HPLC-PDA Simultaneous Determination and Protective Effect of Anemarrhena asphodeloides Against Acute Renal Failure

Abstract

We investigated the protective effects against acute renal failure (ARF) of Anemarrhena asphodeloides (AA) and performed simultaneous analysis of three compounds, neomangiferin (1), mangiferin (2), and isomangiferin (3) in AA using a high-performance liquid chromatography-photodiode array. To measure the protective effect of ARF, the levels of reactive oxygen species (ROS) and glutathione depletion were determined using a kit. HPLC analysis was performed using a Gemini C₁₈ column at 40°C. The mobile phase used gradient elution with 1.0% (v/v) aqueous acetic acid (A) and 1.0% (v/v) acetic acid in acetonitrile (B). The flow rate was 1.0 mL/min. In our assay, AA extract inhibits cisplatin-induced production of intracellular ROS. Pre-incubation of AA extracts (10–200 μg/mL) markedly maintained cell viability compared with controls in the noncisplatin-treated cells. Calibration curves of all compounds showed good linearity (r²  0.9992). Recoveries of the three compounds were 98.9–103.4%. The relative standard deviations of intra- and inter-day precision were 0.07–1.73% and 0.12–1.49%, respectively. The amounts of the three components were 1.22–20.63 mg/g. The AA extract has potential as a therapeutic agent for treatment of ARF. In addition, the established method will help to improve quality control of AA.