Prediction of the Elution Profile of Aromatic Compounds in RP‐HPLC

Abstract

An efficient prediction method for the elution profile was used to separate aromatic compounds such as benzene, toluene, chlorobenzene, o‐xylene, and 1,2‐dichlorobenzene by RP‐HPLC. The retention factor and bandwidth were predicted under linear‐gradient condition with the three retention models. The elution profiles were calculated based on the linear and quadratic equations of retention factor, lnk=lnk _w +Sϕ, lnk=L+Mϕ+Nϕ², k=A+B/ϕ, where ϕ was the vol.% of methanol. The elution profiles were calculated by the Gaussian distribution with obtained retention factor and bandwidth. Two kinds of experiments were performed; one is the isocratic runs to estimate the coefficients of three retention models, and the other is the linear gradient runs that were carried out with same initial mobile phase composition (water/methanol=96/4, vol.%), two final mobile phase compositions (water/methanol=24/76 and 40/60, vol.%), and three gradient times (20, 40, and 60 min). The predicted elution profiles by the three retention models and new prediction method have good agreement with experimental data in the employed gradient conditions. The minimum average errors of calculated and experimental results of aromatic compounds were lower than 3.5% by Bi‐poly equation.

Keywords:

• Retention factor
• bandwidth
• elution profile
• aromatic compounds
• RP‐HPLC

Acknowledgments

The authors gratefully acknowledge the financial support provided by the Center for Advanced Bioseparation Technology. This work was performed in the High‐Purity Separation Laboratory of Inha University, Incheon, Korea.