Optimization of mobile phase condition by using HCI program for extracting luteolin and apigenin from celery

An efficient optimization method was used to separate flavonoid compounds from celery by HCI program software. Two interfering species, tentatively considered the target compounds, were separated successfully by the HCI program with the optimum composition of mobile phase for reversed-phase high performance liquid chromatography separation obtained. The elution profiles were calculated by the plate theory based on the linear and quadratic equations of the retention factor: lnk = lnkw + SF, k = A + B/F, lnk = L + MF + NF2, where F was the volume percentage of the organic modifier. In the binary solvents model, the calculated results of the mobile phase conditions suggested that methanol/water (60/40, v/v) or acetonitrile/water (34/66, v/v) was the optimized component, as was acetonitrile/methanol/water (10/30/60, v/v/v) when changed to a ternary solvents model. Under the experimental conditions, the agreement between the experimental elution profiles and the calculated values of the eluted concentration was relatively good. © 2010 Curtin University of Technology and John Wiley & Sons, Ltd.