High Performance Packed Column for HPLC - Bio HIC column

Significance of the Topic

The Bio-HIC column enables hydrophobic interaction chromatography for the separation and purification of biomolecules such as proteins and peptides. Its hydrophilic polymer support with butyl functional groups offers high resolution, reproducibility and robustness across a wide pH and temperature range, making it essential for both research and industrial laboratories.

Objectives and Overview of the Study

This document provides guidance on the installation, operation and maintenance of the Shim-pack Bio-HIC column. The goal is to help users achieve optimal chromatographic performance, extend column life and prevent common issues such as dead volume, peak broadening and clogging.

Methodology and Instrumentation

• Stationary phase: Shim-pack Bio-HIC Butyl column (100 × 4.6 mm, 4 µm hydrophilic non-porous polymer with butyl bonding).
• System requirements: HPLC or UHPLC capable of up to 20 MPa, operating temperature 10–60 °C, pH 2.0–12.0.
• Consumables: 0.15 mm or smaller internal diameter tubing to minimize band spreading; 0.2–0.5 µm membrane filters for mobile phase and sample; PEEK and stainless steel fittings, nuts and ferrules matched to pressure ratings.

Main Results and Discussion

Adherence to the indicated flow direction and minimizing dead volume during installation are critical for achieving symmetrical peaks and high theoretical plates. Pre-flushing the flow line prevents air or contaminants from impairing performance. Each column ships with a performance report confirming QC-validated metrics. Increases in backpressure and peak splitting typically result from filter clogging or sample particulates; routine filtration and proper sample dissolution prevent these issues. A stepwise cleaning protocol—water flush, NaOH injections, and acetic acid wash—effectively restores column performance when pressure rises or retention shifts occur.

Benefits and Practical Applications

The Shim-pack Bio-HIC column delivers consistent selectivity for biomolecule analysis in quality control, bioprocess monitoring and preparative workflows. Its wide chemical compatibility and robust support facilitate method development and scale-up. The low dead-volume configuration supports high-resolution separations, even at semi-micro flow rates.

Future Trends and Opportunities

Future developments in HIC may involve sub-2 µm particles and monolithic supports for enhanced speed and resolution. Coupling HIC with mass spectrometry and microfluidics could expand its use in proteomics and single-cell analysis. Green solvents and automated cleaning approaches will further improve laboratory efficiency and reproducibility.

Conclusion

Following the recommended installation, operation and maintenance guidelines ensures reproducible performance and extends the lifetime of Shim-pack Bio-HIC columns. Proper tubing selection, routine filtration and periodic cleaning are key to maintaining stable pressure and retention characteristics.