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HSM · Factor of Similarity

HPLC Column Selectivity & Comparison

Two ways to work: Find equivalents & orthogonals — pick a reference column and instantly rank its swap-ready backups (Fs ≤ 3) and its most orthogonal columns (Fs ≥ 10) across all 817 columns; or Compare selected columns — put your column head-to-head against a shortlist you choose, with the Factor of Similarity and the per-term differences that explain why they diverge. Powered by the Hydrophobic-Subtraction Model across 817 real, published columns.

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How HPLC column selectivity is compared

Two reversed-phase columns with the same nominal chemistry (say, "C18") can give very different separations. The published Hydrophobic-Subtraction Model (HSM) explains why by describing each column with five interaction terms, so columns can be compared objectively rather than by trade name.

The five selectivity terms

Retention on a column, relative to a reference, is modelled as the sum of five contributions:

log(k / kref) = η'H − σ'S* + β'A + α'B + κ'C
TermColumn property
HHydrophobicity
S*Steric resistance to bulky solutes
AHydrogen-bond acidity (silanol activity)
BHydrogen-bond basicity
CCation-exchange / ionic interaction (pH-dependent)

Two columns are considered near-equivalent when the differences in these terms are small — expressed as a single column-comparison distance (often written Fs), where a low value means the columns should give similar selectivity.

Worked example

Two C18 phases have nearly identical H (hydrophobicity) but very different C (ionic) terms. For neutral analytes they behave alike, but for a basic, ionisable analyte the difference in C produces a large selectivity gap — so they are not interchangeable in a method that separates ionisable compounds.

Reference: Snyder, Dolan & Carr, J. Chromatogr. A 1060 (2004) 77; USP <621> column-equivalence principles.

Frequently asked questions

What makes two columns "equivalent"?

Small differences across all five HSM terms — a low overall comparison distance. Equivalence is also analyte-dependent: columns that match for neutrals may diverge sharply for acids or bases because of the C (ionic) term.

Can I swap a column in a validated method?

A close HSM match is a strong starting point and is widely used to shortlist alternatives, but any substitution in a validated method still requires experimental verification and appropriate change control.

Why does mobile-phase pH matter so much?

The C (ionic) interaction depends on whether analyte and silanols are charged, which pH controls. Two columns can look equivalent at one pH and very different at another.