2.2. Extraction and HPLC Analysis of Ginsenoside

The ginseng roots were obtained from Jilin, China. The air-dried roots were extracted and lyophilized. To begin, the dried ginseng roots were pulverized into a fine powder using a pulverizer and passed through a 40-mesh screen. Then, 100 g of the powdered ginseng sample was extracted with 70% ethanol, and the solvent of the extract solution was evaporated under vacuum. The resulting dried extract was dissolved in water and subsequently extracted with water-saturated n-butanol. The n-butanol phase was evaporated under vacuum and then lyophilized.

Before the pharmacological evaluation, the ginsenoside was analyzed using High-Performance Liquid Chromatography (HPLC). A 20 μL sample was injected into the column and eluted at room temperature with a constant flow rate of 1.0 mL/min. The mobile phase consisted of acetonitrile (solvent A) and water (solvent B). Gradient elution commenced with 17.5% solvent A and 82.5% solvent B. The elution solvents were then adjusted to 21% A for 20 min, followed by 26% A for 3 min and held for 19 min, 36% A for 13 min, 50% A for 9 min, 95% A for 2 min, and held for 3 min. Finally, the eluting solvents were returned to 17.5% A for 3 min and held for 8 min. The detection wavelength was set at 202 nm. All sample solutions were filtered through a membrane filter with a pore size of 0.2 mm. The content of the constituents was calculated using the standard curves of 32 ginsenosides. The ginsenoside content was measured twice and then averaged.