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HPLC analysis of Phyllanthus amarus samples stored in stability chambers under different conditions and study of the effect on quantification of the phytomarkers phyllanthin and hypophyllanthin

Srivastava P., Raut H. N., Puntambekar H. M, Desai A. C

Acta Chromatographica

2015

Vol. 27, no. 1

147--156

Storage of medicinal plants may cause deterioration of the active principles with time, thus, reducing the efficacy of plants. Therefore, quantification of the active principle is essential before using the crude drug. Phyllanthus amarus (PA) contains lignans, namely, phyllanthin and hypophyllanthin, which shows anti-hepatotoxic activity. In this paper, we highlight the effect of storage conditions on the quantification of bioactive markers by high-performance liquid chromatography (HPLC) analysis in the crude plant material of PA. HPLC analysis of crude PA samples stored for certain period at long-term study (LS, 30 °C and 65% RH), accelerated study (AS, 40°C and 75% RH), and real-time study (RT) conditions was carried out using the LiChroCART Purospher® STAR RP-18 endcapped (250 × 4.6 mm, 5 μm) column along with a Purospher STAR RP 18e (4.0 × 4.0 mm, 5 μm) guard column using methanol:water (70:30) at a flow rate of 0.7 mL min−1 with ultraviolet (UV) detection at 220 nm. The HPLC study indicated that PA samples kept under LS condition are rich in lignan contents as compared to the samples stored under AS and RT study conditions. Therefore, PA should be used fresh to get maximum concentration of active lignans or it should be stored under LS conditions up to 6 months.

Development and validation of RP-HPLC method for simultaneous analysis of andrographolide, phyllanthin, and hypophyllanthin from herbal hepatoprotective formulation

Bhope S. G., Kuber V. V., Nagore D. H., Gaikwad P. S., Patil M. J.

Acta Chromatographica

2013

Vol. 25, no. 1

159--169

A simple, sensitive, and accurate liquid chromatographic method with photodiode array detector was developed for the determination of andrographolide, phyllanthin, and hypophyllanthin. The separation was carried out on a reverse-phase 250 mm × 4.6 mm, 5μ symmetry C8 column (Waters). The gradient was prepared from 0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile:methanol (solvent B) as mobile phase delivered at a flow rate of 1 mL min -1. A linear behavior was observed between observed peak area response, and concentration of analytes was investigated, with good correlation coefficient. The method established was successfully applied to quantify andrographolide, phyllanthin, and hypophyllanthin from the herbal hepatoprotective formulation.