Abstract
A simple, rapid, and sensitive high-performance liquid chromatography (HPLC) method has been developed to simultaneously identify and determine five major active components from Dysosma difformis root, using a Diamonsil octadecyl bonded silica (ODS) C18 column (250 mm × 4.6 mm) and Photo-diode array (PDA) detector at 254 nm. Satisfactory separation was achieved by an isocratic elution with 0.1% aqueous phosphoric acid/methanol (40:60, v/v, pH = 3.2) at a flow rate of 1.0 mL/min. Four flavonoids (quercetin, kaempferol, rutin, and quercitrin) and a lignan (podophyllotoxin) in D. difformis root were well separated on the chromatogram within 24 min. Validation of the reverse phase HPLC (RP-HPLC) method showed good linearity (r2 > 0.9989) of the five compounds, which could be analyzed in relatively wide concentration ranges. The relative standard deviation values for intra-day and inter-day precision were less than 5.1%. The contents of these five active components in D. difformis root of different growth locations were also detected.
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