or has been successfully
conducted.
The CIP validation challenge
The quality of the water used for the final CIP rinse must be the
same quality (or better) as the quality of water used in the
production process. If a production process requires the use of
Water for Injection (WFI), it must be WFI compliant water that is
used in the final CIP rinse. And, as mentioned above, for the CIP
process to be verified and equipment returned to production, it
must be WFI compliant water that exits the vessel at the end of
the final rinse step. In essence, validation of a CIP process can be
summarized as water quality out must equal water quality in.
This is a requirement not only for vessels, but all equipment,
containers and closures that are in contact at any time with
manufactured products.
“Water used for final rinsing should have an equivalent
quality to that of water used for manufacturing drug
products.”
Japanese Pharmacopeia 16 guidelines
4 METTLER TOLEDO White Paper
“The purpose of cleaning validation is to demonstrate
that a particular cleaning process will consistently clean
the equipment to a predetermined limit; the sampling
and analytical test methods should be scientifically
sound and provide adequate scientific rationale to
support the validation”.
U.S. Food and Drug Administration
Although the requirements regarding CIP final rinse water
quality are clear, the challenge to pharmaceutical companies is
that they are responsible for devising and documenting their own
CIP processes. Step-by-step instructions on formulating and
confirming CIP are not described in the pharmacopeia or by
vessel/instrument manufacturers. This is because the cleaning
processes will depend on the chemicals and processes used for
production. These are aspects which are not within the
responsibilities of equipment manufacturers, nor the FDA, EMA,
etc. Pharmaceutical companies must therefore develop their own
CIP processes that are not only effective, but that allow rapid
validation in order to maximize equipment use.
Basic premise of CIP and rinse validation
CIP and rinse validation have been developed to protect product
and production processes by preventing potential chemical or
biological contamination and killing microbes. This can be done
by ensuring that the quality of water exiting the vessel equals the
quality of water entering the vessel and prevent harm to the
product or process system.
How do you create a cleaning process?
A cleaning process must be defined and validated by the user/
owner of the process for the vessel. The system owner must
determine the appropriate cleaning process - not the FDA, not the
USP or any of the other pharmacopeia, and not the instrument
manufacturers.
For a vessel to be cleaned and rinsed using water:
The assumption is that the cleaning waters shall be equal or
better, such that the “quality of (cleaning) water is appropriate
and suitable for intended use”. Decisions to use WFI, Highly
 Purified Water or Purified Water for cleaning and rinsing grab samples are eliminated, resulting in fewer errors, reduced
Pure Steam and CIP

purposes and whether the cleaning water is hot or cold depends
on the application and is determined by the owner. However,
cleaning criteria should not be based on “this is how we have
done it in the past”. Conductivity and TOC, used as real-time,
on-line measurement tools, ensure that the cleaning process is
based on scientific data and can be validated.
User objectives for CIP
Users must monitor water quality during rinsing to verify the
water quality to ensure it is compliant. Cleaning must be verified
prior to the re-introduction of the system back into the process.
The owner must restore the system to production as quickly as
possible to minimize downtime and the risk of contamination.
Why use a portable TOC measurement?
Measurement results from a portable TOC analyzer are obtained
in seconds, not hours. Conductivity and TOC can be checked or
verified simultaneously. Sampling errors or contamination from
testing time and lower costs.
European Pharmacopeia (EP) guidelines for CIP
final rinse
As described in Table 1 (above), the EP (and other
pharmacopoeia) define the quality of water to be used in initial
and final rinses. One methodology to ensure that the proper
water is being used is to measure the quality of the water
entering and exiting the vessel, and ensure that its conductivity
and TOC impurities are at acceptable levels, particularly for final
rinse.
As Purified Water (PW) and WFI are specified (See Table 2), it
therefore follows that water quality must meet or exceed these
specifications per the USP, EP and JP pharmacopeia regulations.

Table 2

Pharmacopeia Regulations for WFI and PW

Attribute Water for Injection Purified Water
(WFI) (PW)
Production Method Distillation Suitable process
US, EU, Japan, WHO US, EU, Japan, WHO
Source Water
drinking water drinking water
Total Aerobic (cfu/100 mL)2 10 NA
Total Aerobic (cfu/mL)2 NA 100
1.3 (Stage 3) 25°C
Conductivity (μS/cm at 25°C) 1.3 (Stage 3) 25°C
EP 5.1 (Stage 1)
TOC (mg/L) 0.5 0.5
Bacterial Endotoxins (EU/mL) 0.25 NA
Nitrates (ppm) – EP only 0.2 0.2
Bacterial Endotoxins (EU/mL) NA < 0.1 mL 0.02 KMnO4
Heavy Metals NA 0.

