Calycosin 7 o β d glucoside

Based on Park et al.’s paper [34 (link)], the profiling of the extract of the dried root of Astragalus membranaceus was analyzed using HPLC. In brief, a YMC ODS-AM (4.6 × 250 mm, 5 μm) column was applied at 30 °C with 0.1% formic acid and acetonitrile as the gradient system of the mobile phase under the Water e2695 series HPLC system (Waters Corporation, Milford, MA, USA). The mobile phase was maintained in 5% acetonitrile for 3 min and increased to 20% for 3 min, and then, up to 28% for 25 min. Then, elution was carried out by increasing the mobile phase to 100% acetonitrile for 4 min with a flow rate of 1 mL/min. The absorbance of the UV detector was measured at a wavelength of 254 nm compared to standard chemicals such as calycosin (99%) and calycosin-7-O-β-D-glucoside (99%) with high purity from ChemFaces (Wuhan, China) and Sigma-Aldrich (St. Louis, MO, USA).

Human SW1353 chondrosarcoma cells (ATCC® HTB-94™) were purchased from the American Type Culture Collection (Rockville, MD, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin and streptomycin and phosphate buffer saline (PBS) were purchased from GIBCO-BRL (Grand Island, NY, USA). Monosodium iodoacetate (MIA), 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT), λ-carrageenan and acetic acid were purchased from Sigma-Aldrich (St Louis, MO, USA). The standard compounds, calycosin, calycosin-7-O-β-D-glucoside and lithospermic acid, were purchased from the Chem Faces (Hubei, China).