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7000RMS Bioburden Analyzer
ISPE
Sep 2016
Real Time Microbial Detection
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Agenda
1 Introduction Thornton and 7000RMS
2 Why On-line Microbial Monitoring
3 7000RMS Technology and Key Features
4 Test Points for 7000RMS
5 Pharmacopeia acceptance of Alternative Methods
6 Validation Guidelines
2
THORNTON – Leading Pure Water Analytics
1964 Dr. Richard Thornton, an MIT professor, founded Thornton Inc.
2001 Acquired by Mettler-Toledo and integrated into Process Analytics Division
Widely considered as the leader in the production of high quality
resistivity / conductivity sensors and the Thornton/Light Curve
Long history of community marketing successful at establishing brand
recognition at key international organizations such as USP (A.
Bevilacqua), EP, JP, FDA, SEMI & PV (J. Cannon) and ASTM (D. Gray)
Strong working relationships at system fabricators
3
THORNTON Innovation Highlights
Long Tradition in Pure Water Analytical Measurements
1989 1948 1964 1992 2000 1996 1994 2003 2005 2008 2010 2007
Multiparameter
2-Channel
Portable TOC Concentric Ti
C/R Cell for
UPW
Sodium and
Silica Analyzers
1st TOC
Sensor
Ozone
Sensor
UniCond®
Multiparameter
4/6 Channel
4000 TOC
2012 2013
1st Integrated
Multiparameter
Instrument
Sanitary C/R
Sensor
Cost Effective
2-Channel
Broad-range 4-e
Conductivity
Sensor
Pure Water pH
Sensor
M300 ½
Channel
M800
Transmitter and
TOC 5000i
Pure Water
Optical DO
Ozone Digital
ISM sensor
2015 2014
Microbial
Analysis
CIP
Monitoring
4
Goals of the Introduction
7000RMS is a "game changer" for microbial detection in pure water
Past: Measuring the bioburden in Purified Water
and Water for Injection has almost exclusively
depended on time-consuming and error-prone
culture-based lab measurements.
Now: Our new 7000RMS analyzer offers
accurate, continuous, on-line determination of
microbial and inert particle contamination in
pharmaceutical water systems.
Introduce the new 7000RMS
(Real Microbiology System)
for Real-Time Bioburden
Detection
5
Value Proposition
7000RMS answers the market need for real-time bacterial detection
Don't miss a bug! With real-time microbial detection.
Don’t Miss A Bug!
Continuous Measurement
Compliant with USP Recommendations/Regulations
Economic On-line Alternative to Sampling
6
A few of our new best friends 7
To shift Highlight
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Agenda
1 Introduction
2 Why On-line Microbial Monitoring
3 7000RMS Technology and Key Features
4 Test Points for 7000RMS
5 Pharmacopeia acceptance of Alternative Methods
6 Validation Guidelines
8
Why On-line Microbial Monitoring?
Off-line method of bioburden
detection:
Unreliable results (human errors)
High risk of false-positive results
Costly
Time consuming
Delay of product release
On-line method of bioburden
detection:
Reliable, real-time results
Allows real-time release of water
Eliminates sampling error
Cost-effective
No delay of product release
9
Drivers for Alternative Methods
Global organizations support development of on-line bioburden analysis
Online Water Bioburden Analysis
Workgroup
- Established as an Industry
workgroup to promote
development and
implementation of online
bioburden analyzers
- Members from Merck,
Novartis, Amgen, Fresinius,
Baxter, P&G, Roche, Sanofi
and Pfizer
OWBA FDA
FDA PAT (Process Analytical
Technologies) Initiative
FDA Aseptic Processing
Guidance - cGMPs
FDA Strategic Plan for
Regulatory Science
FDA Senior Microbiologist
Supports RMMs
10
Traditional Microbial Detection
A colony-forming unit (CFU) is a unit used to estimate the
number of viable bacteria or fungal cells.
Counting with CFU’s requires culturing the microbes and counts
only viable cells.
Duration and temperature of incubation are also critical aspects of
a microbiological test method.
Classical methodologies using high-nutrient media are typically
incubated at 30°–35° for 120 – 168 hours.
Because of the flora in certain water systems, incubation at lower
temperatures (e.g., 20°–25°) for longer periods (e.g., 7–14 days)
can recover higher microbial counts when compared to classical
methods.
Low-nutrient media are designed for these lower temperature and
longer incubation conditions (sometimes as long as 14 and up to
21 days to maximize recovery of very slow-growing oligotrophs or
sanitant-injured microorganisms), but even high-nutrient media can
sometimes increase their recovery with these longer and cooler
incubation conditions.