METTLER TOLEDO White Paper 5

 Figure 2: A possible CIP process showing examples of
The application of TOC and conductivity
required conductivity and TOC levels.
Pure Steam and CIP

measurements for CIP final rinse verification

Cleaning methods employed as part of routine process equipment
Typical Cleaning Phases
maintenance vary nearly as much as manufacturing processes
not recirculated
do (see Figure 2). Recommending monitoring instrumentation Pre-Rinse run time: 3-5 min
and sensors can be simplified by ensuring the sensors used are temp: 30-40 °C

appropriate for the cleaning process. CIP may be limited to an run time: 15-20 min
aqueous rinse, or may require more aggressive cleaning such as Alkaline Wash temp: 70-90 °C
Cond.: 140-200mS
high-strength acidic and caustic solutions. Once the initial
cleaning cycle is performed, a series of water rinses with water run time: 3-5 min
Rinse temp: 30-40 °C
that is suitable for the intended use of the vessel will complete the
cleaning procedure. This range of conditions requires multiple
run time: 15-20 min
sensors for accurate measurement over the complete cleaning Acid Wash temp: 60-80 °C
cycle. Cond.: ~20mS

run time: 3-4 min

Measuring and regulating CIP systems to ensure
process control, achievable cleaning quality and data
to verify the cleaning
“PAT – A Framework for Innovative Pharmaceutical
Development, Manufacturing and Quality Assurance”
states: “Reducing production cycle times by using on-,
in-, and at-line measurements and controls.”
FDA Guidance on PAT
In recent years, the FDA has strongly encouraged the use of the
Process Analytical Technology (PAT) initiative. PAT is a system
for designing, analyzing and controlling manufacturing through
timely measurements of critical quality and performance
attributes of raw and in-process materials and processes with the
goal of ensuring final product quality. In other words, it
advocates use of an engineering focus with in-process
measurement tools and controls while products are being
manufactured.
By designing conductivity measurements into the CIP system,
the concentration of alkaline and acid wash can be monitored
and controlled at the proper concentrations for complete
cleaning. Continuing with this approach, the use of conductivity
and TOC to monitor the final rinse will permit validation of the
water quality and reduce system downtime.
Rinse temp: 30-40 °C
not recirculated
run time: 3-4 min
Rinse with high temp: ~80 °C
quality water Cond.: <2.7 μS (pharma)
TOC: 500ppb (pharma)
Detailed below is a description of how to include conductivity
monitoring into CIP processes and the use of conductivity/TOC
for final rinse validation.
On-line or in-line conductivity measurement has a number of
specific applications in CIP. Conductivity is used to monitor
concentration levels of the cleansing agent to indicate the need
for replenishment. Product water separation in the reverse
cleansing flow (phase separation in the return line) can be
determined quickly and controlled using conductivity.
In addition, conductivity is used to determine the end-point of
the cleaning process, when the caustic or acid solutions have
been totally rinsed away in the process itself, with the sensor
connected to the same transmitter used for the CIP system. And
finally, conductivity is used to verify cleaning chemicals are
removed.

6 METTLER TOLEDO White Paper

 TOC and conductivity measurements have other functions and can be accomplished using the three steps below:
Pure Steam and CIP

benefits, which include verification that the last product batch is

completely removed. Also, both parameters are valuable to
measure and verify that WFI and PW water quality discharged is
equal in quality to the water quality input to the system.
However, limitations of TOC measurement should be noted.
Do not measure TOC with chemicals in process, to measure
concentration of cleaning chemicals or to verify microbial
control.
Selecting the right sensors
Defining the instrumentation required to monitor the CIP cycle
1. D
 etermine the conductivity and chemistry of the cleaning
solution.
2. Select a conductivity sensor appropriate for the cleaning
solution (a 4- or 2-electrode sensor).
3. I f a 4-electrode sensor is required for the cleaning solution,
select the appropriate 2-electrode sensor to:
a. A ct as an intermediate monitoring step prior to initiating
flow to TOC
b. Provide final conductivity verification of the cleaning
process

Table 3: A guideline for the conductivity and TOC transition point monitoring for cleaning and rinsing.

