Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION BY HPLC FOR THE DETERMINATION OF APIGENIN IN JUSTICIA GENDARUSSA

Raghu M. G.* and Pushpa Agrawal

ABSTRACT

A novel, simple, rapid and precise HPLC method for determination of Apigenin in Justicia Gendarussa was developed and validated. The separation of Apigenin was achieved on Thermo Syncronis C18 column (250 × 4.6 mm, 5μm) with an isocratic mobile phase containing acetonitrile and 0.1% trifloroacetic acid in the ratio of 30:70v/v, delivered at a flow rate of 1.0mL/min and the injection volume was 20μL. The eluted compound was detected by DAD detector at the wavelength of 337nm. The validation of the proposed method was carried out for system suitability, specificity, precision, linearity and robustness test as per International Conference on Harmonization (ICH) guideline. The retention time of Apigenin is found to be 27.3min. The diluent peaks were not interfered with Apigenin peak. The peak of Apigenin is pure.The method was found to be linear at the range of 20ppm to 60ppm (r2 =0.999). The method was used to assess the stability of Apigenin in Justicia Gendarussa under various laboratory conditions.

Keywords: HPLC, Apigenin, Justicia Gendarussa.