Shape-Based Peak Identity Confirmation in Liquid Chromatography.

Abstract

Chromatography is used to separate analytes in a mixture. In the absence of a detector providing analyte-specific information, the retention time is used to identify the analyte, typically by comparing the retention time of the unknown with that of a standard. This is the simplest of identification routines, but it remains the most widely used. The challenge is that analyte retention times can and do shift; sole reliance on retention time for identification is not prudent. Retention time is influenced by the total sample concentration, elution of components near the peak of interest, aging of a column, and so on, complicating time window-based identification. We propose a sameness threshold (ST) that is based on numerical comparison of shapes of chromatographic peaks for identification confirmation. We demonstrate shape-based identification confirmation or lack thereof of an unconfirmed peak by comparison with a shape library of standards. In numerous cases we look at, only one analyte in the shape library fits the putative identity. ST can be based on r2, the commonly used coefficient of determination used in regression analysis, or on an "index of width mismatch" that compares the widths of the peaks throughout the available height range.