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