DEVELOPMENT AND OPTIMIZATION OF HPLC-PDA-MS-MS METHOD FOR SIMULTANEOUS QUANTIFICATION OF THREE CLASSES OF FLAVONOIDS IN LEGUME SEEDS, VEGETABLES, FRUITS, AND MEDICINAL PLANTS

Certain plants of the daily human diet are an important source of beneficially healthy flavonoids, possessing significant pharmacological activities against various diseases. Flavonoids display an array of chemical diversity based on relative positioning of benzopyrano moiety and aromatic ring. Due to structural diversity of these molecules, simultaneous quantification of these molecules in plant materials becomes difficult. In the present study, a rapid, simple, and sensitive, High Performance Liquid Chromatography-Photo Diode Array Mass Spectrometry (HPLC-PDA-MS-MS) method was developed for the simultaneous determination of three different classes of flavonoids, namely, isoflavonoids, flavanones, and flavonols in legume seeds, vegetables, fruits, and medicinal plants. The method was evaluated for validation parameters, such as linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), specificity, selectivity, and compound stability. The developed method was applied on extracts of different plant samples for quantification of the flavonoids daidzein, genistein, naringenin, quercetin, kaempferol, and rutin. Method developed in this study can be used in the quality control and standardization of plant extracts as well as herbal drugs and formulations.