11
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Agenda
1 Introduction
2 Why On-line Microbial Monitoring
3 7000RMS Technology and Key Features
4 Test Points for 7000RMS
5 Pharmacopeia acceptance of Alternative Methods
6 Validation Guidelines
12
7000RMS - Features
Instant AFU Counts
Real-time display of microbial
contamination
Easy operation
No sample preparation or reagents
required.
Touchscreen
At-Line or Grab Sample
Wide measurement range
Dependable measurements from 1
cell / 100 mL to 2000 / mL.
Multiple communication options
SCADA connectivity with ModBus TCP
Multiple analog outputs
Ethernet standard RJ45/Wi-Fi capable.
13
7000RMS – Product Specifications 14
More 7000RMS Benefits
Higher product quality at a lower cost The 7000RMS eliminates the need for routine lab
testing, with no sample preparation required. Instant
notification of microbial excursions allows for rapid
remediation, preventing product from being affected.
The reduction in lab measurements and savings in
energy through optimized sanitization cycles results in
markedly lower operating costs
Reliable optical technology Light-induced fluorescence (LIF) and sophisticated
algorithms are used to detect and quantify microbes
and inert particles. Particle sizing is achieved using Mie
scattering. The 7000RMS is also able to measure
organisms that can be missed in growth-based
methods
Easy to use throughout your facility Whether in your RO system, pure water storage or
distribution loop the 7000RMS can be utilized on-line or
off-line. It can also be used in the laboratory for rapid
testing of grab samples. The analyzer's touchscreen
interface displays all critical data and controls in a user-
friendly manner
15
Metabolites Fluorescence Spectra for 7000RMS
By detection of these two metabolites we can identify viable bacteria and do
not need to fluoresce other metabolites
Fluorescence of metabolites with
the 405nm laser used in
7000RMS
(Hill et al, Field Ana. Chem. & Tech, 3(4-5), 221,1999)
405nm Laser in 7000RMS
16
16
Flowchart
Online / sample mode
17
Inside 7000RMS 18
Example of 7000RMS User Interface 19
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Agenda
1 Introduction
2 Why On-line Microbial Monitoring
3 7000RMS Technology and Key Features
4 Test Points for 7000RMS
5 Pharmacopeia acceptance of Alternative Methods
6 Validation Guidelines
20
Bacterial detection as an At-Line measurement
Bring the Microbiology Lab to the System – eliminate sampling !
In-Line
- inside pipe, real-time
- conductivity, pressure, flow
At-Line (Side stream)
- attached to pipe, real-time,
flow to drain
- Microbiological
- TOC, pH, O2, O3
Off-Line
- batch sample
- "Lab" measurements
- minutes, hours, days
- pH, chromatography
Now all 3 USP regulated parameters measured in-line/on-line are available in Thornton's
portfolio!
On-Line
21
Application Example: Storage/Distribution
Validate PW and WFI quality with on-line
Real-time Microbial System – 7000RMS
Reduce false-positive tests from sampling
errors
Reduce lab testing costs and analysis
delays
No delay in product release
Real time microbial detection provides
faster analysis, eliminates delays and
reduces costs
Monitor and trend the water quality thereby
allowing for accurate risk assessment
Reduce testing and insure proper and
timely sanitization
User Objective
Pure Water
or WFI
Why the 7000RMS?
5000TOCi
O3
7000RMS
can also
be installed
after the
purification
unit
22
Application Example: Point of Use Testing
Rapidly validate water at point-of-use
Expand distribution monitoring to include
all Points of Use
Eliminate sampling errors and costs
Sample points-of-use real time
Reduce sample analysis time and eliminate
risk associated with grab samples
See measurement results in seconds
Bring the measurement to the sample
User Objective Why the 7000RMS?
UPW
23
Thornton's Answers to Regulations
Thornton's 5000TOCi
& 450TOC Sensors
USP <643>: established standards
for TOC measurements and the
equipment used to monitor TOC
METTLER TOLEDO
Service
USP <643>, USP <645>, EP 2.2.44
and JP16 - System Suitability
Test, TOC calibration and
conductivity calibration
Thornton's UniCond
Conductivity Sensors USP <645> for in-line or off-line
conductivity testing
Thornton's 7000RMS
Bioburden Analyzer
USP <1231>: monitor UPW at a
frequency that ensures the system is
in control and continues to produce
water of acceptable quality
Now available
on-line
24
To shift Highlight
Click the arrow,
press shift key and
move top or bottom
Agenda
1 Introduction
2 Why On-line Microbial Monitoring
3 7000RMS Technology and Key Features
4 Test Points for 7000RMS
5 Pharmacopeia acceptance of Alternative Methods
6 Validation Guidelines
25
Relevant Sections of the USP
Pharmaceutical Water Monographs
- Purified Water (bulk) Sterile Water for Inhalation
- Water for Injection (bulk) Sterile Water for Irrigation
- Sterile Purified Water (packaged) Bacteriostatic WFI
- Sterile WFI (packaged) Water for Hemodialysis
- Pure Steam
Test Chapters
- 645 Water Conductivity 85 Endotoxins
- 643 Total Organic Carbon 71 Sterility
- 644 Conductivity – planned for the future 791 pH
General Information
- 1230 Water for Health Applications - related to Water for Hemodialysis
- 1231 Water for Pharmaceutical Purposes (contains microbial limits)
- 1233 Instrumentation for Pharmaceutical Water – planned for future?
