A Rapid Microbiological Method RMS™ for Application in Manufacturing

IFPAC 2014, Arlington VA

Bill Puent, J.P. Jiang

Instant BioScan™, USA

January 22, 2014

Outline

• Principle of operation of a real time optical microbial analyzer – RMS™: Real-Time Microbial Monitoring System

• Validation test and feasibility study – BioBall® (bioMérieux) test data – Suggestions for feasibility study

• Application examples – Online water quality monitoring – Surface sampling

Technical Background of RMS™

• RMS™ is a real time waterborne microbial sensor using optics for detections

• RMS™ detects microbes by the following methods

– Mie scattering sensor for measuring the size of particles

– Detection of intrinsic fluorescence of metabolites (NADH and Riboflavin) inside microbial cells to differentiate microbes from inert particles

• Microbial detection is done in real time, no reagent, no sample preparation

Fluorescence Emission Spectra of Metabolites

(Hill et al, Field Ana. Chem. & Tech, 3(4-5), 221,1999)

Applicable to 405nm excitation

RMS™ Principle of Operation

Flow cell of RMS™ and 405nm laser beam Parabolic reflector and reflected light (scattered and fluorescence) Colors of particles under 405nm: Inert particle and microbes

Inert Particles Microbes

RMS™ Principle of Operation

405nm laser

Dichroic beam splitter

Technical Features of RMS™

Non-gr owt h det ect i on met hod di spl ays i n Real -Ti me f or each Par t i cl e:

– I ner t vs. Bi ol ogi c St at us – Particle Size (≥ 0.5 t o > 25 mi cr ons) – Measur es Uni ver sal “Bi o-mar ker s” (NADH and

Ri bof l avi n) i n Bact er i a and Fungi . Sensi t i vi t y: 1 Bi oCount per 100mL. Dynami c r ange f r om 1 cell to 1x10^4 cell/ml

Graphic User Interface of RMS™

RMS™ Internal Validation Test Data

• Test sample: BioBall® from bioMérieux – Quantitative microbial samples

• Acceptance criteria – Adapted from USP<1223>

• Test procedure – Dissolve BioBall® in Water for Injection (WFI) or equivalent

(recommended, for reducing cross contamination or background interference)

– Collect part of the test sample for growth media plate count

– Run test sample through RMS™ and collect data

– Data analysis

Test Data Example: S. Aureas BioBall® (bioMérieux) Data

(Sept. 11, 2013)

RMS™ Validation Test Result Summary: USP<1223> Criteria Matrix

(September 2013)

Color code

pass

fail

Suggestions for Doing a RMS™ Feasibility Study

1) Validation Test – Select acceptance criteria for new method validation test result from published

guideline

• European Pharmacopeia (Ph. Eur.) 5.1.6.

• USP<1223>

• PDA TR33

– Select test samples

• BioBall® products are recommended for their quantitative microbial contents and consistency

– Run comparison study of RMS™ and growth media compendial method

2) Practical Feasibility Tests – Online RMS™ monitoring of water supply line

– Batch sample testing by RMS™

RMS™ Application Examples

• Example 1: Online Water Monitoring

• Example 2: Surface Sampling

EXAMPLE 1: Online Water Monitoring

1-a: RMS™ installation in a purified water line in a pharmaceutical facility

1-b: RMS™ online monitoring of a mineral water bottling operation

Example 1-a: RMS™ Online Monitoring of a Pharmaceutical

Purified Water Line

• A RMS™ analyzer was installed in a purified water line in a pharmaceutical company for online monitoring of water microbiological quality

• RMS™ demonstrated the stability of the water quality

• The time sequence of RMS™ data also displayed the effect of normal activities to the water line

– Usefulness as a process control tool

RMS™ Data Graph: June 16-18

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Purified water BLDG X June 17--June 18 RMS BioCount Shift Start

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Purified water BLDG X June 18 RMS™ BioCount

Example 1-a: Lessons Learned

• Stable RMS™ BioCount data were effective demonstration of the control of the water system

• Sporadic high RMS™ BioCount in June 16 were indication of system cleaning up after a routine annual preventive maintenance sanitization of this water line 2 days prior

– Demonstrating the effectiveness of PM cleaning

• Facilitate economical process verification and control

– Continuous monitoring of bio-burden, treatment when increase of BioCount detected

Example 1-b: RMS™ Online Monitoring of a Mineral Water Bottling Operation

• A RMS™ analyzer was installed in a mineral water bottling plant for online monitoring of water microbiological quality

• During the RMS™ monitoring period of several days, the water source of the bottling operation was switched

–RMS™ was able to detect the difference in water quality between these 2 sources

• The advantage of real time online microbial monitoring was thus demonstrated.

RMS™ Data Graph : April 16—20 (With notation of reported activities in the water plant)

Switched back to water source A (22:30 April 17 – 00:32 April 18) . Signs of gradual

Cleaning of water

Water source A

Switched to water source B

(per 10mL) BioCount

RMS Data Graph : April 22—26 (With notation of reported activities in the water plant)

Signs of gradual cleaning of water

Decreasing height of the bursts

Return to the original condition

of water source A (per 10mL)

BioCount

Example 1-b: Lessons Learned

• Continuous monitoring data from the RMS™

– To demonstrate that a water line is in a state of control

• Real time capability of RMS™

– To detect any change of the condition of the water in the water system, (such as the change of water supply and the composition of new water supply)

– To verify the effect of water system cleansing

EXAMPLE 2: SURFACE SAMPLING

Surface Swabbing in Conjunction with RMS™ Testing

Example 2: Surface Sampling

• Surface sampling and cleaning validation are important part of aseptic operation

–A real-time and sensitive analyzer (such as RMS™) could be beneficial: visualize the sampling results and timely validation

• Application example: RMS™ analyzer

–Aseptic swab: QUANTISWAB® (bioMérieux)

–Analyzer: RMS™ for rinsing the sampling swab

– Sampling result: RMS™ screen in <1 minute

Surface Sampling Photo 1

Surface Sampling Photo 2

Surface Sampling Photo 3

Case Study: Surface Sampling Data Graph

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May 21, 2013 R3 Nordic Gothenborg exhibit Surface sampling test trial bioMérieux QUANTISWAB®/RMS™ RMS™ BioCount (per 10mL)

Water Clean swab

Exhibit area countertop

Student Union countertop

Case Study: Tool for Surface Cleaning Validation

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Control Water Control Swab Surface A_untreated Surface A_cleanx2 Surface A_cleanx3 Surface A_cleanx4

RM

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Dirty plate

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RO water

WFI water Clean QuantiSwab® (in non-aseptic lab)

Caption. The graph shows progressive cleanliness with every additional cleaning procedure applied to the surface. This large surface was tested in duplicate swab tests concurrently. Data displayed represent average values.

Surface Tested: a ceramic plate Sampling Swab: QUANTISWAB® (bioMérieux) Cleaning Procedure: 70% IPA RO water rinse WFI water rinse

Summary

• RMS™ is an optics based real time microbial analyzer for water testing

• A real-time and continuous monitoring tool may be beneficial for economical process control in aseptic facilities

• Online water monitoring and surface sampling are among the potential applications of a RMS™