Simultaneous Determination of the Traditional Herbal Formula Ukgansan and the In Vitro Antioxidant Activity of Ferulic Acid as an Active Compound

Abstract

Ukgansan (UGS), a traditional herbal formula composing seven medicinal herbal plants, has been applied in Asian countries for treating neurosis, insomnia, and irritability. Here, the current study performed a simultaneous determination of the seven marker compounds (liquiritin apioside, liquiritin, ferulic acid, glycyrrhizin, decursin, decursinol angelate, and atractylenolide I) using high-performance liquid chromatography (HPLC), to establish quality control of UGS. A 70% ethanol extract of UGS and a mixture of the seven compounds were separated using a C-18 analytical column on a gradient solvent system of 1.0% (v/v) aqueous acetic acid and acetonitrile. Data were recorded at a UV wavelength of 250 nm for glycyrrhizin; 276 nm for liquiritin apioside, liquiritin, and atractylenolide I; and 325 nm for ferulic acid, decursin, and decursinol angelate. The results exhibited high linearity (correlation coefficient (r2) ≥ 0.9998) and proper precision (0.38–3.36%), accuracy (95.12–105.12%), and recovery (95.99–104.94%) for the seven marker compounds. The amount of the seven marker compounds at the concentrations from 0.190 to 16.431 mg/g. In addition, the current study evaluated the antioxidant effects of UGS by measuring their scavenging activities against the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals using in vitro cell-free systems and observed its antioxidant activity. Among the seven components of the UGS extract, ferulic acid dramatically enhanced the scavenging of ABTS and DPPH radicals compared with other compounds. The concentrations of ferulic acid required for a 50% reduction (RC50) in ABTS and DPPH radicals were 16.22 μM and 41.21 μM, respectively. Furthermore, UGS extract exerted the neuroprotective effect and blocked the inflammatory response in neuronal hippocampal cells and microglia, respectively. Overall, the established method of HPLC will be valuable for improving the quality control of UGS extract, and ferulic acid may be useful as a potential antioxidant agent.

Goat’s Milk Intake Prevents Obesity, Hepatic Steatosis and Insulin Resistance in Mice Fed A High-Fat Diet by Reducing Inflammatory Markers and Increasing Energy Expenditure and Mitochondrial Content in Skeletal Muscle

“…Among the polyphenols found in goat's milk, gallic acid, chlorogenic acid, ferulic acid and catechins are the most abundant [10], which have demonstrated to have antioxidant properties [32][33][34], increase in energy expenditure in mice [35] and humans [36] and anti-inflammatory properties [37]. Thus, the polyphenol content in goat's milk could be directly responsible of the increase in energy expenditure.…”

Similarity of therapeutic networks induced by a multi-component herbal remedy, Ukgansan, in neurovascular unit cells

“…Previously, we identified a few representative chemical compounds included in UGS using the analytical profile of high-performance liquid chromatography (HPLC) in which decursin, glycyrrhizin, and decursinol angelate were predominantly included 18 . Moreover, one of them, namely ferulic acid, shows antioxidant activity.…”

“…In addition, UGS has been reported to improve behavioral deficits and protect neuronal cells from degeneration in animal models [20][21][22] . We also demonstrated that ferulic acid, one of the active compounds of UGS, could play an important role, as an antioxidant, in its therapeutic effects 18 . These previous reports strongly suggest that UGS could exert various therapeutic roles in the brain by targeting diverse cellular components.…”

“…The prepared mixed or single crude herbs were extracted using 70% aqueous ethanol by refluxing at 100 °C for 2 h. The extract was then filtered through a paper filter with a pore diameter of 5 μm and concentrated using a vacuum rotary evaporator system (EYELA N-1000, Bohemia, NY, USA) to give a powder, as described in our previous paper 18 . Voucher specimens have been deposited in extract bank at Korea Institute of Oriental Medicine.…”

“…Ukgansan (UGS), also called Yokukansan or Yi-Gan San, is a traditional oriental prescription composed of 7 medicinal ingredients including Uncaria sinensis (Uncariae Ramulus et Uncus), Atractylodes japonica (Atractylodis Rhizoma Alba), Poria cocos (Poria Sclerotium), Bupleurum falcatum (Bupleuri Radix), Angelica gigas (Angelicae Gigantis Radix), Cnidium officinale (Cnidii Rhizoma), and Glycyrrhiza uralensis (Glycyrrhizae Radix et Rhizoma) 18 . UGS has been approved by the Ministry of Health, Labour and Welfare of Japan for use against pathological conditions such as insomnia, irritability, and neurosis in children 19 .…”