Response Surface Methodology-Based Extraction of Fumaric Acid From Fumaria officinalis L. and Quantification by RP-HPLC.

In the present study, it was aimed to optimize the extraction of fumaric acid from Fumaria officinalis L. by response surface methodology and its quantification by a newly developed and validated reverse phase high performance liquid chromatography method. Chromatographic separations were performed at 40°C with a C18 column. Aqueous o-phosphoric acid solution and methanol were used as the mobile phase by gradient elution. The flow rate was set at 1.0 mL/min, and the injection volume was 20 μL. Fumaric acid was detected at 215 nm with a UV detector. The method was linear in the range of 10.00-200.00 μg/mL. The limits of detection and quantification were 3.02 μg/mL and 9.15 μg/mL, respectively. The relative mean errors were ≤ 3.77%, whereas the relative standard deviation values were calculated as ≤ 0.48%. The conditions to obtain an extract with high fumaric acid content were optimized by central composite design, a type of response surface methodology. The independent variables were extraction time, solvent volume, and temperature for the maceration procedure, where fumaric acid concentration was the response. The experimental model was evaluated statistically and found to fit well, considering the lack-of-fit p value of 0.211 and coefficient of determination 0.9862 at a confidence level of 95%.