Simultaneous identification of rutin, chlorogenic acid and gallic acid in Moringa oleifera by densitometric high-performance thin-layer chromatography method

An HPTLC (high-performance thin-layer chromatography) method was carried out to develop a feasible, simple and accurate method for the estimation of exact concentration of flavonoids in Moringa oleifera. The simultaneous determination of rutin, chlorogenic acid, and gallic acid extracted from different parts (leaf, stem, flower, and root) of M. oleifera was performed and analysis was carried out on glass-backed plates (10 cm × 20 cm) coated silica gel 60 F₂₅₄ (0.2 mm thickness, Merck, Germany) under CAMAG TLC Scanner III with UV densitometrically at λ_max = 254 nm. The mobile phase of ethyl acetate:acetone:water:formic acid with a ratio of 6:3:2:2 (v/v) was used for these flavonoids. The findings of HPTLC gave sharp, symmetrical and well-resolved peaks at Rf values of 0.51 ± 0.02, 0.72 ± 0.04 and 0.83 ± 0.03 with linearity ranging 100–700 ng/spot (r² = 0.9989), (r² = 0.9992), and (r² = 0.9984) for rutin (0.066 ± 0.003%), chlorogenic acid (0.033 ± 0.005%), and gallic acid (0.031 ± 0.005%), respectively. The developed method can also be used as a platform for estimation of complex bioactive(s) bio-synthesized in medicinal or aromatic plants.