ACE 300Å HPLC columns

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ACE 300Å HPLC columns

Excellent peak shape and reproducibility have established ACE HPLC columns as the finest available. This quality is now available for protein chemists desiring the utmost in performance and reproducibility for the separation of peptides, proteins and other high molecular weight biomolecules.

ACE 300Å columns are available in an extensive range of dimensions and particle sizes for use in micro-scale separations, LC-MS analyses, high speed preparative analyses up to process scale.

  • Description
  • Comparison tests
  • When to use
  • Protein & peptide

ACE 300Å Columns for Biotechnology

  • 300Å ultra high purity silica
  • Ultimate protein and peptide application column
  • C18, C8, C4, CN and Phenyl chemistries
  • 3μm, 5μm and 10μm particle sizes
  • Unmatched reproducibility
  • Exceptional chemical stability
Phase Functional group Particle size (µm) Pore size (Å) Surface area (m2/g) Endcapped Carbon load (%)
C18-300 Octadecyl 3,5,10 300 100 yes 9.0
C8-300 Octyl 3,5,10 300 100 yes 5.0
C4-300 Butyl 3,5,10 300 100 yes 2.6
CN-300 Cyano 3,5,10 300 100 yes 2.6
Ph-300 Phenyl 3,5,10 300 100 yes 5.3

Comparison of Leading 300Å 5μm C18 Columns

In order to demonstrate the benefits of ultra-inert phases in biomolecule analysis, several commercially available 300Å pore size reversed-phase columns were tested using three different samples.

  • neutral molecules to measure efficiency,
  • pyridine/phenol to measure silanol activity and
  • antidepressants to measure both silanol activity and metal content

    • These are the same test procedures typically used to evaluate standard pore size columns (eg 100Å) used for the analysis of small molecules in the chemical and pharmaceutical industries. Columns were ranked by efficiency, N, measured at 10% peak height. In addition to measuring overall efficiency, this value also takes into consideration peak tailing usually caused by silanol interactions. The table below summarises the performance of various columns as determined by each test along with an overall ranking based on a combination of all three tests.

ACE 300Å Columns for Peptide and Protein Analyses

The chromatography of biomolecules, in particular peptides and proteins, can be improved by using HPLC columns packed with ultra-inert stationary phases. These columns will have reduced levels of silanol and metal activity to interfere with the separation. In addition, ultra-inert stationary phases perform well even when using low levels of TFA in the mobile phase. Using reduced levels of TFA improves mass spectral detection, in addition to providing a means of increasing selectivity and resolution.


Mobile phase pH is another powerful means for improving selectivity and resolution.
Ultra-inert columns, such as ACE, show no loss in performance at higher pH. Methods developed on ultra-inert columns will be more rugged over time as these columns are more reproducible column-to-column and lot-to-lot.

ACE 300Å Columns for Peptide and Protein Analyses

Chromatographers prefer inert stationary phases for the reversed-phase HPLC of ionic compounds because they minimize the negative effect of silanols on the separation. This results in improved peak shape and reproducibility when separating compounds that contain polar functional groups, especially amines.


A new generation of ultra-inert stationary phases, with extremely low silanol activity, has made it possible to achieve even better peak shape and reproducibility when separating these types of compounds. Scientists working with small molecules have been rapidly adopting this new technology and the recent introduction of wide-pore (300Å) ultra-inert phases makes the benefits of this technology available to those wanting to separate peptides and proteins by reversed-phase HPLC.