- 1644 Theory and Practice of Electrical Conductivity Measurements of Solutions
- 1223 Validation of Alternative Microbiological Methods – 2nd Supplement USP 38
26
USP Recommendation for Microbial Monitoring
7000RMS enables real-time, continuous measurement according to USP
recommendations
USP < 1231 > Water for Pharmaceutical Purposes recommendation:
- Pharmaceutical water systems should be monitored at a frequency that ensures the
system is in control and continues to produce water of acceptable quality.
The general information chapter endorses operating monitoring instruments
continuously in order that historical in-process data can be recorded for
examination
The new USP <1223> promotes and encourages the validation and
development of alternative microbial technologies
27
Revised Chapter for Alternative Microbial
General Information
- 1223 Validation of Alternative Microbiological Methods – 2nd Supplement USP 38
28
European Pharmacopeia Alternative Microbial
Information
- 5.1.6 Alternative Methods for Control of Microbiological Quality
5.1.6. ALTERNATIVE METHODS FOR CONTROL OF MICROBIOLOGICAL QUALITY
The following chapter is published for information.
1. GENERAL INTRODUCTION
The objective of this chapter is to facilitate the implementation and use of alternative
microbiological methods where this can lead to cost-effective microbiological control and
improved assurance for the quality of pharmaceutical products. These alternative methods
may also find a place in environmental monitoring.
The microbiological methods described in the European Pharmacopoeia have been used for
almost a century and these methods - for enumerating and identifying micro-organisms - still
serve microbiologists well. Over the years, these methods have been invaluable to help
control and secure the production of microbiologically-safe pharmaceutical products.
Nevertheless conventional microbiological methods are slow, and results are not available
before an incubation period of typically up to 14 days. Thus the results from the conventional
microbiological methods seldom enable proactive, corrective action to be taken.
Alternative methods for control of microbiological quality have been introduced in recent
years, and some of these methods have shown potential for real-time or near-real-time
results with the possibility of earlier corrective action. These new methods can also offer
significant improvements in the quality of testing.
29
European Medicines Agency EMA 30
• Questions and answers on production of water for injections
by non-distillation methods – reverse osmosis and biofilms
and control strategies.
• This set of questions and answers is intended to provide preliminary
guidance until such time the ongoing revision of Annex I of the GMP guide
is complete.
• 6. What testing should be employed during initial qualification and routine operation sampling? • Testing should be conducted in line with Ph.Eur. Monograph 169 ‘Water for Injections’
• Use of rapid microbiological methods should be employed as a prerequisite to the control strategy to aid with rapid
responses to deterioration of the system.
• Article 23 of Directive 2001/83/EC states: “...the authorization holder must, in respect of the methods of manufacture and
control...take account of scientific and technical progress...”
• Quantitative microbiological test methods – in line with Ph.Eur. 5.1.6 monograph ‘Alternative Methods for control of
Microbiological Quality’.
• Due consideration should be given to employing alternate methods for the rapid quantitative determination of the
contamination levels existing within the water system. The validation of such system should be in line with the above
referenced monograph.
• Use of alternative/ rapid microbiological test methods should be employed as part of the overall control strategy for the
system.
• Taking into account the speed at which organisms can proliferate, the use of rapid microbiological test methods and
systems should be employed in order to improve or increase the probability of early detection and allow timely action to
be taken.
USP Adoption of New Technology
1840 - 1994
Chemistry/Organic tests
are;
- Qualitative,
- Subject to bias
- Off-line.
- Carbon dioxide
- Calcium
- Ammonia
- Chloride
- Sulfate
- Oxidizable
Substances
- Heavy Metals
1840 - 1994 1994 - 1996 > 1996
1994 – 1996
Thornton asked
to prove
instrumentation
versus wet
chemistry
Chemistry tests
- VS.
Conductivity
Instrument
Oxidizable
Substances
- VS.
TOC Instrument
1996 to present
USP Accepted
as Compedial
Tests
Conductivity
Instrument
TOC Instrument
2003 –
2006
Adoption by
EP
JP
ChP
IP
31
USP Adoption of New Technology
1890 - 2016
Bacteria tests are;
- Plate Count
- Qualitative,
- Subject to bias
- Off-line.
