Liquid Chromatography-Mass Spectrometric Method for Simultaneous Estimation of Apigenin and Luteolin from Achillea millefolium Linn.

To develop a rapid, specific, accurate and efficient liquid chromatography-mass spectrometry method and validate as per ICH guidelines for the simultaneous estimation of apigenin and luteolin from Achillea millefolium. The chromatographic separation was achieved by using C₈ column, 150 x 4.6mm i.d., 5μ Hibar Lichrospher, mobile phase containing 0.1% formic acid: acetonitrile (20:80 v/v). The flow rate was 0.4 ml/min. The retention time of apigenin and luteolin was found to be 8.48 min and 8.01 min respectively. Apigenin and luteolin exhibited linear in the range of 10-50 ng/ml. The method was validated for the parameters like system suitability, specificity, linearity, accuracy, precision, limit of detection and limit of quantification. The proposed method was successfully applied for the simultaneous estimation of both the constituents in Achillea millefolium and established a quantitative method for the simultaneous determination of apigenin and luteolin from Achillea millefolium.