HPLC Method Development Training | Course

Description

HPLC Method Development Training | Course

PharmaGuru HPLC Method Development Training equips analysts with practical skills to develop, optimise, and validate robust HPLC methods, covering column chemistry, mobile phase selection (pH, buffers), gradient optimisation, and regulatory guidelines with hands-on experience

Module 1: Getting Started with HPLC Method Development

• How to initiate HPLC method development for assay, related substances, and impurity profiling.
• Understanding regulatory expectations for method scope and target performance.

Module 2: Principles of HPLC Separation Mechanism

• Fundamentals of chromatographic interactions, retention, selectivity, and resolution.
• Role of partitioning, adsorption, ion-exchange, and size exclusion in separation efficiency.

Module 3: Selecting the Right HPLC Mode

• Choosing between reversed-phase, normal-phase, ion-pair, HILIC, and ion-exchange modes.
• Mode selection based on analyte polarity, pKa, functional groups, solubility, and complexity.

Module 4: Selecting HPLC Columns / Stationary Phases

• How to choose the best C18, C8, phenyl, polar-embedded, HILIC, and specialty phases.
• Column selection workflow using analyte structure and separation goals.
• Column screening strategies to increase first-pass success.

Module 5: Method Development for Simple Molecules (<5 Impurities)

• Designing a fast, efficient, and cost-effective method for low-complexity impurity profiles.
• Approach for early screening, mobile phase optimisation, and resolution mapping.

Module 6: Method Development for Complex Molecules (>5 Impurities)

• Structured strategy for multi-impurity separation, including co-elution challenges.
• Gradient design, selectivity tuning, peak tracking, and system suitability mapping.

Module 7: Separating Impurities Along with Isomers & Diastereomers

• Advanced separation approaches for positional isomers, stereoisomers, and diastereomers.
• Use of pH, organic modifiers, and column chemistry to enhance selectivity.
• Real-case strategies for resolving closely eluting or overlapping peaks.

Module 8: Deciding System Suitability Criteria (SST)

• How to define meaningful SST parameters: Rs, tailing, RSD, plate count, retention time.
• Linking SST requirements to method intent and risk assessment.

Module 9: Impurity Calculations

• How to calculate individual impurities, total impurities, unknowns, and % area calculations.
• Relative response factors (RRF): when and how to apply them accurately.
• Acceptance criteria: regulatory expectations.

Module 10: Optimising and Finalising the Routine Method

• Step-by-step workflow: screening → optimisation → robustness → finalisation.
• Setting final gradients, buffer strengths, pH, flow rate, and column temperature.
• Ensuring method ruggedness for QC environments.

Module 11: Method Verification & Development Report Writing

• How to perform method verification, transfer checks, and suitability confirmation.
• Preparing a complete method development report with scientific justification and data summaries.

Module 12: Reducing HPLC Analysis Costs

• Strategies to lower the cost per analysis using:
  • Shorter columns
  • Reduced solvent usage
  • High-efficiency columns
  • Smart gradient design
• Approaches to reduce re-runs, downtime, and column damage.

Module 13: 11+ Real-World Case Studies & Expert FAQs

• Case studies covering:
  • Simple and complex impurity profiles
  • Isomer and diastereomer resolution
  • SST troubleshooting
  • Cost optimisation examples
  • Real R&D and QC industry challenges
• Detailed FAQs with problem–solution formats.
• Bonus: pro tips for handling unexpected chromatographic behaviour