A Method for Converting HPLC Peak Area from Online Reaction Monitoring to Concentration Using Nonlinear Regression

Online HPLC reaction progress monitoring provides detailed data-rich profiles; however, extracting kinetic determine concentrations from peak areas. If the reaction's overall mass balance is known and some analytical trend for all relevant species can be recorded, it is possible to estimate the absolute response factors of all species using a system of linear equations. We delineate a method using the Microsoft Solver plug-in to convert time course profiles to reagent concentrations without analytical standards.

Automated summary

Online HPLC reaction progress monitoring provides detailed data-rich profiles, but it is difficult to determine kinetic concentrations from peak areas. By utilizing a system of linear equations, it is possible to estimate the absolute response factors of all species if the reaction's overall mass balance is known and some analytical trend for all relevant species can be recorded. We propose a method using the Microsoft Solver plug-in to convert time course profiles to reagent concentrations without analytical standards.