eBook: TOC and Microbial Detection Monitoring - 37

ketter
Reduction
Process Control and Risk Reduction

ll, Product Manager, METTLER TOLEDO Process Analytics
7000RMS TAC Team Leader, METTLER TOLEDO Thornton

Peggy Banarhall, Product Manager, METTLER TOLEDO Process Analytics
Kumby Dhliwayo, 7000RMS TAC Team Leader, METTLER TOLEDO Thornton

rement Technique for the 7000RMS

Measurement Technique for the 7000RMS

tablished optical measurement techniques, Laser Induced Fluorescence (LIF) and Mie Scattering,
m in a unique manner to detect microorganisms in high purity waters

* Utilizes two established optical measurement techniques, Laser Induced Fluorescence (LIF) and Mie Scattering,
combining them in a unique manner to detect microorganisms in high purity waters

RMS

Sampling port
for Excited
trial data.
State
CFU reading.

A second 7000RMS was
connected to the same
sampling port with a
T connection feeding
both instruments.

y)

UV

Green Light
(lower energy)

Points
of
Use

Excited State

Blue Light
(high energy)

Heat
Exchanger

Green Light
(lower energy)

Blue Light

Ground State

r Induced Fluorescence): Molecule is excited
er energy level laser light source, then releases
gy by emitting ﬂuorescence

Ground State

Mie Scattering: Molecule absorbs light/photons and
scatters them at different distances based on their size

LIF (Laser Induced Fluorescence): Molecule is excited
to a higher energy level laser light source, then releases
that energy by emitting ﬂuorescence

Blue Light

Mie Scattering: Molecule absorbs light/photons and
scatters them at different distances based on their size

- An AFU spike (likely due to Points Of Use being
ntributing factor
flashed) around 5:30pm
August - 14 August
s a 405 nm laser diode to illuminate
a ﬂow cell
* 7000RMS uses a 405 nm laser diode to illuminate a ﬂow cell
* The diagram
shows a polishing effect not only during
ms pass through the ﬂow cell and
illuminated
by the laserbut
light,
ﬂuorescence
and Mie
* Microorganisms pass through the ﬂow cell and are illuminated by the laser light, ﬂuorescence and Mie scattering occur:
theare
time
of the sanitization
throughout
the time
the scattering occur:
e from NADH and Riboﬂavin water system was quarantined
- Fluorescence from NADH and Riboﬂavin
ng from particles and microorganisms
- Mie scattering from particles and microorganisms
- The lowest AFU reading during these days was
and Mie scattering signals are captured
onAFU/100ml
separate detectors,
and processed
through advanced algorithms
* Fluorescence and Mie scattering signals are captured on separate detectors, and processed through advanced algorithms
3,749
at 3:30pm
on 3 September
and
Mie signals
meet speciﬁc criteria
at same time,
a microorganism
present
- At midnight
on 4 indicates
September
the feed water iswas
re- and an Auto* Fluorescence and Mie signals meet speciﬁc criteria at same time, indicates a microorganism is present and an Auto
performed
in the
nit
(AFU)
is reported
Fluorescent Unit (AFU) is reported
introduced to the AWFI in preparation for use in the
anges
were
made

8 AFU/100ml
n for over 48 hrs

water system the next day. This is when the AFU
spiked to ~85,000 AFU/100ml.

Conclusions
that the bacteria in the water
did not
* Root
cause determination
2)
 The UV lamp installed in the loop was lethaltrial
to theorized that the bacteria in the water did not
SA orfrom
R2A6am
plates
- It was determined after
the data from
the
grow
oop
onbecause:
1
thereviewing
bacteria (formation
of Thymine
dimers in
the on the TSA or R2A plates because:
a were VBNC. USP <1231> "...Howtrial that while biofilmbacteria
might have
a contrib The bacteria were VBNC. USP <1231> "...HowDNA been
that would
not allow any bacteria
onal
limitations
of the growth media
utor to the high AFU counts,
the water
feeding
thewere unable to grow
ever, nutritional limitations of the growth media
s to the
water sysfrom dividing
therefore
they
isfy
AWFI was most likelyon
theR2A
rootorcase
to Athe
high AFUwill continue to be
may not satisfy growth requirements of organisms
weregrowth
loggedrequirements of organisms
TSA).
bacterium
heOfwater
system that originated counts.
from a
in the water system that originated from a
Use flashing)
counted as an AFU until the cell death processpresent
is
a result, traditional cultural* methods
Limitation of plate counttriggered
method and the concentration of metabolite withbiofilm. As a result, traditional cultural methods
etect
fraction
of the biofilm -bacteria
All the results from the
port (see
diagram
med afrom
4:00pm
in sampling
the bacterium
is below
detectable levels. may only detect a fraction of the biofilm bacteria
he
water sample"
1) were 0 cfu.
microbiologist
involved in the
present in the water sample"
September
* The
Process
control benefits
- The passivation and hot water sanitization had been
ineffective in remediating the intermittent contamination events because the customer did not have
true process understanding. By using the 7000RMS
the customer was able to identify that the feed water
was the root cause of their intermittent contamination challenges in 48 days. Future remediation activities will be targeted and effective because of better
process understanding.
50 hr. Sanitization
* Financial benefits and≈ business
justification
- By using the 7000RMS Customer A will be able to
save money by:
Flashing
 eliminating
Point ofcostly,
Use ineffective remediation activities
 reducingports
downtime due to contamination and ineffective remediation activities
Water system
frequenquarantined  cost reduction by optimizing sanitization
Flashing
cy because of continuous, real-time,
reliable
Point
of Use data
Water system
Water system
* Next Steps
ports
uarantine ends
quarantine ends
- The customer plans to purchase 2 - 3 7000RMS
/100 ml
3,749 AFU/100 ml
analyzers in 2020 to install in critical water systems
in their manufacturing facility for True Water System
Surveillance for Better Process Control and Risk
Reduction.

Figure 2

10/11/19 12:22 PM

37

 The UV lamp installed in the loop was lethal to
the bacteria (formation of Thymine dimers in the
bacteria DNA that would not allow any bacteria
from dividing therefore they were unable to grow
on R2A or TSA). A bacterium will continue to be
counted as an AFU until the cell death process is
triggered and the concentration of metabolite within the bacterium is below detectable levels.
* Process control benefits
- The passivation and hot water sanitization had been
ineffective in remediating the intermittent contamination events because the customer did not have
true process understanding. By using the 7000RMS
the customer was able to identify that the feed water
was the root cause of their intermittent contamination challenges in 48 days. Future remediation activities will be targeted and effective because of better
process understanding.
* Financial benefits and business justification
- By using the 7000RMS Customer A will be able to
save money by:
 eliminating costly, ineffective remediation activities
 reducing downtime due to contamination and ineffective remediation activities
 cost reduction by optimizing sanitization frequency because of continuous, real-time, reliable data
* Next Steps
- The customer plans to purchase 2 - 3 7000RMS
analyzers in 2020 to install in critical water systems
in their manufacturing facility for True Water System
Surveillance for Better Process Control and Risk
Reduction.

10/11/19 12:22 PM
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