High-Performance Liquid Chromatographic Quantification and Validation of Luteolin Glycosides from Sonchus brachyotus and Their Peroxynitrite-Scavenging Activity

In Korea, the leaves of Sonchus brachyotus (Compositae), an edible mountainous vegetable, are traditionally used to treat hepatitis and hemorrhage and are known to have diuretic action. The aqueous ethanolic extract of this plant was selected in our screening experiment using the peroxynitrite (ONO2￣)-scavenging assay, and the present study was performed to qualitatively and quantitatively identify the active compounds from S. brachyotus and validate the present high-permormance liquid chromatography (HPLC) coupled with ultraviolet absorption detection method based on accuracy, precision and repeatability. Five phenolic substances including the main compound, luteolin 7-O-β-D-glucuronopyranoside, as well as chlorogenic acid, luteolin 7-O-rutinoside, luteolin 7-O-β-D-glucopyranoside, and luteolin, were found in the aqueous ethanolic extract of S. brachyotus. In the HPLC validation experiment, the linearity of the four compounds was established by R2 values of more than 0.999 within the test ranges, and the recovery rate ranged from 98.2 - 105.3%. Luteolin 7-O-glucuronide was a predominant compound (143 mg/g of extract and 18.3 mg/g of the dry weight of plant material) with a potent peroxynitrite-scavenging effect (IC50, 1.02 ± 0.08 uM). Luteolin and its three glycosides together with chlorogenic acid were qualitatively and quantitatively determined using an HPLC method validated in the present study.