The 4th Workshop on
Recent Issues in Regulated Bioanalysis
Pre-Workshop Short Courses
Short Course 1 (half day): 1pm to 5pm, April 20, 2010
Fundamental of Bioanalysis: Bioanalytical Method Validation (BMV) and Sample Analysis for Chromatographic and Ligand Binding Assays (LBA)
Bioanalytical Method Validation: "History & Definitions"
- Definition of Bioanalytical Method Validation
- Regulatory Guidance on Bioanalytical Method Validation from Crystal City I to Crystal City III
- FDA Guidance for the Industry, Bioanalytical Method Validation (May 2001)
- FDA Good Laboratory Practice for Nonclinical Laboratory Studies: Title 21, Code of Federal Regulations, Part 58
- OECD Principles on Good Laboratory Practice (as revised in 1997)
- Drugs Directorate Guidelines, "Conduct and Analysis of Bioavailability and Bioequivalence Studies"
- Statistics in Bioanalysis
- Statistical Mean
- Standard Deviation and Relative Standard Deviation
- Deviation from Theoretical and Percent Difference
- Correlation Coefficient
- Rejection of Outliers with T-test
- GLP Definitions
- Key Terms & Concepts in Bioanalysis
- GLP Documentation
- Analytical Method
- Data Review
Bioanalytical Method Validation: "Criteria & Concepts"
- The Validation Process and the Validation Goals
- Uncertainty in Quantitative Analysis
- Validation Parameters and Method Validation
- Accuracy and Precision
- Analysis of Variance (ANOVA)
- Assessing Accuracy and Precision
- Stability Tests
- Recovery
- Approaches to Quantification
- Calibration Curve
- Limit of Quantification (LLOQ & ULOQ) and other Key Terms
- Quantitative Relationships in MS Instruments
- Nature of Regression Errors
- Standard Addition
- Defining Acceptance Criteria
- Essential Documentation
- System Suitability and its Importance
- The Steps in a Typical MS Quantitative Analysis
- Routine Sample Analysis
LC-MS/MS & Bioanalysis
- Why LC-MS/MS?
- Advantages & Disadvantages of using LC-MS/MS in Bioanalysis
- Professor Fred McLafferty’s Four S’s of LC-MS Analysis
- Internal Standard
- Importance of Choosing the best Internal Standard
- Analogue vs. Stable Labeled Internal Standards
- Sample Preparation Pros and Con
- Protein Precipitation (PPT)
- Liquid-Liquid Extraction (LLE)
- Solid Phase Extraction (SPE)
- Matrix Effect and Ion Suppression/Enhancement Definitions
- Carryover Definition
Ligand Binding Assays (LBA) & Bioanalysis
- A Special Case: Quantitative Analysis of Therapeutic Bioactive Proteins
- Background and basics
- Yalow & Berson
- Antibodies and Antigens
- Types of Immunoassays
- Competitive Immunoassay
- Non-competitive Immunoassay
- Heterogeneous vs Homogenous Immunoassays
- RIA,EIA,FPIA, KIMS, Lateral Flow
- ELISA
- LBA: Strenghta and Limitations
- Cross-Reactivity Issues
- Testing Formats and Models
- LC-MS/MS vs. LBA
- Types of LBA Protocols
- Reagents vs. Equipment
- Detector System
- Reference Standard
- Calibration Courve & Prozone
- Assay Validation Parameters
- Matrix interference
Short Course 2 (full day): 8am to 4pm, April 21, 2010
Regulated Bioanalysis: Challenges Solutions and Applications
Bioanalytical Method Validation and Sample Analysis: "Common Problems"
- Matrix Effect and Ion Suppression/Enhancement
- Matrix Effect in Electrospray (ESI) LC-MS/MS
- Remedy for Ionization Suppression in SPE-LC-ESI-MS/MS
- Matrix Induced Ionization Suppression in APCI
- Differential Suppression
- The 3 Different Scenarios of Matrix Effect
- Instrument Optimization to Reduce Matrix Effect
- Carry-Over Guidelines
- Impact of Carry-Over in Bioanalysis
- Fighting Carry-Over
- Drug Metabolism & Bioanalysis
- Metabolite Conversion During Sample Extraction
- Metabolite Conversion During Instrumental Analysis
- General Strategy for Method using UV or Fluorescence Detection
- General Strategy for Method using MS or MS/MS
- Study Cases
- Root Cause Analysis during Out of Specification investigation
- Importance of Trend Analysis in GLP Bioanalysis
What's new in Bioanalysis?
- The Purpose of the 3rd AAPS/FDA Bioanalytical Workshop
- Crystal City III Conclusions
- Quality Controls: Conventional and New Approaches
- Evaluating Curve Models: Graphic & Mathematical Comparison
- Factors Contributing to Imprecision and/or Inaccuracy
- Carryover and Contamination Evaluation
- Evaluate Contamination in Validation
- Incurred Sample Re-Analysis (ISR)
- Reproducibility of QC vs. Incurred Samples
- Approach for ISR
- Determination of Metabolites During Drug Development
- Documentation Issues
- Repeat Analyses
- Matrix Effects for MS-Based Assays
- Run Acceptance Criteria
- Stability Recommendations
- Autosampler Re-injection Reproducibility
- Blood Stability
- Method Evolution & Stability Measurements
- Internal Standard Stability
- Validation Topics with No Consensus:
- -70oC vs. - 20oC
- Reference Standard Expiration Date vs. Stock Stability
- Other Validation tests
- Analytical investigations
- Pharmacokinetic repeats
- Challenges in bioanalytical method development
- Impact of metabolites on method development
- Matrix Effects and Hemolysis Effect
- Autosampler stability and re-injection reproducibility
- Acceptance of nonlinear calibration models
- Instruments/Techniques that Can Improve Quality in Regulated Bioanalysis
- Concept Paper/ Recommendations on the Need for a Guideline on the Validation of Bioanalytical Methods
- Metabolite Testing & MIST
- Specificity in Presence of OTC Metabolites
- Use of Variable Injection Volumes
- Anticoagulants
- Cross-Validation Between Different Anticoagulant Salt Forms
