Simultaneous Quantification of Acid chlorogenic, Acid caffeic and Rutin in Herba Siegesbeckiae orientalis by HLPC Method

Herba Siegesbeckiae is a medicinal material derived from the aerial part of Siegesbeckia orientalis, which is used to treat various types of pain such as back, knees, bones, joints, and numbness of limbs. This plant contains several phenolic compounds, including chlorogenic acid (1), caffeic acid (2), and rutin (3). They were chosen as marker compounds to develop a quantification method. In this study, a high-performance liquid chromatography (HPLC) method with a DAD detector was used to simultaneously quantify these three compounds (1-3). The Agilent C18 column (250 x 4.6 mm; 5 µm) was used for the separation of the analytes, and the mobile phase consisted of MeCN - 0.1% phosphoric acid in water with gradient elution. The flow rate was 1.0 ml/min, and the detection wavelength was 320 nm for 1 and 2, and 360 nm for 3. The injection volume was 20 µl. The method was validated according to the guidelines of ICH and AOAC on system suitability, specificity, linearity, precision, and accuracy. The established quantification method was applied to determine the content of compounds 1, 2, and 3 in samples of Herba Siegesbeckiae, which were found to be 0.09-0.82 mg/g, 0.02-0.21 mg/g, 0.07-0.52 mg/g, respectively. This quantification method can be used to assess the quality of Herba Siegesbeckiae and related products.