2nd Edition
Microbial Limit and Bioburden Tests Validation Approaches and Global Requirements,Second Edition
In recent years, the field of pharmaceutical microbiology has experienced numerous technological advances, accompanied by the publication of new and harmonized compendial methods. It is therefore imperative for those who are responsible for monitoring the microbial quality of pharmaceutical/biopharmaceutical products to keep abreast of the latest changes. Microbial Limit and Bioburden Tests: Validation Approaches and Global Requirements guides readers through the various microbiological methods listed in the compendia with easy-to-follow diagrams and approaches to validations of such test methodologies.
Includes New and Updated Material
Now in its second edition, this work is the culmination of research and discussions with technical experts, as well as USP and FDA representatives on various topics of interest to the pharmaceutical microbiologist and those responsible for the microbial quality of products, materials, equipment, and manufacturing facilities. New in this edition is an entire chapter dedicated to the topic of biofilms and their impact on pharmaceutical and biopharmaceutical operations. The subject of rapid methods in microbiology has been expanded and includes a discussion on the validation of alternative microbiological methods and a case study on microbial identification in support of a product contamination investigation.
Substantially updated and revised, this book assists readers in understanding the fundamental issues associated with pharmaceutical microbiology and provides them with tools to create effective microbial contamination control and microbial testing programs for the areas under their responsibility.
I. Microbial Life and Ecology
An Overview of Microbial Life
Microbial Phylogeny
Microbial Taxonomy
Microbial Growth and Survival
Growth Curve
Temperature
Energy Sources
Oxygen
Bacteria
Cell Shape
Mycoplasma
Bacterial Growth and Reproduction
Cell Structures
The Phyla Gram-Positive Bacteria and Proteobacteria
Gram-Positive Bacteria
Proteobacteria
The Gram-Staining Method
KOH Test
Catalase Test
Fungi
Cell Structures
Fungal Growth and Reproduction
Molds
Yeasts
Microorganisms of Interest
Genus Staphylococcus
Staphylococcus aureus
Genus Pseudomonas
Pseudomonas aeruginosa
Genus Burkholderia
Burkholderia cepacia
Genus Ralstonia
Ralstonia pickettii
Genus Comamonas and Genus Stenotrophomonas
Family Enterobacteriaceae
Genus Escherichia
Escherichia coli
Genus Salmonella
Genus Shigella
Genus Serratia
Genus Klebsiella
Genus Bacillus
Bacillus subtilis
Bacillus cereus
Genus Clostridium
Clostridium perfringens
Clostridium sporogenes
Candida albicans
Zygosaccharomyces rouxii
Genus Aspergillus
Aspergillus niger
Genus Penicillium
References
II. Microbial Contamination and Control
Microbiological Contamination
Product Recalls
Nonsterile Products
Microbial Limit Standards
The Preservation of Pharmaceutical Products
Antimicrobial Activity and Efficacy
Types of Preservatives
Alcohols
Benzalkonium Chloride
Benzoic Acid and Salts
Boric Acid and Salts
Chlorhexidine
Cresol
Dowicil 200
Mercurials
Parabens
Phenol
Sorbic Acid Salts
Microbiological Control
Risk Assessment
Objectionable Organisms
Sanitization and Disinfection Practices
Definitions and Types of Chemical Products
Factors in Choice and Use of Disinfectants
Rotation of Disinfectants
Qualification of Disinfectants
In Situ Testing
In Vitro Testing
Expiration Date for Disinfectant Solutions
Sanitizers Used for Equipment Cleaning
Neutralization and Microbial Recovery Studies
Requalification and Change Control
Conclusion
References
III. The USP Microbial Limit Tests
History of the Revision and Harmonization Process
USP Chapter <61>: Microbiological Examination of Nonsterile Products: Microbial Enumeration Tests
Sample Preparation
Total Aerobic Microbial Count
Total Combined Yeasts and Molds Count
Bioburden Tests
Two-Media Bioburden Test
One-Medium, Dual-Temperature Incubation Bioburden Test
TAMC and TYMC Tests via Plate-Count Methods
Pour-Plate Method
Spread-Plate Method
Incubation and Results Calculation
Test Controls
TAMC and TYMC Tests via Membrane Filtration Method
Test Controls
TAMC Test by the Multiple Tube Method
Procedure
Interpretation of the TAMC and TYMC Test Results
USP Chapter <62>: Microbiological Examination of NonSterile Products:
Tests for Specified Microorganisms
Sample Preparation for Direct Inoculation Tests
Test for Absence of Escherichia coli
Test for Absence of Salmonella spp
Test for Absence of Bile-Tolerant Gram-Negative Bacteria
Test for Absence of Pseudomonas aeruginosa
Test for Absence of Staphylococcus aureus
Test for Absence of Candida albicans
Test for Absence of Clostridia
Quantitative Test for Bile-Tolerant Gram-Negative Bacteria
Retesting
References
IV. Pharmaceutical Waters
Types of Water for Pharmaceutical Purposes
Microbial Quality Attributes
Testing of Pharmaceutical Waters
Sampling Program
Sample Collection and Preservation
Bioburden Testing
Recovery Media
Coliform Testing
Identification of Waterborne Microorganisms
Establishing Alert and Action Levels
Validation of Water Systems
Microbial Control and Sanitization
References
V. Environmental Monitoring
Cleanroom Classification
Occupancy State
Routine EM Program
