High-performance liquid chromatographic analysis for quantitation of marker compounds of Artemisia capillaris thunb.

Abstract

Two stable high-performance liquid chromatography (HPLC) methods were developed that could quantitatively analyze 10 major marker compounds of Artemisia capillaris Thunb and could also distinguish among 'Injinho' and 'Myeon-injin' and 'Haninjin'--A. capillaris collected in autumn, A. capillaris collected in spring and A. iwayomogi, which can be misused as 'Injinho' in Korean herbal drug markets. The first HPLC method was a reversed-phase chromatography using a C18 column with an isocratic solvent system of phosphoric acid (0.05%) and acetonitrile at the flow rate of 1.0 mL/min, ultraviolet (UV) detection wavelength at 254 nm and column temperature at 40°C. Calibration and quantitation were made by using acetaminophen as an internal standard (I.S-A) and chlorogenic acid (1) was determined within 20 min. The second HPLC method was a reversed-phase chromatography using a C18 column with a gradient solvent system of phosphate buffer (0.015 M, pH 6) and acetonitrile at the flow rate of 1.0 mL/min, UV detection wavelength at 254 nm and column temperature at 40°C. Calibration and quantitation were made by using ethylparaben as an internal standard (I.S-B) and 3,5-di-O-caffeoylquinic acid (2), 3,4-di-O-caffeoylquinic acid (3), 4,5-di-O-caffeoylquinic acid (4), hyperoside (5), isoquercitrin (6), isorhamnetin 3-O-robinobioside (7), isorhamnetin-3-O-galactoside (8), isorhamnetin-3-O-glucoside (9) and scoparone (10) were determined within 60 min. Pattern recognition analysis of data from the 60 samples classified them clearly into three groups. These assay methods could be applied for QA/QC of A. capillaris and Artemisia iwayomogi.