eBook: TOC and Microbial Detection Monitoring - 35

Real-time Microbial Detection Enables True Water System
Real-timefor
Microbial
Detection
Enables
Water System
Surveillance
Better Process
Control
and RiskTrue
Reduction
Abstract
Overview: METTLER TOLEDO Thornton continues
to be a leader in pure water analytics, manufacturing key products such as conductivity, Total Organic Carbon (TOC) and Ozone for real-time monitoring of pharmaceutical water systems. Real-time
monitoring of microbial (bioburden) activity from
regulated pharmaceutical water systems is today a
reality, offering the means to improve product
quality, reduce risk, increase process understanding and impact product safety.

Case Study
prior to water use so that the user can react immediately to out of specification trends reducing regulatory risk and financial loss.
Objective: Current compendial plate count testing methods allows the assessment of microbial water quality 5-7 days after sampling, limiting
the opportunity for any true understanding of the

actual bioburden profile in the water loop and reducing the ability to react to an out of specification event in
a timely fashion. Using a NEW case study, this poster
will highlight how a company using real-time microbial monitoring dramatically reduced the microbial risk
to their manufacturing process by gaining superior understanding of their water system that was not possible
with the plate count method.

7000RMS Real-time Monitoring Beneﬁts

This poster presentation uses NEW case study data
to discuss the use of a real-time water microbial
analyzer in real world pharmaceutical water systems,
demonstrating their effectiveness in reducing sample
analysis time, better understanding of microbial control activities and the risks associated with compendial plate samples. Results are available in seconds,
enabling the user to visualize water system microbial
activity. This real-time information can be used to
assess the effectiveness of sanitization, provide continuous, real-time information on the health of the
loop and rapid response to microbial excursions.

On-line, real-time microbial analyzers:
* Provide better process control
* Continuously monitor pharma water, enabling faster
response to bioburden excursions
* Assess the effectiveness of sanitization and understanding
the health of a water loop.
* Use the data to demonstrate the limitation of the plate count
method for process control
* Real-time process surveillance ensures water system
control, increasing product safety
* On-line testing enables reduced plate sampling and labbased testing

Real-time water system surveillance increases process understanding so microbial quality is known

Stages of bioﬁlm build up
Bioﬁlm is everywhere, because it's one of the survival
and proliferation mechanisms for bacteria. It can be
particularly problematic in pharmaceutical ultrapure
water systems. The USP deﬁnes bioﬁlm as "a threedimensional structured community of sessile microbial cells embedded in a matrix of extracellular polymeric
substances (EPS)."
Attachment: Bioﬁlm formation starts when
free-ﬂoating or planktonic bacteria ﬁnd an appropriate surface to adhere to and release a slimy
material called extracellular polymeric substance (EPS).
This process takes seconds.

1

Formation: The bacteria continue to release EPS
to strengthen their attachment to the surface, and
form a bioﬁlm structure that is virtually impossible
to remove. This process takes seconds to minutes.

2

Note: Attachment and formation can occur in less than
an hour. Therefore, a WFI and PW water system that is
shut down for at least an hour for maintenance can go
through attachment and formation at multiple sites of the
water system loop.

3

Development/growth: Having established a new
home, the bioﬁlm can start to grow. This stage
can take hours to days.

This is the phase where most bioﬁlm in ultrapure water
exists. The purpose of sanitization and continuous water

A pharmaceutical company was
sues with their water system. Th
possible root cause, so they dec
on a trial basis.
* 7000RMS analyzer was in
Ambient Water for Injection
* The AWFI was passivated 1
* The water loop was sanitiz
with hot water (~80°C)
* Evaluation period: 70 days
15 days on a PW water sy
* The data from the trial was

19 July - 31 July
Baseline: 145,451 AFU/100ml
CFU average: 0 cfu
* The high AFU readings led
ond analyzer to run side-b
* A drop in AFU number was
- The microbiologists invo
that the assumed biofilm
its dispersal phase and
planktonic bacteria

1 August - 14 August (See figure
Baseline: 115,436 AFU/100ml
CFU average: 0 cfu
* A second control 7000RMS
256 AFU/100ml at the MET
installed side-by-side at th
trial 7000RMS for 36 hrs
- Both analyzers were read
- After the side-by-side test
was reinstalled on the sa
TLER TOLEDO facility and
the previous baseline AFU

With confidence in the AFU coun
cess decision to increase the sa
11 August and 8 hrs on 12 Aug

1

2

3

ﬂow is not to eliminate bioﬁlm, but to manage the growth
phase.
Sanitization: weakens the EPS to allow the water ﬂow to
gradually scrape off layers, exposing bacteria to harsh
conditions or diffusing through the EPS to kill bacteria in
the bioﬁlm.
Continuous water ﬂow: keeps planktonic bacteria moving, making it difﬁcult for them to attach to a surface, and
slows bioﬁlm formation or maturation.
USP <1231> 5.3.1 "Although thermal methods control biofilm development by either continuously inhibiting its growth or, in intermittent applications, by
killing the microorganisms within developing biofilms, they are not effective in removing established
biofilms. Killed but intact biofilms can become a nutrient source for rapid biofilm regrowth after the san-

PDA 2019 Poster_7000RMS.indd 1

35

4

5

itizing conditions are removed or halted. In cases
of infrequent thermal sanitizations that allow biofilm
development between treatments, a combination of
routine thermal treatment and periodic supplementation with chemical sanitization may be more effective. The more frequent the thermal sanitization, the
more likely it is that biofilm re-development can be
eliminated."

4

Maturity: A well-established, happy, and healthy
bioﬁlm structure.

Release: Like any living organism, after survival
and growth, bioﬁlm's next stage is proliferation. A
mature bioﬁlm in a water system will intentionally
and strategically release planktonic bacteria into the water
system. The released planktonic bacteria will in turn ﬁnd an
appropriate surface to attach to, and start making new bioﬁlm structures.

5
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