Accucore aq c18 column

A Dionex Ultimate 3000 RSLC system (Thermofisher) combined with an Accucore aQ C₁₈ column (150 mm × 2.1 mm, 2.6 μm, Thermo fisher) was used in the sample separation process, and the separation speed was set at 0.3 μL/min for all the gradients. 1 μL of injection volume and a constant temperature of (25 ± 1) °C were adopted in the column. The eluents were A, water with 0.1% formic acid (v/v) and B, methanol, and the gradient program was as follows: 0–6 min, 8–40% B; 6–8 min, 40–100% B; 8–10 min, 100% B; 10–11 min, 100–8% B; 11–13 min, 8% B.

An ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis of metabolites was performed on an AB SCIEX LC-30A-Triple TOF 5600+ ESI MS system (AB SCIEX, Boston, MA, USA) equipped with a Shim-Pack Velox C18 column (2.7 μm, 150 mm × 2.1 mm, SHIMADZU, Japan), and a Thermo QE Focus instrument (Thermo Fisher Scientific, Waltham, MA, USA) with an Accucore AQ C18 column (2.6 μm, 150 mm × 2.1 mm, Thermo Fisher Scientific, Waltham, MA, USA). High-resolution electrospray ionization mass spectrometry (HRESIMS) data of novel metabolites were acquired by a Thermo Scientific Q Exactive mass spectrometer. Analytical high-performance liquid chromatography (HPLC) and semipreparative HPLC were carried out on an Agilent 1100 series system (Agilent Technologies Inc., Santa Clara, CA, USA) with a YMC-Pack ODS-A column (5 μm, 250 mm × 4.6 mm; 5 μm, 250 mm × 10 mm, YMC CO., LTD., Japan). Medium-pressure liquid chromatography (MPLC) was performed on a Buchi Medium pressure preparative chromatography (BUCHI Labor Technik AG, Switzerland) equipped with a medium-pressure column containing YMC reversed-phase (RP)-C₁₈ silica gel. Gel chromatography was carried out on a gel column with Sephadex LH-20 (40–70 μm; Amersham Pharmacia Biotech AB, Sweden). ¹H, ¹³C, and 2D NMR data were acquired on a Bruker Avance Neo 400 MHz apparatus (Bruker Corporation, Faellanden, Switzerland).