Development of Simultaneous LCâMS/MS Method for the Quantitation of Apigenin, Luteolin and Quercetin in Achillea Millefolium Extract

Introduction Achillea millefolium, family Asteraceae commonly known as Biramjasif is an erect, pubescent, perennial herb, 15-90 cm in height, commonly distributed in the Himalayas. The plant is known to be reported for traditional uses for the treatment of astringent, diaphoretic, anti-spasmodic, etc. The plant is reported to contain salicylic acid, β-sitosterol, inositol, flavonoids, viz. apigenin, luteolin, cosmosin and sesquiterpenic lactone, austricin. Method A simple and rapid liquid chromatography–tandem mass spectrometry method was developed and validated for simultaneous determination of apigenin, luteolin and quercetin in Achillea millefolium extract. The compounds were eluted using Gemini C18, (50 mm x 2.0 mm, 3 μm) with the Isocratic mobile phase consisting of water and 0.1% formic acid, Acetonitrile: Methanol (50: 50 v/v) and 0.1% formic acid, flow rate of 0.400 ml/min. Result The assay exhibited a linear dynamic range of 1.25–5000 ng/ml for apigenin, luteolin and quercetin. The values for Intraday and Interday precision and accuracy were well within the generally accepted criteria for analytical methods. Selectivity, linearity, limit of detection and quantification, accuracy and precision were evaluated. Conclusion The proposed method is more accurate and sensitive can be used for the routine quantification of the apigenin, luteolin and quercetin in the herbal extracts.