Silica gel

The UV spectra were measured on a Shimadzu UV-1700 spectrometer (Shimadzu Corporation, Kyoto, Japan). The IR spectra were measured on a Nicolet 10 Microscope Spectrometer (Thermo Scientific, San Jose, CA, USA). The 1D and 2D-NMR spectra were recorded on Bruker-AC (E)-500 spectrometer (Bruker AM 500, Fällanden, Switzerland) using tetramethylsilane (TMS) as an internal standard. The HR-ESI-MS was determined on a Bruker microTOF-Q instrument (Bruker BioSpin, Rheinstetten, Germany). Column chromatography was performed with silica gel (200–300 mesh; Qingdao Marine Chemical Inc., Qingdao, China), sephadex LH-20 (GE Healthcare), and ODS (50 µm; YMC Co. LTD., Kyoto, Japan). Preparative high performance liquid chromatography (HPLC) separations were performed on a SEP system (Beijing Sepuruisi scientific Co., Ltd., China) equipped with a variable-wavelength UV detector, using a YMC-Pack ODS-A column (250 × 20 mm, 5 μm). Chemical reagents for isolation were of analytical grade and purchased from Tianjin Siyou Co., Ltd., China. Biological reagents were from Sigma Company. Human heptocellular (HepG2), and breast (MCF-7) cancer cell lines were from Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, China.

Optical rotations were determined using an MCP200 automatic polarimeter (Anton Paar, Graz, Austria). Ultraviolet spectra were recorded on a DU 730 nucleic acid/protein analyzer (Beckman Coulter, Brea, CA, USA). IR spectra (film) were measured with a Tensor 27 FT-IR spectrometer (Bruker, Ettlingen, Germany). 1D and 2D NMR spectra were recorded on a Bruker AV-600 spectrometer, δ in ppm rel. to TMS, J in Hz. ESIMS were recorded using an 1290-6420 Triple Quadrupole LC-MS spectrometer (Agilent, Santa Clara, CA, USA). HRESIMS were recorded with an Agilent G6230 TOF mass spectrometer. Silica gel (100–200 mesh, 300–400 mesh, Qingdao Marine Chemical Ltd., Qingdao, China), Sephadex LH-20 (GE Healthcare Bio-sciences AB, Uppsala, Sweden), YMC*GEL ODS-A (S-50 μm, 12 nm) (YMC Co., Ltd., Kyoto, Japan), and reversed-phase HPLC (Rohm and Hass Shanghai Chemical Industry Co., Ltd., Shanghai, China) were used for column chromatography. Biological assays were analyzed using a microplate reader (BioTek Synergy H1, BioTek Instruments, Winooski, VT, USA).

Optical rotations were determined with a Perkin-Elmer 41 polarimeter equipped with a sodium lamp (589 nm). UV, CD, and FT-IR spectra were performed on a Varian Cary 50, a JASCO-810 CD spectrometer, and a Bruker Vertex 70 instruments, respectively. 1D and 2D NMR spectra were recorded on a Bruker AM-400 spectrometer, and the ¹H and ¹³C NMR chemical shifts were referenced with respect to the solvent or solvent impurity peaks. HRESIMS were carried out in the positive ion mode on a Thermo Fisher LC-LTQ-Orbitrap XL spectrometer. X-ray data were collected using a Bruker APEX DUO instrument. Column chromatography was conducted with silica gel (200–300 and 300–400 mesh; Qingdao Marine Chemical Inc., China), Sephadex LH-20 (GE Healthcare Bio-Sciences AB, Sweden), and MCI gel (75–150 μm, Mitsubishi Chemical Corporation, Tokyo, Japan). Semi-preparative HPLC was carried out on a Dionex quaternary system with a diode array detector at a flow rate of 2.5 mL/min using a reversed-phased C₁₈ column (5 μm, 10 × 250 mm, YMC-pack ODS-A).Thin-layer chromatography (TLC) was performed with silica gel 60 F₂₅₄ (Yantai Chemical Industry Research Institute) and RP-C₁₈ F₂₅₄ plates (Merck, Germany).