Quantitative separation of hesperidin, chrysin, epicatechin, epigallocatechin gallate, and morin using ionic liquid...

Recently, an increasing interest has been observed in ionic liquids (ILs) due to their potentialities in various chemical processes. ILs have some unique properties making them excellent additives in CE. In this work a simple, rapid, and reliable CZE method has been developed and validated using 1-butyl-3-methyl imidazolium hexafluorophosphate (BMIM-PF₆ ) ionic liquid as a buffer additive for the determination/separation of five flavonoids including hesperedin, epicatechin (EC), epigallocatechin gallate (EGCG), and morin using photodiode array (PDA) detector. The effect of several parameters such as concentration and pH of the running buffer, applied voltage, and concentration of ionic liquid were optimized. CZE at 25°C with 25 mM borate buffer of pH 9.0 at an applied voltage of 17 kV by adding 17.5 mM of IL was found to be suitable for the separation/determination of all five analytes within 08 min. Validation of the method was performed in terms of linearity, accuracy, precision, and limit of detection and quantification. The calibration curves were plotted in the concentration range of 1-200 μg/mL for all five analytes. The response was linear with R²  = 0.990 for EC, chrysin, and hesperidin, 0.992 for morin, and 0.988 for EGCG. LOD and LOQ were obtained within the range of 0.4-0.5 and 1.4-1.7 μg/mL, respectively. The proposed method showed good reproducibility with RSD of less than 3% for both migration time and peak height. The method was successfully applied for the determination of flavonoids from citrus fruits and tea samples.