Testing Frequency and Sampling Sites
Setting Alert and Action Levels
Test Methods and Equipment
Surface Sampling for Viable Particles
Active Air Sampling for Viable Particles
Active Air Sampling for Nonviable Particles
Microbial Identification Program
Data Analysis
EM During Facility Validation Activities
Room Occupancy
EM of Isolators
Microbial Control in Cleanrooms
References
VI. Bioburden Considerations In Equipment-Cleaning Validation
Biocontamination Control
Disposable and Single-Use Equipment
Equipment-Cleaning Methods
Validation of Cleaning Methods
Sampling Recovery Methods
Swabbing of Equipment
Rinsing of Equipment
Qualification of Sampling Methods
Recovery Study Using the Wet Method
Recovery Studies Using the Dry Method
Effects of Product and/or Cleaning Agent Residue on the Recovery of Microorganisms
Establishing Limits
Execution of Equipment-Cleaning Validation Protocol
Validation of Cleaned Equipment Hold Time
Validation of Dirty Equipment Hold Time
Ongoing Verification of Cleaning
Validation of Holding Time/Shipping Conditions
References
VII. Method Validation and Media Suitability Testing
Suitability Test Design
Representative Challenge Organisms
Maintenance and Preparation of Test Organisms
Preparation of Working Cultures
Validation of Storage Period for Working Cultures
Recovery of Injured Organisms
Suitability Testing By Direct Inoculation/Plating Methods
Validation of Screening for Specified Microorganisms
Modifications to the Direct Inoculation Method
Validation of the TAMC and TYMC Tests
Approach 1
Approach 2
Modifications to the Plate Method
Suitability Testing for Membrane Filtration Methods
Validation of Screening for Specified Organisms
Validation of TAMC and TYMC
Modifications to the Membrane Filtration Method
Suitability of Microbiological Media
Growth Promotion Testing for Microbial Enumeration Media
Growth Promotion Testing for Selective Media
Validation of Rapid Microbiological Methods
The Validation Package
Validation Criteria
Validation of Quantitative Methods
Accuracy
Specificity
Precision
Limit of Quantitation
Linearity
Limit of Detection
Range
Ruggedness
Robustness
Validation of Qualitative Methods
Specificity
Limit of Detection
Ruggedness
Robustness
Accuracy and Precision
Validation of Automated Microbial Identification Methods
Accuracy
Precision
Robustness
Ruggedness
Final Thoughts
Points to Consider
References
VIII. Microbiological Quality of Pharmaceutical and Biopharmaceutical Products and Raw Materials
Microbiological Testing
Raw Materials
Biopharmaceutical Products
Nonsterile Finished Drug Products
USP Chapter <1111>
Testing Frequency
Stability Testing
Water Activity
Measuring Water Activity
The Chilled-Mirror/Dew Point Method
Capacity Sensors
Pharmaceutical Applications for Water Activity
International Harmonization
Looking Ahead
References
IX. Rapid Testing and Alternative Methods in Microbiology
Rapid Method Technology Platforms
Impedance/Conductance Technology
Gas Consumption or Generation
ATP Bioluminescence
The Celsis ATP Bioluminescence Systems
Automated Biochemical Assays
The VITEK® System
The Biolog Systems
Fatty Acid Analysis Using Gas Chromatography
The MIDI System
Enzyme-Linked Immunosorbent Assay (ELISA)
The VIDAS
Analysis of Biomolecules Using Mass Spectrometry
Polymerase Chain Reaction (PCR)
Detection of Microbial Contamination Using PCR Technology
Riboprinting
The Riboprinter®
Fluorescent Labeling Assays
Scan® RDI Microbial Detection
D-Count
Biosensors and Microarrays
Laboratory-on-a-Chip Technology
Barriers to Implementation
Regulatory Climate
Future Trends
Case Study: Genotypically Similar Staphylococci
Contaminant Isolate and Environmental Sampling
Ribosomal Gene Sequencing
Phenotypic Analysis
Genetic Subtyping—PFGE
Results and Reporting
References
X. Biofilms
Biofilm Definition
Biofilm Structure
The Biology of Biofilms
Biofilm Formation
Quorum Sensing
Cell Adhesion
Smooth versus Rough Surfaces
Hydrophobic versus Hydrophilic Surfaces
Electrostatic Charge Properties
Low-Shear versus High-Shear Environments
Biofilm Dispersion
Biofilm Resistance and Phenotypes
Pharmaceutical Production Equipment and Materials Prone to Biofilm Formation
Water Systems
Production Equipment
Ultrafiltration/Diafiltration (UF/DF) Systems
Chromatography Systems
Miscellaneous Parts and Materials
Biofilm Control and Prevention
Heat Treatment
Chemical Treatment
Prevention of Biofilms
Methods for Detection and Recovery of Biofilm Organisms
Qualification of Chemical Sanitization Using Biofilm Cells
Types of Biofilm Reactors
Choosing a Biofilm Reactor
Testing Sanitizers Using the CDC Biofilm Reactor
Setting up the Biofilm Reactor
Exposure of Biofilm to Disinfectant Solution
Harvesting Biofilm Cells
Sanitizer/Disinfectant Efficacy Evaluation
Method Qualification and Test Controls
Testing Sanitizers Using a Static Biofilm Reactor
Industrial Significance of Biofilms
The Future in Biofilm Research
References
XI. Handling Aberrant and Out-of-Specification Microbial Data
Historical Overview of Investigating OOS Results
Out-of-Specification (OOS) Result
Laboratory Investigations
Conducting the Investigation
Retesting and Resampling
Testing for Outliers
Repeat Testing
Concluding the Investigation
Product Lot Disposition
OOS Investigations and FDA Citations
References
INDEX
Biography
Lucia Clontz