Simultaneous determination of β-sitosterol and gallic acid in Nigella Sativa seeds using reverse phase high performance liquid chromatography

Abstract

HPLC determination of β-Sitosterol and Gallic acid in seeds of Nigella sativa was carried out using reversed-phase Grace C18 column (250 mm × 4.6 mm id, 5 µm particle size), with UV detection at 210 nm. The mobile phase for column consisted of 60:40 v/v acetonitrile–water at a flow rate of 0.5 ml/min. The pH of the mobile phase was adjusted to 3.00, with 0.05% ortho-phosphoric acid. The linear response was examined in the concentration range of Gallic acid and β-Sitosterol between 0.5 and 2.5 μg/mL, and linear correlation coefficients of 0.9983 for Gallic acid, and 0.9993 for β-sitosterol, respectively, were observed. The detection limit was 4.8 µg/mL, and the recovery test in the determination of β-Sitosterol and Gallic acid revealed 98–101% for both compounds.