Experimental Design-Assisted Optimization of Chromatographic Method for the Simultaneous Quantitation of Phenolic Compounds in Dried Flowers Extract

This research aimed to develop and validate a reversed phase-high performance liquid chromatography method determine phenolic compounds in dried flowers extract simultaneously. The was divided into two parts: (1) optimization of the separation condition employing Box Behnken design, (2) validation test including assessment for precision, accuracy, applicability High-Performance Liquid Chromatography (HPLC) coupled with Diode Array Detector (DAD). studied factors were flow rate (0.8−1.2 ml min-1), percentage mobile phase at beginning (0−20% B), end (70−100% B) gradient program. It statistically evinced that chromatographic resolutions (Rs>1.0) indicated acceptable protocatechuic acid, p-hydroxybenzoic aldehyde, vanillic p-coumaric ferulic acid. A fast (8.00 min) achieved by applying optimum 1 mL min-1, composition 20% acidified methanol beginning, 100% then performed developed assuring high precision accuracy. Additionally, HPLC-DAD successfully applied three flower extracts revealing reliable routine analyses.