Thermo Scientific Accucore HPLC Columns for Biomolecule Separations

Importance of the Topic

The analysis of peptides and proteins by high performance liquid chromatography is fundamental in biopharmaceutical research, proteomics, and quality control. Rapid, high-resolution methods that maintain sensitivity while minimizing backpressure enhance throughput and data quality in laboratories worldwide.

Objectives and Overview

This document reviews the design and performance of Accucore HPLC columns featuring Core Enhanced Technology. The aim is to demonstrate how 150 Å pore diameter solid core particles and optimized manufacturing processes deliver superior resolution, peak capacity, and reproducibility for biomolecule separations across analytical, micro, and nano formats.

Methodology

Core Enhanced Technology integrates four key elements: solid core particles (2.6 µm diameter), tight particle size distribution, advanced phase bonding for high surface coverage, and automated packing to form uniform beds. The 150 Å pore size is tailored for peptides and proteins, reducing mass transfer resistance and eddy diffusion according to Van Deemter principles. Multiple column formats (analytical 3.0–4.6 mm ID, micro 2.1 mm ID, nano 75 µm ID) support flow rates from 300 nL/min to 1.8 mL/min and temperatures up to 70 °C.

Instrumentation

• Thermo Scientific Accucore 150-C18 and 150-C4 HPLC columns packed with 2.6 µm solid core particles
• Defender guard cartridges for sample protection
• nanoViper fingertight fittings for nanoLC connections (≤ 1000 bar)
• High-pressure stainless steel hardware compatible with ≤ 1000 bar systems
• UV detection (214 nm, 220 nm, 280 nm) and mass spectrometry coupling (e.g., Orbitrap)

Main Results and Discussion

• Peptide separations on Accucore 150-C18 achieve sharper peaks and ~20–25 % higher peak capacity compared to 80 Å phases and fully porous C18 columns, at lower backpressures (~175 bar vs. ~320 bar).
• Retention time reproducibility is excellent (RSD < 0.15 %), enabling reliable quantitation of insulin, ribonuclease A, and synthetic peptides.
• Linear loading capacity up to ~2 µg for peptides and ~25 µg (1200 pmol) for proteins on 150-C4 columns, with minimal loss in resolution.
• Intact protein analyses demonstrate baseline resolution of insulin, cytochrome C, and larger proteins, with MS-compatible formic acid mobile phases and optimized gradients.

Benefits and Practical Applications

• Rapid method development and high throughput due to fast separations and low system backpressure.
• Enhanced sensitivity from narrow peak widths and high peak capacity, critical for trace analysis.
• Robust reproducibility supports routine QC and proteomics workflows.
• Versatile column formats allow seamless scaling from nanoLC to analytical scale.

Future Trends and Opportunities

Ongoing advances may include deeper pore architectures for very large biomolecules, improved phase chemistries for selectivity, and integration with ultra-high resolution mass spectrometry. Further automation in column packing and online sample preparation could streamline workflows, while expansion of nanoViper and microfluidic interfaces will enhance compatibility with emerging proteomic platforms.

Conclusion

Accucore HPLC columns employing Core Enhanced Technology offer a balanced solution for high-resolution peptide and protein separations. The combination of solid core design, precise manufacturing, and flexible formats delivers superior peak capacity, reproducibility, and loading capacity, meeting the demands of modern biomolecular analysis.

Reference

• Thermo Fisher Scientific. Thermo Scientific Accucore HPLC Columns for Biomolecule Separations, 2012.