Quantitative Analysis of Phyllanthus Species for Bioactive Molecules Using High-Pressure Liquid Chromatography and Liquid Chromatography–Mass Spectrometry

Abstract

Phyllanthus amarus Schum. & Thonn. and P. urinaria Linn. are valuable medicinal plants which are widely used in several countries for the treatment of jaundice, diabetes, cancer and other important diseases. The Phyllanthus species are well-known for their important bioactive lignans (phyllanthin, hypophyllanthin and niranthin) and an antioxidant (ellagic acid). Qualitative and quantitative analyses by high performance liquid chromatography (HPLC) and liquid chromatography coupled to mass spectrometry (LC–MS) method revealed the contents of four bioactive molecules in Phyllanthus amarus and Phyllanthus urinaria from district Udupi, Karnataka, India. The phyllanthin, hypophyllanthin and niranthin were eluted out at specific retention time of 16.008, 14.838 and 21.106 min respectively by isocratic method and ellagic acid was eluted out at 28.656 min using gradient method in HPLC. The extracts of P. amarus from three taluks in district Udupi, Karnataka, India showed higher amounts of phyllanthin, hypophyllanthin and niranthin as compared to that of P. urinaria. The ellagic acid was found to be quantitatively higher than the three lignans of P. amarus and P. urinaria. The phytochemical profiling of bioactive compounds in these two species collected from district Udupi showed significant variations in their contents. The quantification of bioactive molecules using HPLC and LC–MS is consistent and reproducible for the future studies on somaclonal variant plants with varied amount of bioactive molecules, and transgenic plants with over-expression of lignans and phenolic compounds which has higher market value in the pharmaceutical industry.