Chromatographic Characterization of Hypophyllantin and Phyllantin in Methanolic Extracts from Three Common Phyllanthus Species in Malaysia.

Phyllanthus amarus, Phyllanthus niruri Linn and Phyllanthus urinaria are small medicinal herbs belonging to the Euphorbiaceae family which is widely distributed in tropical and subtropical countries [1]. The medicinal values possessed by this genus are due to the remarkable secondary metabolites present in order to treat various diseases such as liver cancer [2]. The objective of the study is to investigate the chromatographic profile of hypophyllanthin and phyllanthin from three commonly found Phyllanthus species in Malaysia by utilizing High Performance Liquid Chromatography (HPLC) technique to propose a reference standard. The chemical compounds were isolated from the whole plants of Phyllanthus samples by performing methanol extraction. The extracts then were subjected into reverse-phase chromatography by C-18 column to partitioning hydrophobic-lignan through gradient elution mode. The compounds in the extracts were identified by comparing their peaks with the same HPLC program run by the reference standard. The quantitative analysis for the quantification of both lignans contained in the Phyllanthus extracts showed that percentage of the both compounds ranged from 0.44% to 3.23% for hypophyllanthin and 2.98% to 18.51% for phyllanthin. Phyllanthus niruri Linn presented notable results in both recoveries of the markers. The reversed-phase HPLC method was validated by constructing graphs of linear calibration curve for both reference standards. The correlation coefficient (R2) values obtained were 0.9727 and 0.9257 for hypophyllanthin and phyllanthin, correspondingly. This investigation will aid the identification and authentication of species used in the commercialization of Phyllanthus-based products.

[1] Webster, G. L. Ann Missouri Bot. Gard, 1994, 81:33 – 144.

[2] Sarin, B, Verma, N, Martin, J. P. and Mohanty, A. (2014). The Scientific World Journal, Volume 2014, Article ID 839172, 12 pages.