Development, Optimisation and Validation of a Stability‐Indicating HPLC Method of Achyrobichalcone Quantification using Experimental Designs

Abstract

The new method developed affords evaluation of the quality of achyrobichalcone obtained by isolation, and indicates the stability of the molecule under various stress conditions.

Semi‐preparative isolation and purification of phenolic compounds from Achyrocline satureioides (Lam) D.C. by high‐performance counter‐current chromatography

“…The data were analysed in the LC‐Solution Multi‐PDA software. The quantification of ACB was performed using the following parameters: mobile phase in isocratic mode consisted of a mixture of ACN/H 2 O with orthophosphoric acid 0.16 M (37:63, v / v ), flow rate of 1.2 mL/min, column oven at 33 ± 1°C, and detection wavelength of 367 nm . The concentrations of unidentified peaks observed in the chromatogram were calculated based on the ACB analytical curve (0.1–10.0 μg/mL).…”

“…Equal volumes of the lower and upper phases were evaporated to dryness separately. The residues were diluted with MeOH to 1 mL and analysed by HPLC‐PDA to determine the distribution ratios . The K D was defined as the peak area of compound in the lower phase divided by the peak area of compound in the upper phase ([ A ] low and [ A ] up , respectively).…”

“…Next, 2 mL of each phase consisting of CH 2 Cl 2 /MeOH/H 2 O) of solvent systems (4.0:3.5:2.0; 4.5:3.0:2.0; 3.5:3.5:2.5, and 3.5:4.0:2.0, v / v / v ) . The samples were processed as previously described: equal volumes of the upper and lower phases were evaporated, diluted with MeOH to 1 mL and analysed by HPLC‐PDA to determine the distribution ratios . It is important to point out that, unlike the aforementioned system, the lower phase is now the organic phase, due to the density of CH 2 Cl 2 ( ρ = 1.325 g/mL).…”

“…In this step, the tubes were grouped in two fractions according to the previous HPLC‐PDA analysis: F 1 containing ACB and unidentified compounds, and F 2 containing the flavonoid‐aglycones (QCT, LUT, and 3 O MQ). The solvent was evaporated under reduced pressure and ACB, 3 O MQ, QCT, and LUT concentrations were evaluated by HPLC‐PDA . The concentrations of unidentified compounds in F 1 were determined using the ACB analytical curve.…”

“…Twenty‐four fractions (3 mL each) were collected. The solvent was evaporated under vacuum and the concentration of substances was evaluated by HPLC‐PDA methods …”