Abstract
Phyllanthus amarus is a small herb belonging to the Euphorbiaceae family. It is well known for its potent medicinal properties and widely used worldwide. The chromatographic method was developed for rapid screening of phytochemicals using high-pressure liquid chromatography hyphenated with quadrupole time-of-flight mass spectrometer (HPLC/ESI-QTOF-MS/MS). Fifty two compounds were identified and characterized out of which forty three phenolics and nine lignans were successfully characterized based on retention time, exact mass, molecular formula and their fragmentation patterns. Among them, six bioactive compounds namely phyllanthin, hypophyllanthin, gallic acid, protocatechuic acid, kaempferol 3-O-rutinoside, and quercetin, were further quantified in 3.6min by developing ultra high performance liquid chromatography electrospray tandem mass spectrometry (UPLC/ESI-MS/MS) method in multiple reaction monitoring (MRM) acquisition mode. It was found that the calibration curves for all analytes showed good linearity (R2>0.9986) within the test ranges. The overall limits of detection (LODs) and limits of quantification (LOQs) were 0.18–1.82ng/mL and 0.44–3.82ng/mL, respectively. The relative standard deviations (RSDs) for intra- and inter-day precisions were below 1.12% and 1.92%, respectively. Remarkable differences were observed in contents of the bioactive compounds in all samples. Principle component analysis was used for samples discrimination obtained from different geographical locations. On the basis of PC scores, samples MP and WB were closer and lying in the same quadrant, while UP was much far apart.
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