In practice, gradient slope is the most powerful parameter and almost nobody mentions it, which is why two reports on the same material disagree by a point.
Temperature affects the dynamics of molecular conformation, and if a peptide has proline residues that interconvert on the chromatographic timescale, the peak will split or shoulder at low temperature and collapse at high temperature.
Concretely, tailing factor measures peak shape, and a badly tailing peak spreads into the region where small impurities live, forcing tangent-skim integration that assigns tail area to the main peak.
The caveat is that purity without identity is only half an answer — a high purity does not mean the peak is actually what you think it is.
If you are ranking vendors, specify a method and have all samples tested at the same place.
2This is the first explanation of that which has actually made sense to me. – assay_blank 38 days ago 3Note that the label instructions differ between agents on precisely this point. – sian_llewellyn 3 months ago add a comment