HPLC for Pharmaceutical Scientists is an excellent book for both novice and experienced pharmaceutical chemists who regularly use HPLC as an analytical tool to solve challenging problems in the pharmaceutical industry. It provides a unified approach to HPLC with an equal and balanced treatment of the theory and practice of HPLC in the pharmaceutical industry.

In-depth discussions of retention processes, modern HPLC separation theory, properties of stationary phases and columns are well blended with the practical aspects of fast and effective method development and method validation. Practical and pragmatic approaches and actual examples of effective development of selective and rugged HPLC methods from a physico-chemical point of view are provided.

HPLC for Pharmaceutical Scientists elucidates the role of HPLC throughout the entire drug development process from drug candidate inception to marketed drug product and gives detailed specifics of HPLC application in each stage of drug development.

The latest advancements and trends in hyphenated and specialized HPLC techniques (LC-MS, LC-NMR, Preparative HPLC, High temperature HPLC, high pressure liquid chromatography) are also discussed.

Contents Part I: HPLC Theory and Practice

Introduction

• Chromatography in the Pharmaceutical World
• Chromatographic Process
• Classification
• History of Discovery and Early Development (1903–1933)
• General Separation Process
• Types of HPLC
• HPLC Descriptors

HPLC Theory

• Basic Chromatographic Descriptors
• Efficiency
• Resolution
• HPLC Retention
• Retention Mechanism
• General Column Mass Balance
• Partitioning Model
• Adsorption Model
• Total and Excess Adsorption
• Mass Balance in Adsorption Model
• Adsorption of the Eluent Components
• Void Volume Considerations
• Thermodynamic Relationships
• Adsorption-Partitioning Retention Mechanism
• Secondary Equilibria
• Gradient Elution Principles
• Types of Analyte Interactions with the Stationary Phase

Stationary Phases

• Type of Packing Material (Porous, Nonporous, Monolithic)
• Base Material (Silica, Zirconia, Alumina, Polymers)
• Geometry
• Adsorbent Surface Chemistry
• Surface of Chemically Modified Material
• Polymer-Based Adsorbents
• Stationary Phases for Chiral Separations
• Columns

Reversed-Phase HPLC

• Retention in Reversed-Phase HPLC
• Stationary Phases for RPLC
• Mobile Phases for RPLC
• pH Effect on HPLC Separations
• Effect of Organic Eluent Composition on Analyte Ionization
• Synergistic Effect of pH, Organic Eluent, and Temperature on Ionizable Analyte Retention and Selectivity
• Examples of Applying pH Shift and Analyte pKa Shift Rules
• Effect of Temperature on Analyte Ionization
• Ion-Interaction Chromatography
• Concluding RemarksM

Normal-Phase HPLC

• Theory of Retention in Normal-Phase Chromatography
• Effect of Mobile Phase on Retention
• Selectivity
• Applications

Size-Exclusion Chromatography

• Separation of the Analyte Molecules by Their Size
• Molecular Size and Molecular Weight
• Separation Mechanism
• Calibration
• Columns
• Molecular Weight Distribution
• Effect of Eluent
• Effect of Temperature
• Detectors
• Solving Mass Balance Issues
• Aqueous SEC Applications

LC/MS: Theory, Instrumentation, and Applications to Small Molecules

• Ionization Methods and LC/MS Interfaces
• Mass Analyzers
• Role of Instrumental Parameters on Ionization Efficiency in LC/MS
• Effect of Mobile-Phase Composition on Ionization Efficiency in LC/MS
• MS Interpretation
• Practical Applications

Method Development

• Types of Methods
• Defining the Method
• Method Development Considerations
• Method Development Approaches
• Effect of pH on UV Absorbance
• Analyte pKa—From an Analytical Chemist’s Perspective
• Reversed-Phase Versus Normal-Phase Separations
• Instrument/System Considerations
• Column Testing (Stability and Selectivity)

Method Validation

• Validation Report
• Revalidation
• Assignment of Validation Parameters
• Distinguishing Drug-Related and Non-Drug-Related Degradation Products

Computer-Assisted HPLC and Knowledge Management

• Prediction of Retention and Simulation of Profiles
• Optimization of HPLC Methods
• Structure-Based Tools

Part II: HPLC in the Pharmaceutical Industry

The Expanding Role of HPLC in Drug Discovery

• Applications of HPLC/MS for Protein Identification and Characterization
• Applications of HPLC/MS/MS in Support of Protein Chemistry
• Applications of HPLC/MS/MS in Support of Assay Development and Screening
• Sources of Compounds for Biological Screening
• HPLC/MS Analysis to Support Compound Characterization
• Higher-Throughput Purification Strategies
• ADME Applications
• Fast Serial ADME Analyses Incorporating LC-MS and LC-MS/MS
• Parallel Approaches to Speeding ADME Analyses
• Automated “Intelligent” Metabolic Stability and Metabolite ID

Role of HPLC in Preformulation

• Initial Physicochemical Characterization (Discovery Support)
• Chemical Stability
• Salt Selection
• Polymorphism
• Preformulation Late Stage (Development Support)

The Role of Liquid Chromatography–Mass Spectrometry in Pharmacokinetics and Drug Metabolism

• Tandem-Mass Spectrometry (MS/MS)
• Sample Preparation Using an Off-Line Approach
• Automated Sample Transfer
• Sample Processing Using an On-Line Approach
• Matrix Effect and Ion Suppression
• Regulatory Requirements for LC/MS Method Validation
• Ritalin®: An Application of Enantioselective LC-MS/MS
• GleevecTM (STI571)
• Biomarkers

Role of HPLC in Process Development

• Responsibilities of the Analytical Chemist During Process Development
• HPLC Separation Modes
• Sample Preparation
• HPLC Detectors
• Method Development
• In-Process Monitoring
• Impurity Identification
• Establishment of HPLC Selectivity by Stress Studies
• HPLC Method Validation
• Technology Transfer

Role of HPLC During Formulation Development

• Prerequisite for Analytical Chemists During Formulation Development
• Properties of Drug Substance
• Properties of Excipients
• Impact of Excipients on Degradation of API(s)
• Test Methods for Most Common Dosage Forms in which HPLC Is the Primary Technique
• Forced Decomposition
• Compatibility of Excipients with API(s) (Type and Ratio)
• Mass Balance
• Summary of Assay and Related Substances
• Uniformity of Dosage Units
• Blend Uniformity (BU)
• Cleaning Verification
• Extractables/Leachables
• Dissolution
• Method Development
• Method Validation
• Testing of Samples
• Automation Opportunities
• Implementation of Alternative Technologies
• Challenges and Future Trends

The Role of HPLC in Technical Transfer and Manufacturing

• Prerequisites for Transfer of HPLC Methods
• Types of Technical Transfer
• Different Approaches for Technical Transfer and Manufacturing
• Potential Pitfalls During Technical Transfer and Manufacturing

click here to see books • videos • cd-roms of related interest