Development and Validation of HPTLC Method for Estimation of Podophyllotoxin in Crude Extract, Isolated...

Podophyllotoxin, a naturally occurring cyclolignan isolated from the root and rhizomes of Podophyllum species is mainly used to synthesise etoposide and teniposide, which are used in various diseases. The present work proposes a simple, sensitive, specific, and rapid method to quantify podophyllotoxin using high-performance thin-layer chromatography (HPTLC). Podophyllotoxin was quantified in the roots and rhizomes of Podophyllum hexandrum, isolated podophyllotoxin and marketed mother tincture. A precise and validated densitometric method (R_f 0.41) was developed after resolving it on a silica gel plate using toluene:acetone:formic acid (6.5:5.6:1.4 v/v/v) as the mobile phase. The developed method was validated by linearity, accuracy, precision, range, limit of detection, limit of quantification, specificity and robustness. For podophyllotoxin, there was a linear relationship between standard solution concentration and peak response (area) in the 200-1000 ng spot^-1 range. The interday precision of podophyllotoxin ranged from 194.26 ± 9.302 to 314.06 ± 4.15, with a %RSD of 4.7-1.3. Similarly, the intraday precision ranged from 214.7 ± 19.55 to 443.6 ± 35.84, with a %RSD of 3.5-5.9. The chromatographic results suggest that the developed method can be used for the comparative identification of podophyllotoxin in the ethanolic extract, isolated podophyllotoxin and marketed mother tincture.