High Performance Packed Column for HPLC - Bio-Diol

Importance of the Topic

The efficient separation of biomolecules by high-performance liquid chromatography (HPLC) is essential in biochemical research, pharmaceutical development and quality control. The Shim-pack Bio-Diol series offers dihydroxypropyl-bonded silica columns optimized for hydrophilic interaction chromatography (HILIC), delivering high resolution, reproducibility and column longevity. Proper handling and operation of these columns ensures consistent results, extended service life and minimized downtime.

Objectives and Overview

This document outlines best practices for installation, operation and maintenance of Shim-pack Bio-Diol columns. Key goals include:

• Preserving peak shape and chromatographic efficiency
• Preventing dead volume and sample precipitation
• Maintaining column integrity under specified pressure, pH and temperature limits
• Minimizing clogging and performance drift

Methodology and Instrumentation

Shim-pack Bio-Diol columns are manufactured with silica particles bonded to dihydroxypropyl groups. Particle sizes range from 2 to 5 µm, with pore sizes of 6–30 nm. Key operational parameters include pH 5.0–7.5, maximum temperature 50 °C and mobile phase salt concentration below 0.7 M. Organic modifiers such as acetonitrile or methanol are compatible, noting possible pressure increases.

Used Instrumentation

• Shim-pack Bio-Diol HILIC columns (2–5 µm, 6–30 nm pores)
• UHPLC/Nexlock fittings and PEEK nuts/ferrules rated up to 130 MPa
• Guard columns or UHPLC guard cartridges
• Membrane filters (0.45 µm) for mobile phase and sample filtration
• Standard HPLC or UHPLC systems equipped with low-dead-volume tubing

Main Results and Discussion

Quality control tests confirm uniform chromatography performance across the Bio-Diol series. Recommended installation practices—short tubing, correct flow direction and properly tightened fittings—minimize dead volume and peak tailing. Blockages due to particulates or precipitated salts remain the primary cause of backpressure increases; prevention via in-line filtration and guard columns is critical. Slow valve switching can induce pressure spikes, so sample injection speed should be optimized.

Benefits and Practical Applications

The Bio-Diol columns deliver:

• Consistent retention and peak symmetry for polar analytes
• Flexible use with aqueous buffers and organic solvents
• High pressure tolerance (up to 45 MPa for 2 µm particles)
• Extended lifetime when operated within specified limits

Applications span biomolecule profiling, glycan analysis, metabolomics and QC of biopharmaceuticals.

Future Trends and Possibilities

Advancements in particle technology and bonding chemistries may drive further improvements in resolution and pressure limits. Integration with ultra-high-throughput systems, miniaturized columns and automated equilibration protocols will expand applications in clinical diagnostics and high-volume screening.

Conclusion

Adherence to recommended operating conditions—correct pH, temperature, tubing, filtration and pressure control—ensures optimal performance and lifetime of Shim-pack Bio-Diol columns. Preventative measures against clogging and dead volume are essential for reliable, reproducible separations.

Reference

No external literature references provided in the original manual.