HPLC Tips & Tricks webinars selection in 2024

This webinar is the final part in a three-part series called "A Systematic UHPLC/HPLC Method Development Strategy with Complementary Stationary Phases to Maximize Selectivity and Resolution."

2. Waters Corporation: Chromatographic System Troubleshooting: A Systematic Approach

Best practices for method optimization to enhance your purification efficiency. You will also learn about scaling up methods and discover practical tips to improve workflow efficiency.

4. Phenomenex: LC Pressure 101

Learn about selectivity benefits of phenyl phases during reversed-phase HPLC. Review mobile-phase optimization when using a phenyl column.

Tip 2: Column Care

• Equilibration: Allow the column to equilibrate with the mobile phase for 10–20 column volumes before starting a run.
• Storage: Use an appropriate storage solvent (e.g., 100% organic solvent or a mobile phase with preservatives) to prevent microbial growth.
• Flush Regularly: After using buffer-based mobile phases, flush the column with water followed by an organic solvent like methanol or acetonitrile.
• Avoid Pressure Spikes: Gradually ramp up the flow rate to protect the column.

6. Agilent Technologies: USP 621 Decoded

Troubleshooting HPLC systems and chromatographic issues using a structured workflow.

This webinar will review some of the principles and practices to develop robust RP separation methods showing examples using GLP-1 Agonists.

Tip 3: Sample Preparation

• Filter Samples: Use a syringe filter (0.2 µm or 0.45 µm) to remove particulates that can clog the column.
• Match Solvent Strength: Dissolve samples in a solvent similar to the starting mobile phase composition to avoid peak distortion.
• Use Internal Standards: These help improve quantitation and compensate for variability.

we will cover the fundamentals of how your DAD works. From there we will discuss the various options for DAD method optimization.

This exclusive session is designed specifically to address and help you overcome most pressing maintenance and troubleshooting hurdles in HPLC operation.

We’ll provide an overview of the LC system, from solvent bottles to detector, with a focus on preventive maintenance and keeping systems running at optimal performance.

Tip 4: System Maintenance

• Regularly Clean the System: Flush the injector, pump, and tubing to prevent salt build-up or contamination.
• Monitor Seals and Filters: Check and replace worn pump seals, check valves, and inline filters periodically.
• Avoid Overloading: Use an appropriate sample concentration to prevent column overloading and ensure accurate results.

We’ll discuss some of the most common chromatography issues that we see as technical support specialists.

Learn how to prevent and minimize your LC problems before they occur.

Tip 5: Troubleshooting Common Issues

• Baseline Noise or Drift
  • Check for air bubbles in the detector or mobile phase.
  • Ensure the mobile phase is fresh and fully degassed.
  • Inspect the detector lamp for aging or instability.
• Peak Tailoring
  • Use columns with appropriate stationary phases and well-maintained packing.
  • Ensure no column voids are present.
  • Adjust mobile phase pH for ionizable compounds.
• Low Resolution or Overlapping Peaks
  • Optimize flow rate, gradient, or mobile phase composition.
  • Use a column with smaller particle size or longer length.
• Retention Time Shifts
  • Confirm consistent mobile phase composition and column temperature.
  • Check for leaks in the system or improper mixing in gradient systems.

Get actionable insights, practical tips, and proven strategies to not only overcome obstacles but to significantly enhance your workflow's efficiency.

Modernizing a legacy method can improve accuracy, ruggedness, and sample throughput.

Tip 6: Optimize Method Development

• Gradient Optimization: Start with a wide gradient range to identify peak separation potential, then narrow it for resolution.
• pH Adjustment: Fine-tune mobile phase pH for ionizable analytes to maximize separation.
• Temperature Control: Maintain consistent column temperature to improve reproducibility and resolution.

In this live webinar, we will highlight some distinctions between CAD and other UV-absorbance detection, such as the non-linear response to analyte concentration, and sensitivity to mobile phase impurities.

Understand the importance of knowing your analyte and how they influence analysis and optimization.

Getting your system ready. LC System Start Up and Shut Down. LC-MS System Start Up and Shut Down. On and Off System Column Storage.

7. General Best Practices

• Log System Conditions: Record mobile phase, flow rate, column temperature, and pressure for reproducibility.
• Keep Spare Parts: Maintain spare seals, filters, and lamps to minimize downtime.
• Training: Regularly train operators on best practices and maintenance.