- Sampling errors
- Poor growth
1890 - 2016 2016 to 20?? 20??
2016 – ?
Traditional Plate
Counts
-VS.
On-Line Real Time
Instruments
USP <1223> now
encourages validation
of alternative
microbial methods
Ph.EUR. 5.1.6.
“Alternative Methods
for Control of
Microbiological
Quality”
Accepted by USP
as a Compendial
Test?
On-Line Real Time
Instruments
20??
Accepted by the
other Global
Pharmacopeia’s
32
To shift Highlight
Click the arrow,
press shift key and
move top or bottom
Agenda
1 Introduction
2 Why On-line Microbial Monitoring
3 7000RMS Technology and Key Features
4 Test Points for 7000RMS
5 Pharmacopeia acceptance of Alternative Methods
6 Validation Guidelines
33
Online vs. Plate Counts – Why the difference?
CFU Versus AFU
34
Proposed Methods for Validation – Method 1
Method 1 is the fastest validation approach while minimizing risk for the
customer without requiring live bacteria inoculation.
Method 1: Proposed method for validation utilizing side-by-side testing
Method 1 is likely to be the preferred option for many customers as it:
- does not involve bringing live bacterial cultures into their facility
- provides a direct correlation in their process between the 7000RMS and their current
plate count method
Method 1 compares the direct at-line microbial readings from the 7000RMS to the
laboratory results from standard sampling and plate counting in combination with review
of historical data.
This method does not require the inoculation of a water system with bacterial samples
that maybe non-native or non-existent in the water system.
If the PW and/or WFI distribution systems have been monitored over time and historical
records demonstrate that the system is under microbial control then this side-by-side
validation method eliminates the risk of bacterial introduction.
35
Correlation Sample 1 "Ideal"
Plate Count (CFU) 7000RMS AFU
To validate the alternative method (RMS), the customer needs to demonstrate to
the validation officer the correlation of the RMS values vs. the plate count.
36
Correlation Sample 3 "Wrong"
Alternative methods must show correlation but more important they must not show inferior
(lower) like in sample 3 in the chart below.
- Highlights importance of ensuring good lab technique and sample handling!
Plate Count (CFU) 7000RMS AFU
37
Correlation Sample "USP 1223"
0
10
20
30
40
50
60
70
80
90
100
110
1 2 3 4
Plate Count
7000RMS
Note:
If the bacteria cells in a water
sample are low, you would
typically see 0 CFU with plate
count
If the bacteria in the water
sample is low, you would
typically see proportionally
small difference between the
plate count and RMS AFU
count
The more bacteria in the
sample you will see higher
proportional difference
between the plate count and
RMS AFU count
Bacteria
cells/sample
Plate Count
(CFU)
7000RMS
Biocount
1 0 ≈ 1
10 0 ≈ 10
50 - 100 25 - 40 ≈ 50 - 100
100 - 200 50 - 75 ≈ 100 - 200
Comparison test per USP<1223>
38
Online vs Plate Counts – Why the Difference?
The USP is proactively managing customer expectations that online results
will be different. Help manage this upfront in the sales process!
‘Why do I have higher counts online versus my plate counts?”
USP <1223> “It is important to understand that the cfu has always been an estimation of
microorganisms present, rather than an actual count.”
“Studies on the recovery of microorganisms have demonstrated that traditional plate-count
methods reporting cell count estimates as colony-forming units (cfu) may recover 0.1%–
1% of the actual microbial cells present in a sample.”
“Higher cell counts must not be considered as necessarily indicative of greater risk given
the inherent variability of standard growth methods.”
The CFU (Colony Formation Unit) has always been an estimation of microorganisms
present, rather than an actual count – colony may be from 1 cell or 1,000 cells (plate
count)
Rapid microbiological methods are direct cell count methods, therefore a higher count is
expected
39
Conclusions
Traditional plate count method has been performed for 125 years and is
recognized as only an estimate of the actual microbial cells present
“Alternative microbial detection methods will in most applications report higher
counts”. USP<1223>
“Higher counts should not be interpreted as an increased microbial safety risk
for previously controlled and monitored systems.” USP<1223>
“Any alternative microbial detection methods may be used provided it has been
demonstrated they are equivalent or superior to the compendial Pharmacopeia
method”. FDA Senior Microbiologist
Online vs Plate Counts – Why the difference? 40
Mettler-Toledo Thornton
Thank You!
Thank You!
Merci
Grazie
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ありがとう
Danke
고맙습니다
Gracias
Tak
Terima kasih
Dziękuję
Tack
Cảm ơn bạn
Cпасибо
Obrigado
Go raibh maith agat
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ודה
41