Build robust methods from separation mechanism to transfer.
SEPAX Technologies offers chromatography columns engineered for reliable analytical and preparative separations across pharmaceutical, biopharmaceutical, biotechnology, food, environmental and research laboratories. This page provides technical guidance, applications and specifications to help select the appropriate SEPAX column.
Column selection
Define analyte size, polarity, charge, hydrophobicity, molecular stability and detection needs. Select the separation mechanism first, then phase, particle size, pore size and dimensions.
Particle size
Smaller particles can increase efficiency but generally increase backpressure. Confirm instrument pressure capability and extra-column volume.
Flow rate
Begin within the product and method limits. Optimize linear velocity while monitoring efficiency, pressure and mass-transfer effects.
Gradient
Use gradient scouting when analyte retention spans a wide range. Adjust starting strength, slope, final strength and re-equilibration.
Isocratic
Use isocratic conditions when the analyte retention window is compact and stable. Confirm adequate retention and reasonable run time.
Injection volume
Keep injection volume and solvent strength compatible with column dimensions and starting mobile phase to limit distortion.
Temperature
Temperature can affect viscosity, selectivity and biomolecule stability. Use controlled, reproducible conditions within product limits.
Buffer selection
Choose a buffer with suitable pKa, solubility, detector compatibility and volatility requirements. Prevent precipitation during organic mixing.
pH selection
Select pH according to analyte ionization, separation mechanism and product-specific pH limits.
Sample preparation
Clarify, filter or centrifuge samples as appropriate. Match sample solvent to the method and protect the column from particulates.
Troubleshooting
Change one variable at a time and document pressure, retention, peak shape and resolution responses.
Optimization
Use structured experiments around the critical variables rather than uncontrolled sequential changes.
Scale up
Maintain relevant linear velocity, gradient volume in column volumes, loading relationship and packing chemistry.
Method transfer
Account for dwell volume, extra-column volume, pressure limits, detector cell volume and data rate between systems.
Technical support
For a catalog-specific discussion, contact Akira Analytical Solutions Pvt. Ltd. with the current column, analytes, mobile phase, gradient, flow, temperature and observed chromatogram issue.