Shimadzu Shim-packTM Bio Diol HPLC Column

Importance of the topic

The analysis of biomolecules by high-performance liquid chromatography is critical for applications in biopharmaceutical development, quality control, and proteomic research. Columns with tailored surface chemistry and pore structure, such as the Shim-pack Bio Diol series, enable reproducible separations, improved peak shape, and extended column life.

Objectives and Study Overview

This instruction manual aims to guide users in installation, operation, maintenance, and troubleshooting of Shim-pack Bio Diol columns. Emphasis is placed on maximizing chromatographic performance, ensuring column longevity, and avoiding common sources of peak distortion and pressure issues.

Applied Instrumentation

• Shim-pack Bio Diol columns: dihydroxypropyl-bonded silica phase available in pore sizes of 6, 12, 20, 25, and 30 nm and particle sizes of 2, 3, and 5 µm.
• Guard columns compatible with Bio Diol series in various dimensions (4.6 mm i.d. × 10 mm, 8.0 mm × 30 mm, 20 mm × 50 mm) to protect analytical columns.
• Typical HPLC system components: pump, injector or autosampler, detector, connecting tubing (minimized length and inner diameter to reduce dead volume).

Methodology

• Column Installation: Confirm flow direction arrow; use shortest tubing connections; flush system and column with mobile phase to remove air and contaminants.
• Operating Conditions: Recommended pH range 5.0–7.5 (5.0–7.5 for all but Bio Diol-250); temperature limits up to 50 °C (40 °C for Bio Diol-250); maximum pressure up to 45 MPa for 2 µm particles and 20 MPa for 3–5 µm particles.
• Mobile Phase and Sample Preparation: Use aqueous buffers (≤0.7 M salt) with optional surfactants (≤0.1% Tween 80 or SDS) and denaturants (urea or guanidine hydrochloride). Filter mobile phases (0.45 µm) and samples to prevent column clogging.
• Column Washing and Storage: Rinse with high-salt eluent to remove hydrophobic adsorbates, then flush with water and store in 0.05% sodium azide solution at stable temperature.

Main Results and Discussion

Shim-pack Bio Diol columns demonstrate consistent QC-verified performance, with stable retention times and minimal peak tailing when operated within recommended conditions. Proper handling and flushing protocols prevent dead volume formation and maintain low back pressure.

Benefits and Practical Applications

• Robust separation of proteins, peptides, and other biomolecules in pharmaceutical and research laboratories.
• Extended column lifetime through optimized storage and washing procedures.
• Compatibility with a wide range of aqueous buffers and organic modifiers for method development flexibility.

Future Trends and Potential Uses

Advances may include integration of Bio Diol phases with ultrahigh-pressure systems for faster separations, coupling to mass spectrometry for enhanced proteomic workflows, and development of novel guard cartridge formats for streamlined column protection.

Conclusion

By adhering to the recommended installation, operational, and maintenance guidelines, users can fully exploit the high-performance characteristics of Shim-pack Bio Diol columns, achieving reproducible, high-resolution separations in biopharmaceutical and life-science applications.