Sensor Starting Conductivity Transition Point

4-Electrode Conductivity >500 µS/cm ~500 µS/cm
2-Electrode Conductivity ~500 µS/cm 2 µS/cm

Total Organic Carbon (TOC) 2 µS/cm N/A

Figure 3: An example of a system design for utilization of PID control for conductivity and TOC.

2-electrode 4-electrode
Conductivity Conductivity
Sensor Sensor

Flow
Restrictor

Relay Control Signal

Signal Cable
Sample Flow Line Drain Funnel

METTLER TOLEDO White Paper 7

 An easy to manage solution
Pure Steam and CIP
The instrumentation used for this application should be based on
what is readily available. When conductivity instrumentation is
already in place, the user must ensure sample flow is initiated to
the TOC sensor only when conductivity has lowered to an
acceptable level (the transition point).
This sequence can be easily managed using a METTLER
TOLEDO M800 Transmitter to control two conductivity sensors, a
5000TOCi Total Organic Carbon Sensor, and two solenoid valves
controlled using set-point activated relays on the M800. See
Figure 3 for the schematic of a basic system to provide indication
for a complete cleaning process. In this case, the 4-electrode
sensor monitors the effluent chemicals. As the chemicals are
eventually rinsed to an acceptable level, a 3-way valve redirects
the effluent to a high purity 2-electrode sensor. As the effluent
continues to be rinsed with high purity water, the conductivity
continues to lower. When the conductivity lowers to the transition
point (2 uS/cm, in this case - see Table 3), a second valve
redirects flow to the TOC sensor. The 4- and 2-electrode
conductivity sensors protect the TOC instrument from chemical
attack, and the 2-electrode conductivity sensor and TOC sensor
ensure that the conductivity and TOC limits are met.
The 5000TOCi is well suited for this application because its
automatic flow control compensates for varying inlet pressures,
and additional control functions allow the 5000TOCi to start TOC
measurement as soon as sample flow begins.
The 4-channel M800 transmitter is ideal for monitoring all
necessary sensors and provides set-point and relay functions for
sample flow control throughout the cleaning process. Additional
set-point, relay, and analog signals provide indications to
external control systems.
Mettler-Toledo Thornton, Inc.
900 Middlesex Turnpike, Bldg. 8
Billerica MA, 01821 USA
Phone +1 781 301 8600
Fax +1 781 301 8701
Toll-free +1 800 510 PURE (US & Canada only)
thornton.info@mt.com
Subject to technical changes
© 03/15 Mettler-Toledo Thornton, Inc.
58 087 042 Rev A 03/15
Complete control over CIP and Pure Steam validation
The M800 utilizing multiple conductivity sensors (4-electrode
and 2-electrode) and the 5000TOCi Sensor provide a complete
package to control the cleaning and final rinse validation. For
facilities using an aqueous cleaning and rinse only, and where
there are multiple vessels that must be validated, the Portable
450TOC Analyzer provides the capability to validate the TOC and
conductivity levels of the final rinse at the same time.
For validation of Pure Steam, the M800 plus a UniCond®
conductivity sensor (2-electrode) and 5000TOCi provide a
dependable solution. The Portable 450TOC allows validation of
TOC and conductivity levels of Pure Steam if the facility has
multiple steam condensers or multiple sampling points.
Conclusion
The USP has established the requirements for Pure Steam,
whether used in production or as part of a sterilization or CIP
process. On-line and at-line conductivity and TOC measurements
provide continuous measurements of steam condensate and
hence Pure Steam quality. The verification of CIP final rinse
water quality has been a challenge for the pharmaceutical
industry and can easily delay equipment being put back in
service, resulting in lost production uptime. The regulatory
agencies have issued guidelines for CIP processes, and
pharmaceutical facilities are challenged to meet these
requirements. Using the basic tenet of quality by design, the
inclusion of analytical process controls can reduce delays,
eliminate sampling errors and lower the cost of cleaning and
validating the process.
METTLER TOLEDO Thornton’s portfolio of advanced
transmitters, conductivity sensors and fixed and portable TOC
sensors offers convenience and high performance for the
monitoring and control of all CIP processes.
4www.mt.com/ism-pharma
www.mt.com/thornton
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