ISOLATION AND HPLC METHOD DEVELOPMENT FOR FILIXIC ACID PBP FROM DRYOPTERIS FILIX-MAS

Simple, rapid and specific isolation and HPLC method has been develop for filixic acid PBP in Dryopteris filix-mas extract. HPLC analysis was performed on C18 column using a 90:10 (v/v) mixtures of acetonitrile and methanol as isocratic mobile phase at a flow rate 1.0 ml/min. UV detection was at 254 nm for filixic acid PBP. Filixic acid shown retention time at 4.02 min. Method was validated for accuracy, precision, linearity, specificity and sensitivity in accordance with International Conference on Harmonisation guidelines. Validation studies revealed that the method is specific, accurate, precise, reliable and reproducible. Good linear correlation coefficient (r2 > 0.993) was obtained for calibration plots in the range tested. The limit of detection was 2.25 μg/ml and limit of quantification was 7.53 μg/ml for filixic acid PBP. Intra and inter-day relative standard deviation of precision was less than 1.50 %. Recovery was between 94.57 and 101.05 % filixic acid. The method can be successfully used for quantitative analysis of filixic acid PBP in D. filix-mas for day-to-day studies.