OPTIMIZATION OF SEPARATION OF FLAVONOIDS IN MICELLAR LIQUID CHROMATOGRAPHY USING EXPERIMENTAL DESIGN AND DERRINGER'S DESIRABILITY FUNCTION

Abstract

Derringer's desirability function was used to simultaneous optimization of resolution and analysis time of five flavonoids myricetin, quercetin, hesperetin, naringenin, and apigenin in MLC. The factors studied were the concentration of sodium dodecyl sulfate [SDS], alkyl chain length of the alcohol (N), volume percentage of alcohol (Vm), volume percentage of acetic acid (AcOH) in mobile phase, and flow rate. The experiments were performed according to a face-centered cube half fractional central composite design to map the chromatographic response surface. The chromatographic response functions (CRFs) were formed using two sigmoidal desirability functions and were considered as the responses of design. The qualities of the obtained chromatograms in each experiment were calculated using CRF and model was built by the use of multiple linear regression (MLR) to correlate the obtained CRFs to the experimental factors and their interactions. The developed MLR model showed good descriptive and predictive ability (R2 = 0.9530 and F = 21.88). The optimum mobile phase composition for separation of interested flavonoids using a C18 column was [SDS] = 0.040 mol L−1; 11.2 % v/v butanol, 1.4 % v/v AcOH with flow rate of 1.1 mL/min. The efficiency of prediction of polynomial model was confirmed by performing the experiment under the predicted optimal conditions.