A validated TLC-image analysis method for detecting and quantifying bioactive phyllanthin in Phyllanthus amarus and commercial herbal drugs

Phyllanthus amarus Schum. and Thonn. has long been used for the treatment of liver diseases. The hepatoprotective compound presented in P. amarus was phyllanthin. In this study, the fast determination and quantitation of bioactive phyllanthin in P. amarus and its commercial herbal drugs were developed using a simple thin-layer chromatographic (TLC)- image analysis method. Chromatographic separation of phyllanthin was carried out and the intensity of phyllanthin was examined by TLC-image analysis. The linear equation line showed good relationship for the calibration curve in the various concentrations of phyllanthin. The limits of detection and limits of quantitation were 0.16 and 0.49 µg/spot, respectively. The contents of phyllanthin from fifteen plant materials collected from different locations and twelve commercial herbal drugs determined using the TLC-image analysis method were not significantly different from those determined using the TLCdensitometric method. These results suggest that the proposed TLC-image analysis method can be used as an effective method for quantitating phyllanthin in P. amarus plants and commercial products