Development and validation of Hplc-Uv method for quantification of podophyllotoxin in Podophyllum hexandrum Royle

Abstract

Podophyllum hexandrum Royle, (Podophyllaceae) a critically endangered Himalayan herb is a rich source of a neurotoxin compound-podophyllotoxin, which has antitumour activity and after derivatization to etoposide, etoposide phosphate and teniposide is used for treatment of various malignant conditions. A simple, precise and accurate HPLC method with UV-detection was developed and validated for quantification of podophyllotoxin in Podophyllum hexandrum. The method was validated according to the ICH guidelines which includes the parameters like accuracy, precision, linearity, range, LOD, LOD and robustness. Seperation was achieved on a reverse-phase C18 column using methanol: water: 62: 38 (v/v) as mobile phase in isocratic mode with a flow rate of 0.9 ml/min and detection was done at 280 nm. The method showed high correlation coefficients (R > 0.998) for standard subjected to the entire procedure. Value of RSD % of area for Inter-day and Intra-day precision of the developed method was 0.68 % and 0.66 % respectively. Detection and quantitation limits of the developed method were 0.026µg/ml and 0.106µg/ml respectively.