Applying USP<1223> Guidelines as Part of a Comprehensive RMM Validation

Goals of Validation Architecture

Delivers meaningful and relevant content to end user Can e ectively determine that the performance characteristics of the RMM meet the requirements for the intended application, in comparison to the traditional method (USP<1223>) Harmonizes statistical approach and methods that satisfy related guidelines (e.g. EP, ISO, TR33, JIS...) Methods and results withstand the rigor, scope, and neutrality expected by Regulators
People
Creating the right RMM validation approach begins with the right human resources and connections: Prospective users Engineers Industry thought leaders Scientists Research experts Biostatisticians Microbiologists

Scott Morris Applications Engineering Manager BioVigilant Systems, Inc. BioVigilants Comprehensive Approach
Rapid Micro Methods Validation

Equipment

Testing of an RMM often requires specialized equipment and state of theart facilities; these components should be calibrated and thoroughly characterized for a rigorous assessment. Multimillion dollar purposebuilt bio test facility and automated aerosol test chamber Please refer to PDA Poster entitled, Development and Validation of a Test Chamber by Shinsuke Yamasaki, Azbil Corporation

IMD-A 300 (2) Alternate Methods IMD-A 350 (2) SAS, SMA, MAS Reference Methods Kanomax LSAPC

People

Guidance & Regulations

USP EP ISO

Procedures & Microbes

Equipment

Data & Analysis

Statistics

Test Chamber

Data & Analysis

The goal of the analysis and nal data should be to distill the value of the other ve validation components to clearly demonstrate whether equivalence or better was achieved and deliver meaningful and relevant data to the end user.

Guidance & Regulations

USP<1223> is the benchmark guidance for RMM validation, but other relevant guidance documents should be considered and harmonized for a comprehensive validation:
Part 11 21CFR TR33 RMM JIS 9921 21501 3836 ISO USP EP 13485 14644 1223

Bacillus atrophaeus
1000.00

Biologic Counts or CFU Per Liter

5.1.6

To e ectively apply a Rapid Microbial Method (RMM) technique and leverage its value, the end user must match his or her application and requirements to the bene ts and performance of the RMM. Understanding the general approach to validation as well as the inter-relationships between personnel and their quali cations, guidance and regulations, statistics employed, test equipment, test protocols, and data analysis techniques enhances this evaluation. Through this holistic understanding, more robust dialogue can be fostered, and ultimately, a more successful adoption of an RMM technology may be realized.

100.00

10.00

1.00

0.10 T1 T2 T3 T4 T5

Concentration
IMD-A 300 (#1) IMD-A 300 (#2) IMD-A 350 (#1) IMD-A 350 (#2) SAS MAS SMA

The graph to the left summarizes aerosol chamber results for a Bacillus atrophaeus microbe across ve concentrations. It clearly demonstrates not only the capability of the chamber to produce repeatable and homogeneous results, but it illustrates how the IMD-A systems yield repeatably higher recoveries than the reference methods

Statistics

Procedures & Microbes

Unique procedures for RMM testing are often integral to success, but they should build upon best practices, ensure traceability, and be formalized:
Microbes (ATCC strain #) Bacillus atrophaeus (9372) Corynebacterium afermentans (51403) Escherichia coli (13706) Staphylococcus epidermidis (12228) Micrococcus lylae (27566) Aspergillus niger (9142)

A comprehensive statistical architecture should be created to provide meaningful analysis, and also be detailed and formalized to ensure a repeatable and unbiased approach.

USP<1223>: The Benchmark Guidance for RMM Validation

Prescribed Validation Criteria (and Utilized by BioVigilant) Not Speci ed in USP<1223>
Universal minimum dataset requirements Tiered approach for acceptance criteria Evaluating/validating test environment/collection method Handling anomalous and nonquantitative data results Testing of multiple instruments for validation (reference and alternative instruments) Accommodating and/or circumventing the limitations of reference (traditional) methods
General Themes and Methods for a Robust Statistical Architecture
Tiered Acceptance Criteria Using a primary criteria and alternate criteria for evaluation of statistical signi cance allows for a simpli ed non-inferiority type of approach Parallel Sample Collection All instruments should collect data simultaneously to minimize systematic error and allow pairing of replicates in analysis (e.g. paired T-test) Uniform Dataset Conditioning Consistent rules applied to the conditioning of data and treatment of unsuitable (e.g. TNTC) replicates ensures an unbiased and consistent approach Multiple Alternate and Using multiple instruments creates a more robust evaluation, ensures data has end-user relevance, Reference Instruments and allows for straightforward assessment of Ruggedness Minimum Number of USP<1223> does not specify consistent minimum replicates (if at all), but a universal minimum Replicates allows monolithic testing and also enhances statistical signi cance of the dataset Range of Quantitation De ning an ROQ not only provides guidelines for creating optimal test conditions, it provides a consistent method for ensuring an instrument is operating acceptably for data analysis Appropriate Data Transforms Data transforms [e.g. log10(x)] are useful in statistical analysis, but their suitability should be evaluated and the assumption of data normality avoided

9 8 6
5 12

Design of Experiment: USP <1223> Metrics Instruments (4 x IMD-A) Microbes

Concentrations Replicates

By the Numbers: > 4*108 Microbes aerosolized 2880 Individual samples collected > 1100 Agar plates manually counted 1460 USP<1223> evaluation
34 Tears cried

Validation of an RMM is not something that merely can be claimed; it must be demonstrated.

To help achieve these goals BioVigilant researched and employed the use of some unique statistical tools and methods:

Visit BioVigilants Booth 14 to obtain your copy of IMDA USP<1223> Test Results.

Winsorization Paired TTest Levene Test

Alternative Acceptance Binomial Test Independent Linearity Reference Criteria Precision Transform